bluuebunny/crossref_metadata_2025 huggingface dataset:
The Utility of Caco-2 cells in Isolation of Enteroviruses from Environmental and Clinical Material
Authors: WIECZOREK MAGDALENA , CIĄĆKA AGNIESZKA , WITEK AGNIESZKA , LITWIŃSKA BOGUMIŁA
Published: November 2019

The work presented here demonstrates the utility of Caco-2 cells in the isolation of enteroviruses (EVs) from environmental and clinical materials. Thirty-two samples of cerebrospinal fluid positive in Pan-entero RT-PCR were taken for EV strain isolation in cell culture. Out of the 32 samples analysed, 22 (68.75%) were positive for enteroviruses by isolation in Caco-2 cells, and 10 (31.25%) were positive by isolation in RD cells. High viral titre in clinical specimens resulted in rate increase for isolation in Caco-2 cells and RD cells (87.5% and 50%, respectively). Also, the probability of isolation of enteroviruses from sewage in Caco-2 cells was 20 times higher that in RD cells. We proved that Caco-2 cells were more effective than RD cells in enterovirus isolation, irrespective of the material used in the inoculation process.
Psychrotrophic Lipase Producers from Arctic Soil and Sediment Samples
Authors: RASOL R. , RASHIDAH A.R. , SITI NUR NAZUHA R. , SMYKLA J. , WAN MAZNAH W.O. , ALIAS S.A.
Published: November 2019

Culturable microorganisms were successfully isolated from soil and sediment samples collected in 2011 on the northern coast of Hornsund, West Spitsbergen. A total of 63 single colony isolates from three sampling sites obtained were subjected to temperature dependence study to assess whether they are obligate psychrophilic or psychrotrophic strains. From initial temperature screening, only 53 psychrotrophic isolates were selected that are capable of growing between 4-28 degrees C. The rest that were capable of tolerating higher temperatures up to 37 degrees C were not included in this study. These isolates were chosen for lipase enzyme screening confirmation with the standard plate assay of olive oil and fluorescent dye Rhodamine B. Six lipase positive isolates were also subjected for subsequent lipase enzyme plate screening on tributyrin, triolein, olive oil and palm oil agar. Lipase production by these six isolates was further assayed by using colorimetric method with palm oil and olive oil as the substrate. These isolates with promising lipase activity ranging from 20 U/ml up to 160 U/ml on palm oil and olive oil substrate were successfully identified. Molecular identification by using 16S rRNA revealed that five out of six isolates were Gram-negative Proteobacteria and the other one was a Gram-positive Actinobacteria.
Plant Growth Promotion Rhizobacteria in Onion Production
Authors: ČOLO JOSIP , HAJNAL-JAFARI TIMEA I. , ĐURIĆ SIMONIDA , STAMENOV DRAGANA , HAMIDOVIĆ SAUD
Published: November 2019

The aim of the research was to examine the effect of rhizospheric bacteria Azotobacter chroococcum, Pseudomonas fluorescens (strains 1 and 2) and Bacillus subtilis on the growth and yield of onion and on the microorganisms in the rhizosphere of onion. The ability of microorganisms to produce indole-acetic acid (IAA), siderophores and to solubilize tricalcium phosphate (TCP) was also assessed. The experiment was conducted in field conditions, in chernozem type of soil. Bacillus subtilis was the best producer of IAA, whereas Pseudomonas fluorescens strains were better at producing siderophores and solubilizing phosphates. The longest seedling was observed with the application of Azotobacter chroococcum. The height of the plants sixty days after sowing was greater in all the inoculated variants than in the control. The highest onion yield was observed in Bacillus subtilis and Azotobacter chroococcum variants. The total number of bacteria and the number of Azotobacter chroococcum were larger in all the inoculated variants then in the control. The number of fungi decreased in most of the inoculated variants, whereas the number of actinomycetes decreased or remained the same.
The prevalence of Anaplasma phagocytophilum in questing Ixodes ricinus Ticks in SW Poland
Authors: KIEWRA DOROTA , ZALEŚNY GRZEGORZ , CZUŁOWSKA ALEKSANDRA
Published: November 2019

Ticks constitute important vectors of human and animal pathogens. Besides the Lyme borreliosis and tick-borne encephalitis, other pathogens such as Babesia spp., Rickettsia spp., and Anaplasma phagocytophilum, are of increasing public health interest. In Poland, as in other European countries, Ixodes ricinus, the most prevalent tick species responsible for the majority of tick bites in humans, is the main vector of A. phagocytophilum. The aim of the study was to estimate the infection level of I. ricinus with A. phagocytophilum in selected districts, not previously surveyed for the presence of this agent. Sampling of questing ticks was performed in 12 forested sites, located in four districts (Legnica, Milicz, Lubań, and Oława) in SW Poland. Altogether, 792 ticks (151 females, 101 males, and 540 nymphs) representing I. ricinus were checked for the presence of A. phagocytophilum. The average infection level was 4.3%, with higher rate reported for adult ticks. The highest percentage of infected adults was observed in Milicz (17.4%) and the lowest in Oława (6.8%). The abundance of questing I. ricinus in all examined sites as well as the infection with A. phagocytophilum indicate for the first time the risk for HGA transmission in SW Poland.
Prevalence of EBV Genotypes in Polish, Taiwanese and Arabic Healthy Students and Association Between Genotypes and 30-bp Deletion in the LMP-1 Gene Phylogenetic Analysis
Authors: POLZ DOROTA , PODSIADŁO ŁUKASZ , STEC AGNIESZKA , POLZ-DACEWICZ MAŁGORZATA
Published: November 2019

The aim of this study was to compare the prevalence of EBV genotype and del-LMP-1 in saliva from Polish, Taiwanese and Arabic healthy students. The study group consisted of 56 healthy students; 24 of them Polish, 25 Taiwanese, and 7 Arabic. Typing was carried out using PCR with EBNA-2 primers. A detection of LMP-1 variants was also performed using PCR. EBV DNA was detected in 22 investigated samples (39.3%). Type 1 of the virus was dominant in both Polish and Taiwanese group. Among 62.5% Taiwanese with EBV 1 and 55.6% Polish detected EBV with 30-bp deletion in LMP-1 gene.
Microbial Transformations of 3-methoxyflavone by the Strains of Aspergillus niger
Authors: KOSTRZEWA-SUSŁOW EDYTA , DYMARSKA MONIKA , JANECZKO TOMASZ
Published: November 2019

Microbial transformation of 3-methoxyflavone into 3'-hydroxyflavon-3-yloxymethyl myristate was presented. Six filamentous fungi were used as biocatalysts: a wild strain of Aspergillus niger KB, its four UV mutants (A. niger MB, SBP, SBJ, 13/5) and the strain of Penicillium chermesinum 113. The highest yields were observed for the strains of A. niger KB and A. niger SBP (69.8% and 63.1%, respectively).
Zeaxanthin Biosynthesis by Members of the Genus Muricauda
Authors: PRABHU SUDHARSHAN , REKHA P.D. , ARUN A.B.
Published: November 2019

Zeaxanthin, a C40xanthophyll carotenoid, has potential biological applications in nutrition and human health. In this study we characterized carotenoid composition in 5 taxonomically related marine bacterial isolates from the genus Muricauda. The pigment was characterized using high performance liquid chromatography (HPLC) and mass spectrometry, which confirmed the presence of all-trans-zeaxanthin. Muricauda strains produced zeaxanthin as a predominant carotenoid. M. flavescens JCM 11812(T) produced highest yield (4.4 +/- 0.2 mg L(-1)) when cultured on marine broth at 32 degrees C for 72 h. This is the first report on the presence of zeaxanthin among the majority of species from the genus Muricauda.
The Oral Microbiome in Dental Caries
Authors: STRUŻYCKA IZABELA
Published: November 2019

Dental caries is one of the most common chronic and multifactorial diseases affecting the human population. The appearance of a caries lesion is determined by the coexistence of three main factors: acidogenic and acidophilic microorganisms, carbohydrates derived from the diet, and host factors. Socio-economic and behavioral factors also play an important role in the etiology of the disease. Caries develops as a result of an ecological imbalance in the stable oral microbiom. Oral microorganisms form dental plaque on the surfaces of teeth, which is the cause of the caries process, and shows features of the classic biofilm. Biofilm formation appears to be influenced by large scale changes in protein expression over time and under genetic control Cariogenic microorganisms produce lactic, formic, acetic and propionic acids, which are a product of carbohydrate metabolism. Their presence causes a decrease in pH level below 5.5, resulting in demineralization of enamel hydroxyapatite crystals and proteolytic breakdown of the structure of tooth hard tissues. Streptococcus mutans, other streptococci of the so-called non-mutans streptococci group, Actinomyces and Lactobacillus play a key role in this process. Dental biofilm is a dynamic, constantly active metabolically structure. The alternating processes of decrease and increase of biofilm pH occur, which are followed by the respective processes of de- and remineralisation of the tooth surface. In healthy conditions, these processes are in balance and no permanent damage to the tooth enamel surface occurs.
Bacteriophages as an Alternative Strategy for Fighting Biofilm Development
Authors: PARASION SYLWIA , KWIATEK MAGDALENA , GRYKO ROMUALD , MIZAK LIDIA , MALM ANNA
Published: November 2019

The ability of microbes to form biofilms is an important element of their pathogenicity, and biofilm formation is a serious challenge for today's medicine. Fighting the clinical complications associated with biofilm formation is very difficult and linked to a high risk of failure, especially in a time of increasing bacterial resistance to antibiotics. Bacterial species most commonly isolated from biofilms include coagulase-negative staphylococci, Staphylococcus aureus, Enterococcus faecalis, Enterococcus faecium, Escherichia coli, Proteus mirabilis, Klebsiella pneumoniae, Pseudomonas aeruginosa and Acinetobacter spp. The frequent failure of antibiotic therapy led researchers to look for alternative methods and experiment with the use of antibacterial factors with a mechanism of action different from that of antibiotics. Experimental studies with bacteriophages and mixtures thereof, expressing lytic properties against numerous biofilm-forming bacterial species showed that bacteriophages may both prevent biofilm formation and contribute to eradication of biofilm bacteria. A specific role is played here by phage depolymerases, which facilitate the degradation of extracellular polymeric substances (EPS) and thus the permeation of bacteriophages into deeper biofilm layers and lysis of the susceptible bacterial cells. Much hope is placed in genetic modifications of bacteriophages that would allow the equipping bacteriophages with the function of depolymerase synthesis. The use of phage cocktails prevents the development of phage-resistant bacteria.
Identification of Genomic Heterogeneity among Lactococcus lactis Strains by Plasmid Profiling, PFGE and 16S rDNA Sequence Analysis
Authors: GUNAY-ESIYOK OZLEM , AKCELIK NEFISE , AKCELIK MUSTAFA
Published: November 2019

Lactococcus lactis strains are used commonly as starters, which contribute to desirable flavour and texture properties known as strain-specific, in dairy industry. Genomic heterogeneity of 30 L. lactis strains originating from Turkey and characterized phenotypically were investigated in this study. Plasmid profiling, PFGE and 16S rDNA sequence analyses were performed to determine the genetic variability of strains. High degree of heterogeneity was detected among the L. lactis strains. Plasmid profiles of strains showed that compared to the plasmid free control strains, namely; L. lactis subsp. lactis IL1403 and L. lactis subsp. cremoris MG1614, all tested strains carried one to ten plasmids with molecular size ranging from 1.5 to 41.5kb. The fingerprints of strains obtained by PFGE from digestion with ApaI, SmaI and I-CeuI restriction endonucleases of chromosomal DNA's were compared with each other. All strains out of four were grouped into a large cluster A with at least 44% similarity level. The other four strains formed a minor duster B, distinctively different from major cluster A. PFGE results were confirmed by 16S rDNA sequence analysis and strains included in cluster B were identified as members of different species. These results suggested that morphologic and biochemical methods should be verified by reliable molecular approaches for the purpose of strain typing. Also, PFGE was found suitable to determine genomic differentiations among inter- and intra species.
Clinical and Microbiological Evaluation of Biofilm-Gingival Interface Classification in Patients with Generalized Forms of Periodontitis
Authors: KOWALSKI JAN , GÓRSKA RENATA
Published: November 2019

STUDY OBJECTIVE:
In 2008 a new division of periodontal diseases based on the concept of biofilm-gingival interface (BGI), was presented. The aim of this study was to assess whether the proposed BGI categories could be useful in diagnosing patients with generalized forms of both aggressive (gAP) and chronic (gCP) periodontitis. DESIGN AND KEY METHODS: The study group consisted of 40 individuals (17 with gAP and 23 with gCP). All patients were divided into three groups according to the BGI scale: 4 patients with P1 (pockets deeper than 4 mms with bleeding index below 10%), 20 individuals with P2 (pockets deeper than 4 mms with bleeding index 10-50%) and 16 individuals with P3 (pockets deeper than 4 mms with bleeding index above 50%). The presence of bacteria in subgingival plaque was analyzed using the polymerase chain reaction method.
RESULTS:
Statistically significant differences with respect to all parameters measured were found when patients were grouped according to the BGI scale--they were higher in P3 group. In gAP group A. actinomycetemcomitans and C. rectus were found in a higher numbers. No such differences were observed between P2 and P3 groups.
CONCLUSIONS:
The BGI scale seems to be helpful in effective and quick classifying of patients into adequate clinical subgroups.
Effects of Cryopreservation at -80oC on the Formulation and Pathogenicity of the Obligate Aphid Pathogen Pandora nouryi
Authors: ZHOU XIANG , FENG MING-GUANG , HUANG ZHI-HONG
Published: November 2019

Cryopreservation at -80 degrees C is an alternative to liquid nitrogen storage for Entomophthorales. However, detailed studies about its effects on fungal pathogenicity and formulation are very limited. In the present study, the obligate aphid pathogen Pandora nouryi was formulated as mycelia grown on millet-gel granules after preservation as primary spores at -80 degrees C for 3-18 months, although its ability to produce infectious conidia gradually diminished. The sporulation capacity of this granular formulation was reduced to 18.5 x 10(4) conidia/mg after 18 months of storage, which was still higher than that of mycotized aphids. The half-decline time of sporulation capacity was computed as 13.6 months. The infectivity to the green peach aphid Myzus persicae had no significant decline in 12 months. The ability to yield resting spores within host carcasses remained unchanged, and the probability of resting spore formation increased with the conidial concentrations that infect aphids. Therefore, cryopreservation at -80 degrees C exerted a marginal impact on formulation and pathogenicity of P. nouryi and can substitute for costly liquid nitrogen storage in routine laboratory studies. The potential of the formulation in aphid biocontrol can be maintained although there is a risk of losing fungal sporulation ability in long-term preservation.
Trends in Antifungal Susceptibility of Candida Species – One Year Observation
Authors: GOŁAŚ M. , NETSVYETAYEVA I. , SIKORA M. , PISKORSKA K. , SULIK-TYSZKA B. , SWOBODA-KOPEĆ E.
Published: November 2019

In the past years opportunistic fungal infections have seriously increased, mainly in immunocompromised patients. The aim of the study was to determine the prevalence of yeast-like fungi in invasive candidiasis and to estimate its susceptibility to chosen antifungal agents. One hundred and sixty strains of yeast-like fungi were cultured from various clinical material: samples from lower respiratory tract, blood, the peritoneal cavity and others. The susceptibility tests were established according to the quantitative E-test method. The Candida genus represented the main etiological factor of invasive candidiasis. The predominant species were: C. glabrata (71/160), C. albicans (34/160), C. krusei (17/160), C. tropicalis (14/160). All tested strains were the most resistant to itraconazole. Candida glabrata presented the 100% susceptibility to amphotericin B and caspofungin and was the least susceptible to itraconazole, posaconazole and voriconazole. Candida albicans was the most susceptible species to all antymicotics.
Definitive Assignment by Multigenome Analysis of the Gammaproteobacterial Genus Thermithiobacillus to the Class Acidithiobacillia
Authors: HUDSON COREY M. , WILLIAMS KELLY P. , KELLY DONOVAN P.
Published: November 2019

The class Acidithiobacillia was established using multiprotein phylogenetic analysis of all the available genomes of the genus Acidithiobacillus (comprising Family I, the Acidithiobacillaceae, of the Acidithiobacillales, the order created for Bergey's Manual of Systematic Bacteriology), and for representative genomes of all available bacterial orders. The Acidithiobacillales contain a second family, the Thermithiobacillaceae, represented by Thermithiobacillus tepidarius, and created on the basis of nearest neighbour 16S ribosomal RNA gene sequence similarities. This could not be included in the original multiprotein analysis as no genome sequence for Thermithio bacillus was available. The publication of the genome sequence of Thermithiobacillus tepidarius in 2013 has enabled phylogenetic assessment of this organism by comparative multigenome analysis. This shows definitively that Thermithiobacillus is a member of the class Acidithiobacillia, distinct from the Acidithiobacillus genus, and confirms it to represent a second family within the Acidithiobacillia.
Optimization of Culture Conditions for Exopolysaccharide Production by a Probiotic Strain of Lactobacillus rhamnosus E/N
Authors: POLAK-BERECKA MAGDALENA , WAŚKO ADAM , KUBIK-KOMAR AGNIESZKA
Published: November 2019

The effects of culture conditions on exopolysaccharides (EPS) production by a probiotic Lb. rhamnosus E/N have been studied using the Plackett-Burman design. Process optimization was performed in stationary cultures to maximize the production of EPS. In order to verify the optimal conditions, an analysis was performed of EPS production in fermentation culture. Batch fermentation was carried out at working volume of 2.51. The optimal temperature, pH, carbon source, and nitrogen source conditions were 37 degrees C, pH 5.0, galactose, and yeast extract, respectively. EPS production was improved by 210.28 mg/l in stationary cultures compared to 134.2 mg/l in a control grown on commercial MRS medium. The fermentor experiment showed the possibility of increasing EPS biosynthesis by 175.8%. Our results clearly demonstrate that in the case of Lb. rhamnosus E/N specific culture conditions can enhance EPS production for possible application in the industry.
Application of Microsatellite-Primed PCR (MSP-PCR) and PCR Melting Profile (PCR-MP) Method for Intraspecies Differentiation of Dermatophytes
Authors: CIESIELSKA ANITA , KOZŁOWSKA MAGDALENA , GADZALSKI MAREK , WOREK MARIUSZ , JAWORSKI ADAM , STĄCZEK PAWEŁ
Published: November 2019

In this study, two PCR-based methods (MSP-PCR and PCR-MP) were compared for their abilities to identify intraspecies variations of 23 isolates of Trichophyton rubrum, 78 isolates of Trichophyton interdigitale and 22 isolates of Microsporum canis, obtained mainly from patients in Lódź city. The results allowed to distinguish four types (containing two subtypes) characteristic for T. interdigitale and three types characteristic for T. rubrum using PCR-MP method. Analysis conducted using MSP-PCR with (GACA)4 primer revealed four types for T. rubrum and three types (containing one subtype) for T. interdigitale and with (GTG), primer showed two types (containing one subtype) for T. rubrum and six types (containing one subtype) for T. interdigitale. No differentiation was observed for the M. canis isolates with either method.
Sonication as a Diagnostic Approach Used to Investigate the Infectious Etiology of Prosthetic Hip Joint Loosening
Authors: BOGUT AGNIESZKA , NIEDŹWIADEK JUSTYNA , KOZIOŁ-MONTEWKA MARIA , STRZELEC-NOWAK DAGMARA , BLACHA JAN , MAZURKIEWICZ TOMASZ , MACIAS JULIA , MARCZYŃSKI WOJCIECH
Published: November 2019

The purpose of the study was to evaluate the usefulness of sonication for the diagnosis of prosthetic joint infections (PJIs) by its comparison with periprosthetic tissues (PTs) and synovial fluid (SV-F) cultures. The study groups included 54 patients undergoing exchange of total hip prostheses for so called "aseptic" loosening occurring without clinical manifestations of an accompanying PJI and 22 patients who developed a sinus tract communicating with the prosthesis which was indicative of an ongoing infectious process. Significant positive culture results were obtained among 10 (18.5%) patients with "aseptic" implant failure and in 18 (81.8%) patients who developed a sinus tract. Sonicate-fluid (S-F) yielded bacterial growth in all culture-positive patients with "aseptic" loosening vs. 15 patients with presumed PJIs. There was a concordance in terms of bacterial species isolated from S-F and conventional cultures from individual patients. Coagulase-negative staphylococci were isolated most frequently. Sensitivity of sonication (75%) exceeded that estimated for PTs (69%) and SV-F (45%) cultures. We conclude that identification of causative agents of PJIs which is critical to further therapeutic decisions is aided by the combination of sonication and conventional culture.
Laccase Production by Trameteshirsuta, Characterization, and Its Capability of Decoloring Chlorophyll
Authors: LIU JIAYANG , LIU WENHUA , CAI YUJIE , LIAO XIANGRU , HUANG QINGGUO , LIANG XIAOHUI
Published: November 2019

The present study focused on laccase production, characterization, and its involvement in chlorophyll decolorization. Extracellular laccase, with the highest activity of 11 U/ml on day 8, was efficiently produced from Trametes hirsuta in 5 l bioreactor with optimized media comprising dual carbon sources, glucose and water hyacinth. A laccase was then purified from the supernatant to homogeneity with purification fold of 9.51 and recovery of 39.8% and an estimated molecular mass of 62 kDa by SDS-PAGE. The laccase showed activity at pH 2-6 and temperature 30-80°C and was relatively thermally stable at below 70°C and neutral pH. The laccase was applied to decolorize chlorophyll under different factors: temperature, pH, mediator, metal ions, and enzyme dosage. Other fungal laccases were also found to be able to degrade chlorophyll with rating from 52% to 88% following 1 h treatment with two laccase dosages (5 or 10 U/ml) in the absence of any other mediators at room temperature. These findings may be an important step in developing new, important, and commercially viable industrial applications for laccase enzymes.
Clonal Analysis of Staphylococcus hominis Strains Isolated from Hospitalized Patients
Authors: SZCZUKA EWA , TRAWCZYŃSKI KRZYSZTOF , KAZNOWSKI ADAM
Published: November 2019

Staphylococcus hominis is a part of normal skin flora, but it is also a cause of nosocomial infections. The aim of this study was to investigate the genetic relatedness of 62 strains of S. hominis obtained from hospitalised patients during an 11-year period. For the discrimination of these clinical strains we used repetitive sequence-based PCR method (BOX-PCR) and multiple-locus variable-number tandem repeat analysis (MLVA). BOX-PCR analysis revealed a large genetic diversity among clinical strains and we did not find a predominant clone with the ability to persist in a hospital environment. MLVA is not as discriminatory as BOX fingerprinting and would not be a useful method for epidemiological studies.
Unusual Occurrence of Salmonella Mikawasima in 2012–2013 in the Czech Republic: Part of a Multistate Outbreak?
Authors: MYŠKOVÁ PETRA , KARPÍŠKOVÁ RENÁTA
Published: November 2019

An increased number of cases of salmonellosis caused by the rare serotype Mikawasima was noted in the years 2012 and 2013 in the Czech Republic. A potential on-going outbreak caused by this serotype in European countries has also been reported. Altogether 14 human and 1 environmental isolates from the year 2012 and 11 human isolates from the year 2013 from different locations of the Czech Republic were sent to our laboratory for typing. Macrorestriction analysis together with antimicrobial susceptibility testing and PCR for ESBL and plasmid-mediated quinolone resistance detection were performed to compare our isolates. Twenty-one isolates created two very similar clusters and 5 isolates had different profiles. Twenty-five isolates were fully susceptible to all agents used. One isolate showed resistance to 12 microbial agents and possessed blaTEM, blaSHV, blaOXA, blaCTX-M and qnrB genes.
Disc-diffusion and PCR Detection of Methicillin Resistance in Environmental Airborne Strains of Staphylococcus spp
Authors: WOLNY-KOŁADKA KATARZYNA , LENART-BOROŃ ANNA , KASPROWICZ ANDRZEJ
Published: November 2019

The aim of this study was to assess the species composition of airborne Staphylococcus spp. in public premises, to determine the methicillin resistance of the isolates and the prevalence of mecA gene, determining resistance to β-lactams. In total 65 Staphylococcus strains were isolated from 54 sites. Four strains exhibited phenotypic methicillin resistance, while the presence of mecA gene was found in 11 strains. The results of both assays were compared, showing that the phenotypic tests revealed methicillin resistance only in 36% of the examined samples. This study revealed high species diversity among airborne Staphylococcus spp. population, which consists of multidrug resistant strains.
Could a Step in the Isolation of Mycobacteria from Sputum Samples be Eliminated by New Decontamination Kits?
Authors: KARAKEÇE ENGIN , TERZI HÜSEYIN AGAH , ÇIFTCI İHSAN HAKKI
Published: November 2019

The efficacy of two mycobacterial homogenization, decontamination and concentration kits, Mycoprosafe and Decomics, were compared. A total of 146 sputum samples were examined in this study; 46 (31.5%) mycobacterial isolates were recovered with Mycoprosafe, while 39 (26.7%) mycobacterial isolates were recovered with Decomics. These results, although very preliminary, indicate that Mycoprosafe and Decomics are similar in terms of ease of use and price. However, Decomics offers several advantages, including the need for less equipment, a more rapid detection time, and an easy shipping process; thus, pre-processing can be done anywhere.
Nodulation competitiveness of Ensifer meliloti alfalfa Nodule Isolates and Their Potential for Application as Inoculants
Authors: MAREK-KOZACZUK MONIKA , WIELBO JERZY , PAWLIK ANNA , SKORUPSKA ANNA
Published: November 2019

Alfalfa (Medicago sativa) is a widely cultivated legume, which enters into nitrogen-fixing symbiosis with Ensifer (Sinorhizobium) spp. In this study, an autochthonous rhizobial population of Ensifer sp. occupying alfalfa nodules grown in arable soil was used as the basis for selection of potential inoculants. Alfalfa nodule isolates were identified as Ensifer meliloti by partial 16S rDNA, recA, atpD and nodC nucleotide sequencing. The sampled isolates displayed different symbiotic performance and diversity in the number of plasmids and molecular weight. Isolates that were the most efficient in symbiotic nitrogen fixation were tagged with a constitutively expressed gusA gene carried by a stable plasmid vector pJBA21Tc and used in competition experiments in soil under greenhouse conditions. Two E. meliloti strains LU09 and LU12, which effectively competed with indigenous soil rhizobia, were selected. The metabolic profiles of these selected strains showed differences in the use of carbon and energy sources. In addition, the LU09 strain exhibited bacteriocin production and LU12 mineral phosphate solubilization, which are valuable traits for soil survival. These strains may be considered as potential biofertilizers for alfalfa cultivation.
Agrobacterium-Mediated Transformation of Three Freshwater Microalgal Strains
Authors: SANITHA MARY , RADHA SUDHAKAR , FATIMA ANWAR ALIYA , DEVI SELVARAJU GAYATHRI , RAMYA MOHANDASS
Published: November 2019

Microalgal transformation has gained interest in recent years. Agrobacterium-mediated transformation remains as the most efficient method for the development of transgenic plants and microalgae due to its wide host range, inexpensive procedure and transfer of large segments of DNA. In the present study, three different microalgal species were isolated from freshwater environment and identified based on the morphological characteristics and ITS-2 region amplification. Agrobacterium-mediated transformation was successful for the isolates Chlorella sp., Ankistrodesmus sp and Scenedesmus bajacalifornicus. Gene integration and expression was confirmed by PCR amplification of hptII and GUS histochemical assay. A. tumifaciens contamination was checked by amplification of npt II gene (kanamycin resistant) which lies outside the T-border. Based on GUS assay, transformation efficiencies were found to be 12.25% for Chlorella sp. 2.96% for Scenedesmus bajacalifornicus and 3.5% for Ankistrodesmus sp.
Evaluation of the Time Period for which Real-Time Polymerase Chain Reaction Detects Dead Bacteria
Authors: CHOE HYONMIN , INABA YUTAKA , KOBAYASHI NAOMI , MIYAMAE YUSHI , IKE HIROYUKI , FUJIMAKI HIROSHI , TEZUKA TARO , HIRATA YASUHIDE , SAITO TOMOYUKI
Published: November 2019

Real-time polymerase chain reaction (PCR) is currently widely used for the diagnosis of infections. We evaluated the time after treatment during which real-time PCR can detect dead bacteria. The presence of bacterial DNA was identified by real-time PCR through methicillin-resistant Staphylococcus (MRS)-PCR and universal PCR. Methicillin-resistant Staphylococcus aureus (MRSA), Staphylococcus epidermidis, and Escherichia coli were each killed with alcohol, antibiotics, or heat treatment in vitro. The detection periods of MRS-PCR for MRSA treated by alcohol, vancomycin, linezolid, and heat were found to be less than 16, 8, 12, and 8 weeks, respectively. The detection period of universal PCR for S. epidermidis treated by alcohol, cefazolin, and heat was less than 20, 20, and 4 weeks, whereas that for E. coli was 8, 20, and 4 weeks, respectively. The presence of detectable bacterial DNA in infected arthroplasty patients before and after successful treatment was also assessed by MRS- and universal PCR. MRS-PCR was positive in 6 patients before treatment and all became negative after a mean interval of 20.8 weeks (95% confidential interval, 13.2 to 33.7) after treatment. Universal PCR detected remnant bacterial DNA in 4 patients at a mean of 15.2 weeks (95% CI, 12.4 to 18.0) after treatment and was negative in 7 patients at a mean of 17.3 weeks (95% CI, 10.6 to 24.0) after treatment. Our studies revealed that real-time PCR detects dead bacteria for several weeks, but this capability decreases with time and is likely lost by 20 weeks after treatment.
Compartmentalization in Penicillin G Biosynthesis by Penicillium chrysogenum PQ-96
Authors: KURZĄTKOWSKI WIESŁAW , STANISZEWSKA MONIKA , BONDARYK MAŁGORZATA , GĘBSKA-KUCZEROWSKA ANITA
Published: November 2019

The arrangement of organelles in the sub-apical productive non-growing vacuolated hyphal cells of the high- and the low-penicillin-pro- ducing strains Penicillium chrysogenum was compared using transmission electron microscopy. In the productive cells of the high-yielding strain the endoplasmic reticulum and the polyribosomes with associated peroxisomes are frequently arranged at the periphery of the cytoplasm and around the vacuoles. At the high activity of penicillin G biosynthesis the immuno-label of the cytosolic isopenicillin N synthase is concentrated at the polyribosomes arranged in the peripheral cytoplasm and along the tonoplast as well as around the peroxisomes. On the basis of the obtained results the compartmentalization of the pathway of penicillin G biosymthesis is discussed. The obtained results support the phenylacetic acid detoxification hypothesis of penicillin G biosynthesis.
In vitro study of Biofilm Growth on Biologic Prosthetics
Authors: BELLOWS CHARLES , SMITH ALISON
Published: November 2019

Biologic prosthetics are increasingly used for the repair of abdominal wall hernia defects but can become infected as a result of peri- or early post-operative bacterial contamination. Data evaluating biofilm formation on biologic prosthetics is lacking. The aim of this study was to investigate the influence of different biologic prosthetics on the growth behavior of two different bacterial species and their ability to form biofilms. Methicillin resistant Staphylococcus aureus (MRSA) or Pseudomrnonas aeruginosa were incubated on disks of two biologic prosthetics-human acellular dermis (ADM), and porcine small intestinal submucosa (SIS). The bacteria were allowed to attach to the prosthetics and propagate into mature biofilms for 24 hours at 370C. Images of biofilms were obtained using confocal microscopy and scanning electron microscopy (SEM). The number of viable cells and the biofilm biomass were quantified by colony forming units (CFUs) and crystal violet staining respectively. Analysis of variance was performed to compare the mean values for the different prosthetics. Each biologic matrix had a distinct surface characteristic. SEM visualized mature biofilms characterized by highly organized multi-cellular structures on surface of both biologic prosthetics. Quantification of bacterial growth over time showed that ADM had the lowest CFUs and biofilm biomass at 24 hours post-inoculation compared to SIS for both bacterial strains. MRSA and P. aeruginosa can form mature biofilms on biologic prosthetics but the relative abundance of the biofilm varies on different prosthetic constructs. Biologic material composition and manufacturing methods may influence bacterial adherence.
Bioprotective Potential of Bacteriocinogenic Enterococcus gallinarum Strains Isolated from Some Nigerian Fermented Foods, and of their Bacteriocins
Authors: OLADIPO IYABO C. , SANNI ABIODUN I. , CHAKRABORTY WRITACHIT , CHAKRAVORTY SOMNATH , JANA SAYANTAN , RUDRA DEEP S. , GACCHUI RATAN , SWARNAKAR SNEHASIKTA
Published: November 2019

Enterococcus gallinarum strains isolated from some Nigerian fermented foods were found to produce bacteriocins. The bacteriocins had a broad spectrum of activity against both Gram-positive and negative bacteria. The effects of the bacteriocins and bacteriocinogenic organ- isms on Staphylococcus aureus infections in rats were evaluated. Sprague-Dawley rats were infected with S. aureus MTCC 737 and treated with E. gallinarum T71 and different concentrations of the bacteriocins from E. gallinarum W211 and T71. Staphylococcus aureus infection caused significant upregulation of aspartate aminotransferase and alanine aminotransferase levels in sera of the infected rats. Moreover, gelatin zymography revealed that infected gastric tissues showed elevated matrix metalloproteinase-9 activity. Bacteriocin treatments reduced the MMP-9 activity and inhibited the expressions of both Tumour Necrosis Factor Alpha (TNF-α) and Interleukin-1 Beta (IL-1β) dose dependently, pointing to a potential role of the bacteriocins in attenuating inflammatory responses to Staphylococcus aureus infec- tion. Gastric and GIT damage caused by staphylococcal infection were reduced in the Enterococcus gallinarum T71 and bacteriocin-treated groups also dose dependently. We conclude that these bacteriocins may have useful biomedical applications.
Synthetic Amphibian Peptides and Short Amino-Acids Derivatives against Planktonic Cells and Mature Biofilm of Providencia stuartii Clinical Strains
Authors: OSTROWSKA KINGA , KAMYSZ WOJCIECH , DAWGUL MAŁGORZATA , RÓŻALSKI ANTONI
Published: November 2019

Over the last decade, the growing number of multidrug resistant strains limits the use of many of the currently available chemotherapeutic agents. Furthermore, bacterial biofilm, due to its complex structure, constitutes an effective barrier to conventional antibiotics. The in vitro activities of naturally occurring peptide (Citropin 1.1), chemically engineered analogue (Pexiganan), newly-designed, short amino-acid derivatives (Pal-KK-NH2, Pal-KKK-NH2, Pal-RRR-NH2) and six clinically used antimicrobial agents (Gatifloxacin, Ampicilin, Cefotaxime, Ceftriaxone, Cefuroxime and Cefalexin) were investigated against planktonic cells and mature biofilm of multidrug-resistant Providencia stuartii strains, isolated from urological catheters. The MICs, MBCs values were determined by broth microdilution technique. Inhibition of biofilm formation by antimicrobial agents as well as biofilm susceptibility assay were tested using a surrogate model based on the Crystal Violet method. The antimicrobial activity of amino-acids derivatives and synthetic peptides was compared to that of clinically used antibiotics. For planktonic cells, MICs of peptides and antibiotics ranged between 1 and 256 μg/ml and 256 and ≥ 2048 μg/ml, respectively. The MBCs values of Pexiganan, Citropin 1.1 and amino-acids derivatives were between 16 and 256 μg/ml, 64 and 256 μg/ml and 16 and 512 μg/ml, respectively. For clinically used antibiotics the MBCs values were above 2048 μg/ml. All of the tested peptides and amino-acids derivatives, showed inhibitory activity against P. stuartii biofilm formation, in relation to their concentrations. Pexiganan and Citropin 1.1 in concentration range 32 and 256 μg/ml caused both strong and complete suppression of biofilm formation. None of the antibiotics caused complete inhibition of biofilm formation process. The biofilm susceptibility assay verified the extremely poor antibiofilm activity of conventional antibiotics compared to synthetic peptides. The obtained results showed that synthetic peptides are generally more potent and effective than clinically used antibiotics.
Relationship Between Measles Outbreaks Based on Genetic Analysis of Measles Virus Genomes Detected in Patients in Poland between 2006 and 2012
Authors: MAKÓWKA AGATA , LITWIŃSKA BOGUMIŁA
Published: November 2019

This study describes the molecular characterization of 56 MeV strains obtained from 56 patients in Poland from 2006 to 2012. The C-terminal fragment of nucleoprotein gene was analysed. It has been found out during 2006 and 20012 MeV strains circulating in Poland belonged to genotypes D4, D5, D6 and B3. The D4 strains isolated in Poland were different from any other D4 strain circulating at the same time in Europe, whereas all other MeV strains isolated during 2007-2012 were related to strains from other countries. The present data suggest that after 2006 the MeV strains were imported.
Production of Ligninolytic Enzymes by Cultures of White Rot Fungi
Authors: GÓRSKA EWA B. , JANKIEWICZ URSZULA , DOBRZYŃSKI JAKUB , GAŁĄZKA AGNIESZKA , SITAREK MONIKA , GOZDOWSKI DARIUSZ , RUSSEL STEFAN , KOWALCZYK PAWEŁ
Published: November 2019

Some Basidiomycota were chosen for studies of key ligninases synthesis (25°C, 30 days) in modified medium (shaken or not cultures) with added wheat straw. Liquid Czapek medium with straw yielded a higher amount of laccase than peroxidase, ground straw induced enzyme worse than chopped straw. With peroxidase the reverse dependencies were observed. Laccase of Lentinus edodes synthesized two enzyme isoforms (ca 30 and 16 kDa). In T. versicolor culture active laccase protein with highest molecular mass ca 65 kDa was found. P. sajor-caju yielded three different peroxidase isoforms. Ligninase biosynthesis depended on strain, straw fragmentation extent, culture method and growth medium.
Identification of some Yeasts by Fatty Acid Profiles
Authors: ALOKLAH BASSAM , ALHAJALI ANWAR , YAZIJI SABAH
Published: November 2019

178 yeasts isolated from various foods were identified using ID32 C strip; Saccharomyces cerevisiae (47), Rhodotorula mucilaginosa (19), Candida kefyr (33), Candida krusei (37), Candida lusitaniae (28), Cndida lipolytica (14). The yeasts were harvested, saponified, derivatized, and extracted, and fatty acid analysis was performed by Gas Chromatography Mass Spectrometry. The yeast lipids contained Decanoic acid (C10:0), Luaric acid (C12:0), Myristoleic acid (C14:1), Myristic acid (C14:0), Pentadecanoic acid (C15:0), Palmitoleic acid C16:1), Pamitic acid (C16:0), Heptadecenoic acid (C17:1), Linoleic acid (C18:2), Linolenic acid (C18:3), Oleic acid (C18:1) and Stearic acid (C 18:0). The differences in the composition of these fatty acids could be used to identify the species as an accurate, efficient and relatively rapid method.
KEBERADAAN KEBUTUHAN EXISTENCE, RELATEDNESS, GROWTH, DALAM VIDEO GAME HARVEST MOON BACK TO NATURE
Authors: Budiman Arief
Published: December 2019

ABSTRACT Playing video games is basically an activity where it must have an stimulus energy which is commonly referred to as motivation. Without motivation, player will not be interested in playing a video game with a long duration. The Harvest Moon Back to Nature video game as a video game that was quite successful, which made the video game a legendary video game for the audience's audience as early as 1997. Harvest Moon Back to Nature looks so strong in representating about social life of rural communities, especially one who works as a farmer. It creates interest which in turn encourages researchers to know more about how motivation for human needs is presented by the Harvest Moon Back to Nature video game. This research fundamentally use interpretative analytical descriptive method with narrative approach. Using a view from ERG theory Clayton Alderfer, which according to him motivation is related to basic human needs which consist of existence needs, relatedness needs, and growth needs. In increasing one's motivation, the three basic human needs must be fulfilled wherever possible. From the results of the research conducted using these instruments, it can be concluded that the power of motivation has been presented in the Harvest Moon Back to Nature video game from the fulfillment of existence needs, relatedness needs, and growth needs, both in the context events of farming, raising, and in carrying out other events.Keywords : motivation, existence needs, relatedness needs, and growth needs.
DEFINING THE IDEAL COMPETENCY MODEL FOR THE UNITED NATIONS MILITARY OBSERVERS (STUDY ON THE INDONESIAN MILITARY OBSERVERS)
Authors: Anwarr Syaiful
Published: December 2019

A military observer is a military officer from a certain country who is deployed as part of a United Nations Peacekeeping Mission. The purpose of this study is to define an ideal competency model for the UN military observers of the Indonesian military by considering certain characteristics embedded in superior military observers that distinguish them from the non-superior ones. This study applied several data collecting techniques such as behavioral event interviews, observation, and library study. Two types of analysis, namely: content analysis and discriminant analysis are utilized to create data weighting and to discover the differentiating characteristics. From this study, it can be concluded that there is a very close and significant relationship between the competency and performance of the UN military observers from Indonesia. Those in the superior category do have three strong differentiating competencies, namely: initiative, relationship building, and impacts and influence production. And then this study also found an ideal competency model for a UN military observer, where the model consists of the three competencies plus eight other competencies, namely: team working, self-controlling, local language skill, local culture knowledge, vehicle driving skill, military knowledge and skills, diplomacy and negotiation skills, and administrative skills.

Keywords: Competency, Competency model, Military observer, Recruitment, UN Peacekeeping
THE INDONESIAN NATIONAL ARMED FORCES (TNI) AND FALINTIL- FORÇAS DE DEFESA DE TIMOR LESTE (F-FDTL)’S DEFENSE DIPLOMACY: A WAY OF CONFLICT RESOLUTION IN TIMOR LESTE
Authors: Ambarsari Khory Wandira , Risman Helda
Published: December 2019

Indonesia and Timor Leste have had an irrelevant relationship related to the history of Seroja Operation on 7 December 1975 when the TNI, formerly known as ABRI (Armed Forces of the Republic of Indonesia) carried out a total military operation in order to keep Timor Leste for not separate from Indonesia, while later it caused losses where many TNI and Falintil- Forças De Defesa De Timor Leste (F-FDTL)’s personnel were killed in the war. Time passed, now the relationship between both countries is getting better due to the efforts from the Indonesian and Timor Leste’s parties, especially the national armies, had been done. To solve the conflict between those countries, defense diplomacy is needed. In this article, the writers tend to describe how both armed forces conduct defense diplomacy so Indonesia and Timor Leste’s relationship is getting better, indeed now both countries have done some collaborations in some aspects in the economy and military. Later, the writers will analyze more to find out the best conflict resolution that had been done by both armed forces, and the existence of each State Leaders’ participation. It is clearly stated that by utilizing defense diplomacy through visiting state leaders, having an official meeting, holding military cooperation such as doing exchange troops are some best ways of conflict resolution that can be done by Indonesia and Timor Leste to create a better relationship.

Keywords: Defense Diplomacy, Conflict Resolution, Indonesia National Army (TNI), F-FDTL, Timor Leste