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SIBAL – AJLAB 2024
LACTIC ACID BACTERIA IN THE GENOMIC ERAVI International Symposium on Lactic Acid Bacteria
I Argentinean-Japanese Lactic Acid Bacteria “Tohoku
Forum for Creativity” Meeting

8-9 August 2024
Centro Cultural Eugenio Flavio Virla
TUCUMAN - ARGENTINA

食
料

健
康

SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

SIBAL – AJLAB 2024
LACTIC ACID BACTERIA IN THE GENOMIC ERAVI International Symposium on Lactic Acid Bacteria
I Argentinean-Japanese Lactic Acid Bacteria “Tohoku
Forum for Creativity” Meeting

8-9 August 2024
Centro Cultural Eugenio Flavio Virla
TUCUMAN - ARGENTINASIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

Libro de resúmenes VI International Symposium on Lactic Acid
Bacteria & I Argentinean-Japanese Lactic Acid Bacteria“Tohoku Forum for Creativity” Meeting 2024 /Dr. Lucila Saavedra
Dr. Jean Guy LeBlanc
Dr. Elvira Hebert
Dr. Julio Villena
Dr. Susana Salva
Dr. Guadalupe Vizoso Pinto
Dr. Leonardo Albarracin
Dr. Haruki Kitazawa
Dr. Tomonori Nochi
Dr. Hitoshi Shirakawa
Dr. Jonathan Laiño
Compilación de Jonathan Laiño – 1a edición – San Miguel deTucumán, Tucumán, Argentina – CONICET-CERELA, 2024
Libro digital, PDF

SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

Organizing Committee
VI International Symposium on
Lactic Acid Bacteria
President - Dr. Lucila Saavedra
Vicepresident - Dr. Jean Guy LeBlanc
Secretary - Dr. Elvira Hebert

I Argentinean-Japanese Lactic Acid Bacteria“Tohoku Forum for Creativity” Meeting
Argentina

Japan

Dr. Julio Villena
President

Dr. Haruki Kitazawa
President

Dr. Susana Salva
Vicepresident

Dr. Tomonori Nochi
Vicepresident

Dr. Guadalupe Vizoso Pinto
Dr. Leonardo Albarracin
Secretaries

Dr. Hitoshi Shirakawa
Secretary

SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

Scientific Committee
Dr. Carina Audisio
Dr. Silvia Cazorla
Dr. Silvina Fadda
Dr. Marisa Garro
Dr. Paola Gauffin CanoDr. Luciana Gerez
Dr. Jonathan Laiño
Dr. Carolina Maldonado
Dr. Guillermo Marcial
Dr. Lucia Mendoza
Dr. Sandra Quilodrán Vega
Dr. Cecilia Rodriguez

SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

Welcome Message
Welcome to the VI International Symposium on Lactic Acid Bacteria,
held in conjunction with the I Argentinean-Japanese Lactic Acid Bacteria“Tohoku Forum for Creativity.” This symposium stands as a cornerstone ofinternational collaboration and scientific inquiry, bringing together a diverse
community of researchers, professionals, and students dedicated toadvancing our understanding of lactic acid bacteria.Over the course of thisevent, we will delve into pioneering research and innovative applications thathighlight the significance of these microorganisms in biotechnology, as wellas in human and animal health.
This abstract book showcases the breadth of research in our field,
offering insights into the latest studies and developments.
As you explore these abstracts, we hope you find inspiration and foster
connections that will drive future collaborations and breakthroughs.
Thank you for being a part of this symposium and contributing to thisvibrant scientific event.

SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

Program
Day 1 – Thursday, August 8, 2024

08:30 – 09:30

Opening Ceremony

09:30 – 10:30

SESSION 1 - Chair: Jean Guy LeBlanc

09:30 – 10:30

S1. Effects of the extract of rice bran fermented byfungi and lactic acid bacteria on life-style relateddisease models.
Dr. Hitoshi Shirakawa

10:30 – 11:00

Break

11.00 – 12.00

SESSION 2 - Chair: Guadalupe Vizoso Pinto

11.00 – 11.30

S2A. Microbial therapeutics based on geneticallymodified lactic acid bacteria.
Dr. Fu Namai

11.30 – 12.00

S2B. Microbiota-gut-brain axis and its connection
with mental health.
Dr. Katia Sivieri (on-line)

12.00 – 12.30

Lunch

13.30 – 15.00

Poster Session

15:00 – 17:00

SESSION 3 - Chair: Julio Villena

15:00 – 15:40

S3A. Anti-viral postimmunobiotics from the porcine
small intestine.
Dr. Haruki Kitazawa

15:40 – 16:20

S3B. Fecal microbiota transplantation to improvemicrobiota for curing diarrhea in calves.
Dr. Tomonori Nochi

16:20 – 17:00

S3C. Lactic acid bacteria as silage inoculants:
innovations, probiotics and special challenges forgoat farming.
Dr. Roxana Medina

17:00 – 17:30

Break

ANIMAL HEALTH

Registration and Poster mounting

HUMAN HEALTH

08:00 – 08:30

SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

Program
Day 1 – Thursday, August 8, 2024
17:30 – 18:50

ORAL POSTER PRESENTATIONS - Chair: Alejandra deMoreno

17:50 – 18:00

AH4. Intravaginal administration of pharmabioticphytobiotic capsules with autochthonous lactic
bacteria for the prevention of infections of the bovine
reproductive tract. M.H. Miranda

18:00 – 18:10

AH5. Selection of immunomodulatory lactic acidbacteria from dog’s milk and colostrum.
S. Quilodrán-Vega

18:10 – 18:20

MP2. Nitrogen sources as a key factor regulating theexpression of gad genes in Levilactobacillus brevis
CRL2013. M.P. Urquiza Martínez

18:20 – 18:30

AB2. Effect of selected starter cultures on the
microbiological,
physicochemical,
andsensory
characteristics of cured-fermented sausages madewith llama (Lama glama) meat. L. Nelegatti

18:30 – 18:40

AB8. Physicochemical and microbiological stability,
aroma profile and sensory attributes of Chilto juice
fermented with Lactiplantibacillus paraplantarum
IBFV-10. L. Contreras

18:40 – 18:50

MG3. Proteomic regulation and gene expression ofselenized cells of Fructobacillus tropaeoli CRL2034 ina fermented fruit-based medium. L. Crespo

Biotechnology - Molecular Biology - Genomics

17:40 – 17:50

HH21. Lacticaseibacillus rhamnosus CRL1505 andits postbiotic improve the functional properties ofcirculating neutrophils from immunocompromised
mice and subjected to infectious stress. B. Vasile

Human & Animal Health

17:30 – 17:40

HH17. Oral administration of microencapsulated
Limosilactobacillus fermentum CRL1446 improves themain biomarkers of metabolic syndrome in mice.
C. TomeiSIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

Program
Day 2 – Friday, August 9, 2024
Poster mounting

09:30 – 10:30

SESSION 4 - Chair: Lucila Saavedra

09:30 – 10:30

S4. New insights of lactic acid bacteria through large
scale metagenomic analysis of food microbiomes.
Dr. Paul Cotter (on-line)

10:30 – 11:00

Break

11:00 – 12:00

SESSION 5 - Chair: Elvira Hebert

11:00 – 11:30

S5A. Role of probiotic derived extracellular vesicles ininterkingdom communication.
Dr. Graciela Lorca (on-line)

11:30 – 12:00

S5B. Self-aggregating probiotic bacteria activateenhanced interferon responses via DNA sensing.
Dr. Jorge Gutierrez-Merino (on-line)

12:00 – 13:30

Lunch

13:30 – 15:00

Poster Session

15:00 – 16:00

SESSION 6 - Chair: Fernanda Mozzi

15:00 – 15:30

S6A. Metataxonomic insights on lactic acid bacteria intable olives. Dr. Eugenio Parente (on-line)

15:30 – 16:00

S6B. Genomic characterization of the potential
probiotic
strainLimosilactobacillus
fermentum
CRL2085 isolated from feedlot cattle.
Dr. Francesca Fanelli

16:10 – 17:30

SESSION 7 - Chair: María Pía Taranto

16:10 – 16:50

S7A. Diversity of potassium transporters in lactic acidbacteria and the role of c-di-AMP in potassium
homeostasis regulation.
Dr. Victor Blancato (on-line)

16:50 – 17:30

S7B. Designing computational tools to understandthe bacterial-human cell communications.
Dr. Alexis Salas Burgos

17:30 – 18:50

Closing remarks & award ceremony Best oral &poster presentations - Lucila Saavedra-Julio Villena

19:30

Cocktail Reception

BIOTECHNOLOGY - MOLECULAR BIOLOGY - GENOMICS

09:00 – 09:30

SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

Index
Lecture Sessions
Human Health
S1. Effects of the extract of rice bran fermented by fungi and lactic
acid bacteria on life-style related disease models.
Dr. Hitoshi Shirakawa

18

S2A. Microbial therapeutics based on genetically modified lactic
acid bacteria.
Dr. Fu Namai

19

S2B. Microbiota-gut-brain axis and its connection with mental
health.
Dr. Katia Sivieri (on-line)

20

Animal health
S3A. Anti-viral postimmunobiotics from the porcine small intestine.
Dr. Haruki Kitazawa

21

S3B. Fecal microbiota transplantation to improve microbiota forcuring diarrhea in calves.
Dr. Tomonori Nochi

22

S3C. Lactic acid bacteria as silage inoculants: innovations,probiotics and special challenges for goat farming.
Dr. Roxana Medina

23

Biotechnology – Molecular biology – Genomics
S4. New insights of lactic acid bacteria through large scale
metagenomic analysis of food microbiomes.
Dr. Paul Coter (on-line)

24

S5A. Role of probiotic derived extracellular vesicles in interkingdom
communication.
Dr. Graciela Lorca (on-line)

25

S5B. Self-aggregating probiotic bacteriainterferon responses via DNA sensing.
Dr. Jorge Gutierrez-Merino (on-line)

26

activateenhanced

S6A. Metataxonomic insights on lactic acid bacteria in table olives.
Dr. Eugenio Parente (on-line)

27

S6B. Genomic characterization of the potential probiotic strain
Limosilactobacillus fermentum CRL2085 isolated from feedlot cattle.
Dr. Francesca Fanelli

28

SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

Index
Lecture Sessions
Biotechnology – Molecular biology – Genomics
S7A. Diversity of potassium transporters in lactic acid bacteria andthe role of c-di-AMP in potassium homeostasis regulation.
Dr. Victor Blancato (on-line)

29

S7B. Designing computational tools to understand the bacterialhuman cell communications.
Dr. Alexis Salas Burgos

30

Oral Posters presentations
Human and Animal Health
HH17. Oral administration of microencapsulated Limosilactobacillus
fermentum CRL1446 improves the main biomarkers of metabolic
syndrome in mice. C. Tomei

48

HH21. Lacticaseibacillus rhamnosus CRL1505 and its postbiotic
improve the functional properties of circulating neutrophils from
immunocompromised mice and subjected to infectious stress.
B. Vasile52

AH4. Intravaginal administration of pharmabiotic-phytobiotic
capsules with autochthonous lactic bacteria for the prevention of
infections of the bovine reproductive tract. M.H. Miranda

62

AH5. Selection of immunomodulatory lactic acid bacteria from dog’s
milk and colostrum. S. Quilodrán-Vega

63

Biotechnology – Molecular biology – Genomics
MP2. Nitrogen sources as a key factor regulating the expression ofgad genes in Levilactobacillus brevis CRL2013.
M.P. Urquiza Martínez

69

AB2. Effect of selected starter cultures on the microbiological,
physicochemical, and sensory characteristics of cured-fermented
sausages made with llama (Lama glama) meat. L. Nelegatti

78

AB8. Physicochemical and microbiological stability, aroma profileand sensory attributes of Chilto juice fermented with
Lactiplantibacillus paraplantarum IBFV-10. L. Contreras

84

MG3. Proteomic regulation and gene expression of selenized cells ofFructobacillus tropaeoli CRL2034 in a fermented fruit-based
medium. L. Crespo

98

SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

Index
Posters
Lactic acid bacteria and human health
HH1. Oral pathogen inhibition by selected lactid acid bacteria.
J.R. Naja

32

HH2. Effects of probiotic yoghurt administration in a murine
metastatic breast cancer model under chemotherapy treatment.
M.N. Cerasuolo

33

HH3. Modulation of lung immune response against Klebsiellapneumoniae by orally administered Lacticaseibacillus rhamnosus
CRL1505. R. Serda34

HH4. The intestinal fluids of probiotics fed mice
immunomodulatory properties on innate immune cells.
M.J. Martínez Monteros

35

trigger

HH5. Lacticaseibacillus rhamnosus CRL1505 ameliorates liver
injury and inflammation in poly(i:c) induced acute hepatitis.
M.J. Lorenzo Pisarello

36

HH6. Evaluation of a mixture of selected lactic acid bacteria (LAB)
in a murine model of levodopa induced dyskinesia (LID).
P.A. Yánez

37

HH7. Administration of Lacticaseibacillus paracasei subsp.
paracasei CRL75 modulates humoral and cytokine production
during food allergy. J. Laiño

38

HH8. Oral administration of LP 1518 improves the clinical outcomeof l. amazoniensis infection. J. Arrieta

39

HH9. Lactiplantibacillus plantarum CIDCA83114 as a potential
treatment for hemolytic uremic syndrome. C. Cancino

40

HH10. Probiotics decrease inflammatory bowel damage in aging
mice. A. Vestani41

HH11. Effects of probiotic yoghurt in a murine breast cancer model
under chemotherapy treatment. M. N. Cerasuolo

42

HH12. Lacticaseibacillus rhamnosus CRL1505 peptidoglycan
modulates tissue factor expression and protease activated receptor
1 activation during inflammation-coagulation response triggered bypoly(i:c) in the respiratory tract. H. Zelaya

43

SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

Index
Posters
Lactic acid bacteria and human health
HH13. Effect of riboflavin-overproducing Lactiplantibacillus
plantarum CRL2130 on breast cancer cells under treatment withchemotherapeutic drugs. R. Levit

44

HH14. Effect of probiotics supplementation in re-nutritional diets on
the thymus and intestine of malnourished mice. M.F. Balcells

45

HH15. Evaluation of a riboflavin- overproducing lactic
bacterium in a murine breast cancer model under treatment.
R. Levit

46

acid

HH16. In vivo study of anti-listeria activity of strains of lactic acidbacteria isolated from fermented foods. C. Burgos

47

HH17. Oral administration of microencapsulated Limosilactobacillus
fermentum CRL1446 improves the main biomarkers of metabolic
syndrome in mice. C. Tomei

48

HH18. Lactiplantibacillus plantarum MPL16 activates p38 MAPK
signaling in C. elegans and has a positive impact on longevity andresistance to Enterococcus faecalis infection. M.D. Vera49

HH19. Postbiotic nasal priming accelerates the recovery of innate
immune response in respiratory mucosa-associated lymphoid tissue
in malnourished mice. M. Ivir

50

HH20. Evaluation of lactic acid bacteria on the nutritional,
biochemical and immunological profile of mice fed a high-fat diet.
A. Márquez

51

HH21. Lacticaseibacillus rhamnosus CRL1505 and its postbiotic
improve the functional properties of circulating neutrophils from
immunocompromised mice and subjected to infectious stress.
B. Vasile52

HH22. Functional crackers manufactured with LAB-fermented
chickpea flour prevents oxidative stress in d-galactose induced
aging mice. G. Sáez

53

HH23. Lacticaseibacillus rhamnosus CRL1505 modulates alveolar
macrophages and improve their response to Pseudomonas
aeruginosa strains isolated from cystic fibrosis patients.
L. Arellano-Arriagada

54

SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

Index
Posters
Lactic acid bacteria and human health
HH24. Effect of Lactiplantibacillus plantarum cell free culture on the
respiratory superinfection induced by bacterial pathogens isolated
from cystic fibrosis patients. L. Abán55

HH25. Modulation of the Toll-like Receptor 3-mediated intestinal
immune response by water kefir. S. Dentice Maidana

56

HH26. Effect of Lactiplantibacillus plantarum CRL681 on the TLR4mediated intestinal innate immune response. A. Baillo

57

Lactic acid bacteria and animal health
AH1. Beer bagasse-enriched medium improves biomass productionof probiotic Lactobacillus johnsonii CRL2240. J.D. Babot

59

AH2. Characterization of surface properties and adhesion of
autochthonous lactic acid bacteria isolated from mare's reproductive
tract. A. Vasquetto

60

AH3. Ligilactobacillus salivarius FFIG58 modulates respiratory
immunity and confers long-term protection against Streptococcus
pneumoniae. M. Elean

61

AH4. Intravaginal administration of pharmabiotic-phytobiotic
capsules with autochthonous lactic bacteria for the prevention of
infections of the bovine reproductive tract. M.H. Miranda

62

AH5. Selection of immunomodulatory lactic acid bacteria from dog’s
milk and colostrum. S. Quilodrán-Vega

63

AH6. Effects of Ligilactobacillus salivarius subsp. salivarius A3iob
on honey production and health of Apis mellifera bee hives inArgentina. A.A. Arroyo Guerra

64

AH7. Identification and beneficial characteristics of autochtonous
lactic bacteria with probiotic potential for canine puppies.
N. Carrasco

65

AH8. Mixed inoculation using ferulic acid esterase-producing
lactobacilli improve nutritional, fermentative and functional
properties of corn stover silage. E. Andrada

66

SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

Index
Posters
Metabolism and physiology of lactic acid bacteria
MP1. Growth of lactic acid bacteria strains isolated from silage incorn plant extract media. R. M. Lenz

68

MP2. Nitrogen sources as a key factor regulating the expression ofgad genes in Levilactobacillus brevis CRL2013.
M.P. Urquiza Martínez

69

MP3. Antimicrobial properties of Levilactobacillus brevis strains
isolated from cheese whey of different origins. J.C. Alcocer

70

MP4. Bile acids modify morphology of a pote tial probiotic lactic acidbacterium. C. Moreno

71

MP5. RAMAN spectroscopy, an alternative methodology for theidentification of lactic acid bacteria. A. Ledesma

72

MP6. Monitoring the proteolytic activity of lactic acid bacteria using
Fourier Transform Infrared Spectroscopy. J. Carol

73

MP7. Biotransformation of selenium by lactic acid bacteria under
different stress culture conditions. F.R.E Molina

74

MP8. Aromatic profile of wines produced by mixed cultures ofyeasts and Oenococcus oeni. L.M. Mendoza

75

Applied biotechnology of lactic acid bacteria
AB1. Inhibition of enterohemorrhagic Escherichia coli by
Lactiplantibacillus plantarum CRL681: aggregation, co-aggregation,and electron microscopy studies. A. Baillo

77

AB2. Effect of selected starter cultures on the microbiological,
physicochemical, and sensory characteristics of cured-fermented
sausages made with llama (Lama glama) meat. L. Nelegatti

78

AB3. Lactic acid bacteria isolated from citrus fruits as starter
cultures for the production of fermented lemon juice.
D.A Barrionuevo

79

AB4. Partial characterization of Enterococcus spp. JK06 isolated
from regional salami from Tucumán. J. Gomez

80

AB5. Functional and sensory analysis of Calafate fruit fermented
with patagonian lactic acid bacteria. B. Sede Lucena

81SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

Index
Posters
Applied biotechnology of lactic acid bacteria
AB6. Pleurotus pulmonarius mushroom as novel resources ofprebiotics: effect on the growth and resistance to passage throughthe gastrointestinal tract of lactic acid bacteria. J. Gomez

82

AB7. Inhibition of α-glucosidase by lactic acid bacteria throughcontrolled fermentation of Brassica rapa and Undaria pinnatifida.
B. Martínez Caliari

83

AB8. Physicochemical and microbiological stability, aroma profileand sensory attributes of Chilto juice fermented with
Lactiplantibacillus paraplantarum IBFV-10. L. Contreras

84

AB9. In vitro evaluation of the antagonistic potential of
Ligilactobacillus murinus 26B1 isolated from mouse against
Staphylococcus aureus ATCC 6538P for its application in animal
nutrition. J. Palma85

AB10. Genetic modification of Lactococcus lactis to produce the antiinflammatory protein elafin: evaluation of different expression
systems. R. Levit

86

AB11. Influence of temperature, ph and culture media on
Salmonella enterica serotype enteritidis biofilm formation.
S. López Pérez

87

AB12. Optimization of the freeze-drying process to obtain morestress tolerant Oenococcus oeni starter cultures. E.V. Bentencourt

88

AB13. Sensory analysis of the final prototype of a soy foodfermented with Lacticaseibacillus paracasei subsp. paracasei
CRL207. B.L. Nacchio

89

AB14. Production of mixed fruit juice enriched with probiotic lactic
acid bacteria. F. Escobar

90

AB15. Impairment of Listeria monocytogenes developed on
industrial surfaces by Latilactobacillus curvatus CRL1579
bacteriocin. C. Melián91

AB16. Stability of prebiotic galactooligosaccharides during yoghurt
manufacture and storage. A. Fara

92

SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

Index
Posters
Applied biotechnology of lactic acid bacteria
AB17. Evaluation of the proteolytic profile and antioxidants oflegume flours fermented with selected lactic cultures. A. P. Pezuk

93

AB18. Technological strategy to improve the organoleptic
characteristics of soy food products: fermentation with selected
lactic acid bacteria. G.N. Avila Hael

94

Molecular biology and genomics of lactic acid bacteria
MG1. Genomic characterization of the honeybee-probiotic strain
Ligilactobacillus salivarius subsp. salivarius A3iob. M. Elean

96

MG2. Functional and genomic characterization of Pediococcus
pentosaceus TUCO-3 isolated from cat milk: a potential probiotic
strain to combat infections. S. Quilodrán-Vega

97

MG3. Proteomic regulation and gene expression of selenized cells ofFructobacillus tropaeoli CRL2034 in a fermented fruit-based
medium. L. Crespo

98

MG4. Genomic and proteomic characterization of Lactiplantibacillus
plantarum CRL681 focusing on cell surface and secreted proteins.
A. Baillo

99

乳
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LECTURE SESSIONS

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SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

S1 EFFECTS OF THE EXTRACT OF RICE BRAN FERMENTED BY
FUNGI AND LACTIC ACID BACTERIA ON LIFESTYLE-RELATEDDISEASE MODELS

H. Shirakawa1
1International Education and Research Center for Food and Agricultural Immunology,
Graduate School of Agricultural Science, Tohoku University. Sendai, Japan. E-mail:shirakah@tohoku.ac.jp

Japan produces about 8 million tons/year of brown rice. Rice bran is a byproduct of rice polishing process, and brown rice consists of 10% rice bran (RB).
About 40% of RB is used to produce rice bran oil. RB is also mainly used as livestock
feed and fertilizer; however, the use of approximately 25% of RB is not known, andmost of it is estimated to be discarded. It is widely known that RB is rich inbeneficial substances and has several health benefits. For example, γ-oryzanol,
ferulic acid, and tocotrienol are reported to have blood cholesterol-lowering,angiogenesis-inhibiting, anti-tumor, and hypotensive effects. These non-polar
components are abundantly present in RB oil and are responsible for most of its
beneficial effects. In addition, the residue resulting from the extraction of rice branoil (defatted rice bran) comprises inositol, phytic acid (inositol 6-phosphate), phytin,
and rice bran proteins. These components have been documented to exhibit
antitumor, immunostimulatory, antilipidemic, and antidiabetic effects. Althoughmaximum utilization of RB is desirable, it is not fully utilized to the full extent
because of its tendency to deteriorate and its characteristic odor. To improve theflavor of RB, we developed a novel fermentation method that employing fungi andlactic acid bacteria sequentially. This fermented rice bran (FRB) exhibited an
improved flavor profile and was expected to be enriched in functional components,
which are secondary metabolites produced by microorganisms. The beneficial effectsof FRB have been verified in several animal models of lifestyle-related diseases. FRB
suppressed elevation of blood pressure and improved insulin resistance byenhancing ACE inhibitory activity and elevating adiponectin levels in blood,
respectively, in stroke-prone spontaneously hypertensive rats (SHRSP). FRB also
suppressed the onset and restoration of dextran sulfate sodium (DSS)-induced colitis
in mice and muscular atrophy in streptozotocin (STZ)-induced diabetic rats by
inhibiting NFκB activation. Taken together, these results indicate that FRB may beeffective in preventing lifestyle-related diseases. We anticipate that FRB will be usedin practical applications in the future.

18

SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

S2A MICROBIAL THERAPEUTICS
MODIFIED LACTIC ACID BACTERIA

BASED

ON

GENETICALLYF. Namai1,2, K. Nishiyama1,2, H. Kitazawa1,2, T. Shimosato3,4
University, Aoba, Aramaki, Aoba-ku, 980-8572 Sendai, Japan. 2CFAI, Aoba,
Aramaki, Aoba-ku, 980-8572 Sendai, Japan. 3Department of Biomolecular Innovation,
Institute for Biomedical Sciences, Shinshu University, 399-4598 Nagano, Japan. 4Institutefor Aqua Regeneration, Shinshu University, 399-4598 Nagano, Japan. E-mail:fu.nama.a3@tohoku.ac.jp
1Tohoku

The increasing incidence of inflammatory bowel disease (IBD) in western andrapidly westernizing developing countries is a growing concern. The development ofaffordable alternatives to conventional IBD therapies is a major challenge. In thisstudy, we explored genetically modified lactic acid bacteria (gmLAB) as an approachto IBD treatment. gmLAB can serve as a carrier to deliver target proteins directly tothe intestinal mucosa, as lactic acid bacteria can survive in the intestinal tract. Local
delivery of therapeutic proteins using gmLAB is expected to provide a more efficienteffect while reducing side effects. Here, we constructed a gmLAB that secretes aninterleukin-1 receptor antagonist (IL-1Ra), an inhibitor of IL-1, since theinflammatory cytokine IL-1 is involved in the exacerbation of IBD. IL-1Ra expression
vector was introduced into Lactococcus lactis NZ9000 to generate gmLAB (NZ-IL1Ra).
At the same time, an empty vector was also introduced to generate the control
gmLAB (NZ-VC). The IL-1Ra expression in the bacterial cell and secretion in the
supernatant were detected using Western blot. The secreted IL-1Ra was purified,and its inhibitory activity against IL-1 signaling was evaluated. The results showedthat IL-1Ra secreted by NZ-IL1Ra suppressed IL-2 secretion from IL-1β-stimulated
EL4.NOB-1 cells in a concentration-dependent manner. NZ-IL1Ra was thenadministered orally to mice, and the colon and cecum contents and serum werecollected. The concentration of IL-1Ra in each was determined. The concentration of
IL-1Ra was significantly increased compared to that in the untreated mice. This
indicates that oral administration of NZ-IL1Ra delivers IL-1Ra to the intestinal tract
and blood. To evaluate the efficacy of orally administered NZ-IL1Ra in alleviating IBD
symptoms, we conducted a study using a mouse model of dextran sulfate sodium(DSS)-induced colitis. Mice given 3% DSS ad libitum experienced weight loss anddiarrhea. Oral administration of NZ-VC did not alleviate these symptoms; however,
NZ-IL1Ra significantly accelerated their recovery. Analysis of mesenteric lymph
nodes revealed a significant reduction of CD4+IL-17A+ cells, induced by IL-1
signaling, in the NZ-IL1Ra group compared to the NZ-VC group. In conclusion, we
have successfully constructed a gmLAB that secretes bioactive IL-1Ra with the aimof proposing an approach to IBD. Oral administration of gmLAB transports IL-1Ra tothe intestinal tract of mice and suppresses IL-1 signaling in situ, thereby alleviating
the symptoms of DSS-induced colitis. We hope that this research will contribute to
the improvement of human health.

19

SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

S2B MICROBIOTA-GUT-BRAIN AXIS AND ITS CONNECTION WITH
MENTAL HEALTH
K. Sivieri

20

SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

S3A POSTIMMUNOBIOTICS FROM PORCINE SMALL INTESTINE

H. Kitazawa1,2, Y. Imamura1, H. Yamamuro1, M. Elean3, L. Arce3, F. Namai1,2, K.
Nishiyama1,2, J. Villena3
1Tohoku University, Aoba-ku, 9800 Sendai, Japan. 2CFAI, Sendai, Japan. 3CERELACONICET. Chacabuco 145. 4000 Tucuman, Argentina. E-mail:haruki.kitazawa.c7@tohoku.ac.jp

Infectious diseases are required to be addressed with efficient prevention
strategies to protect livestock and human health. Vaccines and antimicrobials canprotect livestock from infectious diseases, but their development is labor-intensive
and time-consuming. In addition, the drastic increase in antimicrobial-resistantmicrobes make necessary the development of new measures to prevent infections.
Postimmunobiotics, defined as the inanimate microbes and their derived functional
molecules with the ability to modulate the immune system, are a promisingalternative to combat infectious diseases in livestock. We have focused on the studyof postimmunobiotics with the capacity to beneficially modulate anti-viral immune
responses. We have selected a postimmunobiotic candidate derived from theprobiotic strain Ligilactobacillus salivarius FFIG58 strain from our porcine
lactobacilli library using and antiviral immunoassay system developed with theoriginally established porcine intestinal epithelial (PIE) cells. Considering the cholinebinding protein (CbpA) as a functional molecule, a cbpA knockout (ΔCbpA) mutant
from the FFIG58 strain was established. The phenotypes and functional activities ofwild-type FFIG58 and the ΔCbpA strains were compared including their growthability, their surface characteristics by electron microscopy analysis and theiradhesion to PIE cells. The capacity of heat-killed FFIG58 and ΔCbpA strains todifferentially modulate the Toll-like receptor 3 (TLR3)-mediated innate antiviral
immunity and increase the resistance to rotavirus infection was evaluated. In
addition, we studied the interaction of heat-killed wild-type FFIG58 and the ΔCbpA
strains with porcine macrophages, assessing their effect on TLR3-mediatedimmunity including the modulation of intracellular signaling factors and negative
regulators of the TLR signaling pathway. The wild-type FFIG58 strain efficiently
adhered to PIE cells, improved TLR3-mediated immunity and enhanced theresistance to rotavirus infection. The strain was also efficiently phagocyted bymacrophages and positively modulated their response to TLR3 activation. The
immunomodulatory effect of FFIG58 was markedly reduced in the mutant ΔCbpA.
Additionally, the immunomodulatory activities of L. salivarius FFIG58 and the ΔCbpA
mutant were confirmed in vivo using infant mice as a young host model considering
the potential application of this strain to improve the resistance to postweaning viraldiarrhea in piglets. Heat-killed L. salivarius FFIG58 is an interesting candidate to be
developed as a postimmunobiotic with the ability to improve antiviral immunity via
CbpA. It is expected that our research will lead to the expansion of the food and feed
immunological use of postimmunobiotics, which will contribute to the prevention ofviral infectious diseases of livestock. This work was supported by BRAIN-JPJ007097,
JSPS-19H00965, and JSPS-23H00354 grants.

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SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

S3B CROSS FECAL MICROBIOTA TRANSPLANTATION TO TREATDIARRHEA IN PRE-WEANED CALVES
T. Nochi1*, J. Islam1, N. Ohtani2, Y. Shimizu2, J. Yamazaki3, Z. Nakamura4, H. Tanaka2

1Graduate School of Agricultural Science, Tohoku University, Miyagi, Japan. 2Chiba Prefectural

Federated Agricultural Mutual Aid Association, Chiba, Japan. 3Hokkaido Federated Agricultural
Mutual Aid Association, Hokkaido, Japan. 4Okinawa Prefectural Federated Agricultural Mutual
Aid Association, Okinawa, Japan. 468-1, Aoba, Aramaki, Aoba-ku, Sendai, Miyagi 980-8572,
Japan. E-mail: nochi@tohoku.ac.jp

Calf diarrhea is the most common illness in pre-weaned calves. Effective
treatment options to manipulate the gut microbiome environment of calves undercommercial operations are of great importance to improve animal health and reduce
antimicrobial usage. Fecal microbiota transplantation (FMT) involves transplanting
the fecal contents of a healthy donor into a diseased patient to restore gut microbiota
to its original state. FMT has been demonstrated to be an effective treatment for calf
diarrhea in recent times. However, most of the studies on FMT against calf diarrhea
are investigated in a same farm specific manner concerning its safety. However, tointroduce FMT as a nationwide positioning, as well as to better design future
microbial therapeutics for calf diarrhea, we introduced cross-FMT intervention, in
where donors were selected from three different regions within a range ofapproximately 3,000 km across Japan. We collected 180 feces from healthy calves as
donor from three regions of Japan (Hokkaido, Chiba, and Okinawa area, n=60 each),
and among them 24 donors were finally selected as potential donors for FMT basedon 16S rRNA amplicon sequencing and absence of potential pathogens especially
Clostridium perfringens, Cryptosporidium parvum, Coccidia, Salmonella, Rotavirus,
Coronavirus, E. coli as well as BLDV and BLV. From each potential donor, 3 vials (10g feces each) were prepared using the original feces and continued to freeze-dry.
Finally, prepared donors’ feces were sent to three regions and subjected to 72 FMT
trials (n=24 recipients/area). Based on the diarrheal severity, pathogen detection,
16S rRNA amplicon sequencing and fecal metabolomics (CE-TOFMS) results, overall
FMT efficacy is found 76.39% 7 days after FMT. Based on the area, the FMT efficacy
is observed in Hokkaido region (83.34%), Chiba (75.0%) and Okinawa (70.83%).
Comparing the donor efficacy, the donor efficacy is observed in Hokkaido (76.67%),
Chiba (77.78%) and Okinawa (73.34%). Finally, based on the intra-area donor
efficacy, Hokkaido donor showed 100% efficacy in Hokkaido, but 80% for Chiba and
50% for Okinawa. On the contrary, Chiba donors showed similar efficacy (77.78%) in
all three areas, while Okinawa donors showed 60% efficacy in Hokkaido and Chiba,
but 100% efficacy in Okinawa itself. Fecal microbial community analysis based on
16S rRNA amplicon sequencing revealed an increase in the alpha-diversity indicesafter FMT and pairwise PERMANOVA analysis from beta diversity confirmed that
there is significant difference before and after FMT in successful FMT cases.
Furthermore, FMT reduces the gram negative Proteobacteria level as fecal aminoacids (alanine, leucine, valine, isoleucine, glycine, arginine, and glutamic acid),which may be responsible to restore the gut microbial community to healthy stateand lessens the diarrheal severity in successful cases.

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SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

S3C LACTIC ACID BACTERIA AS SILAGE INOCULANTS:INNOVATIONS, PROBIOTICS AND SPECIAL CHALLENGES FOR
GOAT FARMING
R. Medina1,2

1CERELA-CONICET.

Chacabuco 145, Tucumán, Argentina. 2Facultad de Agronomía,
Zootecnia y Veterinaria, Universidad Nacional de Tucumán, Tucumán, Argentina
E-mail: rmedina@cerela.org.ar

Goat production in Argentina is typically small-scale and for local
consumption. It is mainly located in arid and semi-arid regions where overgrazing
during the dry season cause environmental degradation. Therefore, sustainable
farming needs diversified food sources, for which agricultural by-products canrepresent a locally available alternative. Corn stover, as other abundant high-fiber
by-products, are usually challenging to preserve through ensiling. A strategy to
improve its conservation and nutritional properties is using lactic acid bacteria (LAB)
that present ferulic acid esterase (FAE) activity. FAE breaks down esterified sugars ofplant cell walls, like arabinose, to release hydroxycinnamic acids such as ferulic acid(FA), an antioxidant compound. We isolated LAB with FAE activity from goat feces(Lactobacillus johnsonii CRL 2240) and corn silos (Levilactobacillus brevis CRL2239,
Lactiplantibacillus plantarum CRL2241). These strains were selected for studies incorn stover silages, following small farmers’ conditions. An uninoculated (UN) silo
was used as a control for comparison. Fermentative and nutritional profiles,
digestibility, phenolic compounds, antioxidant capacity and metagenomics ofbacterial community were evaluated after 60 days of ensiling. L. plantarum CRL 2241
reduced gas losses and pH, increased the lactic to acetic acid ratio and FA content,
showing characteristics of a first-generation (1G) inoculant with effective FAE
activity. L. johnsonii CRL 2240 reduced the dry matter loss and modified the fiber
composition, acting as a 1G and 3G inoculant. L. brevis CRL 2239 strain increased
acetate content, increased in vitro true digestibility of dry matter and free FA content,
acting as a 2G and 3G inoculant. Metagenomic studies showed that inoculatedsilages had lower diversity and equity at the genus level when compared to UN, while
L. plantarum CRL 2241 and L. johnsonii CRL 2240 strains increased the relative
abundance of Lactobacillus genus. Subsequently, two mixed inoculants were
designed: LPB (CRL 2239 and CRL 2241) and LPBJ (LPB+ CRL 2240). Although both
inoculants modulated the ensiling process and increased the free FA content, theinclusion of L. johnsonii CRL2240 in LPBJ inoculant reduced the dry matter loss,
acid detergent fiber and cellulose content. Furthermore, L. johnsonii CRL 2240
presented probiotic properties in vitro, for which inoculation of this strain in silages
might constitute a probiotic-supply strategy. These can be considered a fourth
generation of inoculants.

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SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

S4 NEW INSIGHTS OF LACTIC ACID BACTERIA THROUGH LARGESCALE METAGENOMIC ANALYSIS OF FOOD MICROBIOMES.
P.D. Cotter1,2

1Teagasc Food Research Centre, Moorepark, Cork, Ireland and the APC Microbiome Ireland,

VistaMilk and Food Health Ireland Researchpaul.cotter@teagasc.ie; paulcotter@seqbiome.com

Centres;

2SeqBiomeLtd.

E-mail:Microorganisms exist along the food chain and impact the quality and safetyof foods in both positive and negative ways. Identifying and understanding the
behaviour of these microbial communities enable the implementation of preventative
or corrective measures in public health and food industry settings. Traditional
culture-dependent microbial analyses are time-consuming and target only specificsubsets of microbes. However, the greater use of culture-independent metagenomic
approaches has the potential to facilitate a thorough characterization of the
microbial communities, and of lactic acid bacteria, in particular along the food chain.
This has been through an in depth investigation of specific foods, such as cheeseand kefir, as well as a broader investigation of a wide range of different food types.
Combining this data in a new curated Food Metagenomic database has allowed avariety of different analyses, including by assembly-based (through generating
metagenome assembled genomes; over 10,000 high and medium quality MAGs) andnon assembly-based, has provided new insights into food associatedmicroorganisms, including many species of Lactic Acid Bacteria, including
information regarding their prevalence, distribution and overlap with gut-associated
taxa.

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SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

S5A ROLE OF PROBIOTIC-DERIVED EXTRACELLULAR VESICLESIN INTERKINGDOM COMMUNICATION
G.L. Lorca
Department of Microbiology and Cell Science. Genetics Institute. Institute of Food andAgricultural Sciences. University of Florida. E-mail: glorca@ufl.edu

We recently showed that Lactobacillus johnsonii N6.2 releases EVs with adistinct composition of proteins, and lipids when compared to whole cells. L.
johnsonii N6.2 is a probiotic bacterium that has been shown to mitigate type 1
diabetes in prone rodents (BBDP rat model) by maintaining euglycemic levels andreducing the inflammatory state. We hypothesized that EVs play a central role indelivering bioactive molecules that may act as mechanistic effectors in immune
modulation. We observed that the addition of EVs to the human pancreatic cell line
βlox5 reduced cytokine-induced apoptosis. The role of EVs on beta cell function was
further evaluated using primary human pancreatic islets. It was found that EVs
significantly induced insulin secretion in the presence of high glucose
concentrations. Through RNAseq analyses, a significant induction of the AHR and 2’,
5’-oligoadenylate synthetase (OAS) pathways were observed. The OAS pathway ispart of an innate immune response activated by viral or bacterial RNA. In mammals,
the OAS family is composed of three enzymatically active enzymes, OAS1, OAS2 andOAS3, all of which were significantly induced in the presence of EVs, but not by
purified membranes from L. johnsonii N6.2. We hypothesized that the activation of
the OAS pathway by specific effector molecules present in L. johnsonii N6.2 secreted
EVs could hinder a viral infection in vivo. I will discuss our approach to study therole of EVs on viral infection using the murine norovirus (MNV-1) as an RNA virus
model of infection.

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SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

S5B SELF-AGGREGATING PROBIOTIC BACTERIA ACTIVATEENHANCED INTERFERON RESPONSES VIA DNA SENSING
S. Solis1; E. Maldonado1; C. Maluquer de Motes1; S. Mukhopadhyay2; J. Gutierrez-Merino1*

1School of Biosciences and Medicine, University of Surrey, UK. 2School of Immunology andMicrobial Sciences, King’s College London, UK. *Corresponding author and speaker. E-mail:j.gutierrez@surrey.ac.uk

The discovery of the human (and animal) microbiome has revolutionized thefield of Medicine and Biological Sciences. To date, we have access to endless data
informing of the microbial diversity present in different organs and body systems,
and how this diversity correlates with many medical conditions. However, we are still
unsure which of the many commensal microbes that reside the host are the main
drivers that restore or protect health from disease. Much littler the information isconcerning the molecules that these key commensals possess to interact with the
immune system. This important question has been addressed by our research team
as we are trying to reveal the molecules that beneficial bacteria utilize to induce hostbeneficial responses. Our recent investigations have shown that certain species oflactobacilli significantly activate the production of type I interferon (IFN-I) cytokines
in macrophages. IFN-I cytokines are essential to confer protection against microbial
infections and auto-immune disorders. For the first time, we have proved that thisIFN-I activation is predominantly driven by cGAS, a molecule that activates thecytosolic sensor STING upon the recognition of bacterial DNA. Furthermore, we haveobserved that lactobacilli encode some surface proteins with the potential to interact
with macrophages for subsequent phagocytosis via non-opsonic scavenger receptors.
Therefore, we are focused on determining the role that these surface proteins play as
a port of entry in macrophages and characterize the IFN-I-mediated intracellularsignalling initiated by cGAS. Elucidating these unknown mechanisms will be
important to inform on how specific molecules of commensals modulate or stimulate
host responses that, in unhealthy individuals, are exacerbated or inhibited. Overall,our studies will provide a better understanding on the molecular crosstalk between
the microbiome and mammalian cells, paving the way for major therapeutic
discoveries.

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SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

S6A METATAXONOMIC INSIGHTS ON LACTIC ACID BACTERIA IN
TABLE OLIVES

E. Parente1, A. Aliano1, R. Pietrafesa1, M. Hidalgo1, A. Ricciardi1
di Scienze Agrarie, Forestali, alimentari ed ambientali. Università degli Studi della
Basilicata, Potenza, Italy. E-mail: eugenio.parente@unibas.it
1ScuolaWith a global consumption of 2,890x103 ton/y (2022/2023, International
Olive Council), table olives are among the most popular vegetable fermented foods.
Production and consumption are concentrated in the Mediterranean area, where firsttraces of olive brining date to 7,000 BP. However, they are increasing in othercontinents, with Peru as leading producer. As for many other vegetablefermentations, removal of the initial microbial contaminants relies only on selection
and washing, and starter cultures are rarely used. Selective factors driving the
dynamics of the microbiota include stressors such as low nutrient content (due toalkali treatment and washing in alkali treated olives), high phenolic and high (6-10%)
NaCl concentration, anaerobic conditions, amensalism and competition. Together
with yeasts, lactic acid bacteria (LAB) are major components of table olives microbialcommunities. Halophilic Pseudomonadota (Celerinatantimonas, Halomonas, Vibrio)
and halophilic and alkaliphilic LAB (HALAB) are also important in alkali treated
Spanish style olives and specialties like Picholine. Amplicon targeted metagenomics,
often combined with metabolomic approaches have recently replaced culture
dependent and low-throughput approaches for the study of the microbiota of table
olives. Publicly available data, when combined in properly annotated databases offer
potential for quantitative meta-analysis: in the framework of the METAolive project,
we combined 11 published studies extracted from the FoodMicrobionet databasewith two unpublished studies from our lab, thus obtaining a dataset with 525
samples, including raw fruits, olives and brines during fermentation, which is
unique in terms of variety and size. We found 24 genera of LAB and HALAB. The
latter (Alkalibacterium, Marinilactobacillus) were confined to alkali treated olives, andwere dominant in the Picholine style. In Spanish style olives large differences exist
between varieties, while in Greek style olives Lactiplantibacillus, together with
Pediococcus and Lentilactobacillus almost invariably coexist and dominate thefermentation, while the occurrence and the abundance of other genera is morevariable. In a study on Nyons olives, LAB prevalence and abundance was very low on
contact surfaces and raw materials. Contamination patterns suggest that thedominance of LAB must be due to dispersal from other sources (reinoculum fromfermentation vessels) combined with strong selective pressure. METAolive is cofunded by Ministero dell’Università e della Ricerca, PRIN 2022, proposal
2022NN28ZZ, and received funding from the European Union Next-GenerationEU PIANO NAZIONALE DI RIPRESA E RESILIENZA (PNRR).

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SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

S6B GENOMIC CHARACTERIZATION OF THE POTENTIALPROBIOTIC STRAIN Limosilactobacillus fermentum CRL 2085
ISOLATED FROM FEEDLOT CATTLE

F. Fanelli1, C. Aristimuño Ficoseco2, M.E.F. Nader-Macías2, G.M. Vignolo2, V. Fusco1, S.
Fadda2
1ISPA-CNR. Via Amendola 122/O, Bari, Italy. E-mail: francesca.fanelli@ispa.cnr.it
1CERELA-CONICET. Chacabuco 145, Tucuman, Argentina. E-mail: sfadda@cerela.org.ar

Limosilactobacillus fermentum is a Gram-positive, heterofermentative species
generally considered safe. It is found in sewage, fermented food, and in human andanimal gastrointestinal tract. Due to its habitat and its resistance to bile salts thisspecies is recognized as probiotic and introduced in intestinal and vaginal tract
diseases treatments. L. fermentum CRL2085 was isolated from feedlot cattle inArgentina, selected for its phenotypic traits and administered in a large-scale survey
to evaluate the modulation of faecal microbiome in feedlot cattle. To gain moreinsight into its probiotic potential we performed a genomic characterization of thisstrain. The genome of L. fermentum CRL 2085 is 1.9 Mb in length, has a GC content
of 52.2 % and a coding density of 86%. The 1957 predicted genes include 34
carbohydrate-active enzymes, with glycosyl transferases and hydrolases being themost abundant. We identified several probiotic genes involved in adhesion,
immunomodulation, temperature, osmotic and oxidative stress responsive genes,
complete pathways for folate and riboflavin biosynthesis and tree putative clusters
for exopolysaccharides production. In silico safety assessment validated the safety ofthis strains, while the presence of numerous CRISPR-Cas element might prevent theacquisition of horizontally transferred antimicrobial genes. Genomic analysis
confirms that L. fermentum has an open genome, undergoing a dynamic evolution,with the acquisition of novel unique genes for adaptation to new environments.
Genome-based phylogeny supported this evidence, as clustering of the strains only
partially correlated with the source of isolation or the geographical origin, thus
indicating that some L. fermentum strains have been recently introduced andtransient in temporary environments. These data confirm the probiotic potential ofthis strains and its safety and encourage its further characterization to revealing new
metabolic attributes that can fully exploit its biotechnological capabilities andoptimize its application as probiotic in commercial feedlot.

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SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

S7A DIVERSITY OF POTASSIUM TRANSPORTERS IN LACTIC ACID
BACTERIA AND THE ROLE OF CYCLIC-DI-AMP IN POTASSIUMHOMEOSTASIS REGULATION
V. S. Blancato1, G. Acciarri1, M. E. Taborra1, F. O. Gizzi1, C. Magni1

1IBR-CONICET-UNR. Suipacha 531, Rosario, Argentina. E-mail: blancato@ibr-conicet.gov.ar

Cyclic-di-AMP, a nucleotide second messenger in bacteria and some Archaea,
regulates diverse metabolic processes including osmolyte uptake, cell wall
homeostasis, antibiotic and heat resistance. Lactic acid bacteria (LAB) possess
enzymes for cyclic-di-AMP synthesis and degradation but vary in their receptors.
Many cyclic -di-AMP interacting partners identified in bacteria are involved inpotassium homeostasis. These include K+ importers or importer gating components(TrkA/KtrA, TrkH, KupA, KupB, and KimA), two-component systems that regulate K+
importer expression (KdpD), a K+ exporter (CpaA), and a riboswitch that regulates K+
importer gene transcription. LAB exhibit diverse combinations of K+ transporters,
there is also variation between strains of the same species. In our laboratory, KupA
and KupB were identified as novel cyclic-di-AMP binding proteins in Lactococcus
lactis, functioning as high-affinity K+ transporters inhibited by cyclic-di-AMP
binding. Kup transporters are widely distributed among various species of
Lactobacillus and Enterococcus and are also found in some species of Streptococcus
and Staphylococcus. However, no Kup potassium transporter is encoded in the
genomes of B. subtilis, L. monocytogenes, S. aureus, or S. pneumoniae. In
Enterococcus faecalis, we demonstrated that Kup and KimA are high-affinity K+
transporters, with KtrA playing a crucial role under stress conditions. Additionally,
we found that the presence of the Kdp system in E. faecalis is strain-dependent,
being more prevalent in clinical strains compared to environmental, commensal, or
food isolates. Our research revealed that kup gene regulation in E. faecalis is affectedby an IS6770 insertion sequence, with a 30% co-occurrence rate in IS6770-positive
strains, suggesting its potential role in E. faecalis adaptation across diverse niches.
These findings highlight the complexity and diversity of cyclic-di-AMP signaling
pathways in bacteria, emphasizing the intricate regulatory networks that enablemicrobial adaptation and survival in different environments. The significance ofpotassium transporters in LAB is especially important, as they have a crucial role inmaintaining cellular homeostasis and ensuring optimal growth and stress response.
This is vital for their application in food production and probiotic development.

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S7B DESIGNING COMPUTATIONAL TOOLS TO UNDERSTAND THE
BACTERIAL-HUMAN CELL COMMUNICATIONS.
A. Salas Burgos

30

乳
酸
菌

LACTIC ACID BACTERIA AND
HUMAN HEALTH

健
康

食
料

SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

HH1 ORAL PATHOGEN INHIBITION BY SELECTED LACTID ACID
BACTERIA

Naja JR1,2, Urquiza MP1, Bulacios GA1, Cataldo PG1, Nader MEF1, Houde V.P2, Hebert EM1,
Saavedra L1 and Minahk C3
1CERELA-CONICET. Chacabuco 145. 4000 Tucumán, Argentina. 2Université Laval, 2420 rue
de la Terrasse, Québec, Canada. 3INSIBIO-UNT-CONICET. Chacabuco 461. 4000 Tucumán,
Argentina. E-mail: johanarnaja@gmail.com

Porphyromonas gingivalis is a Gram-negative anaerobic bacterium that plays
a significant role in the development and progression of periodontitis. This disease
begins when bacteria accumulate on the tooth surface, prompting immune cells to
infiltrate the gingival sulcus (dental plaque). This process leads to inflammation,
destruction of the supporting tissues of the teeth, bone resorption, and ultimately,
tooth loss. Currently, periodontitis is managed through frequent and invasive
periodontal therapy, such as scaling and root planning, to remove dental plaque.
Therefore, alternative treatments are needed, and lactic acid bacteria (LAB) emerge
as a plausible strategy. LAB strains or their metabolites have demonstrated
antibacterial, antioxidant, and antifungal properties, as well as immunomodulatory
capacities against oral pathogens. Consequently, it can be proposed that LAB may
help mitigate the proliferation of periodontopathogens or serve as an adjunct toperiodontal treatments. The objective is to study the antibacterial effect of Cell-freesupernatant (CFS) of two LAB strains Lacticaseibacillus rhamnosus CRL1522 andLactiplantibacillus plantarum CRL1363 against P. gingivalis growth. P. gingivalis(ATCC 33277) was routinely grown in Todd Hewitt broth (THB) enriched with heminand vitamin K (THBe) at 37ºC in anaerobic conditions. L. rhamnosus CRL1522 and L.
plantarum CRL1363 (CERELA Culture Collection) were grown in MRS medium.
Human gingival keratinocytes Htert TIGKs and the macrophage cell line U937 wereused as cell models. The LAB strains under study did not exhibit significant
antibacterial activity against P. gingivalis, although they did slow the pathogen's
growth. However, the cell-free supernatant of L. rhamnosus CRL1522 and L.
plantarum CRL1363 significantly reduced the secretion of IL-6 and IL-8 by the
keratinocytes and TNF-α and IL-6 by the macrophages, which had been induced by
P. gingivalis. More importantly, the activity of Arg-gingipain was markedly reducedby both LAB strains. P. gingivalis KDP112 was used as negative control. The
expression of COX-2, NOD-1, NOD-2, TLR2 and TLR4 was also checked. In
summary, L. rhamnosus CRL1522 and L. plantarum CRL1363 are suitable
candidates for combating the periodontopathogen P. gingivalis.

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HH2 EFFECTS OF PROBIOTIC YOGHURT ADMINISTRATION IN A
MURINEMETASTATICBREASTCANCERMODEL
UNDER
CHEMOTHERAPY TREATMENT
M. N. Cerasuolo1, J. G. J. LeBlanc1, A. de Moreno de LeBlanc1.
1CERELA-CONICET.
Chacabuco 145. C.P. 4000 Tucuman,
mcerasuolo@cerela.org.ar

Argentina.

E-mail:Breast cancer (BC) is the most prevalent type of cancer diagnosed in women
worldwide, and its metastasis are associated with a high mortality and morbidity
rate. Chemotherapy is useful for treating BC metastasis; however, it affects thepatients’ quality of life leading to discontinuation of therapies. In this sense, the usesof probiotics are a promising alternative as adjuvant. Previously, our group designed
a yoghurt made with two strains selected for their immunomodulatory properties.
Considering that the host's immune system plays an important role against cancer
and metastasis; the aim of this study was to evaluate the effects of the probiotic
yoghurt administration in a murine model of BC metastasis undergoing an oral
chemotherapy treatment with capecitabine (CAP). The probiotic yoghurt was
prepared by fermenting milk with Streptococcus thermophilus CRL807 andLactobacillus delbrueckii subspecies bulgaricus CRL864 from the CERELA Culture
Collection. For BC metastasis, 4T1 BC cells were injected into the upper rightmammary gland of 7–8-week-old BALB/c female mice. Tumors were removed after
reaching a volume of 0.07 ± 0.02 cm3, and mice started the treatments 24 hours
after the surgery. Animals were randomly assigned into different groups: i)
Metastasis control without treatment (received daily saline solution); ii) Yoghurt
group administered yoghurt ad libitum; iii) Capecitabine (CAP) group received CAP
for 14 days, followed by a 7-days resting period, and restarted the administration foranother 14 days. i) Yoghurt + CAP group received yoghurt ad libitum and thedescribed cyclic treatment with CAP. After the second cycle of CAP, mice wereeuthanized and samples were collected. Blood cell counts, small intestine and lunghistology and serum cytokines were evaluated. Results showed that CAP treatment
decreased the metastatic areas in the lungs, while this effect was maintained in micethat received probiotic yoghurt, which by itself also was associated with a decrease ofmetastasis in some animals. CAP treatment was associated with 57 % mortality rate,
which was prevented in mice that received yoghurt. Yoghurt administration reduced
CAP side effects, improved the blood cell counts, and decreased intestinal mucositis.
CAP treatment was associated to decreases of most serum cytokines evaluated; andyoghurt administration increased the regulatory cytokine IL-10 levels in these mice.
Yoghurt administration by itself maintained cytokines levels similar to the control
but increased levels of IL-10. In conclusion, the probiotic yoghurt has the potential
to be administered as immune adjuvant by decreasing undesirable side effects inpatients under chemotherapy without affecting the primary treatment.

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HH3 MODULATION OF LUNG IMMUNE RESPONSE AGAINST
Klebsiellapneumoniae
BY
ORALLYADMINISTERED
Lacticaseibacillus rhamnosus CRL1505

R. Serda1, S. Dentice Maidana2, L. Albarracín2, M. Elean2, M.A. Jure1, J. Villena2.
1National University of Tucuman, Tucuman, Argentina. 2CERELA-CONICET, Tucuman,
Argentina. E-mail: jcvillena@cerela.org.ar

Orally administered Lacticaseibacillus rhamnosus CRL1505 enhances innate
respiratory immunity, providing increased protection against respiratory viruses andStreptococcus pneumoniae. However, the capacity of the CRL1505 strain to improve
respiratory immunity against Gram-negative bacterial infections has not beenstudied before. The aim of this work was to evaluate whether L. rhamnosus CRL1505
was able to beneficially regulate the respiratory innate immune response andenhance the resistance to hypermucoviscous KPC-2-producing Klebsiellapneumoniae of the sequence type 25 (ST25). Six-week-old male BALB/c mice weretreated with the CRL1505 strain via the oral route (108 CFU/mouse/day) during 5
consecutive days and on day 6 nasally challenged with K. pneumoniae ST25 strains
LABACER 01 or LABACER 27 (107 CFU/mouse). Mice without lactobacilli treatment
and infected with ST25 strains were used as controls. Bacterial cell counts, lunginjuries and the respiratory and systemic innate immune responses were evaluated
2 days after the bacterial infections. Both K. pneumoniae strains were able tocolonize the lungs of mice while only LABACER 01 was detected in blood samples.
LABACER 01 and LABACER 27 increased the levels of albumin and LDH in bronchoalveolar lavages (BAL) that were used as markers of the alteration of the alveolarcapillary barrier and cellular toxicity, respectively (p<0.05). The LABACER 01 strain
induced a higher lung damage than LABACER 27. The results also showed that K.
pneumoniae ST25 strains increased the levels of TNF-α, IL-1β, IL-6, IFN-γ, IL-17, KC
and MPC-1 (p<0.01) in the respiratory tract and blood, as well as the numbers of
BAL neutrophils and macrophages (p<0.05). Mice orally treated with L. rhamnosus
CRL1505 had significantly lower K. pneumoniae counts in their lungs (p<0.05) as
well as reduced levels of BAL albumin and LDH (p<0.05) than controls. The
treatment with the CRL1505 strain avoided the dissemination into blood of K.
pneumoniae LABACER 01. In addition, reduced levels of inflammatory cells (p<0.05),
cytokines and chemokines in the respiratory tract and blood (p<0.01) were found in
CRL1505-treated mice compared to infected controls. Furthermore, higher levels of
IFN-γ and the regulatory cytokines IL-10 and IL-27 (p<0.05) were found in the
respiratory tract and blood of CRL1505-treated mice than controls. These results
demonstrate the ability of orally administered L. rhamnosus CRL1505 to improve theresistance against K. pneumoniae infection and to help with the control of
detrimental inflammation in the lungs. Although further mechanistic and clinicalstudies are necessary, L. rhamnosus CRL1505 can be proposed as a candidate toimprove patients’ protection against hypermucoviscous KPC-2-producing strains
belonging to the ST25, which is endemic in the hospitals of the North of Argentina.

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SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

HH4 THE INTESTINAL FLUIDS OF PROBIOTICS FED MICETRIGGER IMMUNOMODULATORY PROPERTIES ON INNATE
IMMUNE CELLS

Martínez Monteros MJ1, Vestani A1, Perdigón G1, Maldonado Galdeano C1,2, Cazorla SI1,2
1CERELA-CONICET, Chacabuco 145. 4000 Tucumán-Argentina. 2Cátedra de Inmunología
FacultaddeBioq.,
Qca.
y
Fcia,
UNT.
Tucumán-Argentina.
E-mail:mariajosemartinezmonteros@gmail.com

In the past decade, it has been proposed that the antimicrobial activity is only
one of multiple functions of the antimicrobial peptides. We have previously
demonstrated that oral administration of probiotics increases the number of Paneth
cells, the 5-α-defensine and the antimicrobial activity of the intestinal fluids against
pathogen bacteria. Therefore, our aim was to analyzed whether oral administrationof probiotics can modulate the function and survival of key innate-immune cells.
Murine peritoneal macrophages and bone marrow derived DC were cultured at 37◦C
and 5% CO2 with LPS, medium alone or the intestinal fluids from mice fed by 7 and
5 days upon a conventional diet or the supplementation with L. casei CRL 431 or L.
paracasei CNCM I-1518, respectively. Twelve hours later, significant increases on
the costimulatory molecules CD80 and CD86 were detected on the surface of theantigen presenting cells (APC) stimulated with the intestinal fluids of the mice fed
with the probiotics (p≤0.05) was observed by flow cytometry and confocal assay.
Additionally, significant increases in IL-6 and INF-γ were found in the supernatantsof the APC stimulated with the intestinal fluids of animals fed with the probiotics, incomparison to the absence of stimuli. Most important, IL-10 counterbalance theincrease of the aforementioned proinflammatory cytokines. Intestinal fluids fromprobiotics fed mice significant promote the migration of macrophages andlymphocytes comparing to those observed in the presence of the chemotactic
medium alone (***p<0.001), as was observed in an in vitro wound-healing andtranswell migration assay. Finally, intestinal fluids of probiotics fed animals enables
the survival of the macrophages upon exposition to UV, decreasing the number ofcells upon early and late apoptosis (p≤0.05). These results point out the oral
administration of Lc 431 and Lp1518 reaches beyond their intestinal antimicrobial
effect, modulating the function and survival of key innate-immune effector cells toenhance the clearance of pathogen microbes and the resolution of inflammatory
process.

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HH5 Lacticaseibacillus rhamnosus CRL1505 AMELIORATESLIVER INJURY AND INFLAMMATION IN POLY(I:C) INDUCED ACUTE
HEPATITIS

M.J. Lorenzo Pisarello1, M. Elean1, L. Albarracín1, R.E. Serda1, F. Namai2, Y. Suda3, H.
Kitazawa3, J. Villena1
1CERELA-CONICET. 4000, Tucuman, Argentina. 2Tohoku University, 980-8572, Sendai,
Japan. 3 Miyagi University, 980-8572, Sendai, Japan. E-mail: jcvillena@cerela.org.ar

The immunomodulatory strain Lacticaseibacillus rhamnosus CRL1505
regulates the innate antiviral immune responses in the intestinal and the respiratory
tracts, allowing a higher resistance to viral challenges and protecting against the
inflammatory-mediated tissue damage. The beneficial effects of the CRL1505 strain
were not investigated in the context of viral-induced liver inflammation. This work
aimed to evaluate the potential protective effect of L. rhamnosus CRL1505 in murine
model of acute hepatitis induced by the Toll-like receptor 3 (TLR3) viral mimetic,poly(I:C). Male 6-week-old BALB/c mice fed L. rhamnosus CRL1505 in the drinking
water for 5 consecutive days (108 cells/mouse/day). Mice without lactobacilli
treatment were used as controls. On day 6, the CRL1505-treated and the untreatedcontrol mice received an intraperitoneal injection of 50 μg of poly(I:C) in 100 μl of
PBS. Two days after the TLR3 activation, the liver damage, the serum hepatic
enzymes ALT and AST as well as the expression of interferons (IFNs) (IFN-β, IFN-γ,
IFN-λ1, IFN-λ2/3), antiviral factors (Mx1, OAS1, RNAseL, IFITM3), inflammatory
mediators (TNF-α, IL-1β, IL-6), and regulatory cytokines (IL-10, IL-27) in the livertissue were determined. The intraperitoneal administration of poly(I:C) induced
inflammatory-mediated liver tissue damage, characterized by elevated levels of ALT
and AST (p<0.05), increased inflammatory cell infiltration and upregulation of IFNs
(p<0.05), and the pro-inflammatory factors TNF-α, IL-1β, IL-6 (p<0.05) compared tobasal levels. The supplementation of L. rhamnosus CRL1505 resulted in a notablereduction in pro-inflammatory mediators (p<0.05), accompanied by a significant
elevation in IFNs, IL-10 and IL-27 (p<0.01), as well as Mx1, OAS1, RNAseL and
IFITM3 (p<0.05). The differential regulation in the expression of inflammatory andanti-inflammatory factors induced by the CRL1505 strain was clearly reflected in thedecrease of poly(I:C)-induced liver damage, which was observed in the improved
levels of transaminases. The results of this work demonstrate for the first time that
L. rhamnosus CRL1505 can exert immunomodulatory and protective effects in theliver in the context of TLR3-induced acute hepatitis. The variations in the immune
factors evaluated suggest that the CRL1505 strain would have the ability to enhanceliver antiviral defenses and reduce inflammatory-mediated liver damage, making it apromising tool to prevent or reduce the severity of viral hepatitis.

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SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

HH6 EVALUATION OF A MIXTURE OF SELECTED LACTIC ACID
BACTERIA (LAB) IN A MURINE MODEL OF LEVODOPA INDUCEDDYSKINESIA (LID)
P. A. Yánez1, A. de Moreno de Leblanc1 J. G LeBlanc1.
1CERELA-CONICET.
Chacabuco
145.
4000
pyanez@cerela.org.ar

Tucumán,Argentina.

E-mail:Parkinson's disease is the second most common neurodegenerative disease
and levodopa is the most widely used drug to treat its signs and symptoms.
However, after 5-6 years of use it is associated with complications, such as
dyskinesia, that affect patients' quality of life. The possibility that changes in theintestinal microbiota may be associated with these effects is being increasingly
studied, and probiotics are being studied as complementary treatments. Previously,
in our group, the neuroprotective effect of a mixture (MIX) of three LAB(Streptococcus
thermophilus(St.)
CRL808,afolate-producing
strain,Lactiplantibacillus plantarum CRL2130, a riboflavin overproducing strain, and St.
CRL807, a strain with immunomodulatory properties) was demonstrated in a murine
parkinsonism model. The aim of this work was to evaluate the potential of thisbacterial MIX on the adverse effects associated with levodopa treatment (especiallydyskinesia), through a murine model of Levodopa Induced Dyskinesia (LID).
C57BL/6 male mice were used as model. After establishing parkinsonism throughinjections of the neurotoxic drug 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP) with intraperitoneal injections of levodopa-benserazide from day 21 wereadministered with the purpose of inducing dyskinesia. During this period ofdyskinesia establishment some groups of animals received the MIX in order toanalyze the effect during their development. At day 42, mice that did not receive LAB
were separated into two groups that either received or not the bacterial MIX toevaluate the effect once dyskinesia was established. A Motor tests were conducted
throughout the model and abnormal involuntary movements (AIMs) of both axial andlimbs were evaluated at the end of the experimental period. Mice were sacrificed andserum and brain samples were taken for cytokine determinations. The parkinsonism
model was confirmed with the motor tests. Mice received levodopa-benserazide
completed the tests in shorter times than parkinsonism control animals. being thegroup that received LAB MIX from the beginning the one that presented the best
results. However, prolonged administration of levodopa was associated with thepresence of AIMs, being the MPTP/LEVO group the one that presented the highest
score. On the other hand, mice received the LAB MIX from the beginning of themodel were the ones with the lowest score, similar to the healthy control animals.
Serum cytokines showed the highest levels in the parkinsonism control group. The
administration of levodopa decreased most cytokines without significant differenceswith the other experimental groups. Similar results were observed at the brain level,
highlighting that mice received the MIX after the model was established increased IL10, modulating the immune response at the brain level. In conclusion, the selected
LAB MIX has potential to be used as an adjuvant to levodopa, reducing its side
effects.

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SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

HH7 ADMINISTRATION OF Lacticaseibacillus paracasei subsp.
paracasei CRL75 MODULATES HUMORAL AND CYTOKINEPRODUCTION DURING FOOD ALLERGY
J. Laiño1*, M.H. Zelaya2, M.P. Taranto1.
1CERELA-CONICET, Chacabuco 145, CP 4000, San Miguel de Tucumán, Tucumán,
Argentina. 2Instituto de Bioquímica Aplicada - U.N.T, Balcarce 747, San Miguel de Tucumán,
Tucumán, Argentina. *E-mail: jlaino@cerela.org.ar

In general, food allergies are not exclusive to one type of food, but rather manyof them have cross-reactions with other foods and/or pollen. Some examples ofallergenic foods are shellfish, fish, eggs, soy, peanuts, strawberries, and cow's milk,
among others. On the other hand, lactic acid bacteria (LAB) have been demonstrated
to be useful in improving immune responses in many different diseases. Their
beneficial effect on consumer health is widely reported, as their ability to release
and/or produce specific compounds (nutraceuticals) with positive outcomes for thehuman or animal host. Many exopolysaccharides (EPS) produced by beneficial LAB
are powerful immunomodulatory molecules. Previously, it was demonstrated that
Lacticaseibacillus (L.) paracasei subsp. paracasei CRL75 has a potent
immunostimulatory effect and produces functional EPS. The aim was to evaluate theability of L. paracasei CRL75 to modulate the immune response during food allergy
induction by regulating the humoral immune response and related cytokines. The
immunomodulatory capacity of L. paracasei CRL75 was evaluated in vivo in an OVAinduced intestinal allergy mouse model. Six-week-old BALB/c mice were divided into
3 groups: a) Control (C): non-sensitized animals received a conventional balanced
diet and water ad libitum; b) OVA: animals sensitized to OVA by bi-weekly
intraperitoneal injection; c) L. paracasei CRL75 (CRL75): OVA-sensitized animals
supplemented with L. paracasei CRL75 in drinking water simultaneously to OVA
challenge. Two weeks after second injection, the intestinal allergy was induced byfeeding an egg white diet for 7 days. Samples were obtained to evaluate: a) cytokines:
IL-4, IL-10, IFN-γ, TNF-α, and IL-6; b) OVA-specific antibodies (OD450nm): IgE, IgG,
IgG1, and IgG2a; c) bacterial translocation to the liver and spleen. Results
demonstrated that administration of L. paracasei CRL75 induced the production of
IL-10 (pg/ml, C: 89.18±8.11; OVA: 176.3±18.7; CRL75: 371.99±25.49, p<0.0001)
and IFN-γ, with a significant decrease in IL-4 levels (pg/ml, C: 71.43±18.1, OVA:
941±94.77; CRL75: 545.49±66.92, p<0.0001). Regarding humoral response, L.
paracasei CRL75 induced a significant decrease of OVA-specific IgE levels in serum(OD450nm, C: 0.064±0.017; OVA: 0.717±0.085; CRL75: 0.449±0.085, p<0.001), but
increased OVA-specific IgG1 and IgG2a compared to the OVA group. No bacterialtranslocation was detected in any experimental group. In conclusion, L. parcasei
CRL75 is able to regulate the allergic immune response when administeredsimultaneously with allergy induction. L. paracasei CRL75 developed a Th-1
response, decreasing Th-2 response, with reduced levels of OVA-specific IgE and IL4. L. paracasei CRL75 constitutes a promising option for designing functional foods
with beneficial effects on food allergies.

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SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

HH8 ORAL ADMINISTRATION OF LP 1518 IMPROVES
CLINICAL OUTCOME OF L. amazoniensis INFECTION

THE

J. Arrieta1, A. Vestani1, EM Vélez1,2, C. Maldonado1,2, S. Cazorla1,2
Chacabuco 145. 4000 Tucumán, Argentina. 2Cátedra de Inmunología
Facultad de Bioquímica, Química. y Farmacia, UNT. Tucumán-Argentina. E-mail:jarrieta@cerela.org.ar
1CERELA-CONICET.

Depending on the virulence factors of the parasites and the immune response
established by the host, a wide spectrum of disease known as leishmaniasis canappear. One of them, cutaneous leishmaniasis, causes serious disability leaving
patients permanently scarred. Neither reliable nor safe vaccine is available. Thus,
there is an urgent need for development effective prophylactic and/ or therapeutic
approaches against leishmaniasis. It has been proposed that intestinal dysbiosis
may cause changes in inflammatory responses and progression of leishmaniasis.
Since probiotics contribute to maintain the intestinal homeostasis, we decided toinvestigate the effect of oral supplementation of Lacticaseibacillus paracasei (Lp)
CNCM I-1518 on the L. amazoniensis infection. Balb/c mice received the bacterium:
ten days before the infection (GI); the day of the infection (GII); or at the time thesymptoms appeared (GIII). Since the times indicated, all the animals received theprobiotics in the drinking water until they were sacrificed. GIV-Infected controls
received a conventional diet along all the experience (GIV). Mice were subcutaneous
infected in their footpads with 106 stationary-phase promastigotes of L.
amazoniensis. We observed that only the administration of the probiotics before theinfection was able to significant decrease the progressive footpad swelling caused by
the parasite infection (p<0.05). By contrast no significant differences were observedfor GII and GIII with respect to infected control animals which developed anulcerated dermal lesion. Three months after the infection mice were sacrificed andtheir large intestine, footpads and draining lymph nodes to the site of infection wereaseptically excised for parasite load and intestinal and skin microbiota analysis.
Unfortunately, no differences were observed in the parasite load neither in thefootpad nor in the draining lymph nodes of the animals. By plate count agar on
specific selective media, we observed significant increases in the count of total
anaerobic bacteria and lactobacilli of GI and GV of large intestine homogenizedsamples. Interestingly, increases UFC of Enterococcus and Streptococcus were
determined in the cutaneous lesions from GI in comparison with the contralateral
healthy skin microbiota from the same individuals and from GIV (p<0.01). Thesefindings represent the first steps to understand the potential effects of probiotics
supplementation in modulating the leishmaniasis. However, further studies arenecessary to explore the mechanisms by which Lp CNCM I1518 modify the clinicalcourse of Leishmania spp. infection.

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SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

HH9 Lactiplantibacillus plantarum CIDCA83114 AS A POTENTIAL
TREATMENT FOR HEMOLYTIC UREMIC SYNDROME

C. Cancino1, Y. Carpintero Polanco1, A. Bernal1, F. Sosa1, V. Ramos1, A. Abraham2, G.
Garrote2, M. Palermo1, R. Fernández Brando.
1IMEX-CONICET-Academia Nacional de Medicina, José Andrés Pacheco de Melo 3081. 1045
Buenos Aires, Argentina. 2CIDCA-CONICET-UNLP-CIC, Calle 47 esq. 116. 1900 La Plata,
Buenos Aires, Argentina. E-mail: cdcancino@gmail.com

Shiga toxin (Stx)-producing Escherichia coli (STEC) O157:H7 is a foodborne
pathogen, which can lead to the life-threatening Hemolytic Uremic Syndrome (HUS).
There is no treatment to reduce HUS outcome up to date. Growing evidence supports
the use of probiotics to treat and prevent intestinal diseases. Lactiplantibacillus
plantarum CIDCA83114 (CIDCA83114) is a probiotic strain isolated from milk kefir.
We have previously demostrated that CIDCA83114 is able to reduce thepathogenicity of STEC in vitro by inhibiting its growth, Stx production and activity,
adhesion to Caco-2 cells by exclusion as well as modulating the expression ofcytokines such as IL8 and TGFᵦ. The aim of this work was to evaluate the preventiveor therapeutic capacity of CIDCA83114 on the development of the characteristic
symptoms of HUS in a murine model by intragastric inoculation of a STEC strain at
weaning. Three protocols were evaluated: pre (inoculating mice 24h before and thesame day as STEC), post (inoculating mice daily with CIDCA83114 after STEC
infection) and pre-post (PP, combining both treatments). Mice and their food were
weighed daily. Blood samples were taken at 72h to determine the leukocyte formula
and urea levels as parameters of HUS outcome. Feces samples were collected at 48
and 72h, and then plated onto MacConkey agar to test the excretion of STEC and
MRS agar for lactobacilli excretion. Mortality tended to be reduced and/or delayedwith the probiotic treatments (median survival: STEC and pre=72h, post and
PP=96h, Log Rank test p=0.17), accompanied with an incremented food consumption
and mice weight at 72h (mean weight±SD: STEC=6.6±0.2; pre, post and PP=7.7±0.7,
Welch’s ANOVA p=0.13), as indicators of growth, well-being and an attenuation of
the digestive disease. Moreover, urea levels of pre and PP-treated mice showed atendency to decrease compared to post-treated and STEC mice (mean±SD [mg/dl]:
pre: 99.8±16.7, PP: 80.9±13.1, post: 121.2±39.9, STEC: 103.6±15.4; Kruskal-Walis
p=0.053) suggesting a reduction of the renal failure characteristic of HUS. Also,
STEC excretion in feces was significantly lower in mice treated with CIDCA83114 at
48h (mean±SD [UFC/mg]: STEC=2.4±0.8⋅109, pre=2.8±1.5⋅104, PP=6.0±2.7⋅106;
Kruskal-Walis p<0.05). In conclusion, CIDCA83114 accelerated STEC excretion andtended to reduce kidney damage and weight loss along with a higher foodconsumption, and to alleviate its lethality in mice. These results encourage us tokeep testing the ability of this and other LAB strains as potential treatments to
reduce HUS outcome after STEC infection.

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SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

HH10 PROBIOTICS DECREASE INFLAMMATORY BOWEL DAMAGE
IN AGING MICE

A. Vestani1, J.G. Arrieta1, E.M. Velez1,2, C. Maldonado Galdeano1,2, S.I. Cazorla1,2.
1CERELA-CONICET. Chacabuco 145. 4000 Tucumán, Argentina. 2Catedra de Inmunología
Facultad de Bioquímica, Química y Farmacia, UNT. Tucumán-Argentina. E-mail:avestani@cerela.org.ar

Inflammatory bowel disease (IBD) is a chronic, multi-causal condition
characterized by progressive structural and functional damage of the gastrointestinal
tract. Evidence from recent epidemiological data suggests that the patient population
with IBD is chronologically aging. There are several treatments to IBD including
immunomodulators, aminosalicylates, antibiotics, among others. However, many
patients do not respond to such therapies or lose responsiveness over time.
Probiotics arise as adjuvants to these conventional therapies. Balb/c mice from 120
days old were feeding during 60 days with the probiotics (1x108) Lacticaseibacillus
paracasei CNCM I-1518 (Lp1518) or a Mix of probiotic bacteria (Bio-Kult Protexin) in
the drinking water. Supplementation was administered in intervals of 10 days;
followed by 5 days of rest, in which animals received only a conventional diet. An
intestinal inflammation was induced by two subcutaneous injections of
7.5mg/kg/day of indomethacin (I) on the last two days of the experiment.
Experimental groups are as follow: G1: indomethacin (I); G2: Lp1518+I; G3: Mix+I.
Controls groups are: G4: normal control (NC); G5: Lp1518; G6: Mix. Controls
animals received 2 injection of PBS. Mice were euthanized by cervical dislocation andblood, intestinal fluids and small and large intestinal were taken. We observed asignificant body weight loss in (I) compared to (NC) (p<0.05). In contrast, mice withindomethacin-induced inflammation and supplemented with the probiotics showed
an increase in body weight compared to their pre-indomethacin injection values.
Interestingly, while (I) mice showed a significant reduction in their hematocrit(p<0.05), Lp1518+I and Mix+I presented similar values than NC mice. In
hematoxylin-eosin staining of small intestine sections, damage of the intestinearchitecture was observed. (I) animals showed dilated crypts, villi that did not run
parallel and an important shortening of the intestinal villi, compared to NC and
Lp1518+I and Mix+I mice (p<0.05). Finally, Lp 1518 supplementation restore thelarge intestine microbiota shifts induced by indomethacin, increasing the total
lactobacilli and anaerobes population. These results highlight probiotics as aneffective supplement to face aging-related disorders in IBD, thereby helping to avoid
adverse clinical outcomes among older community dwellers.

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SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

HH11 EFFECTS OF PROBIOTIC YOGHURT IN A MURINE BREASTCANCER MODEL UNDER CHEMOTHERAPY TREATMENT
M. N. Cerasuolo1, J. G. LeBlanc1, A. de Moreno de LeBlanc1.
1CERELA-CONICET.
Chacabuco 145. C.P. 4000. Tucuman,
mcerasuolo@cerela.org.ar

Argentina.

E-mail:Breast cancer (BC) remains the most commonly diagnosed cancer globally inwomen. Surgical removal of the tumor is still the best treatment; however, adjuvant
treatments are usually required to eliminate possible remaining tumor cells.
Chemotherapy is the most common treatment in these cases, which is not specificfor tumor, affecting other highly replicative cells and the patients’ quality of life. In
this sense, probiotics have been proposed as candidates, not only to decrease theseverity of non-desired side effects, but also because their immune modulating
properties. Previously our group developed a probiotic yoghurt with two starter
bacteria selected for their immunomodulatory properties demonstrated in models ofintestinal inflammation and colon cancer. The aim of this study was to evaluate theeffects of probiotic yoghurt administration in a murine model of breast cancer underoral chemotherapy treatment with capecitabine (CAP). The probiotic yoghurt was
prepared by fermenting milk with Streptococcus thermophilus CRL807 andLactobacillus delbrueckii subspecies bulgaricus CRL864. For BC induction, 4T1 cells
were injected into the upper right mammary gland of 7–8-week-old BALB/c female
mice. When tumors reached a diameter of 0.3 ± 0.1 cm, mice started receiving
different treatments. Animals were randomly assigned into different groups: i) a BC
control without treatment received unfermented milk ad libitum; ii) Yoghurt group
received yoghurt ad libitum; iii) CAP group received unfermented milk and daily oral
CAP administration during 14 days; iv) Yoghurt + CAP group received yoghurt ad
libitum and CAP. Tumor volume and body weight was assessed during the wholeexperiment. After 14 days, mice were euthanized and samples were collected. Bloodcell counts, small intestine histology, and serum cytokines were evaluated. Results
showed that probiotic yoghurt by itself decreased the tumor growth. When it wasadministered in mice treated with CAP, there was a decrease in side effects without
affecting the anti-tumor treatment. Mice that received the probiotic yoghurt showed
less intestinal inflammation, reduced weight loss, and a higher survival rate. Serumcytokines showed that yoghurt administration was associated with a modulation of
the immune response increasing the levels of IL-10 (a regulatory cytokine). In
conclusion, probiotic yoghurt administration did not interfere with the cancer
chemotherapeutic treatment, was able to reduce its side effects and modulated thehost immune response, making it an ideal candidate for adjunct therapies.

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SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

HH12 Lacticaseibacillus rhamnosus CRL1505 PEPTIDOGLYCANMODULATES TISSUE FACTOR EXPRESSION AND PROTEASEACTIVATED RECEPTOR 1 ACTIVATION DURING INFLAMMATIONCOAGULATION RESPONSE TRIGGERED BY POLY(I:C) IN THE
RESPIRATORY TRACT
H. Zelaya1,2, L. Arellano-Arriagada2, K. Fukuyama3, K. Matsumoto3, G. Marranzino2,4, F.
Namai3, S. Salva2, S. Alvarez1,2, G. Agüero1, H. Kitazawa3, and J. Villena2,3
1Institute of Applied Biochemistry, Tucuman University, Tucuman, Argentina. 2Laboratory of
Immunobiotechnology, CERELA-CONICET, Tucuman, Argentina. 3Graduate School of
Agricultural Science, Tohoku University, Sendai, Japan. 4 Universidad del Norte Santo
Tomás de Aquino (UNSTA), Tucuman, Argentina. E-mail: jcvillena@cerela.org.ar

Previously, we demonstrated that nasally administered viable or non-viable
Lacticaseibacillus rhamnosus CRL1505 as well as its peptidoglycan (PG-Lr1505),
beneficially modulates TLR3-mediated inflammation-coagulation relationship inmice. In this work, to deep in the possible mechanisms underlying this positiveeffect, we investigated the capacity of PG-Lr1505 to modulate the tissue factor (TF)
expression and protease activated receptor 1 (PAR1) activation as part of immunocoagulative response triggered by the viral pathogen-associated molecular patternpoly(I:C) in the respiratory tract. Adult BALB/c mice were nasally treated with PGLr1505 (8 µg/mL in 50 µL of PBS) for two days. Treated and untreated control mice
were then nasally challenged with poly(I:C) (250 µg/day). Mice received three dosesof poly(I:C) with a 24 h rest period between each administration. The immunocoagulative response was studied after the last administration of poly(I:C). The
challenge with poly(I:C) significantly increased blood and respiratory proinflammatory mediators and induced hemostatic changes in plasma as well as TF,
TF pathway inhibitor (TFPI), and thrombomodulin (TM) expressions in the lungs.
However, PG-Lr1505-treated mice differentially modulated the hemostaticparameters in plasma and lungs. The study of TF in lungs samples and in specificlung myeloid populations (CD45+, Gr1+, Gr1low, Gr1high, F4/80+ and, F4/80+MHCII+
cells), showed that poly(I:C) significantly increased TF expression in all the lung cell
populations evaluated, in both experimental groups. No significant differences wereobserved between the groups for the distinct myeloid populations, except for
F4/80+TF+ (p<0.01) and F4/80+MHCII+TF+ (p<0.05) lung cells, which weresignificantly lower in the PG-Lr1505 group than controls. Considering thet PARs
activation by coagulation proteases contributes to the increase of inflammation, we
evaluated the activation of the thrombin receptor PAR1 in lungs. Poly(I:C) induced asignificant increase in PAR1 activation in total lung samples, in lung CD45+, Gr1+,
Gr1low, and Gr1high neutrophils as well as in F4/80+, and F4/80+MHC-II+ cells fromboth groups. However, PG-Lr1505-treated mice evidenced a trend toward decreased
total PAR1 activation in the lung (p=0.057). These results indicate that the
peptidoglycan from L. rhamnosus CRL1505 is able to regulate TF expression
modulating the procoagulant state which could contribute to less PAR1 activation inlungs in the context of the activation of TLR3 signaling pathways, contributing to abeneficial modulation of inflammation-hemostasis crosstalk. The CRL1505
peptidoglycan could be used as a nasally administered postbiotic to reduce
inflammation-coagulation alterations during the course of respiratory viral
infections.

43SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

HH13
EFFECTOFRIBOFLAVIN-OVERPRODUCING
Lactiplantibacillus plantarum CRL2130 ON BREAST CANCERCELLS UNDER TREATMENT WITH CHEMOTHERAPEUTIC DRUGS
R. Levit1, M. N. Cerasuolo1, J. G. LeBlanc1, A. de Moreno de LeBlanc1.
1CERELA-CONICET. Chacabuco 145. 4000 Tucumán, Argentina. E-mail: rlevit@cerela.org.ar

Breast cancer (BC) is the deadliest cancer in women. Patients with early BC
are normally treated with chemotherapeutic agents that produce undesirable side
effects such as gastrointestinal toxicity that can cause the discontinuation or delaysof therapy. Therefore it is necessary to explore new strategies that may improve
oncological therapies and/or attenuate side effects. A riboflavin-overproducing
strain, Lactiplantibacillus (L.) plantarum CRL2130, was selected because of its antiinflammatory properties shown in mucositis and colitis models. The aim of thisstudy was to evaluate L. plantarum CRL2130 in vitro using a murine BC cell line
exposed to chemotherapeutic drugs. The effect of immune cells stimulated with L.
plantarum CRL2130 was also analyzed. Murine BC cell line 4T1 was cultured in thepresence of bacterium cell extract and then treated with capecitabine (CAP) or 5Fluorouracil (5-FU), evaluating viability by MTT assay, and the production of reactive
oxygen species (ROS) by using 2’,7’–dichlorofluorescin diacetate dye. L. plantarum
CRL725 (strain from which CRL2130 was derived that does not overproduce
riboflavin) and commercial riboflavin were used as controls. To study the
immunomodulatory potential, spleen cells from BALB/c mice (n=9) fed by gavage forthirteen days with L. plantarum CRL2130 (108 CFU/mL) were cultured and theconditioned medium (CM) obtained was evaluated on the viability of tumor cells
exposed to CAP. The levels of cytokines (IL-10, TNF-α and IL-6) in CM were also
determined by cytometric bead array. The results showed that L. plantarum
CRL2130 extract significantly (p<0.05) decreased the viability of tumor cells even in
the absence of CAP (71±3 %), and had a synergetic effect in its presence (56±2 %). L.
plantarum CRL2130 extract also exerted an antioxidant effect by a significantly(p<0.0001) reduction in the production of ROS both in presence (76±1 %) and inabsence (68±1 %) of 5-FU. CM from splenocytes from mice that received L. plantarum
CRL2130 decreased the viability of tumor cells even in the absence of CAP, without
interfering with the effect of this drug. The cytokines analysis showed in the CM alower level of the pro-inflammatory cytokine TNF-α than in the CM of splenocytes
from mice that did not receive the bacterium. In conclusion, L. plantarum CRL2130
showed an anti-tumor effect in the in vitro model of BC without interfering with thechemotherapeutic treatment. This beneficial effect is, at least in part, due to its
antioxidant properties (associated with the production of riboflavin) and its
immunomodulatory potential.

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SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

HH14 EFFECT OF PROBIOTICS SUPPLEMENTATION IN RENUTRITIONAL DIETS ON THE THYMUS AND INTESTINE OF
MALNOURISHED MICE

MF Balcells1,2, E Velez1,3, S Cazorla1,3, G Perdigón1, S Feliu4 y C Maldonado1,3*
1CERELA-CONICET, Chacabuco 145. 4000 Tucumán-Argentina. 2 Cátedra de Microbiología yParasitología, Nutrición. UNSTA. Tucumán-Argentina.3Cátedra de Inmunología Facultad de
Bioq., Qca. y Fcia, UNT. Tucumán-Argentina. 4Cátedra de Nutrición, Facultad de Fcia. y Bioq.
UBA. C.A.B.A. Buenos Aires-Argentina. E-mail: cmaldo@cerela.or.ar

Malnutrition is a major public health problem, affecting millions of people inour country and around the world. Nutritional imbalances compromise the immune
system causing various degrees of immunodeficiency, affects the intestinal barrier,especially the intestinal epithelial cells (IEC) and the thymus, increasing thesusceptibility to acute infections and leading to the development of more severe
forms of disease. Studies have shown that the addition of nutrient recovery diets
allows for partial or even full restoration of the thymus functionality. Our group has
demonstrated that probiotic supplementation recovers the intestinal barrier,histological alterations, and the mucosal and systemic immune functions eventhough nutrient intake remained insufficient. Our aim was to study the effect ofnutritional recovery diets supplemented with different strains of probiotics in amouse malnutrition model. Adult Balb/c mice were divided into eight groups: A) NC:
Normal Control; B) DSN: Malnutrition Control; C) DSN+F: DSN+ conventional food;D) DSN+M: DSN+Milk; E) DSN+Lp: DSN+ Lacticaseibacillus Paracasei 1518; F)
DSN+PM: DSN+ commercial probiotic MIX; G) DSN+F+M: DSN+ conventional food +Milk; H) DSN+F+Lp: DSN+ conventional food + Lp 1518; and I) DSN+F+PM: DSN+
conventional food + commercial probiotic MIX. Samples of thymus and IEC were
cultured to measure IL-6, IFN-γ, TNF-α, IL-12 and IL-10 levels. Also T lymphocyte
population was analyzed in thymus. Fatty acid profile was analyzed in serum
samples. DSN group showed an elevated proinflammatory cytokines level in Thymus
and IEC culture supernatant. Re-nutrition diets in all malnourished micesignificantly decreased levels of proinflammatory cytokines and increased IL-10
values compared to DSN mice, especially in DSN mice receiving Lp and PM. Renutrition diet with probiotic supplementation recovers LT CD4+ and LT CD4-/CD8population in thymus. PM supplementation increased the percentage of LT CD4+and Lp normalized the percentage of the LT CD4+/CD8+ population. Re-nutrition
diet in combination with Lp or PM improve the percentage of some omega 3 fatty
acids (alpha linolenic, EPA, DHA) and the omega 6 family (Linoleic and arachidonic)
and oleic, diminished in DSN mice. Our research revealed that the re-nutrition dietwith probiotics promote a better recovery of the immune system and improved fatty
acid profile compared to re-nutrition diet alone. These findings suggest that the
combination of a re-nutrition diet with probiotics may be an effective strategy to
improve health and lifespan in malnourished individuals.

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SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

HH15 EVALUATION OF A RIBOFLAVIN- OVERPRODUCING LACTIC
ACID BACTERIUM IN A MURINE BREAST CANCER MODEL UNDER
TREATMENTR. Levit1, M. N. Cerasuolo1, J. G. LeBlanc1, A. de Moreno de LeBlanc1.
1CERELA-CONICET. Chacabuco 145. 4000 Tucuman, Argentina. E-mail: rlevit@cerela.org.ar

Breast cancer (BC) is the most common cancer in women and is the leading
cause of cancer-related death worldwide. Chemotherapy is widely used for thetreatment of BC; however, it produces adverse effects in patients. The search for
adjuvant agents that minimize the toxicity of oncological drugs without reducing
their anti-tumor efficacy has become an important issue. Previously,
Lactiplantibacillus (L.) plantarum CRL2130, a riboflavin- overproducing strain,
showed an anti-tumor effect in an in vitro model of breast cancer without interferingwith the chemotherapeutic treatment. The aim of this study was to evaluate theeffect of L. plantarum CRL2130 in an in vivo model of breast cancer under treatment
with capecitabine (CAP). BALB/c mice were injected with 4T1 cells into a mammary
gland. When the tumor reached a diameter of 0.4 ± 0.1 cm, mice received an oral
treatment with CAP (cyclically for two weeks with one week of rest) and wereseparated into groups that received: L. plantarum CRL2130, L. plantarum CRL725(strain from which CRL2130 was derived that does not overproduce riboflavin) (100µl/day for one week, concentration of 1x108 CFU/ml), commercial riboflavin (in aconcentration equivalent to that produced by L. plantarum CRL2130), or
physiological solution (FS). Other groups received only the bacteria, the riboflavin or
FS. The tumor volume and weight of the mice were measured every two days. Blood
samples were taken to measure serum cytokine levels by flow cytometry. Animals
that received L. plantarum CRL2130 not only did not inhibit treatment with CAP but
also improved tumor volume decrease. These animals also presented a better generalcondition with less weight loss associated with CAP treatment. Mice that received
CAP and L. plantarum CRL2130 showed a modulation of systemic cytokines modified
by both tumor growth and CAP treatment. In conclusion, the administration of L.
plantarum CRL2130 was associated with an anti-tumor effect in the BC model.
Additionally, the riboflavin- overproducing strain was able to reduce some sideeffects associated with chemotherapy without affecting its primary anti-tumor
activity.

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SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

HH16 IN VIVO STUDY OF ANTI-LISTERIA ACTIVITY OF STRAINSOF LACTIC ACID BACTERIA ISOLATED FROM FERMENTED FOODS
C. Burgos1, B. Vasile1, P. Castellano1*, S. Salva1*.
1CERELA-CONICET.
Chacabuco
145.
4000
cburgos@cerela.org.ar

Tucuman,Argentina.

E-mail:Listeria monocytogenes is an intracellular foodborne pathogen which causes
the severe disease listeriosis in immunocompromised individuals. Macrophages playa dual role during L. monocytogenes infection by both promoting dissemination of L.
monocytogenes from the gastrointestinal tract and limiting bacterial growth upon
immune activation. Some strains of lactic acid bacteria (BAL) are able to modulate
the mucosal immune system and increase resistance to infections. This study
evaluated the effect of oral administration of 12 LAB isolated from fermented foods
on the functionality of peritoneal macrophages and resistance to L. monocytogenes
infection in a murine model. First, safety and immunobiotic potential of the selectedstrains were evaluated. Different groups of 6-week-old C57BL/6 mice were fed witheach of the isolated bacilli (108 CFU/mL/day) for 7 days. On the eighth day, both
treated and untreated control mice were evaluated for hepato/splenomegaly andphagocytic activity of peritoneal macrophages by flow cytometry. The results showedthat none of the bacilli studied induced hepato/splenomegaly. More importantly, thephagocytic capacity of peritoneal macrophages was significantly higher in mice
preventively treated with two strains of bacilli (SC411 and SC076) compared to thecontrol group (p<0.05). Then, a murine model of listeria infection was achieved. On
the eighth day, both treated mice and untreated control mice received one
intraperitoneal
doseof
cyclophosphamide(150mg/kg)
that
inducedimmunosuppression. On day 11, mice were infected orally with L. monocytogenes(109 UFC/mice). The infection resistance and innate immune response were
evaluated after the listerial challenge. The untreated group showed a highsusceptibility to listerial infection, an impaired innate immune response in peritoneal
lavage and a decrease of phagocytic cells. Interestingly, SC411 and SC076
treatments significantly increased peritoneal lavage neutrophils and macrophages,
blood neutrophils and peroxidase+ cells with respect to the untreated mice. This, inturn, significantly reduced the translocation of the pathogen to liver and spleencompared with the untreated mice. In conclusion, SC411 and SC076 lactobacilli
strains may be particularly suitable for food-based applications to reinforce the hostimmune system and induce a more efficient local innate immune response against L.
monocytogenes.

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SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

HH17
ORAL
ADMINISTRATIONOFMICROENCAPSULATED
Limosilactobacillus fermentum CRL1446 IMPROVES THE MAIN
BIOMARKERS OF METABOLIC SYNDROME IN MICE
C. Tomei1, A. Márquez1, M. Russo1, E. Andrada1, R. Medina1, P. Gauffin-Cano1.
1CERELA-CONICET.
Chacabuco
145.
4000
Tucumán,Argentina.
ctomei@cerela.org.ar

E-mail:Metabolic syndrome (MS) is one of the most relevant health problems in theworld due to the increased consumption of high-fat diets. The probiotic
Limosilactobacillus fermentum CRL 1446 has been scientifically described for its
ability to improve biomarkers of MS. Microencapsulation has become an appropriatetechnological alternative to protect probiotic microorganisms from drastic conditionsof the gastrointestinal tract or the technological process. The work aim was toevaluate the effectiveness of oral administration of microcapsules containing
Limosilactobacillus fermentum CRL1446 (CRL1446) on improving nutritional,
biochemical, and inflammatory parameters in a mice pre-clinical model in MS. Adult
male C57BL/6 mice were fed for 15 weeks and divided into the following groups(n=6). (a) Control group (Cont; mice fed a standard diet [SD]; (b) Metabolic Syndrome(MS; mice fed a high-fat diet [HFD]); (c) MS + Bacteria-free empty microcapsules(Mic); (d) MS + microcapsules containing CRL1446 strain (MicPro; mice fed a HFDplus a daily microcapsule dose in drinking water [1x109 CFU/day/mouse]). Body
weight was measured weekly. At the end of the study, animals fasted for 12 hours
and were subsequently anesthetized and sacrificed. The adiposity index wasdetermined. The following metabolic and inflammatory parameters were evaluated.
(a) Lipidic profile: total cholesterol (TC), high-density lipoprotein cholesterol, lowdensity cholesterol (LDL), and triglycerides (TG) by enzymatic methods. (b) Glycemic
profile: plasma glucose levels (enzymatic methods), Insulin (ELISA Test), and Oralglucose tolerance tests (OGTT). (c) Pro-inflammatory profile: TNF-α and IL-6
cytokines (flow cytometry) and (d) Leptin levels (ELISA Test). The results show thatthe administration of MicPro did not influence the decrease in body weight or
adiposity index. However, metabolic parameters were significantly improved. MicPro
induces a significant reduction in TG, TC, and LDL. Likewise, we observed adecrease in glucose and insulin, which allowed significant improvement in OGTT.
Also, a significant reduction in the levels of leptin and pro-inflammatory cytokines
was observed in response to the treatment. This probiotic strain maintains its
hypoglycemic and hypolipidemic properties after the microencapsulation process,which could facilitate their incorporation into different food matrices.

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SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

HH18 Lactiplantibacillus plantarum MPL16 ACTIVATES P38
MAPK SIGNALING IN C. elegans AND HAS A POSITIVE IMPACT ON
LONGEVITY AND RESISTANCE TO Enterococcus faecalis
INFECTION

M.D. Vera1, J. Sacur1,3, H. Kitazawa3, J. Villena 2,3* y M.G. Vizoso Pinto 1,3*.
1Infection Biology Lab, INSIBIO (CONICET-UNT) and Faculty of Medicine of the National
University. Av. Gral. Roca 2070. 4000 Tucumán, Argentina. 2 CERELA-CONICET. Chacabuco145. 4000Tucumán, Argentina. 3Tohoku University, Aoba-ku, 9800 Sendai, Japan. 3CFAI,
Sendai, Japan. E-mail: mgvizoso@fm.unt.edu.ar

Lactiplantibacillus plantarum strains are good probiotic candidates. Research
on Caenorhabditis elegans provides compelling evidence that probiotics canpositively influence lifespan and health through various mechanisms, including
immune modulation. Our objective was to study L. plantarum MPL16 and CRL1506
for their probiotic properties, with special focus on innate immune modulation,
lifespan increase, and resistance to Enterococcus faecalis infection. We fed C. elegans
with probiotics or E. coli Op50 as control and monitored the survival rate daily for 25
days. For immune modulation assays, we extracted RNA from the lactobacilli fed
worms after 24h and performed a RT-qPCR for genes involved in microbialassociated molecular patterns recognition (tlr-1), the PMK-1 pathway (p38 mitogenactivated protein kinase analog pathway, pmk-1), effector proteins (lysozymes: lys-1,
lys-3, lys-5, lys-8), and the tight junction protein claudin-like in Caenorhabditis (clc1), mainly expressed in the epithelial cells of digestive tube. To evaluate theprotective effect of probiotics on the susceptibility to E. faecalis infection, we fed
worms during 24h with L. plantarum MPL16 or CRL1506 and then challenged themwith E. faecalis IBL102. Nematodes at the L4 stage were extracted from NGM plateswith E. coli Op50 using a washing solution. New NGM plates were seeded with 50 µlof a 16-hour culture of strains MPL16 and CRL1506, or Op50 as control. We placed
15 to 20 nematodes per plate and determined survival every 24h and assessedbacterial count to test colonization for 7 days. We examined intestinal distention
using optical and transmission electron microscopy. Feeding C. elegans with L.
plantarum MPL16 or CRL1506 increased worms’ lifespan by approximately 20%.
Both strains increased the expression of the effector lys-5, significantly. L. plantarum
MPL16 enhanced also tir-1, pmk-1 and clc-1 expression. E. faecalis infection reduced
worms’ survival and affected their offspring. Pre-treatment of nematodes with
CRL1506 or MPL16 delayed the onset of infection-related lethality (L50) by 24h and
48h, respectively, compared to the control group. MPL16 treatment resulted in a 1
log reduction in enterococci CFU/mL and restored reproductive capacity affected byenterococcal infection. Both strains increased longevity and improved resistance to
E. faecalis infection but L. plantarum MPL16 was better at controlling infection. This
strain also modulated the expression of genes involved in the p38 MAPK signaling
pathway analog and clc-1. The upregulation of clc-1 has been related to astrengthening of tight junctions, which may improve the gut barrier function. Thesefindings in C. elegans offer promising insights that could be applicable to otherorganisms, including humans, although further research is needed to confirm theseeffects in higher organisms. These results lead us to conclude that L. plantarum
MPL16 is a promising candidate for developing probiotic supplements, and thatmodulating innate immunity pathways, such as the PMK-1 pathway, and improving
the gut barrier function are part of their mechanisms of action.

49

SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

HH19 POSTBIOTIC NASAL PRIMING ACCELERATES THE
RECOVERY OF INNATE IMMUNE RESPONSE IN RESPIRATORYMUCOSA-ASSOCIATED LYMPHOID TISSUE IN MALNOURISHED
MICE
M. Ivir1, B. Vasile1, F. Gutiérrez1, S. Avarez1*, S. Salva1*.
1CERELA-CONICET. Chacabuco 145. 4000 Tucuman, Argentina. E-mail: hivir@cerela.org.ar

Lymphoid tissue of the upper respiratory tract is responsible for
immunosurveillance of inhaled respiratory pathogens. The hosts immunosuppressedby malnutrition are known to be particularly susceptible to Streptococcus
pneumoniae. The nasal priming of malnourished mice with the peptidoglycan (PG)
from Lacticaseibacillus rhamnosus CRL1505 (Lr) is as effective as viable strain forimproving systemic and respiratory immune response. But the impact of thesetreatments on mucosa-associated lymphoid tissue is unknown. Hence, the immune
stimulation of nasopharynx-associated lymphoid tissue (NALT) and cervical lymph
nodes (CLN) induced by Lr or PG on the resistance to infection with a respiratory
pathogen (S. pneumoniae) was studied in malnourished mice under repletion
treatments. The cytokine profiles that induced the innate immune response in theinfectious process were investigated. Weaned Swiss mice were malnourished with aprotein-free diet (PFD) for 21d. Malnourished mice received a balanced conventional
diet (BCD) during 7d (BCD group) or BCD with nasal supplementation with Lr (108cells/mouse/d) or PG (8 µg/mouse/d) during the last 2d of treatment (Lr or PG
groups). Malnourished control mice (MNC) received PFD while the well-nourishedcontrol group (WNC) consumed BCD. On d8, all groups were infected with S.
pneumoniae (107 cells/mouse). Before infection, MNC showed a significant decreaseof the total cells, T and B lymphocytes in NALT and CLN as well as the macrophages,
myeloid and dendritic cells in NALT. In addition, the MNC showed an increase of
NALT Gr-1+ cells % and the microbial load in nasal washes. BCD treatment was notable to normalize these parameters. However, the Lr and PG groups improved thetotal cells, B and T cells counts in NALT and CLN. Challenge with S. pneumoniae
increased the numbers of neutrophils and macrophages and TNF-α, INF-γ, IL-6, IL17
and IL-10 levels in nasal washes. The values were lower in MNC than in WNC.
However, unlike the BDC group, Lr and PG groups showed values of NALT
phagocytes, and lymphocytes, T CD4+ cells in CLN and NALT similar to WNC mice.
Moreover, IL-10 and TNF-α levels in nasal washes were higher in Lr or PG groups.
NALT is a target for postbiotics administration to improve respiratory immunity in
immunocompromised malnourished hosts.

50

SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

HH20 EVALUATION OF LACTIC ACID BACTERIA ON THE
NUTRITIONAL, BIOCHEMICAL AND IMMUNOLOGICAL PROFILE OF
MICE FED A HIGH-FAT DIET
A. Márquez1, C. Tomei1, E. Andrada1,, M. Russo1, R.Medina1,, P. Gauffin-Cano1.
1CERELA-CONICET.
Chacabuco
145.
4000
Tucumán,Argentina.
amarquez@cerela.org.ar

E-mail:Currently, the use of probiotic lactic acid bacteria (LAB) as nutritional therapy
are a promising option to reverse the metabolic markers that characterize dietinduced obesity (DIO). The objective of this work was to comparatively evaluate theadministration of 3 strains of LAB on body weight gain (BWG), biochemical, andinflammatory parameters in mice with DIO. C57BL/6 mice were fed for 10 weeks
different diets and divided into the following groups (n=6): Control: standard diet(SD); Obese (Ob): high-fat diet (HFD); Ob+CRL1446: HFD + Limosilactobacillus
fermentum CRL1446; Ob+CRL1449: HFD + Lactiplantibacillus plantarum CRL1449
and Ob+CRL1472: HFD + Lactiplantibacillus plantarum CRL1472. The dose of LAB
used was 108 CFU/day/mouse. BWG was evaluated weekly. After 10 weeks offeeding, the livers were extracted for histological analysis, and the following
determinations were made in plasma: cytokines by CBA, and glucose and lipid
profile by enzymatic methods. Cardiovascular risk indices (CRI) were calculated thefollowing: Total cholesterol-HDL/HDL; LDL/HDL and Triglycerides/HDL. The results
were compared with the Ob group. BWG decreased by 20, 14, and 15% in
Ob+CRL1446, Ob+CRL1449, and Ob+CRL1472 groups, respectively. HFD
significantly increased TNF-α, MCP-1, and IL-6, while no significant differences wereobserved in IL-10 levels, demonstrating a pro-inflammatory effect of this dietcompared to SD. Supplementation with the different strains tended to restore thepro-inflammatory cytokines; however, this difference was not significant concerning
the Ob group. The animal groups that received LAB showed a lower fat infiltration in
the hepatocytes, and the histology of the livers of CRL1446 group was similar to theControl group. Levels of glucose values decreased in groups fed with CRL1446 (36%),
CRL1449 (34%) and CRL1472 (22%). CRI of treated groups was similar to theControl group. Interestingly, although total cholesterol and LDL cholesterol
values did not change notably, the administration of the LAB increased HDL
values in obese animals, thus improving atherogenic indices. In conclusion, theadministration of LAB significantly improves the metabolic and inflammatory
markers present in obesity, with a strain-dependent effect. Limosilactobacillus
fermentum CRL1446 turned out to be the most suitable to be used as an adjuvant in
the nutritional treatment of this pathology.

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SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

HH21
Lacticaseibacillus
rhamnosus
CRL1505ANDITS
POSTBIOTIC IMPROVE THE FUNCTIONAL PROPERTIES OF
CIRCULATING NEUTROPHILS FROM IMMUNOCOMPROMISED MICE
AND SUBJECTED TO INFECTIOUS STRESS
B. Vasile1, M. Ivir1, F. Gutiérrez1, S. Avarez1, S. Salva1*.
1CERELA-CONICET.
Chacabuco
145.
4000
Tucuman,bvasile@cerela.org.ar

Argentina.

E-mail:Polymorphonuclear neutrophils are the first line of defense against pathogens.
Blood neutrophils number depends on the balance between bone marrow
granulopoiesis and apoptosis in peripheral tissues. Many current chemotherapeutic
drugs have cytotoxic side effects that impair the life quality of the patients
conditioning their treatment. We previously showed that both Lacticaseibacillus
rhamnosus CRL1505 and its cell wall (CW) were able to improve bone marrow
emergency myelopoiesis and protection against respiratory pathogens in mice
undergoing chemotherapy. In this work we studied the functional properties ofcirculating neutrophils from immunocompromised mice fed with strain CRL1505 or
its CW, and subjected to infectious stress. Adult Swiss-mice were orally treated with
L. rhamnosus CRL1505 or CW during 16 consecutive days. On day 6, treated anduntreated mice received one intraperitoneal dose of cyclophosphamide (Cy150mg/kg). On day 9, mice were infected with Streptococcus pneumoniae (107UFC/mice). The microbicidal activity, expression of integrins and formation of
extracellular trap of neutrophils (NETs) were evaluated after the pneumococcal
challenge. The Cy group showed a decrease of number of peroxidase positive cells inblood, a low expression of CXCR4 and CD62L in Gr1+Ly6G+Ly6C- cells in bonemarrow. The reduced neutrophil activation was consistent with reduced NETs
formation in Cy-treated mice. However, L. rhamnosus CRL1505 and CW treatments
were effective to significantly increase lung neutrophils and macrophages, bloodneutrophils and peroxidase+ cells with respect to the Cy group. Besides, thetreatment with CW was more effective than L. rhamnosus CRL1505 to decrease
retention signals in the BM cells. Furthermore, mice treated with CRL strain and
CW showed a higher expression of SYTOX positive neutrophils compared to the Cy
group, evidencing a recovery of the ability to form NETs. In conclusion, L. rhamnosus
CRL1505 and its cell wall are able to modulate neutrophil homeostasis, key pieces
for the resolution of a respiratory infectious process. In this way, the scientific bases
are provided for future technological developments of products intended for thispurpose.

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SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

HH22 FUNCTIONAL CRACKERS MANUFACTURED WITH LABFERMENTED CHICKPEA FLOUR PREVENTS OXIDATIVE STRESS IN
D-GALACTOSE INDUCED AGING MICE

G. Sáez1, G. Zárate1,2.
1CERELA-CONICET. Chacabuco 145. 4000 Tucumán, Argentina. 2Universidad de San Pablo
Tucumán, Argentina. E-mail: gzarate@cerela.org.ar

Legumes are economically relevant food crops worldwide. In addition toproviding nutrients such as carbohydrates, proteins, dietary fiber, minerals andvitamins, pulses also contain diverse phytochemicals, which confer numerous
physiological and health benefits. The consumption of pulses has been associatedwith the prevention of many non-communicable diseases such as obesity, metabolic
syndrome, diabetes, cardiovascular disease, among others. In view of their benefits,
the researches on novel applications of legumes have significantly raised in the lastyears. Fermentation has been proposed as an effective biotechnological strategy for
improving nutri-functional properties of legumes as it has proven to remove
antinutrients with the simultaneous release of bioactive compounds. In previous
studies we selected 2 strains of lactic acid bacteria (LAB) isolated from legumes for
improving by fermentation the legume flours quality. In the present study functional
crackers manufactured with LAB-fermented chickpea flour were fed to D-galactose
induced aging mice in order to assess the prevention of oxidative stress. With thisaim chickpea flour was fermented for 24 h at 37°C with Lactiplantibacillus plantarum
CRL 2211 and Weisella paramesenteroides CRL 2182 (≅ 106 CFU/g). Fermented andunfermented (control) doughs were incorporated to crackers and fed C57/6BL micefor 9 weeks. The oxidative stress associated to aging was induced by a daily
subcutaneous injection of 150 mg/kg of D-galactose to mice during the whole test
period. The animals were euthanized and oxidative damage to lipids and proteins
were determined on livers, kidneys and brains by spectrophotometric methods(TBARS and DNPH, respectively) whereas relevant groups of the colonic microbiota
were determined by qPCR. Long administration of D-galactose, significantly
increased malondialdehyde and carbonyl groups in the organs and decreased the
population of lactobacilli, bifidobacteria and bacteroides in the colon. Consumptionof fermented crackers did not produced any structural alteration of the organs andprevented lipid peroxidation and the oxidative damage of proteins in the livers andbrains. These effects could be due to the high concentration of phenolic compounds
and the antioxidant activity observed in the aqueous extracts of fermented crackers.
In addition, fermented crackers prevented the decrease of colonic lactobacilli andbifidobacteria. Our results demonstrate the potential benefits of including fermented
legume flours in the development of new functional foods useful to protect the hostagainst physiological alterations related to chronological senescence.

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HH23 Lacticaseibacillus rhamnosus CRL1505 MODULATESALVEOLAR MACROPHAGES AND IMPROVE THEIR RESPONSE TO
Pseudomonas aeruginosa STRAINS ISOLATED FROM CYSTICFIBROSIS PATIENTS

L. Arellano-Arriagada1, M. Elean2, J.N. Argañaraz Aybar3, L. Albarracín1, S. Orosco4, J.C.
Valdéz3, H. Kitazawa2, N. Gobbato3, J. Villena1
1Tohoku University, 980-8572, Sendai, 2CERELA-CONICET. 4000, Tucuman, Argentina.
Japan. 3University of Tucuman. Argentina. 4Niño Jesus Children Hospital, SIPROSA,
Argentina. E-mail: jcvillena@cerela.org.ar

Lacticaseibacillus rhamnosus CRL1505 can modulate respiratory immunity
improving the resistance to respiratory virus and Streptococcus pneumoniae.
However, its beneficial effect in the context of Gram-negative bacteria-mediatedrespiratory infections was not investigated in depth. Previously, we isolated
pathogenic Pseudomonas aeruginosa (Pa) from cystic fibrosis (CF) patients andshowed that the multi-resistant (PaR) strain had a higher capacity to induce lungdamage and inflammation than the antibiotic-sensitive (PaS) strain. This work
evaluated whether nasally administered L. rhamnosus CRL1505 can modulate theresponse of alveolar macrophages to pathogenic PaS and PaR strains. Adult BALB/cmice (6-weeks-old) were nasally primed with viable (LV1505) or heat-killed (HK1505)
L. rhamnosus CRL1505 (108 CFU/mouse/day) during 2 consecutive days while
control mice received no treatment. On day 3, alveolar macrophages were isolated
from mice, cultured (105 CFU/well) and stimulated in vitro with LPS ((50 ng/ml),
PaS, or PaR (106 CFU/well). One day after the challenges, the levels of TNF-α, IL-1β,
IL-6, IFN-γ, IL-10, IL-27, CCL2, CXCL2, and CXCL10 were determined in alveolar
macrophages supernatants by ELISA kits. The results showed that the challenge ofalveolar macrophages significantly increased the levels of all the inflammatory
factors in cells stimulated with LPS, PaS and PaR although the concentrations of
TNF-α, IL-1β, IL-6, CCL2, and CXCL10 were higher in the LPS group (p<0.05) thanin groups infected with PaS or PaR. Alveolar macrophages obtained from LV1505treated mice produced higher levels of IFN-γ, IL-6, and IL-27 (p<0.05) and lower
levels of TNF-α, CCL2, CXCL2, and CXCL10 (p<0.05) compared to controls, for thethree LPS, PaS and PaR challenges. No differences were observed between LV1505treated and control macrophages when IL-1β and IL-10 were analyzed. Of note,when HK1505 was evaluated, it was observed that its effect was comparable to theobserved for LV1505, except for IFN-γ levels that were higher in the LV1505 groupthan in HK1505. The results presented here show that the nasal priming with L.
rhamnosus CRL1505 can modulate alveolar macrophages response to pathogenic P.
aeruginosa strains isolated from CF patients. Interestingly, the non-viable CRL1505
strain was as efficient as the viable bacteria to modulate alveolar macrophages´function, which could be an important advantage for its application in patients withcompromised immune system or lung functionality, such as patients with CF.

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HH24 EFFECT OF Lactiplantibacillus plantarum CELL FREE
CULTURE ON THE RESPIRATORY SUPERINFECTION INDUCED BY
BACTERIAL PATHOGENS ISOLATED FROM CYSTIC FIBROSISPATIENTS

L. Abán1, S. Orosco2, J.N. Argañaraz Aybar3, L. Albarracín4, A. Venecia5, L. Perret6, S. Ortiz
Mayor7, K. Nishiyama8, J.C. Valdéz3, H. Kitazawa8, J. Villena4, N. Gobbato3
1CONICET-Salta-Jujuy (Argentina), 2Niño Jesus Children Hospital (Argentina), 3University of
Tucuman (Argentina), 4CERELA-CONICET (Argentina), 5Maternity and Gynecology Hospital(Argentina), 6Tucuman Ministry of Health (Argentina), 7Centro de Salud Hospital (Argentina),
8Tohoku University (Japan). E-mail: nadiagobbato@hotmail.com

Lactiplantibacillus plantarum ATCC10241 cell free supernatant (LpCFS) can
reduce the lung damage and inflammation induced by pathogenic Pseudomonas
aeruginosa (Pa) strains isolated from cystic fibrosis (CF) patients, in mice.
Considering that the co-infection with Streptococcus milleri group (SMG) can enhance
the respiratory damage in Pa-infected CF patients, this work aimed to investigate the
LpCFS ability to modulate lung inflammation and damage induced by the respiratory
superinfection with Pa and SMG. A multi-resistant (PaR) and an antibiotic-sensitive(PaS) Pa strains, with the capacities to induce respiratory infections of different
severities were used. Adult BALB/c mice (6-week-old) were nasally challenged with
PaS or PaR (106 CFU/mouse), for 2d. After 5d, animals were infected with SMG (104
CFU/mouse) for 3d. Nebulization of mice with LpCFS started 1d after SMG challenge
and was performed for 1d, 3d or 5d, 5 minutes by day using a compressor/nebulizer
connected to a hermetically sealed plastic container. LpCFS was obtained from the
ATCC10241 strain cultured in MRS medium for 16h at 37°C, centrifugated for 30
min at 30,000g at 10°C and filtered through a millipore filter (0.22 µm). Micenebulized with PBS were used as controls. Results showed that the Pa-SMG
superinfections induced significantly higher lung damage, inflammation andbacterial colonization for both PaR and PaS, than the previously reported for
infections induced by PaR and PaS alone. The PaR-SMG superinfection was more
severe that the induced by PaS-SMG. The nebulization with LpCFS for 5dsignificantly reduced PaR and PaS loads in lungs (p<0.05) and the inflammatorymediated tissue damage as demonstrated by the lower levels of elastase release andleukocytes counts in broncho-alveolar fluid (BAL) samples (p<0.05) and thehistological examination. Nebulization with LpCFS for 1d did not induce changes in
the resistance of mice against the Pa-SMG superinfection, while 3d treatment wasless efficient that the 5d LpCFS administration. This work demonstrated that theseverity of Pa respiratory infection is exacerbated by SMG in mice and that thetreatment with LpCFS can reduce the lung damage and inflammation induced by thesuperinfection. Although further clinical and mechanistic studies are necessary, theresults show that the LpCFS could be an adjuvant alternative to treat Pa-SMG
respiratory superinfections in CF patients.

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HH25 MODULATION OF THE TOLL-LIKE RECEPTOR 3-MEDIATEDINTESTINAL IMMUNE RESPONSE BY WATER KEFIR

S. Dentice Maidana1, M. Elean1, L. Albarracín1, R. Ortiz Moyano1, F. Namai2, Y. Suda3, H.
Kitazawa2, J. Villena1
1CERELA-CONICET. 4000, Tucuman, Argentina. 2Tohoku University, 980-8572, Sendai,
Japan. 3 Miyagi University, 980-8572, Sendai, Japan. E-mail: jcvillena@cerela.org.ar

Kefir, a fermented beverage containing lactic acid bacteria (LAB), has been
associated to beneficial effects on the host´s health. The previous works examining
the impact of kefir on the immune system focused on milk kefir or the
exopolysaccharides and LAB strains derived from it, while water kefir has not been
evaluated. Furthermore, studies have focused on kefir's ability to modulate immune
system hemostasis and exert anti-inflammatory effects, while its specific action on
antiviral immunity has not been investigated. Then, the aim of this work was to
examine the potential immunomodulatory effects of water kefir on the intestinal
innate antiviral immunity mediated by Toll-like receptor (TLR)-3. Male 5-week-old
BALB/c mice fed water kefir ad libitum, diluted 1:5, 1:10, or 1:20 in the drinking
water, for 6 consecutive days. Mice without water kefir treatment were used ascontrols. On day 7, the treated groups and the untreated control mice received an
intraperitoneal injection of 100 μl of PBS containing 30 μg of the TLR3 agonist
poly(I:C). Two days after the TLR3 activation, the intestinal damage and the innate
immune response were studied. The intraperitoneal administration of poly(I:C)
induced inflammatory-mediated intestinal tissue damage, characterized by increased
inflammatory cell infiltration and upregulation of interferons (IFNs) (IFN-β, IFN-γ)
and pro-inflammatory mediators (TNF-α, IL-1β, IL-15, IL-6). The histological analysis
of small intestinal samples showed that mice receiving water kefir 1:5 exhibited
reduced damage and a lower inflammatory cell infiltration. Kefir treated mice hadsignificantly lower levels of serum LDH, AST, and ALT (p<0.05). In addition, lower
levels of TNF-α, IL-15, and IL-6 (p<0.01), and higher concentrations of IFN-β, IFN-γ
and IL-10 (p<0.05) were found in intestinal and serum samples of water kefir 1:5treated mice compared to controls. The treatment with 1:10 of water kefir reduced
intestinal damage and modulated cytokines but its effect was significantly lower than
the 1:5 treatment. The administration of water kefir 1:20 did not modify theparameters evaluated compared to control mice. These results indicate that water
kefir exert its immunomodulatory effects in a dose dependent manner. The in vivo
studies allow to speculate that water kefir can induce two beneficial effects on theintestinal TLR3-mediated immune response: the enhancement of antiviral defenses
and the protection against the inflammatory-mediated tissue damage. These
protective effects of water kefir require further exploration to understand how water
kefir, or its specific molecules/strains can influence the immune response and to
determine the extent of its protection against a real viral challenge.

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HH26 EFFECT OF Lactiplantibacillus plantarum CRL681 ON THE
TLR4-MEDIATED INTESTINAL INNATE IMMUNE RESPONSE
A. Baillo1, M. Elean1, R. Ortiz Moyano1, S. Fadda1, J. Villena1
Chacabuco
145.
4000
Tucuman,jcvillena@cerela.org.ar

1CERELA-CONICET.

Argentina.

E-mail:Previously, we demonstrated that the oral administration of Lactiplantibacillus
plantarum CRL681 increase the resistance to enterotoxigenic Escherichia coli
infection and that this protective effect was related to its capacity to modulate
intestinal immunity. In this work, we aimed to further characterize theimmunomodulatory capacities of the CRL681 strain evaluating its effect on the Tolllike receptor (TLR)-4-mediated intestinal innate immune response. Female 5-weekold BALB/c mice were fed L. plantarum CRL681 in the drinking water for 5
consecutive days (108 cells/mouse/day). Mice without lactobacilli treatment wereused as controls. On day 6, the CRL681-treated and the untreated control micereceived an intraperitoneal injection of 60 μg of lipopolysaccharide (LPS) in 100 μl of
PBS. On day 6 (before LPS challenge) and 2 days after the TLR4 activation, thephagocytosis of peritoneal macrophages (PMs) and the levels of TNF-α, IL-6, IFN-γ
and IL-10 in intestinal fluid and serum samples were determined. The CRL681
increased the phagocytic activity of PMs (p<0.05) while no significant differenceswere detected in the levels of intestinal and serum cytokines when CRL681-treatedand control mice were compared at basal conditions. The intraperitoneal
administration of LPS induced an increase in the PMs phagocytic activity as well as
in the levels of TNF-α, IL-6, IFN-γ and IL-10 in both experimental groups. However,mice treated with the CRL681 strain had levels of PMs phagocytic activity (p<0.05),
intestinal and serum IL-6 and IFN-γ (p<0.01) as well as intestinal IL-10 (p<0.05) thatwere significantly higher than controls. In a second set of experiments, mice wereorally treated with the CRL681 as described before. On day 6, these mice andcontrols were sacrificed, the PMs were collected, cultured and stimulated in vitro
with LPS. Twenty-four hours after the challenge, the levels of TNF-α, IL-1β, IL-6, IFNγ, IL-12, CSF2, CSF3, CCL2, CCL8, IL-27, and IL-10 were determined in culture
supernatants. Results showed that PMs from CRL681-treated mice were able toproduce significantly higher levels of IFN-γ, TNF-α, IL-6, CSF3, and IL-27 (p<0.05),
and lower levels of IL-1β, CSF2, CCL2, and CCL8 (p<0.05) than PMs from control
animals. No differences were detected in the levels of IL-12 and IL-10 when the twogroups were compared. The results show that L. plantarum CRL681 can exert
immunomodulatory effects in the gut in the context of TLR4-induced inflammation
by modifying the cytokine profile produced by peritoneal macrophages. The
variations in the immune factors evaluated suggest that the CRL681 strain can
increase immune defenses and regulate inflammation induced by TLR4 activation,
making it a promising tool to prevent or reduce the severity of not only pathogenic E.
coli infections, but also infections induced by other Gram-negative bacteria.

57

乳
酸
菌

LACTIC ACID BACTERIA AND
ANIMAL HEALTH

健
康

食
料

SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

AH1 BEER BAGASSE-ENRICHED MEDIUM IMPROVES BIOMASSPRODUCTION OF PROBIOTIC Lactobacillus johnsonii CRL2240
J. D. Babot, M. Obregozo, E. Andrada, R. Medina, J.G. Le Blanc
CERELA-CONICET. Chacabuco 145. 4000 Tucumán, Argentina. E-mail: jbabot@cerela.org.ar

Ferulates play an important role in inhibiting ruminant digestion of plant cell
walls. Therefore, the ferulic acid esterase-producing Lactobacillus johnsonii CRL2240
has been studied as a silage inoculant and goat probiotic. In order to perform farmscale experiments, a cost-effective conservation strategy must be developed, forwhich biomass production is the first step. Cultures of this strain in MRS or LAPTg
broths normally reach sub-optimal counts for a conservation process (2 × 108
CFU/mL). Therefore, the aim of this work was to design a low-cost, animal proteinfree medium that allows a higher biomass yield of L. johnsonii CRL2240. To this end,
protein or fiber-rich industrial by-products were used. L. johnsonii CRL2240 was
inoculated into different culture media, incubated at 37 °C, and CFU/mL and pH of
the cultures were determined at regular intervals. The following culture media andincubation conditions were tested: 1) LAPTg broth, static, free pH; 2) MM broth(containing texturized soybean flour), static, free pH; 3) MM broth, with stirring (75rpm), free pH; 4) MM broth, with stirring (75 rpm), pH fixed at 6.50; 5) MM broth +FeSO4, static, free pH; 6) BSG broth (containing beer bagasse) with 1.5% yeast
extract, static, free pH; 7) BSG broth with 1% yeast extract, static, free pH; and 8)
BSG broth with 1% yeast extract, centrifuged, static, free pH. Counts reached 2 ×
108 CFU/mL and pH 4.3 at 8 h when biomass was produced in LAPTg broth, while
growth in MM broth in a bioreactor with stirring or without it led to similar counts
but lower pH (3.8) for the latter condition. Similar results were obtained by adding
FeSO4 to MM broth, although cultures reached the stationary phase after only 4 h ofincubation. Remarkably, counts rose to around 1 × 109 CFU/mL and pH reached 3.8
after 6 h in BSG with 1.5% yeast extract. To further lower the cost of the broth, yeast
extract concentration was reduced to 1% without affecting biomass production.
Finally, eliminating excess beer bagasse flour by centrifugation after autoclaving
BSG broth led to similar cell counts and pH results. In conclusion, a low-cost,animal protein-free broth where L. johnsonii CRL2240 reaches counts four times
higher than LAPTg broth (1 × 109 vs 2 × 108 CFU/mL), and in a shorter incubation
time, was designed. Moreover, a valorization of a residue of the beer industry is
described.

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AH2 CHARACTERIZATION OF SURFACE PROPERTIES ANDADHESION OF AUTOCHTHONOUS LACTIC ACID BACTERIAISOLATED FROM MARE'S REPRODUCTIVE TRACT

A. Vasquetto1, J.A. Silva1, H. Mouguelar2, N. Pereyra3, J. Aguilar4, M.E.F. Nader-Macías5,
M.S. Pellegrino1.
1UNRC, FCEFQyN, Microbiology and Immunology Dept. -INCIVET-CONICET, Route 36 Km
601. 5800 Córdoba, Argentina. 2UNRC, FAyV, Anatomy Department. 3UNRC, FAyV, Animal
Pathology Dept. 4UNRC, FAyV, Animal Production Dept. 5CERELA-CONICET, Chacabuco
154. 4000 Tucumán, Argentina. E-mail: mpellegrino@exa.unrc.edu.ar

Endometritis is an inflammatory disease of the endometrium and is the mostcommon cause of subfertility in mares, causing great economic losses in equine
breeding. Since there are currently no established protocols for the treatment and/or
prevention of this disease, alternative and innocuous preventive measures, including
probiotic formulas with beneficial autochthonous Lactic Acid Bacteria (LAB), areproposed. The aim of this work was to evaluate surface-adhesive properties of LAB
isolated from mare’s reproductive tract. Twenty-nine LAB were evaluated for: *biofilm
formation, using the microplate method with crystal violet staining andspectrophotometric quantification; *exopolysaccharide production (EPS), where LAB
were inoculated in MRS with different carbohydrates (glucose, sucrose and skimmedmilk), seeded in spots and their phenotype (mucoid/ropy) observed; *in vitro
adhesion of LAB to equine epithelial cells (EEC), for which EEC and LAB
suspensions were co-incubated in equivalent proportions at 37°C for 1 h, transferred
to smears, stained with Haematoxylin-Eosin and the Percentage of Adhesion (PA)calculated by microscopic observation; *and adhesion and permanence of LAB in invivo assay, in which mare's vagina was inoculated with three preselected LAB (1010
CFU/ml): Pediococcus pentosaceus, Enterococcus hirae and Weissella cibaria, and at
24-96 hours post-inoculation, vaginal swabs taken for LAB count in MRS agar
medium and vaginal biopsies evaluated by scanning electron microscopy. All the
LAB studied showed to form biofilm on MRS broth, in higher amount on media
without surfactant. Forty-eight percent of the LAB produced EPS, most of them witha ropy phenotype in all the condition, except two strains with mucoid phenotype insucrose-added medium. Twenty-six LAB strains exhibited medium to high PA.
Referred to the in vivo inoculation assay, an increase, although not significant(p>0.05), was observed in CFU LAB numbers (~4.4 log10 CFU/ml) at day 4 postinoculation. After one day of inoculation, an increase in the number of bacteria wasobserved, with similar morphology to those inoculated, on the surface of epithelial
tissue in the treated group. The results obtained support the characterization ofthese native LABs, for their selection and subsequent inclusion in the design of ahomologous tract specific probiotic formula to prevent equine endometritis.

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AH3
Ligilactobacillus
salivarius
FFIG58RESPIRATORYIMMUNITYAND
CONFERSPROTECTION AGAINST Streptococcus pneumoniae

MODULATESLONG-TERM

M. Elean1, L. Arellano-Arriagada2, L. Albarracín1, F. Namai2, H. Kitazawa2, J. Villena1
1CERELA-CONICET. 4000, Tucuman, Argentina. 2Tohoku University, 980-8572, Sendai,
Japan. E-mail: jcvillena@cerela.org.ar

Ligilactobacillus salivarius FFIG58 was isolated from the intestine of wakamefed pigs. This strain was selected based on its ability to modulate the innate immune
responses triggered by the activation of Toll-like receptor (TLR)-3 or TLR4 signaling
pathways in the intestinal mucosa. This work aimed to evaluate whether nasallyadministered L. salivarius FFIG58 can modulate the innate immune response in the
respiratory tract and confer long-term protection against the respiratory pathogen
Streptococcus pneumoniae. Infant mice (3-weeks-old) were nasally primed with L.
salivarius FFIG58 (108 CFU/mouse/day) during 2 consecutive days and thenstimulated with the TLR3 agonist poly(I:C) for three days. Five (t1) or thirty (t2) days
after the last poly(I:C) administration mice were infected with pneumococci serotype
6B (106 CFU/mouse). Animals without lactobacilli treatment and challenged withpoly(I:C) and S. pneumoniae were used as controls. Experiments with the nonimmunomodulatory strain L. salivarius FFIG79 were performed in parallel forcomparison. Mice were sacrificed 2 days after the pneumococcal challenge toevaluate the resistance to the infection and the respiratory innate immune response.
Results of t1-experiments demonstrated that L. salivarius FFIG58 could enhance theprotection against the secondary pneumococcal infection as shown by the reduced
lung and blood bacterial cell counts (p<0.05) and the lower levels of the biochemicalparameters of broncho-alveolar lavages that evaluate lung damage (p<0.05)
compared to control and FFIG79-treated mice. The effect of the FFIG58 strain was
related to its ability to modulate the alveolar macrophages-mediated respiratory
immune response. L. salivarius FFIG58 improved the ability of alveolar macrophages
to produce IL-6, IFN-γ, IFN-β, TNF-α, IL-27, CCL2, CXCL2, and CXCL10 (p<0.05) in
response to the pneumococcal challenge compared to controls and FFIG79-treatedmice. Of note, when the resistance to secondary pneumococcal infection and theinnate immune response were evaluated in t2-experiments, the effect of the FFIG58
was comparable to that observed in t1-experiments. The results presented in thiswork show that the nasal priming of infant mice with L. salivarius FFIG58 protected
the animals against secondary pneumococcal infection for at least 30 days after the
stimulation with poly(I:C), The long-term phenotypic changes in alveolar
macrophages induced by the FFIG58 strain suggest that nasally administeredimmunomodulatory lactobacilli would be able to stimulate trained immunity in the
respiratory tract.

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AH4 INTRAVAGINAL ADMINISTRATION OF PHARMABIOTICPHYTOBIOTIC CAPSULES WITH AUTOCHTHONOUS LACTIC
BACTERIA FOR THE PREVENTION OF INFECTIONS OF THE
BOVINE REPRODUCTIVE TRACT
MH Miranda, MEF Nader Macías.
CERELA-CONICET.
Chacabuco
hmiranda@cerela.org.ar

145.

4000

Tucuman,Argentina.

E-mail:The postpartum bovine reproductive tract is susceptible to infections that aretreated with antibiotics and hormones, or a combination of them. The growing
problem of the transmission of resistance to antimicrobials requires to reduce theiruse and look for alternative therapies, such as probiotics and phytobiotics. The
objective of this work is to advance in the design of pharmabiotics/phytobiotics
formulas with bovine beneficial autochthonous lactic acid bacteria (BBALB) for theprevention of reproductive tract infections. Hard gelatin capsules containing six
different spray-dried BBALB strains (3x1010 CFU), individually or combined withphytoderivatives (Malva and Lapacho) were administered intravaginally to cows
(n=30). Two doses were applied with an interval of 15 days, before the probable dateof delivery, and two after the postpartum. The modification of the cultivable
autochthonous microbiota from vaginal washings and the permanence/colonizationof the inoculated BBALBs were evaluated, and the safety of the designed formulas
was determined through nutritional-clinical and hematological-biochemicalparameters in blood and serum. All the females of the different experimental groups(EG) remained healthy before parturition (5±0.0 RS). During the postpartum period,
two cows from the BBALB MG (mammary strains group) and one Control showedsigns of reproductive infections. The females of the different EG maintained orslightly decreased their postpartum weight (304.5 ± 4.95 KgLW). All the animals
showed normal hematocrit (36.44±1.08%), and leukocyte formula within
bibliographic reference values (RV) in the BBALB Vg, BBALB VG (Vaginal
strains)+VE (Vegetal extracts) and BBALB MG+VE groups, no significant differenceswere obtained with Control. Glycemia was lower than RV=40-88.2 mg/dl, with nosignificant differences between EG and Control at the same sampling time. Metabolic
parameters (glycemia, proteinemia, albuminemia and C-reactive protein) werenormal in all the animals throughout the protocol. The cultivable microbiota of thebovine vagina was slightly modified after the administration of the capsules. Total
aerobic mesophylls increased slightly in all the EGs after parturition.
Enterobacteriaceae remained at 2.50±0.05 logUFC/ml during the assay in all EGs,
except in BBALB Vg+VE. Lactic bacteria increased at postpartum (BBALB
Vg:2.15±0.07, BBALB Vg+VE:2.00±0.02 and BBALB MG:1.91±0.05 logUFC/ml). The
results indicate that the intravaginal administration of pharmabiotic/phytobiotic
capsules is safe and do not produce adverse effects when administered to pre- andpostpartum cows.

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AH5 SELECTION OF IMMUNOMODULATORYBACTERIA FROM DOG’S MILK AND COLOSTRUMLACTICACID

S. Quilodrán-Vega1, C. Muñoz1, L. Albarracín2, J. Toledo1, J. Villena2.
1University of Concepción, Concepción, Chile. 2CERELA-CONICET, Tucuman, Argentina.
E-mail: squilodran@udec.cl

Probiotic microorganisms are an alternative to prevent and treat infectious
enteropathies in dogs. Previously, we isolated lactic acid bacteria (LAB) from canine
milk and colostrum and characterized potential probiotic strains according to theirfunctional properties, including their resistance to gastrointestinal conditions,inhibitory effect against dog´s pathogens (Escherichia coli, Salmonella andClostridium perfringens), and intestinal adhesion. In this study we aimed to further
characterize dog´s LAB strains assessing their immunomodulatory activities. For
these studies, experiments were performed in vitro in the canine macrophage cell line
(DH82 cells) and in vivo in murine models of E. coli or Salmonella intestinal
infections. For in vitro experiments, canine macrophages (106 cells/well) were
stimulated for 12 hours with the different canine LAB strains using four doses (106,107, 108 or 109 CFU/mL). Negative controls without LAB treatment were included.
The expressions of the pro-inflammatory cytokines TNF-α and IL-8, and the patternrecognition receptors TLR2 and NOD2 were determined by real-time PCR. Among thestudied LAB, Lactiplantibacillus plantarum TUCO-16 (from canine colostrum) and
Lacticaseibacillus rhamnosus TUCO-17 (from canine milk) strains induced asignificant increase in the expression of TNF-α, IL-8, and TLR2 (p<0.05) in canine
macrophages, with the higher dose evaluated. These two strains were selected forstudies in mice. Female 5-week-old BALB/c mice were orally treated with strains
TUCO-16 and TUCO-17 (108 cells/mouse/day) for five consecutive days. On day 6,
mice were challenged with a mouse adapted enterotoxigenic E. coli (ETEC) K88 strain(109 cells/mouse) or Salmonella typhimurium (107 cells/mouse). Two days after thechallenges the resistance to the infections and the innate immune responses were
evaluated. The oral administration of TUCO-16 and TUCO-17 strains to micesignificantly augmented their resistance to pathogenic E. coli or Salmonella intestinal
infections. Both canine strains reduced intestinal damage and pathogen counts inthe liver and spleen (p<0.05) and avoided their dissemination into the bloodstream.
These protective effects were related to the ability of TUCO-16 and TUCO-17 strains
to augment the production of IFN-γ, IFN-β, IL-6, and IL-10 (p<0.05) and reduce TNFα, KC, MCP-1, and IL-15 (p<0.05) in the intestinal mucosa. L. plantarum TUCO-16
and L. rhamnosus TUCO-17 are potential probiotic candidates for improving
intestinal health in dogs, particularly for their ability to inhibit the growth of Gramnegative pathogens common in gastrointestinal infections and modulate the animal’s
immune response. Further studies are required to effectively demonstrate thebeneficial effects of TUCO-16 and TUCO-17 strains in dogs.

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AH6 EFFECTS OF Ligilactobacillus salivarius subsp. salivarius
A3iob ON HONEY PRODUCTION AND HEALTH OF Apis mellifera
BEE HIVES IN ARGENTINA

A.A. Arroyo Guerra1, J. Villena2, M.C. Audisio 1,3,4
2 CERELA-CONICET. Chacabuco 145. 4000 Tucumán, Argentina. 3 Fac.
4
de Cs. Exactas. Fac. de Ingeniería. UNSa. Av. Bolivia 5150. 4400 - Salta, Argentina. E mail:arroyoguerraagustin@gmail.com
1 INIQUI-CONICET.

The honeybee (Apis mellifera) plays a key role in the pollination of crops andwild plants globally, significantly contributing to ecosystem balance. In Argentina,
their importance is noteworthy, positioning the country as the third-largest honeyproducer and second-largest exporter worldwide. However, beehive health isthreatened by the Varroa sp. mite, which feeds on bees' fat bodies and spreads
various infectious agents, weakening colonies and reducing honey production. This
study, analysed the effects of Ligilactobacillus salivarius subsp. salivarius A3iob, astrain isolated from the gut of a worker bee, in experimental hives under field
conditions. The strain was administered monthly to each hive from June to October
2022. Five hives per group received the following treatments: 1) viable bacteria at aconcentration of 109 cfu/mL and 2) thermal-treated bacteria (98ºC for 10 minutes).
These groups were compared to a control group treated with sterile MRS broth. The
parameters studied were i) honey production, ii) abdomen weight, and iii) incidenceof Varroa sp. Statistical evaluation was carried out using a non-parametric method(Kruskal-Wallis), considering significant differences as those with a p-value ˂ 0.05.
Hives treated with viable bacteria produced significantly more honey (14.2 ± 6.06 kg,
p=0.0381) than the control hives (3.55 ± 2.42 kg). The thermal-treated group showedsimilar performance to the control group (5.80 ± 3.9 kg). Additionally, a significant
increase in abdomen weight was observed in bees treated with live bacteria (58.70
mg ± 16.33) compared to the control group (40.6 mg ± 12.5, p=0.0003). The thermaltreated group showed similar results to the control (48.79 mg ± 14.08). Regarding
varroa infestation rates, no significant differences were found between the treatedgroups at the end of the trial (p= 0.1399). However, the viable bacteria group had ahigher incidence of the parasite (2.20% ± 3.36) compared to the thermal-treated(1.20% ± 1.26) and control groups (0.68% ± 1.05). These results indicate that A3iob
improves honey production and abdomen weight in treated bees. The hives with theviable bacteria produced considerably more honey (14.2 ± 6.06 kg, p=0.0381) thanthe control group (3.55 ± 2.42 kg). The abdomen weight of bees treated with live
bacteria was significantly higher than the control group (58.70 mg ± 16.33,
p=0.0003). However, the higher varroa incidence in the live bacteria group suggests
the need for further studies to better understand these interactions.

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AH7 IDENTIFICATION AND BENEFICIAL CHARACTERISTICS OF
AUTOCHTONOUS LACTIC BACTERIA WITH PROBIOTIC POTENTIALFOR CANINE PUPPIES

N. Carrasco, R. Levit, C. Aristimuño Ficoseco, A. de Moreno de LeBlanc, J.G LeBlanc, M.E.F.
Nader-Macias.
CERELA-CONICET.
Chacabuco
145.
4000
Tucumán,Argentina.
E-mail:ncarrasco@cerela.org.ar

Probiotics are frequently applied for the restoration of the autochtonous
microbiota and prevention of infections, being of main importance the host andmucosa of isolation. For the selection process of new homologous probiotic
microorganisms, functionality and technological aspects must be evaluated. This
work is aimed to evaluate different beneficial properties and to identify lactic acidbacteria (LAB) isolated from mother's milk and faeces of different dog breeds, inorder to advance in the design of a homologous LAB probiotic formula to restore thedog gut microbiota and protect them from infections. 100 different strains wereisolated from dogs: 17 from mother's milk, 83 from faeces, and were evaluated in the
production of beneficial enzymes (protease, lipase, amylase, cellulase and feruloyl
esterase) and H2O2 production. Thirty strains were selected for sharing beneficial
properties and genetically identified. The ability to adheres to intestinal cells wasevaluated in 10 strains. Protease was determined in agarized skim milk, lipase in
MRS-milk cream, amylase in MRS-starch agar medium (revealed with Indole),
feruloyl esterase in 1% in methanol (w/v) ethylferulate 1g/L added to MRS agar
without glucose, cellulase in MRS-carboxymethyl cellulose agar (revealed with lugol).
H202 production was detected using the tetramethyl benzidine-MRS agar (TMB-MRS)
plate method. Genetic identification was performed by sequencing the 16s RNA gene.
The ability of LAB strains to adhere to the gut epithelium was assessed in vitro using
the Caco-2 human colon carcinoma epithelial cell line. The survey of beneficial
enzymes showed 24% strains had protease activity, 20% cellulase activity and 19%
feruloyl esterase activity, 39% produced H202 (12% weak, 9% strong and 18% verystrong). The genetic identification indicated: 4 Lactobacillus johnsonii, 2Ligilactobacillus salivarius, 2 Lactiplantibacillus plantarum, 2 Streptococcus
thoraltensis, 2 Pediococcus acidilactici, 7 Enterococcus (E.) canintestini, 4 E. faecalis,
6 E. faecium and 1 E. durans. 10 selected LAB strains evidenced adhesion ability
close to 70%. This work supports the selection of LAB strains with the best
characteristics, and to define those that share properties, in a way to decide theiroptimal combinations, to advance in the design of probiotic formulas withhomologous strains for the health of canine puppies.

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AH8 MIXED INOCULATION USING FERULIC ACID ESTERASEPRODUCINGLACTOBACILLI
IMPROVE
NUTRITIONAL,FERMENTATIVE AND FUNCTIONAL PROPERTIES OF CORNSTOVER SILAGE

E. Andrada1,2, N. Suarez1,2, M. C. Abeijón-Mukdsi1, R. Medina1,2.
1CERELA-CONICET. Chacabuco 145. 4000 Tucumán, Argentina. 2Facultad de Agronomía,
Zootecnia y Veterinaria, Universidad Nacional de Tucumán, Tucumán Argentina. E-mail:rmedina@cerela.org.ar

Mixed inoculants combine different species of lactic acid bacteria to potentiate
its effects, and currently dominate the silage market. Ferulic acid esterase producing(FAE+)-inoculants can potentially enhance the fiber digestibility by ruminants,
therefore enhancing their productive performance. Up to date, mixed FAE+
inoculants are scarcely studied. We have previously studied three FAE+ lactobacilli
that induced positive and different fermentative and nutritional modifications in cornstover when used as single-strain inoculants. In this work, we aimed to evaluate theeffects of the mixed inoculation of these strains. Therefore, an uninoculated control(UN) group was compared to the inoculation of two formulae: Lactiplantibacillus
plantarum CRL2241 and Levilactobacillus brevis CRL2239 (Mix LPB), or these strains
plus the addition of Lactobacillus johnsonii CRL2240 (Mix LPBJ). Inoculants wereapplied (106 CFU/g fresh matter) and vacuum-sealed. In 60-day corn stover silos,
fermentative (soluble carbohydrates, organic acids and ethanol), nutritional (fiber
digestibility) and functional (total phenolic compounds, concentration ofhydroxycinnamic acids, antiradical activity) parameters were determined. Likewise, ametagenomic analysis of the bacterial community was carried out. The studies
showed that both mix formulae reduced dry matter losses (2% LPB vs UN), andincreased the free ferulic acid content (10X vs UN) and antioxidant activity of cornstover. Metagenomic analysis revealed a strong lactobacilli dominance in the
inoculated silages. When compared to LPB, LPBJ further reduced dry matter loss
(4% vs UN), and cellulose content (2% vs UN). In conclusion, beneficial modifications
were induced by mixed inoculants when considering their fermentative, nutritionaland functional parameters.

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METABOLISM AND PHYSIOLOGYOF LACTIC ACID BACTERIA

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MP1 GROWTH OF LACTIC ACID BACTERIA STRAINS ISOLATEDFROM SILAGE IN CORN PLANT EXTRACT MEDIA

R. M. Lenz1, M. C. Audisio1,3,4, C. Ibarguren2,3.
1INIQUI-CONICET. 2Fac. de Cs. de la Salud. 3Fac. de Cs. Exactas. 4Fac. de Ingeniería. UNSa.
Av. Bolivia 5150- 4400 - Salta, Argentina. E-mail: rominalenz22@gmail.com

Biological additives, mainly lactic acid bacteria, are one of the best options forcontrolling and improving the silage fermentation process. In silage, the watersoluble carbohydrates are fermented under anaerobic conditions, creating an acidic
environment that inhibits the proliferation of undesirable microorganisms. The aimof this work was to determine the growth capacity of lactic acid bacteria strains,
isolated from corn and sorghum silages, in a corn plant extract medium, todetermine their potential as silage inoculants. We worked with 6 out of 28 isolates
obtained from sorghum and corn silages samples, selected mainly for their acidifying
capacity. These strains were phylogenetically characterized as Lactiplantibacillus
plantarum R3EA, L. plantarum F1M, Pediococcus pentosaceus R1M1, P. pentosaceus
A1B2, P. pentosaceus B2M and Limosilactibacillus reuteri AUM1. A corn extract
medium (MEM) was used as culture medium, obtained by maceration of a chopped
fresh corn plant in sterile distilled water (10% w/v) for 10 min, with subsequent
filtration and sterilization in an autoclave (121°C, 15 min). Growth of each strain was
evaluated by inoculating (1% v/v) a suspension in PBS buffer. These suspensions
were obtained by recovering of the pellet of a fresh overnight culture in MRS broth,centrifuging and double rinsing with PBS buffer, and then resuspending in the same
buffer. The cultures were incubatedat 37°C under microaerophilic conditions. The
viability of each strain was determined by plate counting using MRS agar at 0, 2, 4,
6 and 24 h. Based on these results, a strain of each genus was selected.
Subsequently, the growth of L. plantarum R3EA, P. pentosaceus A1B2 and L. reuteri
AUM1 in a mixed culture in MEM was evaluated, with and without addition of
glucose (0.2, 0.5, 1% w/v) as additional carbon source. All strains maintained their
viability in MEM. Regarding the growth of each individual strain, the cultures
maintained a viability ca. 7-7.5 log cfu/ml during 24 h, except for L. plantarum F1M,
which recorded a viability of ca. 5.5 log cfu/ml at 24 h. In the case of mixed cultures,
the best growth profile was observed in the MEM + 0.5% w/v glucose, with a viabilityof ca. 7.6 log cfu/ml after 24 h, while in media without added glucose they ca. 6.5-7
log cfu/ml. These assays allowed the selection of 3 over 6 strains with suitableproperties as potential inoculants for corn silage, according to their capacity fordevelopment in a medium with reduced nutrients. Furthermore, the addition, of analternative carbon source to the plant substrate could optimize silage fermentation
process.

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MP2 NITROGEN SOURCES AS A KEY FACTOR REGULATING THE
EXPRESSION OF GAD GENES IN Levilactobacillus brevis CRL 2013
M.P. Urquiza Martínez, P.G. Cataldo, G.A. Bulacios, J.R. Naja, L. Saavedra, E.M. Hebert.
CERELA-CONICET.
Chacabuco
145.
4000
Tucumán,Argentina.
E-mail:purquiza@cerela.org.ar

Gamma-aminobutyric acid (GABA) is a non-protein amino acid that primarily
functions as an inhibitory neurotransmitter in the central nervous system. Interest
in GABA-enriched foods has surged in recent years due to its antihypertensive, antiinflammatory and antidepressant effects. Given the low natural abundance of GABA
in food products, microbial fermentation emerges as a highly promising approach for
producing this valuable bioactive compound. The conversion of glutamate to GABA
in lactic acid bacteria (LAB) is carried out by the GAD system, which consists of thetranscriptional regulator (GadR), the glutamate:GABA antiporter (GadC) and the
enzyme glutamate decarboxylase (GadB and/or GadA). Levilactobacillus brevis CRL
2013, an efficient producer of GABA, harbors two gad genes encoding glutamate
decarboxylase enzymes, gadA and gadB. Notably, gadB is located adjacent to gadC,with the transcriptional regulator gadR identified upstream of gadC. To elucidate theregulation of GABA production in L. brevis, we analyzed the transcriptionalorganization of the gad genes and evaluated the effect of different nitrogen sources ina chemically defined medium (CDM) on GABA production. Transcriptional analyses
of gad related genes in L. brevis CRL 2013 identified two transcriptional units; onecomprising the gadR and gadC genes, and the other encompassing gadC, gadB, andgltX genes. To further investigate the physiology of GABA production, we optimized a
CDM and supplemented it with glutamate to analyze its impact on GABA production.
While glutamate supplementation alone did not activate GABA synthesis in CDM, theaddition of nutritional factors beyond the natural amino acids present in CDM, suchas yeast extract (YE), significantly enhanced GABA production. YE contains peptides,
nucleotides, proteins, amino acids, sugars, and various trace elements. Then, todetermine whether nitrogenous components activate GABA synthesis in L. brevis, weanalyzed growth, pH, and GABA production in CDM containing different amounts(0.5% to 5%) of casitone (digested casein) and vegetable peptone. In 2% casitone,
GABA production was similar to that obtained in CDM supplemented with 1% YE,
where maximum production was observed. Although 2% vegetable peptone also
showed GABA production, the levels were not comparable to those observed with
Casitone. Transcriptional analysis confirmed a substantial increase in gadB and
gadR expression upon casitone supplementation, further supporting their role inactivating the GABA production pathway.

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MP3 ANTIMICROBIAL PROPERTIES OF Levilactobacillus brevis
STRAINS ISOLATED FROM CHEESE WHEY OF DIFFERENTORIGINS

J.C. Alcocer1,2, R.M. Lenz1 N. Álvarez1, C. Coronel1, H. Cristóbal1, C. Ibarguren1,2, M.C.
Audisio1,3,4.
1INIQUI-CONICET. 2Fac. de Cs. de la Salud. 3Fac. de Cs. Exactas. 4Fac. de Ingeniería. UNSa.
Av. Bolivia 5150. 4400 - Salta, Argentina. alcocerjimena20@gmail.com

Industrial cheese whey (ICW) and artisanal cheese whey (ACW) are
differentiated by the use of lactic bacteria cultures and commercial rennet in theformer, and natural rennet or “panchera” in the latter. The aim of this work was toevaluate the antimicrobial spectrum of previously isolated strains from ICW and
ACW, and to characterize them phylogenetically. A total of 23 isolates (5 strains from
ICW and 18 from ACW) were grown in MRS broth, with glucose replaced by lactose(MRSL), at 37°C for 24h, under microaerophilic conditions. Then, a screening was
carried out using the agar diffusion technique against indicator strains(Staphylococcus aureus, Bacillus cereus, Listeria monocytogenes and Salmonella sp.),to select those strains with the best antimicrobial activity. Subsequently, the effect of
direct contact of cell-free supernatants (CFS) of two strains of Lactobacillus sp., at pH
4 and after the neutralization with 1M NaOH, on the viability of a cell-suspension of
S. Typhimurium ATCC14028 was evaluated. The viability of the pathogenic strain
was determined at 0, 1, 2 and 24 h by plate count in BHI agar medium. A
suspension of the pathogen in BHI broth was used a pathogen growth control.
Finally, the isolates with the best antimicrobial profile were selected andcharacterized phylogenetically by sequencing the 16S rRNA subunit (Macrogen,Seoul, Korea) and performing an online search for similarities using the BLAST
program in GenBank. The agar diffusion technique showed that the strains with an
inhibition zone ≥ 6 mm against the indicators tested were those identified as LS91
and LS94 from ICW and ACW, respectively. On the other hand, the direct contact
with S. Typhimurium ATCC14028, at acidic pH. LS91 produced a total inhibition
after 24 h, while LS94 generated a decrease of 4 log orders (ca. 6 log cfu/mL) during
the 24 h of contact. On the other hand, the SLC at neutral pH together with thecontrol maintained a viability between 7 and 9 log cfu/mL. The analysis of the 16S
rRNA subunit revealed that LS91 and LS94 exhibited 97% and 99% homology with
Levilactobacillus brevis, respectively. In conclusion, the isolated strains from ICW
and ACW presented antimicrobial activity, mainly due to the production of organic
acids. The phylogenetic identification allowed to corroborate that the isolates
correspond to bacterial species that are present in artisanal cheeses and cheese
whey, and are usually used in industrial fermentations.

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MP4 BILE ACIDS MODIFY MORPHOLOGY OF A POTENTIALPROBIOTIC LACTIC ACID BACTERIUM

C. Moreno1, N. Gómez1, A. Ledesma1,2, M. P. Taranto3, A. Y. Bustos1,4,5
Ruta Nacional 9 Km 1125, Villa El Zanjón. 4206 Santiago delEstero, Argentina. 2Facultad de Ciencias Exactas y Tecnología, Universidad Nacional deSantiago del Estero. Av. Belgrano (s) 1912. 4200 Santiago del Estero, Argentina. 3CONICETCERELA. Chacabuco 145. 4000 Tucumán, Argentina. 4Facultad de Agronomía yAgroindustrias, Universidad Nacional de Santiago del Estero. Av. Belgrano (s) 1912. 4200
Santiago del Estero, Argentina. 5Facultad de Ciencias Sociales, Humanidades y de la Salud,
Universidad Nacional de Santiago del Estero. Av. Belgrano (s) 1912. 4200 Santiago delEstero, Argentina. E-mail: yaninabustos17@gmail.com
1CIBAAL-UNSE-CONICET.

Bile acids (BAs) are the main endogenous modulators of the composition andmetabolic activity of the intestinal microbiota. In the present work, the effect ofconjugated [taurodeoxycholic (TDCA)] and free [cholic acid (CA) and deoxycholic acid
(DCA)] BAs, on the survival, structural and surface properties of Lentilactobacillus(L.) parabuchneri CB12 was evaluated applying viability assays and ScanningElectron Microscopy (SEM). Our results evidenced that free BAs were more toxic
than conjugated, with CA being significantly more harmful than DCA). SEM showedthat untreated bacterial cells exhibit well-defined morphology characteristic withshort rod shape and a smooth surface. No bacterial aggregates were observed. With
TDCA addition, the cell surface appeared rougher and irregular with somedepressions. Granular precipitate corresponding to the DCA released by the action of
the CB12-bile salt hydrolase enzyme was observed. Treatment with free BAs such as
DCA and CA caused significant alterations at lower concentrations. Thus, bacterial
cells treated with 2 mM DCA, and 0.05 mM CA showed significant surface
distortions and many bacteria presented depressions and fold formation on thesurface. In general, cells were longer than control cells and interrupted septa as
signal of incomplete cell division were also observed. SEM micrographs showedbacterial aggregates, which would be related to modifications in their surface
properties by exposure to BAs. Remarkably, exposure to CA revealed pore-like
depressions. In general, the magnitude of the effects founded was dependent to thetype of BA and its concentration, being more evident in the presence of CA even at
low concentrations, which would explain its greater inhibitory effect. This work
provides useful information about the effects of BAs on lactic acid bacteria strains,and it would allow the development of strategies to positively modulate thecomposition of the microbiota.

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MP5 RAMAN SPECTROSCOPY, AN ALTERNATIVE METHODOLOGYFOR THE IDENTIFICATION OF LACTIC ACID BACTERIA

A. Ledesma1,2, J. Carol1, Y. Bustos1,3,4
Ruta Nacional 9 Km 1125, Villa El Zanjón. 4206 Santiago delEstero, Argentina. 2Facultad de Ciencias Exactas y Tecnología, Universidad Nacional deSantiago del Estero. Av. Belgrano (s) 1912. 4200 Santiago del Estero, Argentina. 3Facultad deAgronomía y Agroindustrias, Universidad Nacional de Santiago del Estero. Av. Belgrano (s)1912. 4200 Santiago del Estero, Argentina. 4Facultad de Ciencias Sociales, Humanidades yde la Salud, Universidad Nacional de Santiago del Estero. Av. Belgrano (s) 1912. 4200
Santiago del Estero, Argentina. E-mail: carolpazjuanjose@gmail.com
1CIBAAL-UNSE-CONICET.

Conventional methods of bacterial identification usually rely on biochemical
and molecular tests, which are often time-consuming and expensive. In this context,
Raman spectroscopy can be a valuable and attractive tool in bacterial identification,
because it allows obtaining single-cell spectra within a population and describes theDNA, protein and lipid composition of the cell. The sample is illuminated from theincident laser, it scatters the light with the vibrations of the molecular bonds
providing information which defined the identity of cell kind, and it allows
discriminations and classification of microorganisms at level of a single cell. For thisreason, the objective of this work was to obtain Raman spectra of a group of 17 lactic
acid bacteria (LAB), previously isolated by our group and identified by MatrixAssisted Laser Desorption Ionization-Time of Flight (MALDI-TOF) Mass Spectrometry(MS) and 16S gene sequence. For this purpose, cells from a stationary phase culture
were resuspended in 100 mM sodium phosphate buffer (pH 7.4) before processing.
The spectra were obtained using a confocal Raman spectrometer with a 532 nm
laser excitation source. A single bacterial cell was focused using a 100 X objective.
Each spectrum consisted of an average of 10 measurements per strain, with an
exposure time of 5 seconds in the range of 100 to 2000 cm-1. The profile ofcharacteristic bands attributed the major cell components (proteins 1660 and 533
cm−1, carbohydrates 980 cm−1, lipids 887, cm−1 and nucleic acid 1081 and 391 cm−1)
observed for each analysed strain, allowed us to differentiate the three groups ofgenera identified as Leuconostoc mesenteroides, Lentilactobacillus parabuchneri and
Lactiplantibacillus plantarum. For the last genus three subgroups were differentiated.
This could be due to differences between the molecular composition at the strainlevel. In addition, statistical treatment of spectra through principal component
analysis and partial least squares discriminant confirms it. In conclusion, Raman
spectroscopy served to identify certain LAB, at least, at the genus and species level,
positively correlating with the results obtained through genetic sequencing and mass
spectrometry. However, it is still necessary to obtain more readings per cell to
confirm if this methodology allows differences in bacteria at the strain level.

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MP6 MONITORING THE PROTEOLYTIC ACTIVITY OF LACTIC ACID
BACTERIAUSING
FOURIER
TRANSFORM
INFRARED
SPECTROSCOPY

J. Carol1, M. Ibarra1, A. Ledesma1,2, Y. Bustos1,3,4.
1CIBAAL-UNSE-CONICET. Ruta Nacional 9 Km 1125, Villa El Zanjón. 4206 Santiago delEstero, Argentina. 2Facultad de Ciencias Exactas y Tecnología, Universidad Nacional deSantiago del Estero. Av. Belgrano (s) 1912. 4200 Santiago del Estero, Argentina. 3Facultad deAgronomía y Agroindustrias, Universidad Nacional de Santiago del Estero. Av. Belgrano (s)1912. 4200 Santiago del Estero, Argentina. 4Facultad de Ciencias Sociales, Humanidades yde la Salud, Universidad Nacional de Santiago del Estero. Av. Belgrano (s) 1912. 4200
Santiago del Estero, Argentina. E-mail: carolpazjuanjose@gmail.com

During fermentation of milk to produce cheese or yogurt, the microorganisms
used as starters progressively modify the casein structure and chain length, giving
rise to oligopeptides and free amino acids. The study of the changes induced by thecomplex proteolytic system of microorganisms requires procedures that, in general,are not very sensitive. In fact, there are few routine instrumental methods for the
study of conformational changes of complex proteins. Thus, the objective of thiswork was to monitor the proteolytic activity on casein of the lactic acid bacteria (LAB)
strains Lactiplantibacillus (L.) plantarum CB2, Lentilactobacillus (L.) parabuchneri
CB7 and L. plantarum CB11, by Fourier transform infrared spectroscopy. In
addition, total proteins and free amino acids were quantified using the Bradford and
OPA methods, respectively. For this purpose, cell suspensions prepared fromstationary phase cultures of the strains were incubated with 5 mg/mL casein at 37°C and samples were taken at baseline, 16 and 20 hours. In the supernatants, theintensities of the amide I and II bands of the proteins were recorded between 1500
and 1700 cm-1, which refer to the tension and bending of the carbonyl and aminegroups. Comparison of the spectra obtained with the control casein showed thatboth the amide I and amide II bands presented notable differences after incubation
with the different strains. The results showed a significant decrease, proportional tothe incubation time, in the intensity of both bands for casein incubated with L.
plantarum CB2 and L. parabuchneri CB7 strains. Remarkably, with L. plantarum
CB11 at 20 h, the intensity of the amide II band increases and shifts to higher
wavelengths, indicating possible formation of other compounds. In relation to protein
quantification, CB2 and CB7 strains decreased the initial casein concentration by
21.13 and 43.43 %, respectively. On the other hand, L. plantarum CB11 showed nosignificant protein consumption. Amino acid quantification shows an increase for thethree strains evaluated, with a higher value observed for CB11. From these tests it
can be concluded that infrared spectroscopy served as a highly sensitive
methodology to study the degree of hydrolysis of three LAB, whose proteolytic activity
could serve to increase the nutritional value of fermented foods in terms of peptides
and amino acids content.

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MP7 BIOTRANSFORMATION OF SELENIUM BY LACTIC ACID
BACTERIA UNDER DIFFERENT STRESS CULTURE CONDITIONS
F.R.E Molina1, L.G. Ruiz Rodríguez1, R.R. Raya1, F. Mozzi 1
1CERELA-CONICET.
Chacabuco
145.
4000
Tucumán,fmolina@cerela.org.ar

Argentina.

E-mail:Selenium (Se) is an essential micronutrient for most living organisms. Se isingested through food consumption of animal and plant sources, which absorb thismetalloid from the soil. Se concentration, redox state, and bioavailability are highlyvariable and depend on the geographic region. Se is known for its antioxidant, antiinflammatory, and antiviral activities and its deficiency in humans is associated withviral infections, male infertility, different types of cancer, and aging. Some lactic acidbacteria (LAB) can biotransform inorganic Se (iSe) into seleno-amino acids andseleno-nanoparticles (SeNPs), which are more bioavailable than inorganic Se salts.
This work aimed to evaluate the ability of Weissella cibaria 25 and Lactiplantibacillus
paraplantarum CRL 2051 to grow and bioaccumulate Se in the presence of thismetalloid under different stress culture conditions. Both strains were grown in MRS
with sodium selenite (5 ppm) as iSe for 24 h under the following culture and stressconditions: i) 30 and 37 °C, ii) pH 4.0, 5.5, and 6.6, iii) 0.05% (v/v) H2O2, and iv) 4
and 6% (w/v) NaCl. Microbial growth was determined by OD600; SeNPs formation was
positively visualized by the reddish color of the colonies on MRS agar after
incubation at 30 °C for 48 h. The intracellular bioaccumulation of iSe was indirectly
determined in the culture medium supernatant by the di-amino naphthalene
colorimetric method. The results showed that the presence of Se did not significantly
affect the microbial growth of W. cibaria 25 and L. paraplantarum CRL 2051 underthe majority of the assayed conditions. LAB colonies, typically white, presented a
reddish color in the presence of Se indicating their ability to reduce sodium selenite
to elemental Se (Se0) and form SeNPs. Both strains CRL 25 and CRL 2051 showed
the highest microbial growth (∆DO: 0,40 and 0,57), and iSe intake (97 and 76%),
respectively, when cultured at 30 °C and pH 6.6. A higher temperature, lower pH,
and the presence of H2O2 and NaCl caused a decrease in Se accumulation andmicrobial growth in both strains. At pH 4.0 and 5.5 the intracellular Se decreased
13% in both cases. The addition of 6% NaCl affected Se accumulation the most in
both LAB, being L. paraplantarum the most sensitive strain. Under oxidative stressculture conditions, a decrease of 31 and 47% intracellular Se in W. cibaria and L.
paraplantarum, respectively, was recorded. In summary, both assayed strains could
grow in the presence of and biotransform Se under different stress culture
conditions; however, they could not cope with the presence of H2O2. In most of thestudied conditions, Se helped to counteract the detrimental effect caused by theassayed stress factors.

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MP8 AROMATIC PROFILE OF WINES PRODUCED BY MIXEDCULTURES OF YEASTS AND Oenococcus oeni

L.M. Mendoza1, M. Flores2, C. Belloch2
1CERELA-CONICET. Chacabuco 145. 4000 Tucumán, Argentina. 2IATA-CSIC. Avda. Agustín
Escardino 7. 46980 Paterna, Valencia, España.
E-mail: lmendoza@cerela.org.ar

During wine-making process, the alcoholic fermentation conducted by yeasts
is the main reaction transforming grape sugars into ethanol. However, the malolactic
fermentation (MLF) is a critical step to obtain high quality wines. The MLF consistsin the conversion of malic acid into lactic acid and it is desirable because decreases
total acidity and improves the stability and sensorial characteristics of wines. Amongwine lactic acid bacteria, Oenococcus oeni is the best adapted species and is almost
exclusively used for the induction of MLF. In addition, it was descripted that somestrains of O. oeni are able to produce several enzymes (glycosidases, esterases,
phenolic acid decarboxylases, citrate lyases) that modify the wine aroma profile. The
aim of this study was to evaluate the potential to produce aroma volatile compounds
by different strains of O. oeni in combination with wine yeasts. Three strains of O.
oeni (X2L, ST and Sb10) were evaluated in mixed cultures with Saccharomyces
cerevisiae Cf8, Hanseniaspora vineae mc1 and Metschnikowia pulcherrrima T2.
Bacteria were inoculated at 24 h of alcoholic fermentation and at the end of thisfermentation in synthetic wines. Samples were incubated at 25 and 15 °C for 14 and25 days, respectively. Volatile compounds were analyzed using a gas chromatographwith a quadrupole mass detector. Regardless of the studied strain, the inoculation of
O. oeni modified the concentrations of esters, alcohols and diacetyl in comparison towines produced without MLF, being their content higher at 25 °C than 15 °C. The
differences between the two temperatures could be related to better bacterial growthat 25 °C. O. oeni X2L showed the greatest modifications in the aromatic profile withhigh 2-phenylethanol and aldehyde concentrations but low ethyl hexanoate content,
whereas strains ST and Sb10 increased ethyl octanoate concentration. Based onthese results, it could be indicated that each O. oeni strain showed a differential
effect on wine volatile composition. Further studies with grape must are needed toknow whether O. oeni inoculation and the observed changes in volatile compounds
improve wine aroma.

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APPLIED BIOTECHNOLOGY OF
LACTIC ACID BACTERIA

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AB1 INHIBITION OF ENTEROHEMORRHAGIC Escherichia coli BY
Lactiplantibacillus plantarum CRL681: AGGREGATION, COAGGREGATION, AND ELECTRON MICROSCOPY STUDIES
A. Baillo1, S. Fadda1, J. Villena1,
1CERELA-CONICET.
Chacabuco
abaillo@cerela.org.ar

145.

4000

Tucuman,Argentina.

E-mail:Lactiplantibacillus plantarum CRL681 (Lp681) inhibits the growth ofenterohemorrhagic Escherichia coli (EHEC) in various matrices. The cell surface is
crucial to compete with pathogens for specific binding sites. Therefore, good selfaggregation and co-aggregation with pathogens are desirable features in potentiallyprobiotic strains. This study compared the inhibitory capacity of three
Lactiplantibacillus plantarum strains (Lp681, Lp1506 and a mutant of Lp1506)
against EHEC in a chemically defined medium (CDM), in addition to evaluating theirability to self-aggregate and co-aggregate with EHEC. Finally, the interaction of
Lp681 with E. coli was observed by scanning electron microscopy (SEM) andtransmission electron microscopy (TEM). Growth curves were performed for eachstrain in 100 mL of CDM, inoculating 106 CFU/mL of the strains and 104 CFU/mL of
EHEC, incubated at 30°C for 48 hours. Samples were taken at different times todetermine pH and viability. Aggregation assays were performed by inoculating 4 mL
of CDM with a concentration of 108 CFU/mL for each bacterium and incubated at
30°C for 24 hours without shaking. For co-aggregation assays, 2 mL of each LAB
culture with 108 CFU/mL was mixed with 2 mL of the EHEC culture with 108
CFU/mL, incubated at 30°C without shaking for 6 hours. In both assays, opticaldensity (OD) was measured at 600 nm. The results showed that Lp681 inhibits
EHEC growth at 24 hours of incubation. No viable counts were observed at 48 hours
when EHEC were co-cultured with Lp1506. No inhibitory activity was detected for
LpMut. In self-aggregation assays, significant differences were observed at 6 hours of
incubation, with Lp681 achieving 80% self-aggregation, compared to 27% and 36%
for Lp1506 and LpMut, respectively. At 24 hours, Lp681, Lp1506 and LpMut
reached values between 92% and 74%, with Lp681 and Lp1506 being the most selfaggregating. In coaggregation tests with EHEC, Lp681 showed significantly higher
values than the other strains, reaching 49% at 6 hours. SEM and TEM images
showed that Lp681 surrounds EHEC cells, evidencing cell lysis with disruption of
the cell membrane and cell wall. The results show that L. plantarum CRL681 caninhibit EHEC in a chemically defined medium by cell lysis. In addition, it has highself-aggregation and co-aggregation abilities, suggesting its potential to bind toepithelial mucosa and prevent the adhesion of pathogens such as EHEC.
Lipoteichoic acid is important for aggregatory and inhibitory capacity, as seen bycomparing strain Lp1506 and its mutant. L. plantarum CRL681 proved to be more
efficient than Lp1506 in inhibiting EHEC and coaggregating, making it a strain ofinterest for probiotic applications.

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AB2 EFFECT OF SELECTED STARTER CULTURES ON THE
MICROBIOLOGICAL,PHYSICOCHEMICAL,ANDSENSORY
CHARACTERISTICS OF CURED-FERMENTED SAUSAGES MADE
WITH LLAMA (Lama glama) MEAT
L. Nelegatti1, A. Baillo1, M. Torino1, S. Fadda1
Chacabuco
145.
4000
sfadda@cerela.org.ar

1CERELA-CONICET.

Tucuman,Argentina.

E-mail:Llama meat (LM) is a viable alternative to beef and pork in the Andean
regions. Its high protein and low fat content make it a promising product for the
innovative food market. The formulation of specific starter cultures (SC) for thefermentation of LM constitutes an interesting biotechnological strategy for its
comprehensive utilization. This approach will facilitate the diversification andpreservation of this raw material while obtaining additionally products with added
value. The objective of this work was to produce fermented sausages with LM usingSC, composed of previously selected Lactic Acid Bacteria (LAB) and a CoagulaseNegative Staphylococcus (CNS). Three batches (B) were obtained: B1 (control) withspontaneous fermentation; B2 inoculated with L plantarum CRL681 and S. carnosus
F833; and B3 with L. sakei CRL2243 and S. vitulinus GV318. Handmade curedsausages of the salamín type were produced and evaluated at 0 (initial time), 3(fermentation), and 17 days (end of maturation) through microbiological,
physicochemical (pH, water activity -aw-, lactic acid, color, and free amino acid -FAAcontent), and sensory analysis. The results confirmed the competence of LAB and
CNS in the meat curing environment, as at 17 days and in all three batches, LAB
reached a count of approximately 9 log CFU/g, while CNS reached between 7-9 log
CFU/g. Additionally, during the fermentation stage (3rd day), a significant decreasein pH was observed in the batches with SC compared to the control, demonstrating ahigher acidifying capacity of the LAB strains of the SC. B2 recorded the highest lactic
acid production at the end ripening (7.78 g/g). The typical drying process of theseproducts was evidenced by a decrease in aw, from 0.94 to 0.85±0.01 in B2 and B3,
while B1 showed a slightly smaller decrease (aw 0.89). The color parameters (L*, a*,and b*) also varied at the final time. The increase in L* and a* in the 3 batchessuggests brighter and more intensely red-colored sausages. The more pronounced
decrease in b* in B2 and B3 resulted in products with less yellow hue. Increase in
FAA was observed due to various metabolic processes such as proteolysis. However,
the higher content of FAA in B2 and B3 stands out. The hygienic quality of all final
products was adequate, with no pathogenic bacteria detected. Sensory analysis withtrained judges revealed that sausages with SC scored higher in overall uniformity,
spice aroma, spicy, acidic, and umami flavors, and chewiness. On the other hand, B1
recorded higher scores in less favorable attributes such as greasy texture, rancidity,
moldy and strange odor, bitter taste, and pasty mouthfeel. This work demonstrates
that the use of selected SC produces safer and improved quality Llama meat
fermented sausages.

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AB3 LACTIC ACID BACTERIA ISOLATED FROM CITRUS FRUITS AS
STARTER CULTURES FOR THE PRODUCTION OF FERMENTEDLEMON JUICE

D.A Barrionuevo1,2, F. Escobar1, A.S Isas1, F.R.E Molina1, C.P Van Nieuwenhove1,3
Chacabuco 145, 4000, Tucumán, Argentina. 2Facultad de Bioquímica
Química y Farmacia-UNT, Batalla de Ayacucho 471, S.M. de Tucumán, Argentina.
3Fundacion
Miguel Lillo, Miguel Lillo 251, S.M. de Tucumán, Argentina. E-mail:dbarrionuevo@cerela.org.ar

1CERELA-CONICET.

Lemon (Citrus limon L.) is widely cultivated in Tucumán, representing one of
the main economies of the region. The juice possesses bioactive compounds ofinterest for the prevention of diseases such as obesity, diabetes, and cardiovasculardisorders, with a high content of polyphenols (mostly flavonoids) and vitamin C.
Lactic acid bacteria (LAB) isolated from fruits can adapt to the fruit matrix and be
used as starter cultures to obtain new fermented beverages with longer shelf life. The
objective of this work was to genetically characterize five LAB strains isolated from
citrus fruits and evaluate their potential use as starter cultures into a pasteurizedlemon juice (LJ: 2.0° Brix; pH 4.6 adjusted with sodium carbonate). For
identification, a fragment of the 16S rRNA gene was amplified by PCR and using a
specific primer (plb16) from the bacterial DNA, then purified and sequenced.
Afterwards, sequences were compared with others deposited in the GenBank
database. The LAB were identified as Leuconostoc citreum (three strains) andWeissella bombi (two strains). Subsequently, Lc. citreum M5C1 and W. bombi M6C2
were selected as starter cultures and inoculated at 2% (v/v) in LJ. Fermentation was
carried out at 30°C for 48 h, then stored at 4°C for 28 days. Aliquots of samples werecollected during fermentation (0; 24; and 48 h) and at the end of shelf life (28 days)
to evaluate bacterial growth (by plating on agar media), pH, total phenolic content(TPC) and antioxidant activity (TEAC, FRAP and DPPH methods). After fermentation,
both strains achieved 1 log unit growth (Lc. citreum M5C1: from 7.4 ± 0.12 to 8.79 ±0.10; W. bombi M6C2: 6.90 ± 0.13 to 8.55 ± 0.08 CFU/mL), remaining constant after
28 days at 4ºC. Despite the microbial growth, the pH of the juices slightly increased
from 4.6 to 4.8-5.0 after fermentation, possibly attributed to citric acid metabolism.
Both strains increased TPC around 40-50% after 48 h of fermentation (p< 0.05), witha consequent increase in the antioxidant activity. The viability of both LAB remained
at similar values after storage. However, there was a significant decrease on TPC andantioxidant activity (p< 0.05). The results demonstrated the ability of L. citreum
M5C1 and W. bombi M6C2 to tolerate the citrus matrix and increase thefunctionality of the beverages after fermentation. The uses of autochthonous starter
cultures allow greater adaptation to adverse substrates and enhances the functional
properties of lemon juice.

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AB4 PARTIAL CHARACTERIZATION OF Enterococcus SPP. JK06
ISOLATED FROM REGIONAL SALAMI FROM TUCUMÁN

J. Gomez, F. Gharzia, K. Gianni de Carvalho
PROIMI-CONICET. Avenida Belgrano y Pasaje Caseros. 4000 Tucuman, Argentina. E-mail:ka20_04@hotmail.com

The food industry is always looking preservatives that are highly effective,
long-lasting, and nowadays, of natural origin. Additionally, those capable ofinhibiting various pathogens such as Listeria monocytogenes, Salmonella spp., andEscherichia coli are preferred. In this regard, bacteriocins produced by lactic acidbacteria (LAB) represent an effective, natural method that can also provide benefits
when supplied together with the producing strain, acting as a probiotic. This work
aimed to identify an LAB isolate from regional salami from Tucumán, determine its
safety and probiotic characteristics, evaluate the production of antimicrobial
compounds, and assess its application in a food model for biopreservation. The LAB
isolate named JK06 was identified by 16S rRNA gene sequencing and Gram staining.
The presence of virulence factors was evaluated using phenotypic assays. The ability
to grow on MRS agar with bile and resistance to the gastrointestinal tract (GIT) were
evaluated at pH values of 1.5, 2, 2.5, and 3 (gastric juice, GJ) for 120 min and at pH
8 (enteric juice, EJ) for 24 h. Cell-free supernatants (CFS) were evaluated for theirinhibitory capacity against Listeria innocua, L. monocytogenes, E. coli, Enterococcus
faecalis, E. faecium, Pseudomonas aeruginosa, Staphylococcus aureus, Salmonella
enterica, and S. typhimurium. The active compound causing inhibition wasdetermined by neutralization, heating to 100°C, and treatment with proteinase K,
catalase, and lipase. The inhibitory capacity of selected LAB and/or bacteriocins on
L. monocytogenes Scott A was evaluated during hamburger storage for 15 days at
4°C. Based on the position of isolate JK06 in various phylogenetic trees obtained, it
was concluded that the LAB belongs to the Enterococcus genus. Enterococcus sp.
JK06 was found to be non-hemolytic and did not exhibit lipase or gelatinase activity.
Additionally, it was capable of growing in MRS medium supplemented with 3 g/L ofbile salts. The ability of Enterococcus sp. JK06 to withstand simulated GIT conditions
was evaluated, and no viable cell count was observed under the most extreme
conditions (pH = 1.5 and 2) after 30 min of exposure to simulated GJ. However, thestrain recovered its viability when treated with simulated EJ for 1 h. It was also
observed that the inhibitory activity of the CFS was unaffected by heat treatment orby the action of lipase and catalase but was affected by proteinase K. As for the foodmodel, it was observed that after 5 days of storage at 4°C, there was a complete
elimination of Listeria, while in control hamburgers, the count remained stable (10³CFU/mL). The active compound responsible for antimicrobial activity is of a protein
nature, possibly a bacteriocin. Given the results observed throughout the study, it
can be concluded that Enterococcus sp. JK06 and/or its bacteriocin have highpotential for use as a probiotic or food additive, respectively.

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AB5 FUNCTIONAL AND SENSORY ANALYSIS OF CALAFATE FRUITFERMENTED WITH PATAGONIAN LACTIC ACID BACTERIA

B. Sede Lucena1, L. Crespo2, O. Ordoñez1, J. Burini3, M. Bruzone3, F. Mozzi2, M. Pescuma1de Investigación y Extensión Forestal Andino Patagónico (CIEFAP), Ruta 259 Km
16,24 - CC 14 (9200) Esquel, Chubut, Argentina.2CERELA-CONICET. Chacabuco 145. 4000
Tucumán, Argentina. 3Centro de Referencia en Levaduras y Tecnología Cervecera (CRELTEC),
Instituto Andino Patagónico de Tecnologías Biológicas y Geoambientales (IPATEC), CONICET
Universidad Nacional del Comahue, Quintral 1250 (CP8400), San Carlos de Bariloche,
Argentina. bsede@correociefap.org.ar
1CentroCalafate (Berberis microphylla) is a native Patagonian barberry used inculinary and medicinal practices. This study aimed to analyze the ability of lactic
acid bacteria (LAB) isolated from calafate fruits and flowers to modify the phenolic
compound concentration and antioxidant capacity of calafate juice. Additionally, theinhibition of metabolic syndrome-related enzymes by the fruit juices was tested, andthe sensory attributes of the selected fermented juice were analyzed. The total
phenolic compounds in juice supernatants were determined using the FolinCiocalteu method, and antioxidant activity was measured by four methods (ABTS•+,
DPPH, FRAP, and ILP). The inhibition of α-glucosidase, α-amylase, and lipase wasexpressed as a percentage relative to the control (enzyme activity without juice
supernatants) based on the total phenolic compound concentration in the juice’s
supernatants (mg GAE/ml). Sensorial analysis was conducted by the trained sensory
panel of IPATEC according to ASBC methods. Leuconostoc citreum F2,
Lacticaseibacillus paracasei B4, Fructobacillus fructosus B7, and Latilactobacillus
curvatus B34 grew in the calafate juices (1.33 ± 0.03-2.61 ± 0.30 log CFU/ml at 24h). F. fructosus B7 consumed glucose and fructose the most (2.30 ± 0.45 g/L and3.73 ± 0.44 g/L, respectively) producing the natural sweetener mannitol (3.89 ± 0.77g/L). All juices exhibited antioxidant capacity although, the juice fermented with L.
paracasei B4 showed the highest total phenolic compound concentration (2662.58 ±344.51 mg GAE/100 ml) and antioxidant capacity as measured by ABTS•+(38916.42± 2157.52 µmol TE/100 ml). The juices inhibited metabolic syndrome-related
enzyme activities, with the lowest IC50 values for α-glucosidase observed in juices
fermented by F. fructosus B7 and L. paracasei B4 (0.56 ± 0.10 and 0.64 ± 0.05 mg
GAE/ml, respectively), and for α-amylase in juices fermented by L. curvatus B34 andL. paracasei B4 (0.34 ± 0.01 and 0.37 ± 0.06 mg GAE/ml, respectively). The nonfermented juice IC50 values were 0.47 ± 0.05, 0.77 ± 0.02 and 0.20 ± 0.02 for αamylase, α–glucosidase and lipase, respectively. Based on antioxidant and enzyme
inhibitory activity, the L. paracasei B4 fermented juice was selected for sensory
evaluation and compared with the non-fermented one. Both juices had a fruity
aroma, but the L. paracasei B4 fermented juice had distinctive lactic acid anddiacetyl notes. Both samples were sweet and fruity, with the L. paracasei B4 juice
also having higher acidity and lactic acid content. L. paracasei B4 could be suitablefor producing a functional fermented calafate juice.

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AB6 Pleurotus pulmonarius MUSHROOM AS NOVEL RESOURCES
OF PREBIOTICS: EFFECT ON THE GROWTH AND RESISTANCE TO
PASSAGE THROUGH THE GASTROINTESTINAL TRACT OF LACTIC
ACID BACTERIA

J. Gomez1, A. Ledesma1,2, M. Taranto3, A. Bustos1,4,5
1CIBAAL-UNSE-CONICET. Ruta Nacional 9 Km 1125, Villa El Zanjón. 4206 Santiago delEstero, Argentina. 2Facultad de Ciencias Exactas y Tecnología, Universidad Nacional deSantiago del Estero. Av. Belgrano (s) 1912. 4200 Santiago del Estero, Argentina. 3CONICETCERELA. Chacabuco 145. 4000 Tucumán, Argentina.4Facultad de Agronomía yAgroindustrias, Universidad Nacional de Santiago del Estero. Av. Belgrano (s) 1912. 4200
Santiago del Estero, Argentina. 5Facultad de Ciencias Sociales, Humanidades y de la Salud,
Universidad Nacional de Santiago del Estero. Av. Belgrano (s) 1912. 4200 Santiago delEstero, Argentina. E-mail: yaninabustos17@gmail.com

Prebiotics improve host health by promoting the growth and metabolic activityof beneficial strains such as probiotic lactic acid bacteria (LAB). Due to theirnumerous benefits, the search for and characterization of new prebiotics is an areaof growing interest. In this sense, the Pleurotus (P.) pulmonarius mushroom is rich in
non-digestible carbohydrates that could act as prebiotics favoring the growth andactivity of beneficial bacteria. The objective of this work was to extract potential
prebiotic oligosaccharides from P. pulmonarius and evaluate their efficiency tomodulate the growth and resistance of probiotic LAB applying simulated
gastrointestinal tract (SGIT) conditions. For this, the relative growth rate (RGR),
prebiotic activity (PA), prebiotic index (PI) and survival through SGIT of selected LAB
strains in the presence of mushroom oligosaccharides was evaluated. The strains
were inoculated in MRS broth supplemented with fungal carbohydrates as the only
carbon source or with glucose or commercial FOS as controls. Our results showedthat fungal carbohydrates exert a positive effect on the growth of selected LAB strain
compare to glucose and FOS. So, Levilactobacillus (L.) brevis ATCC 14869,
Lactiplantibacillus (L.) plantarum CB11 and Lentilactobacillus (L.) parabuchneri CB12presented RGR and PA > 1. On the other hand, Bifidobacterium longum DSM 20219,
and Limosilactobacillus reuteri ATCC 23272 did not grow in the presence of fungal
carbohydrate or FOS. A PI of 0.47-0.55 was obtained indicating that the extract was
selectively utilized by beneficial microorganisms in relation to glucose. After gastricpassage, no significant difference was observed compared to the initial count. Afterincubation in intestinal juice, the colony-forming capacity of L. brevis and L.
plantarum CB11 decreased approximately 0.5 log units when glucose was used as acarbon source, while in the presence of fungal oligosaccharides, viability wasmaintained at initial levels of 8 log CFU/mL. Our results indicate that components of
P. pulmonarius could stimulate the growth and resistance of certain probiotic strains
during the SGIT passage, which would allow its use in the design of efficient
symbiotic prototypes.

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AB7 INHIBITION OF Α-GLUCOSIDASE BY LACTIC ACID BACTERIATHROUGH CONTROLLED FERMENTATION OF Brassica rapa AND
Undaria pinnatifida

B. Martínez Caliari1, R. Parada2, F. Sosa1,2, M Vallejo1, K. Gianni de Carvalho3
de Biotecnología Bacteriana, Fac. de Cs. Naturales y Cs. de la Salud (SedeTrelew) -Univ. Nac. de la Patagonia San Juan Bosco. 9 de julio 25, (9100) Chubut, Argentina.
2Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires,
Argentina. 3 PROIMI-CONICET. Avenida Belgrano y Pasaje Caseros. 4000 Tucuman,
Argentina. E-mail: ka20_04@hotmail.com
1 Laboratorio

Type 2 diabetes mellitus (T2DM) is a metabolic disorder associated withinsulin resistance and dysfunction of the hormone-producing beta cells of thepancreas. T2DM is increasing in prevalence worldwide and today represents 95% ofdiabetes cases, becoming a serious public health problem, especially in developing
countries. Fruits and vegetables are natural sources of active ingredients that
promote health. Specifically, brassicas serve as a model for determining theinfluence of biochemical processes during fermentation on the contribution of
metabolites related to nutrition and health. Previous studies have demonstrated anincrease in bioactive compounds in fermented vegetables and the functional
properties exhibited by seaweed, particularly Undaria pinnatifida. This study aimed
to develop new functional foods through two-step controlled fermentations using
isolated autochthone strains of the genus Leuconostoc spp. and Lactiplantibacillus
spp, whose consumption can inhibit α-glucosidase activity, an enzyme linked toT2DM. The inhibitory effects of extracts from spontaneous and controlled
fermentation of Brassica rapa (Chinese cabbage) and U. pinnatifida were assayed.
The fermentation process was conducted at 18 °C for 30 days. Then, samples were
dehydrated, and water and methanol extracts were prepared using a solid-to-liquidratio of 1:10 w/v. Alpha-glucosidase activity was assayed using maltose as a
substrate, and the amount of glucose liberated was measured using the glucose
oxidase-peroxidase method (GOD-POD). The reaction rate of α-glucosidase activity
and enzyme inhibition by water and methanol extracts was analyzed by MichaelisMenten kinetics using Lineweaver-Burk plots. The Michaelis constant (Km) andmaximal velocity (Vmax) were determined by the Lineweaver-Burk plots equation. No
significant difference was detected between the Vmax results of control,
spontaneous, and controlled fermentation in both types of extracts. On the other
hand, Km values mM were 1.94, 2.95, and 7.98 for control, spontaneous, andcontrolled fermentation, respectively, when water extracts were assayed. In the caseof methanol extracts, Km values mM were 3.12, 3.27, and 7.04 for control,
spontaneous, and controlled fermentation, respectively. These results suggest thatmetabolites from the fermentation of Chinese cabbage and U. pinnatifida exertcompetitive inhibition of α-glucosidase activity, with an enhanced effect in controlled
fermentation. Controlled fermentation could thus be a potential method fordeveloping anti-diabetic foods that can improve diabetes management.

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AB8 PHYSICOCHEMICAL AND MICROBIOLOGICAL STABILITY,AROMA PROFILE AND SENSORY ATTRIBUTES OF CHILTO JUICEFERMENTED WITH Lactiplantibacillus paraplantarum IBFV-10
L. Contreras1, E. Salguero1, M.I. Isla1,2, S. Torres1,2.
1INBIOFIV, CONICET-UNT. S. Martin 1545. 4000 Tucumán, Argentina. 2FCN e IML, UNT.
Miguel Lillo 205. 4000 Tucumán, Argentina. E-mail: storres@csnat.unt.edu.ar

The fruits of chilto (Solanum betaceum), a plant native to South America, arenatural functional foods rich in health-beneficial bioactive compounds (phenolics,
flavonoids, anthocyanins, and carotenoids). However, fresh chilto fruits and theirproducts, such as juice, have a short shelf life. Producing fermented chilto juice withprobiotic lactic acid bacteria offers a promising alternative to extend shelf life andenhance its functional, nutritional, and organoleptic qualities. This study aimed toassess the microbiological and physicochemical stability, sensory properties, andaroma compounds profile of fermented chilto juice using previously selected
Lactiplantibacillus paraplantarum IBFV-10, a potentially probiotic strain
autochthonous from chilto fruits. Fermented chilto juice (FJ) was prepared byfermenting (24 h; 37 °C) pasteurized juice (64 °C; 30 min) with IBFV-10 strain andthen stored for 30 days at 8°C. Unfermented pasteurized chilto juice (CJ) stored
under the same conditions was used as a control. The microbiological quality ofjuices during storage was studied by investigating the presence of total mesophiles,
fungi, yeasts, enterobacteria, and lactic acid bacteria (LAB). Also, thephysicochemical parameters titratable acidity, pH, total soluble solids, browningindex, and color were evaluated in both juices. The aroma characterization wasinvestigated by GC–MS, and a sensory evaluation was conducted through pairwise
comparison analysis and acceptability testing (7-point hedonic scale) using
untrained panels. The FJ showed no growth of contaminating microorganisms
during the 30-day storage period at 8°C. In contrast, the CJ exhibited contamination
with various bacteria, including total mesophiles, LAB, and enterobacteria. The
physicochemical parameters of the FJ were maintained throughout refrigerated
storage. Contrarily, the CJ experienced a decrease in the pH values, an increase in
the titratable acidity and the browning index, and color changes, all indicative of
deterioration. The fermentation altered the volatile profile of chilto juice, resulting inchanges in aroma compounds and sensory differences compared to the CJ. Certain
aroma compounds that are characteristic of chilto fruits (methyl butanoate, ethyl
butanoate, methyl hexanoate, (Z)-3-hexen-1-ol, and eugenol) decreased after thefermentation, while others like ethyl acetate and acetic acid, increased. In line with
this, the sensory analysis showed different perceptions of sensory attributes between
the FJ and the CJ. However, panelists showed satisfactory acceptability of bothjuices, with no significant preference between them, expressing willingness toconsume and purchase the FJ product. This study demonstrates that fermenting
chilto juice with L. paraplantarum IBFV-10 boosts the shelf life of the juice. Further,
distinctive new organoleptic characteristics of the FJ could promote its acceptanceas a functional product by the non-dairy probiotic food market.

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AB9 IN VITRO EVALUATION OF THE ANTAGONISTIC POTENTIAL
OF Ligilactobacillus murinus 26B1 ISOLATED FROM MOUSE
AGAINST Staphylococcus aureus ATCC 6538P FOR ITSAPPLICATION IN ANIMAL NUTRITION

J. Palma1, I. Sandoval1, A. Llorente2, C. García3, S. Fadda4, C. Mercado3, C. Soto3
1Posgrado, FESC Cuautitlán, UNAM, San Sebastián Xhala, Cuautitlán Izcalli 54714, México.
2Ingeniería y Tecnología, FES Cuautitlán, UNAM, San Sebastián Xhala, Cuautitlán Izcalli
54714, México.3 Ciencias Biológicas, FES Cuautitlán, UNAM, San Sebastián Xhala,
Cuautitlán Izcalli 54714, México. 4CERELA-CONICET. Chacabuco 145. 4000 Tucumán,
Argentina. Email: llorente@unam.mx

Due to their ability to produce various metabolites with antagonistic activity,
lactic acid bacteria (LAB) are being focus of study for their probiotic potential for thecontrol of pathogens of sanitary concern. In this project, the objective was todemonstrate the antagonistic activity of Ligilactobacillus murinus 26B1 (L. murinus)
isolated from the mouse intestinal tract, through bacterial inhibition tests and in situ
zymography, against Staphylococcus aureus (S. aureus), one of the main causal
agents responsible of foodborne diseases. L. murinus 26B1 was isolated from theintestinal contents of lactating mouse. Molecular characterization (16S rRNA) was
performed, and the antagonistic activity of its culture supernatants was evaluated.
Proteins were quantified by the Bradford method, by means of the Spot on the Lawn
technique and in situ zymography, inhibitory and lytic activity were evaluated,
respectively; the relative molecular mass was determined (SDS-PAGE). The biofilm
formation capacity of both strains, L. murinus and S. aureus ATCC 6538P, wasevaluated by crystal violet staining. In compliance with the requirements for new
probiotics, established by FAO-WHO-WHOSA (2022), the identification results (16S
rRNA) showed that strain 26B1 belongs to the genus Ligilactobacillus and species L.
murinus. Using the spot of the lawn technique, inhibition zones of up to 9 mm wereobtained with viable L. murinus cells against S. aureus. Zymography with M.
lysodeikticus revealed a lytic band around 108 kDa, whereas three bands wereidentified with S. aureus, with relative molecular masses of 108±4.80, 92±0.22, and23±1.11 kDa. The amino acid sequence of the 23±1.11 kDa protein with lytic activity
was obtained by peptide fingerprinting (UHPLC). The inhibition kinetics determined a
MIC (minimal inhibitory concentration) was 0.07±0.01 mg/mL and a reduction of 3
log CFU/mL of S. aureus at 24. L. murinus showed moderate biofilm production at
4°C and 25°C, and strong biofilm production at 37°C, whereas S. aureus exhibited
moderate biofilm production at 4°C and strong biofilm production at 25°C and 37°C.
Applying the FAO-OMS-OMSA criteria (2022) our results confirm that L. murinus
26B1 strain from murine origin was identified at the genus and species level; its
antagonistic potential against S. aureus ATCC 6538P and the 23±1.11 kD bandshowed similarity to a protein fragment with aminopeptidase function.

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AB10 GENETIC MODIFICATION OF Lactococcus lactis TO
PRODUCETHE
ANTI-INFLAMMATORYPROTEIN
ELAFIN:EVALUATION OF DIFFERENT EXPRESSION SYSTEMS
R. Levit1, A. Aucouturier2, P. Langella2, L. Bermúdez-Humarán2, A. de Moreno de LeBlanc1,
J. G. LeBlanc1.
1CERELA-CONICET. Chacabuco 145. 4000 Tucumán, Argentina. 2Université Paris-Saclay,
INRAE, AgroParisTech, Micalis Institute, 78350, Jouy-en-Josas, Francia. E-mail:rlevit@cerela.org.ar

Lactococcus (L.) lactis is the model lactic acid bacteria (LAB) for genetic
modification for the production of heterologous proteins of therapeutic interest. Ourgroup constructed two strains of L. lactis capable of expressing and administering
the anti-inflammatory protein elafin using two different expression systems. In thefirst, L. lactis LL-thy-elafin produces elafin through the NICE (Nisin-Inducible
Controlled Expression) system, a system based on the autoregulation of nisinbiosynthesis. In this strain, the elafin expression cassette (under the control of thenisin-inducible promotor) is located on the bacterial chromosome and it was
modified to make it thymidine auxotrophic, contributing to its safety. In the second,
L. lactis LL-elafin produces elafin trough the SICE (Stress-Inducible Controlled
Expression) system, a system based on a vector carrying an expression cassette
under the transcriptional control of a stress-inducible promotor. In this system, theexpression of the protein of interest is induced after the administration of LAB to thehost in which it finds different conditions than in culture. The aim of this study wasto evaluate and compare the anti-inflammatory potential of L. lactis LL-thy-elafinand L. lactis LL-elafin in vitro using cancerous and non-cancerous intestinal celllines, and in vivo in a murine model of intestinal mucositis (IM). In vitro: the effect of
the recombinant strains on the efficacy of 5- Fluorouracil (5-FU) was evaluated with
Caco-2 and IEC-18 cells exposed to 5-FU and/or the recombinant bacteria andmeasuring viability by MTT assay. In vivo: BALB/c mice were injected daily with 5FU to induce IM and orally administered with the recombinant strains (108
CFU/mL). L. lactis carrying the empty plasmid vector (LL-empty) and the respective
wild-type strains (LL) were used as controls. Diarrhea onset, small intestinemorphology and histopathology, and serum cytokines levels were assessed. In vitro
assays demonstrated that the elafin-producing strains enhanced the cytotoxic effectsof 5-FU on colon cancer cells (Caco-2) and improve the survival of non-cancerous
intestinal epithelial cells (IEC-18) in the presence of 5-FU. In vivo assays showed thatelafin-producing strains were able to reduce the onset of diarrhea, with an increasein the villus height/crypt depth ratio, and a reduction of intestinal inflammation.
Recombinant strains reduced serum levels of pro- inflammatory cytokines withincreases of the anti-inflammatory cytokine IL-10. In conclusion, although both
systems were effective, L. lactis expressing elafin with the SICE system showed thebest results. This system has the additional advantage that the presence ofregulatory genes is not required nor does not require chemical induction before their
use.

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AB11 INFLUENCE OF TEMPERATURE, PH AND CULTURE MEDIAON Salmonella enterica SEROTYPE ENTERITIDIS BIOFILM
FORMATION
S. López Pérez1, C. Melián1, P. Castellano1.
1CERELA-CONICET.
Chacabuco
145.
slopez@cerela.org.ar

4000

Tucuman,Argentina.

E-mail:Biofilms are defined as complex microbial communities established on a wide
range of surfaces and embedded in a self-produced matrix of extracellular polymeric
substances. Adhesion of pathogenic and/or spoilage microorganisms to equipment
materials and biofilm development constitute a potential chronic source of microbial
contamination threatening the safety and quality of food products, resulting in foodborne disease and economic losses. Particularly, Salmonella enterica serotype
Enteritidis one of the most important agents of foodborne disease in several
countries forms biofilms to evade the action of traditional disinfectants,
environmental stress and the host's immune system. Food products including
contaminated meat and eggs have caused outbreaks of human salmonellosis. In
addition, biofilm formation is a multi-step process starting with attachment to asurface then formation of micro-colony that leads to the formation of threedimensional structure and finally ending with maturation followed by detachment.
This process depends on an interaction among bacterial cells, the attachment
surface and environmental conditions. In this context, the aim of this study was todetermine the biofilm production capacity of S. enterica serotype Enteritidis at two
different temperatures of incubation, pH and four culture media. The pathogenic
strain was isolated from poultry and identified by using conventional standard
biochemical and serological tests. 200 µl of a culture (16 h of incubation) were added
to each well of the microplate. The following culture media were used separately:
triptein soy broth (TSB) + 0.5% yeast extract (YE), Luria-Bertani (LB) both with pH
7.0 and TSB + 0.5% YE + 0.5% NaCl at pH 5.5. Two temperatures were tested, 25 ºC
to promote the expression of the components of the extracellular matrix and 10 ºC as
the temperature of meat and poultry processing lines. Microplates incubated at 25ºC were examined at 24 and 48 h and those incubated at 10 ºC at 3 and 6 days.
Biofilm formation was determined by crystal violet staining and classified as weak,moderate or strong based on the established cut-off line. In microplates incubated at
25 ºC, S. enterica biofilm formation was greater in the TSB+YE+NaCl medium, withweak and moderate formation at 24 and 48 h, respectively. At 10 ºC, a greater
increase in pathogen biofilm formation was detected in all culture media at 6 days of
incubation, with moderate formation profiles. At 3 days, a similar biofilm formation
pattern was only observed in TSB+YE+NaCl and LB media. The contribution to the
Knowledge of factors that influence in the formation and dispersion of S. enterica
serotype Enteritidis biofilms could allow the design of prevention strategies to avoid
microbial contamination, controlling an important public health problem.

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AB12 OPTIMIZATION OF THE FREEZE-DRYING PROCESS TOOBTAIN MORE STRESS TOLERANT Oenococcus oeni STARTER
CULTURES
E.V. Bentencourt1, R.R Raya1, L.M. Mendoza1
Chacabuco
145.
4000
ebentencourt@cerela.org.ar

1CERELA-CONICET.

Tucuman,Argentina.

E-mail:Malolactic fermentation (MLF) is an important process in wine production, as
it contributes to improving the quality and organoleptic characteristics of wine by
reducing acidity and increasing microbiological stability. Oenococcus oeni is the best
species adapted to winemaking conditions and it is almost exclusively used as an
MLF starter culture. Therefore, achieving success in this fermentation has led toincreased emphasis on developing methods for the production and conservation ofstarter cultures. Freeze-drying is widely used in the production of starter cultures;
however, this process can cause damage to the cell membrane, denaturation ofproteins and DNA, resulting in a significant decrease in cell survival. For this reason,
cryoprotectants are used to enhance cell viability. The objective of this study was toevaluate the viability of O. oeni strains pre-treated with glutathione (GSH) or aminoacids as stress protective agents and later subjected to freeze-drying process using
different cryoprotectants. Two strains of O. oeni (X2L and ST), native to the NOA
region, were cultured at 30°C in modified MRS medium (5 g/L fructose and 4 g/L
DL-malic acid) with and without GSH or cysteine and glutamate (5 mM). Cells were
harvested at the end of the exponential phase (OD 0.9) at 600nm, washed withphosphate buffer, and treated with three types of cryoprotectants: sucrose (10%),
sodium glutamate (2.5%), and glutamic acid (2.5%), suspended in phosphate buffer.
Vials were previously frozen overnight to freeze-drying. Rehydration was carried out
using the same cryoprotectants. Cell counts were performed before and after freezedrying as well as at 7 days, 1, 4, and 8 months, to determine if cells remained viable
over time. When sodium glutamate and glutamic acid at 2.5% were used as
cryoprotectants, the initial cell concentration was 10^9 CFU/ml, and cell viability
was maintained throughout the 8 months evaluated. However, when cells were
freeze-dried with 10% sucrose, cell survival dropped by two logarithmic units after
freeze-drying. Later, cells were inoculated into wine to assess cell viability andmalolactic activity. In synthetic wine, both strains of O. oeni showed higher viability
and malic acid consumption when glutamic acid was used as the cryoprotectant andGSH as the protective agent against stress wine. The results of this study confirmed
glutathione has a positive effect on growth of O. oeni contributing to biomass
preparation of MLF starter cultures and in combination with glutamic acid moreresistant freeze-dried cells were obtained. Therefore, the combination GSH andglutamic acid could be used to optimize biomass production and enhance the qualityof O. oeni starter cultures used for MLF in winemaking.

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AB13 SENSORY ANALYSIS OF THE FINAL PROTOTYPE OF A SOY
FOOD FERMENTED WITH Lacticaseibacillus paracasei subsp.
paracasei CRL207
B.L. Nacchio1, G.N. Avila Hael1, O.A. Garro2, R.B. Medina1,3, M.S. Garro1.
1CERELA-CONICET.
Chacabuco 145. 4000 Tucuman, Argentina. 2INIPTA-CONICETUNCAUS. Cte Fernandez 755. 3700 Pcia R S Peña, Chaco, Argentina. 3FAZyV, Universidad
Nacional de Tucumán, Avda. Pte. N. Kirchner 1900. 4000 Tucumán, Argentina. E-mail:mgarro@cerela.org.ar

Soy foods have a low acceptance among consumers; this could be due to theorganoleptic characteristics of soybean, especially the strange flavors and odors (“offflavors”). The use of fermentation with lactic acid bacteria confers to the matrix
various properties, from the nutritional, functional and organoleptic point of view. In
previous works it was determined that the Lacticaseibacillus paracasei subsp.
paracasei CRL207 strain conferred favorable characteristics to soybean products
and the aroma was studied sensorially, this study allowed obtaining descriptors tocarry out a more detailed sensorial study of the product in its final phase. Sensory
analysis involves an important phase in the final stage of the prototyping of a new
food. The aim of the present work is to determine if the inclusion of flavorings
improves the organoleptic profile of a food prototype fermented with lactic acidbacteria in a soybean matrix. A semi-trained panel selected the flavorings that wereadded to the food prototypes (strawberry and orange). The selected additives werethen moved on to a further prototyping stage where a more detailed analysis was
carried out with a panel of 50 untrained consumers who were given three codedsamples, two of which were added with flavorings and as a control a sample without
additives was given. Acceptability and association were evaluated in the survey using
a “check all that apply” (CATA) type survey. The results showed that the samples
with the addition of strawberry and without flavoring were the ones that theparticipants liked the most, obtaining a score greater than 4 considering theattributes of aroma, flavor and overall appreciation, while the sample added withflavoring was the one that obtained a lower score (4). Regarding the association: theparticipants described the sample without flavorings by selecting both positive andnegative terms (cereals, nuts and unpleasant), the sample with added strawberry
flavor was associated with descriptors with positive connotations such as sweet,
yogurt and caramel; finally, the sample with orange flavor was described as sour,vinegar, soy and citrus, in this last case the selected terms also have a mixed
connotation. In conclusion, we can say that based on the results obtained, thestrawberry flavored sample is the one that obtained a better performance since it
obtained a good acceptability score and was described with positive terms for
consumers.

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AB14 PRODUCTION OF MIXED FRUIT JUICE ENRICHED WITH
PROBIOTIC LACTIC ACID BACTERIA
1,4 F. Escobar, 1,2 D.A. Barrionuevo, 1 S.A. Isas,4 B.J. Pérez, 1,3 C.P. Van Nieuwenhove.
1 Centrode Referencia para Lactobacilos (CERELA)-CONICET, Laboratorio de Ecofisiología
Tecnológica, Chacabuco 145, San Miguel de Tucumán, 4000, Tucumán, Argentina. ²
Facultad de Bioquímica Química y Farmacia-UNT, Batalla de Ayacucho 471, San Miguel deTucumán, Argentina. 3 Fundación Miguel Lillo, Miguel Lillo 251, San Miguel de Tucumán,
4000, Tucumán, Argentina. 4 Facultad de Ciencias Naturales e IML, Miguel Lillo 205, San
Miguel de Tucumán, 4000, Tucumán, Argentina. Email: fescobar@cerela.org.ar

The consumption of functional foods has acquired great interest over theyears, as they provide health benefits beyond the basic nutritional characteristics offresh foods. Probiotics are live microorganisms that, when administered in adequate
amounts, confer a health benefit to the host. Among functional foods, probiotic foods
and drinks are in great demand due to their close relationship with the prevention ofmultiple diseases. Fermented fruit-based juices are one of the most active functional
food categories. They provide several bioactive compounds and could be used forprobiotic delivery as an alternative to dairy foods, useful for those consumers whoare vegan or intolerant to dairy products. The aim of this study was to evaluate amixed fruit juice as a vehicle for three commercial probiotic strains:
Lactiplantibacillus plantarum CRL 972 (ATCC 14917), Lacticaseibacillus casei CRL
1110 (ATCC 393) and Lacticaseibacillus rhamnosus CRL 1227 (ATCC 11443). The
juice was obtained by mixing pomegranate (20% v/v), blueberries (10% v/v),
blackberries (10% v/v) juices, and water (60% v/v), 6º Bx, pH 4.4 adjusted withsodium carbonate. Juices were pasteurized at 65°C during 15 min, then immediately
cooled. Probiotic strains were individually inoculated at 2% (v/v) (107 CFU/mL), thenincubated at 37°C for 48 h, further refrigerated at 4°C for 28 days. Viable cell count,pH, total phenolic content (TPC) and antioxidant capacity by three different methods(FRAP, DPPH and TEAC) were measured in the formulated juices during
fermentation and cold storage. L. plantarum CRL 972 grew after 48 h of fermentation(up to 108 UFC/mL) and maintained at a high viable cell count of 107 UFC/mL after
28 days at 4ºC. The viability of L. casei CRL 1110 was around 10⁷ CFU/mL during
fermentation and shelf life; whereas L. rhamnosus CRL 1227 decreased near one log
unit after 28 days of cold storage (from 10⁷ CFU/mL to 106 CFU/mL). The TPC
significantly increased after shelf life in fermented juice by L. casei CRL 1110 (p˂0.05). Antioxidant activities were similar in all juices (with or without probiotic
strains) and times assayed. No yeast, mold or Enterobacteriaceae were found inprobiotic juices. Results show that the mixed fruit juice could be used as analternative functional drink, with high antioxidant capacity and preserved
polyphenol content, for delivering probiotic strains.

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AB15 IMPAIRMENT OF Listeria monocytogenes DEVELOPED ON
INDUSTRIAL SURFACES BY Latilactobacillus curvatus CRL1579
BACTERIOCIN

C. Melián1, D. Ploper2, R. Chehín2, G. Vignolo1, P. Castellano1
1CERELA-CONICET. Chacabuco 145. 4000 Tucuman, Argentina. 2 IMMCA-CONICET-UNTMinisterio de Salud Pública, Gobierno de Tucumán. Pje. Dorrego 1080. 4000 Tucuman,
Argentina. E mail: cmelian@cerela.org.ar

Under environmental stress, Listeria monocytogenes can form biofilms byattaching to a variety of surfaces like stainless steel (SS), polytetrafluoroethylene(PTFE) or glass. Biofilm protected pathogens from sanitization procedures, allowing
them to survive and persist as a chronic source of contamination, increasing theirresistance to antimicrobials and sanitizers. The use of eco-friendly approaches
involving microorganisms and their metabolites has recently emerged as alternative
anti-biofilm strategies. Lactic acid bacteria (LAB) are known for their ability to
produce bacteriocins, which are “friendly” antimicrobial agents. Particularly, thebacteriocin lactocin AL705 produced by Latilactobacillus (Lat.) curvatus CRL1579
showed high effectiveness at inhibiting Listeria in different meat systems. In
addition, at sub-inhibitory concentrations was able to control L. monocytogenes
FBUNT biofilm formation by disruption of quorum sensing through a signal molecule
inactivation. On this basis, the aim of this study was to evaluate the ability of Lat.
curvatus CRL1579 and its bacteriocin lactocin AL705 to displace L. monocytogenes
FBUNT biofilm formed SS and PTFE industrial surfaces at 10 °C under static andcontinuous flow conditions. The bacteriocin was purified from an overnight cultureof Lat. Curvatus CRL1579 in MRS by fractional precipitation with ammonium sulfate,solid phase chromatography and subsequent evaporation of solvents by rotary
evaporator. The minimal inhibition concentration (MIC) was determined. L.
monocytogenes FBUNT biofilms were formed on SS and PTFE coupons during 3 and6 days at 10 °C under static and continuous flow conditions using LB as a culture
medium. The inhibitory effect of Lat. curvatus and lactocin AL705 was evaluated bydisplacement assay and analyzed by confocal laser scanning microscopy (CLSM).
Partially purified bacteriocin (800 UA/mL) effectively inhibited L. monocytogenes
preformed biofilm through displacement strategy, reducing the pathogen by 5.54 ±0.26 and 4.74 ± 0.05 log cycles at 3 and 6 days, respectively. The bacteriocinproducer decreased the pathogen biofilm by 2.84 log cycles. CLSM allowed tovisualize the ability of lactocin AL705 to reduce L. monocytogenes FBUNT biofilm
under static and hydrodynamic flow conditions. A greater effect of the bacteriocin
was found at 3 days independently of the surface matrix and pathogen growth
conditions at 10 °C. As a more realistic approach, biofilm displacement strategy
under continuous flow conditions showed a significant loss of biomass, mean
thickness and substratum coverage of pathogen biofilm. These findings highlight theantibiofilm capacity of lactocin AL705 and their potential application in foodindustries.

91SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

AB16 STABILITY OF PREBIOTIC GALACTOOLIGOSACCHARIDESDURING YOGHURT MANUFACTURE AND STORAGE

A. Fara1, J. Palacios1; A. Montilla2, G. Zárate1,3.
1CERELA-CONICET. Chacabuco 145. 4000 Tucumán, Argentina. 2Instituto de Investigación
en Ciencias de la Alimentación (CIAL-CSIC), Madrid, España. 3Universidad de San Pablo
Tucumán, Argentina. E-mail: gzarate@cerela.org.ar

Prebiotics, are substrates selectively utilized by host microorganisms
conferring a health benefit. Among the targets for prebiotics, the intestinal
microbiota is the most relevant due to its crucial role in physiology and well-being.
Since diet greatly influences the composition and metabolism of the intestinal
microbiota, positive microbiome modulation by functional food ingredients, such as
prebiotics, has become a strategy for reducing disease risk and fortifying
homeostasis. At present, most widespread prebiotics are carbohydrates, particularly
oligosaccharides such as fructans (inulin and fructo-oligosaccharides), galactooligosaccharides (GOS), and lactulose. In particular, GOS have attracted the interest
from researchers and food industry because of the presence of galactose-based units
with relative structural similarity to human milk oligosaccharides. GOS derived fromlactose, lactulose (OsLu) and lactitol (LOS) are synthetized by transgalactosylation
catalyzed by microbial β-galactosidases (β-gal). In previous studies we reported thesynthesis and characterization of GOS by the β-gal from Acidipropionibacterium
acidipropionici LET 120 (PAB-GOS) and Lactobacillus bulgaricus CRL 450 (LAB-GOS).
GOS produced by both strains were scarcely digested by the gastrointestinal tractand demonstrated prebiotic effects on recognized probiotic microorganisms (CRL 431
and LGG). In the present study we manufactured a yoghurt supplemented with 2%
PAB-GOS or LAB-GOS before and after fermentation of free-lactose milk with S.
thermophilus CRL 1598 and L. bulgaricus CRL 450. The growth of the starter
cultures (by plate count method), acidification of milk (by pH measurement) andconsumption of GOS and monosaccharides (by HPLC) were determined at intervals
during 8 h of fermentation and 28 days of storage at 4 °C. PAB-GOS remained
unaltered throughout fermentation, while the viability of starter culture was similar
to that observed in the control yoghurt. When LAB-GOS were added to milk priorfermentation, CRL 1598 grew like the control while CRL 450 reached 0.5 log
CFU/mL more than the control and reached a lower pH (4.81 vs 4.93). Growthduring fermentation was mainly due to monosaccharides consumption whereas lessthan 10% of LAB-GOS were consumed. During cold storage, GOS were notsignificantly hydrolyzed whereas pH decreased in a similar manner in both control
and GOS supplemented yoghurts and starters remain higher than 107 CFU/mL. Our
results show the feasibility of obtaining a reduced-lactose yogurt enriched in stableprebiotic GOS. The manufacture of a prebiotic yoghurt with probiotic starters would
further increase the functional properties of the product.

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AB17 EVALUATION OF THE
ANTIOXIDANTS OF LEGUME
SELECTED LACTIC CULTURESPROTEOLYTIC PROFILE AND
FLOURS FERMENTED WITH

A. P. Pezuk1, L. R. M. Juarez1, G. Zárate1,2, M. S. Garro1.
1CERELA-CONICET. Chacabuco 145. 4000 Tucumán, Argentina. 2Universidad de San PabloTucumán. Gral. José de San Martín 435. 4000 Tucumán, Argentina. E-mail:ppezuk@cerela.org.ar

Currently, the production of legumes represents an important economic
support for the Northwest region of Argentina, particularly soybeans, chickpeas andbeans, due to their high nutritional value and vegetable protein content. The
fermentation with lactic acid bacteria is an attractive strategy to improve thenutritional, sensory and techno-functional characteristics of legume flours withapplications in novel foods. The objective of this work was to evaluate the effect oflactic fermentation on the proteolysis, antioxidant activity and the in vitro
digestibility of legume flours. The microorganisms used were: Limosilactobacillus
fermentum CRL 251, Lacticaseibacillus paracasei subsp. paracasei CRL 207 andLactiplantibacillus plantarum CRL 2211. Pastes were made with the individual
legume flours: soybean paste (PS), bean paste (PP), kabuli chickpea paste (PG) and a
mixture (1:1:1 ratio) of the three flours (PSGP). Pastes containing 65% of moisture,
were inoculated at 1% rate (~106 CFU/g) with each strain and incubated at 37°C for
24 h, uninoculated pastes were used as control. Bacterial growth during
fermentation was followed by the plate dilution count method (log CFU/g of sample)
and pH. The concentration of soluble amino acids in the pastes was measured by theo-phthaldialdehyde method (OPA), antioxidant activity by the DPPH method and thequantification of in vitro protein digestibility (IVPD) was carried out through amultienzyme complex. The assayed strains showed a significant microbial growthand increase of acidification in all pastes, compared to the controls. All thefermentation increased the soluble amino acids contents particularly 40% in PS and
PP, 30% in PSGP and 20% in PG, compared to their controls. The antiradical activity
showed the greatest increase in PSGP (30% in the three strains compared to thecontrol) in contrast to the individual pastes (20% compared to the control). The
strains CRL 251 (in PS) and CRL 207 (in PG) did not show significant variations withrespect to the control. Regarding the IVPD%, values above 20% IVPD were observedin all treatments compared to their controls, except in PG fermented with CRL 251
which was significantly lower compared to the other treatments. In this work it wasdemonstrates that the three strains grew and acidified the vegetable matrices at
37°C for 24 h, observing an improvement in the antioxidant, proteolytic activity anddigestibility of the flours. Our results demonstrate the potential of selected lactic acidbacteria to improve the properties of legume flours by increasing their antioxidant
activity, the availability of peptides and amino acids for the consumer, promoting
their use as starters for the formulation of products derived from legumes.

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AB18
TECHNOLOGICALSTRATEGYTO
IMPROVETHE
ORGANOLEPTIC CHARACTERISTICS OF SOY FOOD PRODUCTS:FERMENTATION WITH SELECTED LACTIC ACID BACTERIA
G.N. Avila Hael1, B.L. Nacchio1, O.A. Garro2, R.B. Medina1, M.S. Garro1.
1CERELA-CONICET.
Chacabuco 145. 4000 Tucumán, Argentina. 2INIPTA-CONICETUNCAUS. Cte Fernandez 755. 3700 Pcia R S Peña, Chaco, Argentina. E-mail:mgarro@cerela.org.ar

The production of plant-based foods is currently expanding. Soybean is a
widely cultivated, legume rich in vegetable protein (35–40%), lipids (19–22%) andother important nutrients (isoflavones, minerals and dietary fiber). However, its
consumption is limited by the beany flavors (off flavors). The use of selected lactic
acid bacteria (LAB) emerges as a technological strategy to modify the soy matrix,
increasing its nutritional value, and improving the organoleptic characteristics. LAB
that metabolize citrate produce compounds such as acetate, diacetyl, acetaldehyde
and acetoin, which determine the flavor of many fermented foods. The objective of
this work was to evaluate the production of organic acids, diacetyl (DA) and acetoin(AC) in fermented soybean products added with different sources of citric acid.
Aqueous soy extract (EAS) and soy pastes (PS) were used with the addition of
strawberry, orange peel or pulp, and/or sodium citrate. In total, 6 PS matrices and 5
EAS matrices were evaluated without and with additions of the different citratesources. Each matrix was individually inoculated at 2% with Lacticaseibacillus
paracasei subsp. paracasei CRL 207 (strain selected for its ability to utilize citrate)and incubated at 37°C for 8 h in EAS and 16 h in PS. PS and EAS without and withaggregates uninoculated were used as controls. Organic acids were determined by
HPLC, and volatile compounds by GC. The main acids detected in all samples werelactic, citric and acetic. It was shown that the soybean matrix contains citric acid
(EAS and PS without aggregates: 1.24 and 6.03 mg acid/g of sample, respectively).
The addition of orange or strawberry to EAS produced a slight increase in the initialcitric acid concentration, being higher with strawberry (EAS+strawberry: 1.79 mg
citric acid/g of sample). After 8 h of incubation was production of lactic acid wasobserved in all EAS sample. In EAS + commercial citrate presented the highest
concentration of lactic acid (12.63 mg lactic acid/g of sample). In all PS citrateconsumption was observed, detecting high consumed in PS + strawberry (95%). In
this sample, the highest production of lactic acid (9.64 mg acid/g of sample) was
obtained. DA and AC (DA-AC) production was detected in all fermented PS (138.4
mM to 219.29 mM), while in EAS these compounds were detected at very low
concentrations (4.68 mM to 17.85 mM). In soy matrices added orange peel, wasobserved the highest values of the DA-AC. In conclusion, a strategy to improve theorganoleptic characteristics of soy products consisting in ferment soybean paste
added of oranges or sodium citrate inoculated with L. paracasei subsp. paracasei
CRL 207 for 16 h at 37 °C.

94

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MOLECULAR BIOLOGY AND GENOMICSOF LACTIC ACID BACTERIA

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MG1 GENOMIC CHARACTERIZATION OF THE HONEYBEEPROBIOTIC STRAIN Ligilactobacillus salivarius subsp. salivarius
A3iob

M. Elean1, A. Arroyo Guerra2, L. Albarracín1, C. Audisio2, J. Villena1.
1CERELA-CONICET. Chacabuco 145. 4000 Tucuman, Argentina. 2INIQUI-CONICET-UNSA,
Salta, Argentina. E-mail: carina.audisio@exa.unsa.edu.ar, jcvillena@cerela.org.ar

Previous studies demonstrated the beneficial effects of the probiotic strain
Ligilactobacillus salivarius A3iob on honeybee (Apis mellifera) colonies. This
bacterium is able to enhance protection against bee pathogens. Furthermore, hives
from honeybees treated with the A3iob strain displayed significant increases inhoney production. The present study aimed to assess the genomic characteristics ofthe probiotic L. salivarius A3iob to understand its ability to improve bee´s health.
The comparative genomic analysis was performed with the A3iob genome and the
genomes of probiotic L. salivarius strains from human, porcine and chicken origin as
well as bacteria isolated from the bee´s gut including Bombella intestine, Bombellaapis, Bifidobacterium lactis, Apilactobacillus kunkeei, Enterococcus durans,
Apilactobacillus micheneri, Apilacobacillus timberlakei, Apilactobacillus quenuiae,
Apilactobacillus apinorum and Apilactobacillus waqarii. The analysis included theexamination of metabolic and functional genes related to adhesion, production of
bioactive compounds, modulation of the host's immune system, and antimicrobial
substances. The total size of the A3iob genome is 2.05 Mb with a 34.6% GC. In silico
studies revealed that L. salivarius A3iob possesses genes for glycosyltransferases(GT) from the families GT2, GT4 and GT similar to B. apis and B. intestinalis andglycosylhydrolases (GH) from the families GH1, GH2, GH13, GH36, GH65 andGH177 similar to A. kunkeei, E. durans and bifidobacteria isolated from bee
intestine. It was also observed that the A3iob strain has a unique genetic profile witha high number of secretion system genes and adhesion genes including the ones
coding for the SecA2/Y2 system, the mucus biding proteins MucBP1, MucBP2 and
MucBP3 and a pilli cluster (pilA, SpaA, SpaB, sortaseA) that was described only infive strains of the L. salivarius species and in the intestinal bee-derived strain E.
durans EDD2, indicating its potential for successful colonization in the bee
gastrointestinal tract. Additionally, the A3iob strain showed the presence of
exopolysaccharide biosynthesis clusters (EPS1 and EPS2) described in the probiotic
L. salivarius UCC118 (LSL_0977-LSL_0997 and LSL_1547-LSL_1574, respectively),
which have been associated to adhesive and immunomodulatory effects. Genes
related to oxidative stress response (thioredoxin and NrdH-redoxin system) weredetected, indicating that the A3iob strain may help bees to maintain a healthy
intestinal redox balance. In addition, the bacteriocin genes abp118A and abp118B
were found in the A3iob genome. The genomic characterization of the probiotic strain
L. salivarius A3iob performed in this work provides some clues about the genetic
mechanisms underlying the probiotic properties of the strain, paving the way for
future research aimed at improving bee´s health and productivity in the face ofenvironmental challenges.

96SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

MG2 FUNCTIONAL AND GENOMIC CHARACTERIZATION OF
Pediococcus pentosaceus TUCO-3 ISOLATED FROM CAT MILK: A
POTENTIAL PROBIOTIC STRAIN TO COMBAT INFECTIONS

S. Quilodrán-Vega1, M. Elean2, L. Albarracín2, C. Muñoz1, J. Toledo1, J. Villena2.
1University of Concepción, Concepción, Chile. 2CERELA-CONICET, Tucuman, Argentina. Email: squilodran@udec.cl

Diarrheal pathologies in cats are the most common cause of consultation inveterinary clinics. The most common bacteria that affect pets are Gram negative
pathogens. Usually, there is no investigation of the diarrhea causative agent, so themost common treatment consists of the administration of broad-spectrum
antibiotics, which leads to the selection of antibiotic-resistant microorganisms andintestinal dysbiosis in cats. Therefore, it is necessary to find alternatives to preventor reduce diarrhea in cats. In this work, lactic acid bacteria were isolated from breast
milk samples of cats and potential feline probiotic strains were selected based on
their ability to inhibit pathogenic bacteria including Escherichia coli, Salmonella
enterica, and Klebsiella pneumoniae. Among the studied strains, Pediococcus
pentosaceus TUCO-3 stood out because of its remarkable capacity to inhibit thegrowth of E. coli ATCC25922, S. enterica ATCC13076, and K. pneumoniae
ATCC13883, as well as enterohemorrhagic E. coli isolates. In vitro studies showedthat P. pentosaceus TUCO-3 has resistance to pH=3, pancreatine and bile salts,
indicating its potential to survive gastrointestinal conditions. The TUCO-3 strain issusceptible to most antibiotics assessed and revealed no hemolysin activity. In
addition, the complete genome of the TUCO-3 strain was sequenced using theIllumina Miseq platform. P. pentosaceus TUCO-3 identity was determined via averagenucleotide identity (ANI), obtaining an identity value of 98,96% compared withstrains of the P. pentosaceus species. The annotation of the genome was performed
with the NCBI Prokaryotic Genome Annotation Pipeline. The total size of the TUCO-3
genome is 1,9 Mb with a 37% GC content and a total of 1,996 coding sequences.
Several genes consistent with features of probiotic microorganisms are detected in
the TUCO-3 genome, including a MucBP gene associated with adhesion andcolonization (NVZ01419.1), and two putative genes coding for class IIa bacteriocins:
pediocin A and a bacteriocin with 69.7% identity to hiracin JM79 (NVZ00664.1). The
production and secretion of these bacteriocins by the TUCO-3 strain could
contribute to maintaining intestinal health in cats by inhibiting the growth ofpathogenic bacteria such as Clostridium perfringens, a major cause ofgastrointestinal infections. In addition, a vitamin B2 biosynthetic cluster was found;however, the ribDG gene has a premature termination codon, suggesting that the B2
vitamin de novo synthesis pathway is likely incomplete. Additional phenotypic tests
are necessary to evaluate vitamin B2 production. Screening for the virulence factor
genes in the genome of TUCO-3 showed no hits using the Virulence Factor Database(VFDB), confirming the safety of the tested strain. P. pentosaceus TUCO-3 ispotential probiotic candidate for improving intestinal health and resistance toinfections in cats.

97

SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

MG3 PROTEOMIC REGULATION AND GENE EXPRESSION OFSELENIZED CELLS OF Fructobacillus tropaeoli CRL2034 IN A
FERMENTED FRUIT-BASED MEDIUM

L. Crespo1, F. Mozzi1, M. Pescuma2.
1CERELA-CONICET. Chacabuco 145. 4000 Tucumán, Argentina. 2CIEFAP-CONICET. Ruta
Nacional 259 Km 16, Esquel, Chubut, Argentina. E-mail: lcrespo@cerela.org.ar

Selenium (Se) is an essential micronutrient for humans. Some lactic acidbacteria (LAB) can grow on fruit substrates and biotransform selenite into Senanoparticles and Se-amino acids. These LAB are considered an interestingalternative to formulate new fermented fruits-based beverages fortified with Se. This
work aimed to compare the growth and the differential protein and gene expressionof Fructobacillus tropaeoli CRL2034 grown in MRSf (fructose 20 g/L) with selenized(Se+) and control (Se-) cells grown in a fruit-like medium (SP). Se+ and Se- cells weregrown in MRSf and SP for 6 h (exponential growth phase). The protein differential
expression was analyzed by LC-MS/MS (CEQUIBIEM-CONICET), while theexpression of those genes likely related to Se metabolism was analyzed by RT-qPCR.
When Se- cells were grown in SP, 146 proteins were differentially expressed; 118
were upregulated and 28 were downregulated, while the Se+ cells differentiallyexpressed 228 proteins; 152 upregulated, and 76 downregulated compared withcells grown in MRSf. Venn diagram showed that 125 proteins were deregulated
independently of the Se growth condition, being 103 proteins deregulated in Se+
cells. When F. tropaeoli CRL2034 was grown in SP, two important enzymes involvedin the synthesis and incorporation of seleno-amino acids into proteins weredifferentially expressed. SerC showed a fold chance of 0.82 and 0.92 times, whilemethionyl tRNA synthetase increased its expression 1.58 and 2.00 times in Se- andSe+ cells, respectively. Moreover, two glutathione reductases showed higher
expression in the Se+ cells (1.84 and 1.49 times) than in Se- cells (1.00 and 1.24
times), while its gene expression increased in Se+ cells between 0.22 and 0.38 units
with respect to MRSf grown cells. Mannitol 2-dehydrogenase (MDH), an enzyme
involved in fructose metabolism, showed a negative deregulation trend with a foldchange of 0.89 and 0.76 times in the Se+ and Se- cells, respectively. Consistently,
mdh expression decreased by 0.48 and 0.60 units in cells grown in SP with respect
to MRSf grown cells. Malolactic enzyme showed a positive deregulation of 162.86 and148.96 times for Se+ and Se- cells, respectively. Similar results were observed in its
gene expression which increased by 1.23 and 1.46 units compared to the cells grownin MRSf. This fact was probably due to the presence of malic acid in the SP medium.
Proteomic and gene expression analysis indicated that the fruit-like medium
influenced the regulation of proteins involved in cellular processes. Moreover, an
increase in the expression of proteins related to Se metabolism was observed in Se+
cells with respect to Se- cells grown in SP, compared with cells grown in MRSf.

98

SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

MG4 GENOMIC AND PROTEOMIC CHARACTERIZATION OF
Lactiplantibacillus plantarum CRL681 FOCUSING ON CELL
SURFACE AND SECRETED PROTEINS
A. Baillo1, L. Albarracín1, J. Villena1, S. Fadda1
1CERELA-CONICET. Chacabuco 145. Tucuman, Argentina. E-mail: sfadda@cerela.org.ar

Lactiplantibacillus plantarum CRL681 (Lp681) can inhibit the growth ofpathogenic Escherichia coli both in food matrices and in the intestinal mucosa. In
addition, this strain can modulate intestinal innate immune responses. Consideringthat the bacterial surface and secreted factors have been associated with theinteraction of beneficial lactobacilli with the host as well as in their abilities to inhibit
pathogens, in this work, we performed genomic and proteomic studies tocharacterize the cell surface and secreted proteins of Lp681. In a first set of studies
an in silico analysis of cell surface and secreted factors was performed using thecomplete genome sequence of Lp681. The extracellular proteins in the genome wereidentified by bioinformatic analysis considering their subcellular location (secreted or
expressed on the surface), their type of anchorage and the secretory mechanism(proteins with signal peptides). A total of 277 proteins were detected in Lp681 whichwere classified into eight groups: C-terminal anchored proteins (with and withoutcleavage sites) (10), N-terminal anchored proteins (with and without cleavage sites)(138), lipid-anchored proteins, proteins anchored to the wall by the LPxTG domain(45), secreted proteins with cleavage sites (55) and proteins secreted by minor
pathways (7). Among the surface proteins, factors involved in adhesion were detected
including MubB2, MucBP2, MucBP5, MucBP-MubB2-YGX, MucBP-MubB2-YGX,
PgrA, and proteins of adhesion to collagen and fibronectin. Among the secreted
proteins, genes coding for resistance to gastrointestinal conditions and adhesion(cbh, treA, celB, eno2) were detected. In a second set of experiments, proteinssecreted by Lp681 in the presence and absence of enterohemorrhagic E. coli (EHEC)
O157:H7 in a chemically defined medium were studied. The cell-free supernatant
proteins were precipitated with trichloroacetic acid, while the surface cells proteins
were obtained by scraping bacteria with trypsin (5 μg). The samples were analyzed
by mass spectrometry and the identified proteins were analyzed by bioinformatics. A
total of 113 and 533 proteins were identified in the exoproteome and the surface of
Lp681, respectively, in the presence of EHEC. Interestingly, the most abundant
proteins belonged to the categories of membrane and cell wall biogenesis(hydrolases), and translation (ribosomal proteins). In addition, proteins withmoonlighting functions were found. The results show that Lp681 possesses several
surface adhesion factors that would allow its interaction with the intestinal mucosa
as well as to coaggregate with pathogenic E. coli. In addition, Lp681 secrete proteinsthat would exert an antimicrobial effect on EHEC through the generation of reactive
oxygen species and cell lysis by membrane permeabilization.

99

SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

Author index
L. Abán

HH24

M.C. Abeijón-Mukdsi

AH8

A. Abraham

HH9

G. Acciarri

S7A

G. Agüero

HH12

J. Aguilar

AH2

L. Albarracín

HH3

HH5

HH23 HH24 HH25 AH3MG2 MG4
J.C. Alcocer

MP3

A. Aliano

S6A

S. Alvarez

HH12 HH19 HH21

N. Álvarez

MP3

E. Andrada

HH17 HH20 AH1

L. Arce

S3A

L. Arellano-Arriagada

HH12 HH23 AH3J.N. Argañaraz Aybar

HH23 HH24

C. Aristimuño Ficoseco

S6B

J. Arrieta

HH8

A.A. Arroyo Guerra

AH6

A. Aucouturier

AB10

M.C. Audisio

AH6

G.N. Avila Hael

AB13 AB18

J.D. Babot

AH1

A. Baillo

HH26 AB1

M.F. Balcells

HH14

D.A. Barrionuevo

AB3

C. Belloch

MP8

L. BermúdEz-Humarán

AB10

A. Bernal

HH9

E.V. Betencourt

AB12

V.S. Blancato

S7A

M. Bruzone

AB5

G.A. Bulacios

HH1

AH8

AH7
MG1
MP1

AB14

MP2

MP3

MG1

AB2

MG4

AH5

MG1

SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

Author index
C. Burgos

HH16

J. Burini

AB5

A.Y. Bustos

MP4

C. Cancino

HH9

J. CarolMP5

Y. Carpintero Polanco

HH9

N. Carrasco

AH7

P. Castellano

HH16 AB11 AB15

P.G. Cataldo

HH1

MP2

S.I. Cazorla

HH4

HH8

M.N. Cerasuolo

HH2

HH11 HH13 HH15

R. Chehín

AB15

L. Contreras

AB8

C. Coronel

MP3

P.D. Cotter

S4

L. Crespo

AB5

H. Cristóbal

MP3

MP5

MP6

AB6

MP6

HH10 HH14

MG3

A. de Moreno de LeBlanc HH2

HH6

S. Dentice Maidana

HH3

HH25

M. Elean

S3A

HH3

HH11 HH13 HH15 AH7
HH5

HH23 HH25 HH26 AH3

MG2
F. Escobar

AB3

AB14

S. Fadda

S6B

HH26 AB1

F. Fanelli

S6B

A. Fara

AB16

S. Feliu

HH14

R. Fernández Brando

HH9

M. Flores

MP8

K. Fukuyama

HH12

V. Fusco

S6B

C. García

AB9

M.S. Garro

AB13 AB17 AB18

O.A. Garro

AB13 AB18

AB10

AB2

AB9

MG4

MG1

SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

Author index
G. Garrote

HH9

P. Gauffin-Cano

HH17 HH20

F. Gharzia

AB4

K. Gianni de Carvalho

AB4

F.O. Gizzi

S7A

N. Gobbato

HH23 HH24

J. Gomez

AB4

J. Gomez

AB6

N. Gómez

MP4

F. Gutiérrez

HH19 HH21

J. Gutierrez-Merino

S5B

E.M. Hebert

HH1

M. Hidalgo

S6A

V.P. Houde

HH1

C. Ibarguren

MP1

M. Ibarra

MP6

Y. Imamura

S3A

A.S. Isas

AB3

M.I. Isla

AB8

J. Islam

S3B

M. Ivir

HH19 HH21

L.R.M. Juarez

AB17

M.A. Jure

HH3

H. Kitazawa

S2A

AB7

MP2

MP3

AB14

S3A

HH5

HH12 HH18 HH23 HH24 HH25

AH3
J. Laiño

HH7

P. Langella

AB10

J.G. LeBlanc

HH2

HH6

HH11 HH13 HH15 AH1

A. Ledesma

MP4

MP5

MP6

AB6

R.M. Lenz

MP1

MP3

R. Levit

HH13 HH15 AH7

AB10

A. Llorente

AB9

S. López Pérez

AB11

AH7

AB10

SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

Author index
G.L. Lorca

S5A

M.J. Lorenzo Pisarello

HH5

C. Magni

S7A

E. Maldonado

S5B

C. Maldonado Galdeano HH4

HH8

C. Maluquer de Motes

S5B

A. Márquez

HH17 HH20

G. Marranzino

HH12

B. Martínez Caliari

AB7

HH10 HH14

M.J. Martínez Monteros HH4
K. Matsumoto

HH12

R. Medina

S3C

C. Melián

AB11 AB15

L.M. Mendoza

MP8

C. Mercado

AB9

C. Minahk

HH1

M.H. Miranda

AH4

F.R.E. Molina

MP7

A. Montilla

AB16

C. Moreno

MP4

H. Mouguelar

AH2

F. Mozzi

MP7

S. Mukhopadhyay

S5B

C. Muñoz

AH5

B.L. Nacchio

AB13 AB18

M.E.F. Nader

S6B

Z. Nakamura

S3B

F. Namai

S2A

L. Nelegatti

AB2

K. Nishiyama

S2A

T. Nochi

S3B

M. Obregozo

AH1

O. Ordoñez

AB5

HH17 HH20 AH1

AH8

AB13 AB18

AB12

AB3

AB5

MG3

MG2
HH1

AH2

AH4

AH7

S3A

HH5

HH12 HH25 AH3

S3A

HH24

SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

Author index
S. Orosco

HH23 HH24

S. Ortiz Mayor

HH24

R. Ortiz Moyano

HH25 HH26

N. Ohtani

S3B

J. Palacios

AB16

M. Palermo

HH9

J. Palma

AB9

R. Parada

AB7

E. Parente

S6A

M.S. Pellegrino

AH2

G. Perdigón

HH4

N. Pereyra

AH2

B.J. Pérez

AB14

L. Perret

HH24

M. Pescuma

AB5

A.P. Pezuk

AB17

R. Pietrafesa

S6A

D. Ploper

AB15

S. Quilodrán-Vega

AH5

V. Ramos

HH9

R.R. Raya

MP7

A. Ricciardi

S6A

L.G. Ruiz-Rodríguez

MP7

M. Russo

HH17 HH20

L. Saavedra

HH1

J. Sacur

HH18

G. Sáez

HH22

A. Salas Burgos

S7B

E. Salguero

AB8

S. Salva

HH12 HH16 HH19 HH21

I. Sandoval

AB9

B. Sede Lucena

AB5

R. Serda

HH3

HH14

MG3

MG2
AB12

MP2

HH5

SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

Author index
Y. Shimizu

S3B

T. Shimosato

S2A

H. Shirakawa

S1

J.A. Silva

AH2

K. Sivieri

S2B

S. Solis

S5B

F. Sosa

HH9

F. Sosa

AB7

C. SotoAB9

N. Suarez

AH8

Y. Suda

HH5

M.E. Taborra

S7A

H. Tanaka

S3B

M.P. Taranto

HH7

MP4

J. Toledo

AH5

MG2

C. Tomei

HH17 HH20

M. Torino

AB2

S. Torres

AB8

M.P. Urquiza

HH1

J.C. Valdéz

HH23 HH24

M. Vallejo

AB7

C.P. Van Nieuwenhove

AB3

B. Vasile

HH16 HH19 HH21

A. Vasqueto

AH2

E.M. Vélez

HH8

A. Venecia

HH24

M.D. Vera

HH18

A. Vestani

HH4

HH8

G. Vignolo

S6B

AB15

J. Villena

S3A

HH3

HH5

HH12 HH18 HH23 HH24 HH25

HH26 AH3

AH5

AH6

M.G. Vizoso Pinto

HH18

H. Yamamuro

S3A

HH25

AB6

MP2

AB14

HH10 HH14

HH10

AB1

MG1 MG2 MG4

SIBAL – AJLAB 2024: Lactic Acid Bacteria In The Genomic Era

Author index
J. Yamazaki

S3B

P.A. Yánez

HH6

G. Zárate

HH22 AB16 AB17

M.H. Zelaya

HH7

HH12