bluuebunny/crossref_metadata_2025 huggingface dataset:
Influence of flow velocity on the removal of faecal coliforms in horizontal subsurface flow constructed wetland
Authors: Lohay W. S. , Lyimo T. J. , Njau K. N.
Published: October 2012

In order to determine the influence of flow velocity on the removal of faecal coliforms (FC) in constructed wetlands (CWs), removal rate constants of FC (kFC) were studied at various flow velocities (u). Membrane filtration technique was used during analysis. Values of kFC were determined using Reed's equation of pathogen removal; the results were compared with the plug flow equation. According to Reed's equation, kFC values ranged from 1.6 day−1 at a velocity of 4 m/day to 34.5 day−1 at a velocity of 42.9 m/day. The removal rates correlated positively with flow velocity (r = 0.84, p < 0.05). On assuming a plug flow equation, removal rates constants ranged from 0.77 to 11.69 day−1; a more positive correlation (r = 0.93, p < 0.05) was observed. Optimum removal rate constants were observed for the velocity ranging 36 to 43 m/day. Generally, the increase of flow velocity improved FC removal rate constants: implying that pathogen removals are influenced by diffusion of the microorganisms into the biofilms on CW media. The velocity dependent approach together with the plug flow equation is therefore proposed for incorporation in the design of CW in a tropical climate where temperature variations are minor.
HIV-1 resistance profile in plasma and peripheral blood lymphocytes in a group of naive patients
Authors: Siljic Marina , Salemovic Dubravka , Jevtovic Dj. , Pesic-Pavlovic Ivana , Zerjav Sonja , Nikolic Valentina , Stanojevic Maja
Published: October 2012

Transmitted HIV-1 drug resistance (TDR) is a persisting problem, even though
the prevalence of primary resistance may remain stable or start to decline.
Proviral DNA detectable in peripheral blood mononuclear cells (PBMCs) is a
reservoir of drug resistant viral variants and could be an alternative marker
to viral RNA for the detection of drug resistance mutations. The aim of this
study was to compare the HIV-1 resistance profile between plasma viral RNA
and proviral DNA in a group of untreated patients. Thirty-one HIV-1
seropositive patients without prior ARV treatment were included in the study.
The presence of non-polymorphic drug resistance mutations was identified in
10 cases in proviral DNA and in 11 cases in plasma according to different
scoring systems. Our results show a similar resistance profile between plasma
RNA and proviral DNA, but with some discordances present. The sequencing of
proviral DNA could provide useful additional information with regard to
primary resistance.
First case of visceral Leishmaniosis/HIV coinfection in Nis - southeastern Serbia
Authors: Marjanovic G. , Miladinovic-Tasic Natasa , Gabrielli Simona , Otasevic Suzana , Popovic-Dragonjic Lidija , Kocic Branislava , Arsic-Arsenijevic Valentina , Tadic Ljiljana , Cancrini Gabriella
Published: October 2012

Visceral leishmaniosis (VL) has emerged as an important opportunistic
parasitosis associated with human immunodeficiency virus (HIV) infection. The
aim of this paper is to report the first case of Leishmania/HIV coinfection
in a patient from Nis (Southeastern Serbia). Microscopical examination of
Giemsa-stained bone marrow (BM) smears show the presence of Leishmania spp.
amastigotes based on their morphological characteristics. In spite of the
parasitological finding, the serological test applied gave negative results.
Molecular analyses confirmed the infection and allowed us to identify the
leishmania species as Leishmania infantum (100% identity). VL/HIV coinfection
has important clinical, diagnostic and epidemiological implications. In fact,
the failure of serological tests is expected in this condition, and the
application of molecular diagnostics to the blood may offer, apart from an
easy and non-invasive diagnostic opportunity, the possibility of warning
about the risk of possible nosocomial infections.
Successful isolation of Helicobacter pylori after prolonged incubation: A case report of prolonged incubation for H. pylori
Authors: Babic Tatjana , Miljkovic-Selimovic Biljana , Kocic Branislava , Nagorni A. , Ristic Ljiljana
Published: October 2012

The culture of Helicobacter pylori from a gastric biopsy is the ?gold
standard? in the diagnosis of H. pylori infection. However, the primary
isolation of H. pylori from gastric biopsies is rather demanding. The
duration of incubation for the isolation of H. pylori has been recommended to
be five to seven days: in the present case, we found that a prolonged
incubation period allowed the successful isolation of H. pylori from a
patient with ulcus ventriculi. Biopsies were placed directly into transport
medium and processed for culture within two hours. On day 14, one suspected
H. pylori-like colony appeared on one of the plates. The isolate was
confirmed to be H. pylori based on its typical colony morphology, negative
Gram stain, and positive urease, catalase and oxidase tests. The isolate,
requiring 14 days recovery, later exhibited the normal growth characteristics
of H. pylori strains, indicating its unusually long incubation requirement
was a temporary predicament.
Detection of drug-resistant mycobacterium tuberculosis strains isolated in Serbia by the GenoType MTBDRsl assay
Authors: Zivanovic Irena , Vukovic Dragana , Dakic Ivana , Stefanovic Gordana , Savic Branislava
Published: October 2012

The new GenoType MTBDRsl assay (Hain Lifescience) detects mutations most
frequently associated with resistance to fluoroquinolones (FLQ),
aminoglycosides/cyclic peptides (AG/CP), and ethambutol (EMB) and thus,
enables rapid identification of extensively drug-resistant (XDR)
Mycobacterium tuberculosis strains. A set of 19 multidrug-resistant (MDR)
strains isolated in Serbia in 2011 was tested by the MTBDRsl assay and by
conventional drug susceptibility testing (DST). The sensitivity and
specificity of the MTBDRsl assay were as follows: 100% for both for FLQ; 100%
for both for AG/ CP; and 58.3% and 85.7%, respectively, for EMB. The
sensitivity for detection of XDR strains was 100%. Mutations in the gyrA,
rrs, and embB genes established in local M. tuberculosis strains resistant to
the respective drugs have been reported as the most prevalent in other
studies as well. We recommend the MTBDRsl assay as a screening test for the
preliminary detection of XDR-TB cases in Serbia, but not as a replacement of
the conventional second-line DST.
Sphingomonas paucimobilis as a biofilm producer
Authors: Gusman Vera , Medic Deana , Jelesic Zora , Mihajlovic-Ukropina Mira
Published: October 2012

The aim of this study was, for the first time in our country, to identify the
capability of isolates of Sphingomonas paucimobilis to form a biofilm. In the
3-month period from January 1st to March 31st 2010, a total of 2630 samples
of drinking water were microbiologically examined in the Institute of Public
Health of Vojvodina, Serbia. From all examined samples of drinking water,
non-fermentative Gram-negative oxidase positive bacilli were identified in
113 samples (4.30%). The bacteria isolates were identified as Sphingomonas
paucimobilis (4 isolates), based on analysis by the automated VITEK 2 Compact
system; biofilm formation was examined according to the modified method of
Stepanovic et al. (2000). All 4 Sphingomonas paucimobilis strains tested
showed a strong biofilm-producing ability. Considering the potential
pathogenic features of Sphingomonas paucimobilis, the presence of these
strains in drinking water distribution systems is not desirable. Therefore,
adequate biofilm degradation and management of drinking-water distribution
networks that will guarantee microbiologically safe drinking water is
recommended.
Humoral immune response to campylobacter jejuni in patients with enterocolitis and Guillain-Barré syndrome
Authors: Ristic Ljiljana , Kocic Branislava , Babic Tatjana , Apostolski S. , Spasic Mirjana , Miljkovic-Selimovic Biljana
Published: October 2012

Campylobacter jejuni is one of the most important causes of diarrheal disease
worldwide. In addition, it can cause neurological post-infectious sequels,
such as Guillain-Barr? syndrome (GBS). Humoral immune response to C. jejuni
was monitored in patients with C. jejuni enterocolitis, GBS patients and
healthy persons, by ELISA. Statistical significance between patients with
enterocolitis and healthy persons, as well as among GBS patients and healthy
controls, was proven. Statistical significance in IgA among the examined
groups was also noticed. The highest values of IgM were found in the
patients with GBS, while the highest values of IgG were found in those with
enterocolitis. C. jejuni is a significant cause of antecedent infection in
GBS. ELISA techniques can be considered a reliable method in determining the
presence of serum antibodies in patients with enterocolitis caused by C.
jejuni, as well as in patients with GBS.
In vitro antibacterial activity of garlic and synergism between garlic and antibacterial drugs
Authors: Cirkovic Ivana , Jovalekic M. , Jegorovic B.
Published: October 2012

nema
Campylobacter and helicobacter in the etiology of gastrointestinal diseases
Authors: Miljkovic-Selimovic Biljana , Kocic Branislava , Babic Tatjana
Published: October 2012

The order Campylobacterales comprises two genera: Campylobacter and
Helicobacter, with a widespread distribution in both humans and animals. They
are Gram-negative, spiral, helical and microaerophilic bacteria, with an
optimal growth temperature of 37?C for H. pylori and 42?C for C. jejuni
strains. While Helicobacter pylori are restricted to humans, other
helicobacter species can be found in different mammals and occasionally in
humans. Several Campylobacter species are recognized as human pathogens,
while distinct species are pathogenic only occasionally, in children, the
elderly and immunocompromised patients. Campylobacters and helicobacters are
well adapted to the living conditions inside the gastrointestinal tract,
where they can cause diseases as a consequence of inflammation. In addition,
they are related to certain extraintestinal diseases, post-infectious
sequels, malignancy and autoimmunity. Different clinical presentations of
human disorders may be the consequences of the diversity in host immune
response, bacterial genome, endotoxin activity as well as specific bacterial
virulence factors.
Screening for the presence of biosynthetic genes for antimicrobial lipopeptides in natural isolates of Bacillus sp.
Authors: Stankovic S. , Mihajlovic Sanja , Draganic V. , Dimkic I. , Vukotic G. , Beric Tanja , Fira Dj.
Published: October 2012

A collection of 205 natural isolates of Bacillus was tested for the presence
of genes for biosynthesis of antimicrobial lipopeptides, iturin, surfactin,
fengycin and bacillomycin D. For the detection of iturin producers by PCR
screening, we used forward ITUP1-F and reverse ITUP2-R primers which are
capable of detecting a 2-kb region that includes the intergenic sequence
between the ituA and ituB genes. A 675-bp fragment from the gene sfp from B.
subtilis encoding 4?-phosphopantetheinyl transferase involved in the
biosynthesis of surfactin was targeted for amplification by using primers P17
and P18. Other two pairs of primers were BACC1F and BACC1R for bacillomycin D
and FEND1F and FEND1R for potential fengycin producers, respectively. The
results of the screening showed that the majority of tested strains had more
than one biosynthetic operon, since 81% possessed the genes for bacillomycin
D production, 54% for surfactin, 38% for iturin and 25% for fengycin
production.
Determination of staphylococcal enterotoxins in cheese by immunoenzyme assays
Authors: Jankovic Vesna , Djordjevic Vesna , Lakicevic Brankica , Borovic Branka , Velebit B. , Mitrovic Radmila
Published: October 2012

Staphylococcal food poisoning is one of the most common foodborne diseases
resulting from the ingestion of staphylococcal enterotoxins (SEs) preformed
in foods by enterotoxigenic strains of coagulase-positive staphylococci
(CPS), mainly Staphylococcus aureus. The presence of enterotoxigenic strains
of coagulase-positive staphylococci in raw milk during the production process
leads to the contamination of products and outbreaks of alimentary
intoxication. The problem of Staphylococcus aureus in cheese remains
significant on a global level. Domestic cheese contaminated with
enterotoxigenic staphylococci can result in the formation of enterotoxin,
which can produce foodborne illness when the product is ingested. Due to
microbiological contamination, microbiological criteria are tools that can be
used in assessing the safety and quality of foods. In order to avoid
foodborne illness, the Serbian Regulation on General and Special Conditions
for Food Hygiene (Official Gazette of RS, No. 72/10) provides microbiological
criteria for staphylococcal enterotoxins in dairy products.
Quantification of viable spray-dried potential probiotic lactobacilli using real-time PCR
Authors: Radulovic Zorica , Mirkovic N. , Bogovic-Matijasic Bojana , Petrusic Milica , Petrovic Tanja , Manojlovic Verica , Nedovic V.
Published: October 2012

The basic requirement for probiotic bacteria to be able to perform expected
positive effects is to be alive. Therefore, appropriate quantification
methods are crucial. Bacterial quantification based on nucleic acid detection
is increasingly used. Spray-drying (SD) is one of the possibilities to
improve the survival of probiotic bacteria against negative environmental
effects. The aim of this study was to investigate the survival of spray-dried
Lactobacillus plantarum 564 and Lactobacillus paracasei Z-8, and to
investigate the impact on some probiotic properties caused by SD of both
tested strains. Besides the plate count technique, the aim was to examine the
possibility of using propidium monoazide (PMA) in combination with real-time
polymerase chain reaction (PCR) for determining spray-dried tested strains.
The number of intact cells, Lb. plantarum 564 and Lb. paracasei Z-8, was
determined by real-time PCR with PMA, and it was similar to the number of
investigated strains obtained by the plate count method. Spray-dried Lb.
plantarum 564 and Lb. paracasei Z-8 demonstrated very good probiotic ability.
It may be concluded that the PMA real-time PCR determination of the viability
of probiotic bacteria could complement the plate count method and SD may be a
cost-effective way to produce large quantities of some probiotic cultures.
Comparative analysis of the potential probiotic abilities of lactobacilli of human origin and from fermented vegetables
Authors: Petrovic Tanja , Dimitrijevic Suzana , Radulovic Zorica , Mirkovic N. , Rajic Jasmina , Obradovic D. , Nedovic V.
Published: October 2012

In this study, twelve strains of Lactobacillus plantarum derived from
spontaneously fermented vegetables (carrots and beetroot) and traditionally
prepared sauerkraut were compared for their potential probiotic abilities
with seven Lactobacillus rhamnosus strains of human origin. The tested
strains were investigated for some technological properties and in vitro
functional characteristics for potential probiotic strains. Selection
probiotic criteria included the ability of the strains to withstand
conditions similar to the digestive tract, antimicrobial activity against a
wide range of intestinal pathogens and sensitivity to antibiotics. The total
acidity in milk was generally higher in intestinal strains in relation to
plant strains. The ability of the tested strains to survive simulated gastric
conditions showed a greater resistance of the human strains at a low pH 2.5
and the presence of pepsin, while in the presence of bile salts and
pancreatin, some intestinal strains were more sensitive compared to plant
strains. A wide spectrum of antimicrobial activities was observed in all
tested strains. Most of the plant strains were resistant to aminoglycosides
and vancomycin but sensitive to ampicillin and penicillin, while only some
intestinal strains were resistant to these drugs.
The possibility of using macrophytes in lake Palic sediment remediation
Authors: Petricevic J. , Gujanicic V. , Radic D. , Lalevic B. , Bozic M. , Rudic Z. , Raicevic V.
Published: October 2012

Lake Palic is a typical, shallow Pannonian plain lake, with thick sediment
layers and high content of phosphorus and nitrogen. The thick layers are the
result of accelerated eutrophication, and cause biodiversity loss and
disruption of the ecosystem. Numerous methods can be used to overcome the
problem of accelerated eutrophication, (sediment removal, phosphorus
inactivation etc.). However, these methods have many deficiencies. Lately,
using macrophytes as a way to resolve this problem is becoming more and more
popular. The aim of this work was to examine the germination possibilities of
white mustard (Sinapis alba L.), alfalfa (Medicago sativa L.), oats (Avena
sativa L.) and lettuce (Lactuca sativa L.) on Lake Palic?s sediment, as a
prescreening test for their suitability for further phytoremediation. The
results show that plant species can germinate and grow in early stages in
such substrates, so they can be recommended for sediment phytoremediation.
Phenazines producing pseudomonas isolates decrease Alternaria tenuissima growth, pathogenicity and disease incidence on cardoon
Authors: Josic Dragana , Protolipac Katarina , Starovic Mira , Stojanovic Sasa , Pavlovic Snezana , Miladinovic M. , Radovic Svetlana
Published: October 2012

Phenazines, secondary metabolites of fluorescent Pseudomonas, represent a
group of heterocyclic nitrogen-containing compounds showing a broad spectrum
of antibiotic properties. Phenazines producing fluorescent Pseudomonas
species are studied extensively for their application in plant disease
management. In this study, we examined the antifungal activity of different
indigenous Pseudomonas isolates (Q16, B25 and PS2) against the
phytopathogenic fungus Alternaria tenuissima, which had infected cardoon
(Cynara cardunculus L., Asteraceae). An in vitro experiment demonstrated the
antifungal activity of selected indigenous isolates. In addition, an in vivo
experiment under gnotobiotic conditions showed suppression of C. cardunculus
disease caused by A. tenuissima. The quantification of phenazines revealed
significant amounts of phenazine-1-carboxylic acid (PCA) and
2-hydroxy-phenazine-1-carboxylic acid (2-OH-PCA). PCR analysis confirmed the
presence of PCA genes in all examined indigenous Pseudomonas isolates. Based
on our results, we assume that these Pseudomonas isolates have potential in
controlling plant diseases caused by A. tenuissima.
Effects of burial depth on seed germination and seedling emergence of Mexican oaks: A glasshouse experiment
Authors: Flores-Cano J. , Badano E.I. , Flores J.
Published: October 2012

Despite the interest in restoring oak forests in Mexico, very little is known
about their regeneration ecology. We assessed the influence of acorn burial
depth on seed germination and seedling emergence for eight Mexican oak
species (Q. affinis, Q. castanea, Q. coccolobifolia, Q. laeta, Q. mexicana,
Q. polymorpha, Q. tinkhamii and Q. viminea). We performed a glasshouse
experiment in which acorns were buried at five soil depths (0, 2, 4, 6 and 8
cm). After four months, acorn germination and seedling emergence were
recorded. Buried acorns showed higher germination and seedling emergence than
unburied ones, but burial depth also influenced these responses. The optimum
burial depth for seedling emergence of each species was 2, 4, 6 and 8 cm
depth for four species (Q. castanea, Q. mexicana, Q. tinkhamii, and Q.
viminea); 2 and 4 cm for Q. laeta; as well as 2, 4 and 8 cm for Q.
coccolobifolia.
Roncus meledae n.sp. and Neobisium oculatum n.sp., from the island of Mljet, Dalmatia (Neobisiidae, Pseudoscorpiones)
Authors: Curcic B.P.M. , Makarov S.E. , Radja T. , Ilic B.S. , Antic D.Z.
Published: October 2012

Two species of troglobitic false scorpions, Roncus meledae n. sp. and
Neobisium oculatum n. sp. (Neobisiidae, Pseudoscorpiones), are described from
inside underground habitat, i.e. the Jama Na Zutim Kokom Cave, the Island of
Mljet, Dalmatia, Croatia. The analyzed pseudoscorpions prove to differ from
all other congeners. It is possible that the subterranean pseudoscorpions
analyzed represent relicts of an old north tropical faunal pattern of the
Mediterranean.
The effect of fluoride on the serum level of calcium in the rat (Rattus norvegicus)
Authors: Focak M. , Haskovic E. , Suljevic D.
Published: October 2012

The effect of fluoride on the calcium level in serum was analyzed in the
laboratory rat Rattus norvegicus. The control group consisted of 10, and the
experimental group of 15 animals. In the experimental group, fluoride at a
concentration of 3 mg/100 g body weight of rats was intramuscularly injected
into the musculus gluteus maximus. The concentration of calcium was measured
by the CPC method. The average serum calcium concentration was 2.46 mmol/l,
with female rats having higher values of serum calcium than male rats.
Fluoride caused the reduction of calcium concentration in serum (p<0.05); the
reduction was significantly expressed in female rats (p<0.000).
Microscopic investigation of trichomes of wild-growing Satureja montana L.
Authors: Marin Marija , Ascensão Lia , Marin P.D.
Published: October 2012

Considering the importance of S. montana L. as a medicinal plant, due to the
essential oils produced in its glandular trichomes, and the lack of data
about the leaf ultrastructure and micromorphology, we have carried out a
comprehensive study of trichomes distributed in the vegetative and
reproductive organs. Micromorphological and ultrastructural investigations of
the glandular trichomes of Satureja montana L. and autofluorescence analyses
of their secreted products were carried out using scanning electron
microscopy (SEM), transmission electron microscopy (TEM), confocal laser
scanning electron microscopy (CLSM) and light fluorescence microscope. The
leaves, stem and calyx bear numerous nonglandular, unbranched trichomes and
two types of glandular trichomes - peltate and capitate. Ultrastructural
analyses showed the presence of mitochondria, plastids and proliferations of
the granular endoplasmatic reticulum in secretory phases of the heads of
glandular trichomes. Strong red autofluorescence of the lipophilic and
hydrophilic secreted material was observed with CLSM. Green-yellow
autofluorescence of secreted material were noticed in peltate and capitate
trichomes observed with a fluorescence microscope.
Life cycle impact assessment of biodiesel using the ReCiPe method
Authors: Kiss Ferenc , Boskovic Goran
Published: October 2012

This paper presents the life cycle impact assessment (LCIA) results of
biodiesel produced from rapeseed oil. The functional unit (FU) is defined as
3750 km of distance traveled by a truck fuelled with biodiesel. The reference
flow is 1000 kg of biodiesel. The LCIA method used in the study is the ReCiPe
method. At midpoint level the ReCiPe method addresses environmental issues
within 18 impact categories. Most of these midpoint impact categories are
further converted and aggregated into 3 endpoint categories (damage to human
health, damage to ecosystem diversity, damage to mineral resource
availability). The total impact of biodiesel?s life cycle was estimated at
540 Pt/FU. The damage to ecosystem diversity (1.48E-04 species?year/FU), the
damage to human health (7.48E-03 DALY/FU) and the damage to mineral resource
availability (8.11E+03 US$/FU) are responsible for 63%, 27% and 10% of the
total negative impact in the life cycle of biodiesel, respectively. The
results have revealed that only 4 impact categories are responsible for most
of the impacts within the specific endpoint categories. These are impacts
associated with global warming (3000 kg CO2 ekv./FU), particulate matter
formation (12.4 kg PM ekv./FU), agricultural land occupation (6710 m2a./FU)
and fossil fuel depletion (21168 MJ/FU). Greenhouse gases emitted in the life
cycle of biodiesel (mainly N2O, CO2) are responsibly for 56% of the damage
caused to human health and for 16% of the damage caused to ecosystem
diversity. Airborne emissions which contribute to particulate matter
formation (NOx, NH3, PM, SO2) are responsible for 43% of the damage caused to
human health. Agricultural land occupation is responsible for 82% of the
damage caused to the ecosystem diversity. Damage to mineral resource
availability is almost entirely related to the depletion of fossil energy
sources. The production chain of biodiesel and the combustion of biodiesel
are responsible for 69% and 31% of the total impact of biodiesel?s life
cycle, respectively. The negative impact of the production chain is mainly
related to biodiversity loss due to agricultural land occupation (38%) and
the life cycle impacts of mineral fertilizers used in the production of
rapeseed (47%). The environmental impact of biodiesel can be reduced by
increasing the yield of rapeseed with more efficient use of fertilizers and
optimization of agro-technical processes.
Biomechanical and computational evaluation of two loading transfer concepts for pancarpal arthrodesis in dogs
Authors: Rothstock Stephan , Kowaleski Michael P. , Boudrieau Randy J. , Beale Brian S. , Piras Alessandro , Ryan Mark , Bouré Ludovic , Brianza Stefano
Published: October 2012

Objective—To evaluate 2 plate designs for pancarpal arthrodesis and their effects on load transfer to the respective bones as well as to develop a computational model with directed input from the biomechanical testing of the 2 constructs.

Sample—Both forelimbs from the cadaver of an adult castrated male Golden Retriever.

Procedures—CT imaging was performed on the forelimb pair. Each forelimb was subsequently instrumented with a hybrid dynamic compression plate or a castless pancarpal arthrodesis plate. Biomechanical testing was performed. The forelimbs were statically loaded in the elastic range and then cyclically loaded to failure. Finite element (FE) modeling was used to compare the 2 plate designs with respect to bone and implant stress distribution and magnitude when loaded.

Results—Cyclic loading to failure elicited failure patterns similar to those observed clinically. The mean ± SD error between computational and experimental strain was < 15% ± 13% at the maximum loads applied during static elastic loading. The highest bone stresses were at the distal extent of the metacarpal bones at the level of the screw holes with both plates; however, the compression plate resulted in slightly greater stresses than did the arthrodesis plate. Both models also revealed an increase in bone stress at the proximal screw position in the radius. The highest plate stress was identified at the level of the radiocarpal bone, and an increased screw stress (junction of screw head with shaft) was identified at both the most proximal and distal ends of the plates.

Conclusions and Clinical Relevance—The FE model successfully approximated the biomechanical characteristics of an ex vivo pancarpal plate construct for comparison of the effects of application of different plate designs.
Ultrasonographic assessment of the thyroid gland and adjacent anatomic structures in Indo-Pacific bottlenose dolphins (Tursiops aduncus)
Authors: Kot Brian C. W. , Ying Michael T. C. , Brook Fiona M. , Kinoshita Reimi E. , Cheng Sammy C. H.
Published: October 2012

Objective—To evaluate the use of ultrasonography for thyroid gland assessment in healthy Indo-Pacific bottlenose dolphins (Tursiops aduncus), describe the ultrasonographic appearance of the thyroid gland and adjacent anatomic structures, and identify potential associations between variations in thyroid gland morphology and demographic features in this species.

Animals—18 captive Indo-Pacific bottlenose dolphins.

Procedures—1,404 ultrasonographic examinations of the thyroid gland and adjacent anatomic structures (eg, cervical lymph nodes, musculature, and vasculature) were performed during the > 3-year study period. Shape, echogenicity, and homogeneity of thyroid glands were assessed, and glands were categorized into morphological configurations on the basis of results of 2-D and 3-D ultrasonographic evaluation. Associations between demographic factors and thyroid gland morphology were assessed.

Results—Thyroid lobes appeared elliptical or fusiform in the transverse scan plane and round to oval in longitudinal scan planes; morphologically, glands comprised 2 lobes joined by an isthmus or a roughly diamond-shaped structure located on the ventral surface of the trachea. Major blood vessels and cervical lymph nodes were identified. Thyroid parenchyma was typically uniform and homogeneous, with echogenic reticulations and well-defined borders. Thyroid glands were hypoechoic or isoechoic relative to the sternocephalicus muscle; echogenicity was greater in adolescents than in adults. Thyroid gland volume differed between sexes, between sexually mature and immature dolphins, and among age groups and was positively correlated with body length and weight.

Conclusions and Clinical Relevance—Ultrasonography provided a reliable and repeatable method for evaluation of thyroid glands and adjacent anatomic structures in live dolphins.
Tissue distribution of enrofloxacin after intramammary or simulated systemic administration in isolated perfused sheep udders
Authors: López Cadenas Cristina , Fernández Martínez Nélida , Sierra Vega Matilde , Diez Liébana Maria J. , Gonzalo Orden Jose M. , Sahagún Prieto Ana M. , García Vieitez Juan J.
Published: October 2012

Objective—To determine the tissue distribution of enrofloxacin after intramammary or simulated systemic administration in isolated perfused sheep udders by measuring its concentration at various sample collection sites.

Sample—26 udders (obtained following euthanasia) from 26 healthy lactating sheep.

Procedures—For each isolated udder, 1 mammary gland was perfused with warmed, gassed Tyrode solution. Enrofloxacin (1 g of enrofloxacin/5 g of ointment) was administered into the perfused gland via the intramammary route or systemically via the perfusion fluid (equivalent to a dose of 5 mg/kg). Samples of the perfusate were obtained every 30 minutes for 180 minutes; glandular tissue samples were obtained at 2, 4, 6, and 8 cm from the teat base after 180 minutes. The enrofloxacin content of the perfusate and tissue samples was analyzed via high-performance liquid chromatography with UV detection.

Results—After intramammary administration, maximun perfusate enrofloxacin concentration was detected at 180 minutes and, at this time, mean tissue enrofloxacin concentration was detected and mean tissue enrofloxacin concentration was 123.80, 54.48, 36.72, and 26.42 μg/g of tissue at 2, 4, 6, and 8 cm from the teat base, respectively. Following systemic administration, perfusate enrofloxacin concentration decreased with time and, at 180 minutes, tissue enrofloxacin concentrations ranged from 40.38 to 35.58 μg/g of tissue.

Conclusions and Clinical Relevance—By 180 minutes after administration via the intramammary or systemic route in isolated perfused sheep mammary glands, mean tissue concentration of enrofloxacin was greater than the minimum inhibitory concentration required to inhibit growth of 90% of many common mastitis pathogens in sheep. Use of either route of administration (or in combination) appears suitable for the treatment of acute mastitis in sheep.
Mechanical and morphological properties of trabecular bone samples obtained from third metacarpal bones of cadavers of horses with a bone fragility syndrome and horses unaffected by that syndrome
Authors: Symons Jennifer E. , Entwistle Rachel C. , Arens Amanda M. , Garcia Tanya C. , Christiansen Blaine A. , Fyhrie David P. , Stover Susan M.
Published: October 2012

Objective—To determine morphological and mechanical properties of trabecular bone of horses with a bone fragility syndrome (BFS; including silicate-associated osteoporosis).

Sample—Cylindrical trabecular bone samples from the distal aspects of cadaveric third metacarpal bones of 39 horses (19 horses with a BFS [BFS bone samples] and 20 horses without a BFS [control bone samples]).

Procedures—Bone samples were imaged via micro-CT for determination of bone volume fraction; apparent and mean mineralized bone densities; and trabecular number, thickness, and separation. Bone samples were compressed to failure for determination of apparent elastic modulus and stresses, strains, and strain energy densities for yield, ultimate, and failure loads. Effects of BFS and age of horses on variables were determined.

Results—BFS bone samples had 25% lower bone volume fraction, 28% lower apparent density, 18% lower trabecular number and thickness, and 16% greater trabecular separation versus control bone samples. The BFS bone samples had 22% lower apparent modulus and 32% to 33% lower stresses, 10% to 18% lower strains, and 41 % to 52% lower strain energy densities at yield, ultimate, and failure loads, compared with control bone samples. Differences between groups of bone samples were not detected for mean mineral density and trabecular anisotropy.

Conclusions and Clinical Relevance—Results suggested that horses with a BFS had osteopenia and compromised trabecular bone function, consistent with bone deformation and pathological fractures that develop in affected horses. Effects of this BFS may be systemic, and bones other than those that are clinically affected had changes in morphological and mechanical properties.
In vitro cytoprotective effects of acetylsalicylic acid, carprofen, meloxicam, or robenacoxib against apoptosis induced by sodium nitroprusside in canine cruciate ligament cells
Authors: Waldherr Katrin , Zurbriggen Andreas , Spreng David E. , Forterre Simone
Published: October 2012

Objective—To determine whether incubation of cruciate ligament cells with acetylsalicylic acid, carprofen, meloxicam, or robenacoxib provides protection against apoptosis induced by sodium nitroprusside (SNP).

Sample—Explants of cranial (CCL) and caudal (CaCL) cruciate ligaments from eight 1-day-old Beagles.

Procedures—Primary cultures of CCL and CaCL cells were created via enzymatic dissociation of cruciate explants. Purified cell cultures were incubated for 2 hours without (controls) or with 1 of 3 concentrations of 1 of 4 NSAIDs (10, 100, or 200 μg of acetylsalicylic acid/mL; 0.1, 1, or 10 μg of carprofen/mL; 0.1, 1, or 10 μg of meloxicam/mL; or 0.1, 1, or 10 μg of robenacoxib/mL) and subsequently incubated for 18 hours with 1 of 3 concentrations of SNP in an attempt to induce mild, moderate, or severe cytotoxic effects. Cell viability and apoptosis were analyzed via a cell proliferation assay and flow cytometry, respectively. Prostaglandin E2 concentrations were measured via an ELISA.

Results—Cytoprotective effects of NSAIDs were dependent on the extent of SNP-induced apoptosis and were greatest in CCL and CaCL cell cultures with moderate SNP-induced cytotoxic effects. Preincubation with an NSAID improved cell viability by 15% to 45% when CCL and CaCL cells were subsequently incubated with SNP. Carprofen (10 μg/mL) had the greatest cytoprotective effects for CCL and CaCL cells. Incubation with NSAIDs resulted in a nonsignificant decrease in PGE2 production from SNP-damaged cells.

Conclusions and Clinical Relevance—Results indicated that carprofen, meloxicam, and robenacoxib may reduce apoptosis in cells originating from canine cruciate ligaments.
Associations among serum N-terminal procollagen type III concentration, urinary aldosterone-to-creatinine ratio, and ventricular remodeling in dogs with myxomatous mitral valve disease
Authors: Hezzell Melanie J. , Boswood Adrian , Chang Yu-Mei , Moonarmart Walasinee , Elliott Jonathan
Published: October 2012

Objective—To assess relationships among serum N-terminal procollagen type III concentration, urinary aldosterone-to-creatinine concentration ratio (UAC), and clinical variables in dogs with myxomatous mitral valve disease (MMVD) and healthy dogs.

Animals—162 dogs with MMVD and 24 healthy control dogs of comparable age and body weight.

Procedures—Blood and urine samples were collected from each dog. Dogs with MMVD underwent echocardiography and ECG. Ventricular diameter measurements were normalized for body weight. Serum N-terminal procollagen type III and urinary aldosterone concentrations were measured via radioimmunoassay. Each dog was examined on 1 to 3 occasions. Examinations were repeated at approximately 6-month intervals.

Results—Serum N-terminal procollagen type III concentration decreased with increasing severity of MMVD and was negatively associated with age and left ventricular end-diastolic and end-systolic diameters. The UAC increased with prior percentage change in left ventricular end-diastolic diameter per month, subsequent percentage change in left ventricular end-systolic diameter per month, and treatment with diuretics and was negatively associated with age. Both UAC and serum N-terminal procollagen type III concentration were higher in Cavalier King Charles Spaniels than in other breeds when other measured variables were controlled for.

Conclusions and Clinical Relevance—In dogs with MMVD, echocardiographic indicators of left ventricular remodeling appeared to be associated with a decrease in serum concentration of a marker of collagen type III turnover and an increase in urinary aldosterone concentration.
Investigation of hydration processes of the equine hoof via nuclear magnetic resonance microscopy
Authors: Sugimoto Miki , Kuwano Atsutoshi , Ikeda Shuntaro , Kume Shin-ichi , Yoshihara Eiru , Wada Shinya
Published: October 2012

Objective—To examine the distribution of water in hoof wall specimens of horses via nuclear magnetic resonance (NMR) microscopy and determine changes in water distribution during hydration.

Sample—4 hoof wall specimens (2 obtained from the dorsum and 1 each obtained from the lateral quarter and lateral heel regions) of the stratum medium of healthy hooves of 1 horse.

Procedures—Equine hoof wall specimens were examined via NMR microscopy. Proton density–weighted 3-D images were acquired. Changes during water absorption were assessed on sequential images.

Results—The inner zone of the stratum medium had higher signals than did the outer zone. Areas of high signal intensity were evident in transverse images; these corresponded to the distribution of horn tubules. During water absorption, the increase in signal intensity started at the bottom of a specimen and extended to the upper region; it maintained the localization pattern observed before hydration. The relationship between the local maximal signals in areas corresponding to the horn tubules and minimal signal intensities in areas corresponding to the intertubular horn was similar and maintained approximately a linear distribution.

Conclusions and Clinical Relevance—Based on the premise that signal intensity reflects water content, hydration in the equine hoof wall during water absorption occurred concurrently in the tubules and intertubular horn, and there was maintenance of the original water gradients. This technique can be applied for the assessment of pathophysiologic changes in the hoof wall on the basis of its hydration properties.
Plasma 25-hydroxyvitamin D3 concentrations in Hermann's tortoises (Testudo hermanni) exposed to natural sunlight and two artificial ultraviolet radiation sources
Authors: Selleri Paolo , Di Girolamo Nicola
Published: October 2012

Objective—To determine the effect of various UVB radiation sources on plasma 25-hydroxyvitamin D3 concentrations in Hermann's tortoises (Testudo hermanni).

Animals—18 healthy Hermann's tortoises.

Procedures—Tortoises were exposed to sunlight in an outdoor enclosure located in the natural geographic range of Hermann's tortoises (n = 6 tortoises) or a self-ballasted mercury-vapor lamp (6) or fluorescent UVB-emitting lamp (6) in an indoor enclosure for 35 days. Plasma samples were obtained from each tortoise on the first (day 0) and last (day 35) days of the study, and concentrations of 25-hydroxyvitamin D3 were determined. Amount of UVB radiation in enclosures was measured.

Results—Mean ± SD plasma 25-hydroxyvitamin D3 concentrations for tortoises exposed to the mercury-vapor and fluorescent lamps were significantly lower on day 35 (155.69 ± 80.71 nmol/L and 134.42 ± 51.42 nmol/L, respectively) than they were on day 0 (368.02 ± 119.34 nmol/L and 313.69 ± 109.54 nmol/L, respectively). Mean ± SD plasma 25-hydroxyvitamin D3 concentration for tortoises exposed to sunlight did not differ significantly between days 0 (387.74 ± 114.56 nmol/L) and 35 (411.51 ± 189.75 nmol/L). Mean day 35 plasma 25-hydroxyvitamin D3 concentration was significantly higher for tortoises exposed to sunlight versus those exposed to mercury-vapor or fluorescent lamps. Sunlight provided significantly more UVB radiation than did the mercury-vapor or fluorescent lamps.

Conclusions and Clinical Relevance—Plasma 25-hydroxyvitamin D3 concentrations differed between tortoises exposed to sunlight and those exposed to artificial UVB sources. Exposure to sunlight at a latitude similar to that of the natural geographic range is recommended for healthy and calcium-deficient tortoises.
Assessment of the long-term effect of vaccination on transmission of infectious bovine rhinotracheitis virus in cattle herds hyperimmunized with glycoprotein E–deleted marker vaccine
Authors: Ampe Bart , Duchateau Luc , Speybroeck Niko , Berkvens Dirk , Dupont Alain , Kerkhofs Pierre , Thiry Etienne , Dispas Marc
Published: October 2012

Objective—To assess long-term effects and risk factors for the efficacy of hyperimmunization protocols against infectious bovine rhinotracheitis (IBR) during a longitudinal field study of dairy and dairy-beef mixed farms.

Animals—Approximately 7,700 cows from 72 farms.

Procedures—Farms were assigned to 3 treatment groups (hyperimmunization groups [HIGs] 1 and 2, which were hyperimmunized with glycoprotein E [gE]–deleted marker vaccines, and a nonintervention group [NIG]). Cattle in HIG 1 were initially vaccinated with an attenuated vaccine, whereas cattle in HIG 2 were initially vaccinated with an inactivated-virus vaccine. Cattle in both HIGs received booster inoculations with inactivated-virus vaccines at 6-month intervals. The risk for gE seroconversion was compared among experimental groups via a shared frailty model with a piecewise constant baseline risk to correct for seasonal and secular effects.

Results—Risk for gE seroconversion significantly decreased over time for the HIGs, compared with the NIG. Seasonal changes in the risk of gE seroconversion were detected, with a higher risk during winter periods, compared with grazing periods. No significant difference was detected between HIGs 1 and 2. The only significant risk factor was the number of buildings for cattle on a farm; the higher the number of buildings, the lower the risk for gE seroconversion. Prevalence of IBR decreased over time in both HIGs but remained constant or increased in the NIG.

Conclusions and Clinical Relevance—Hyperimmunization via repeated administration of attenuated and inactivated-virus gE-deleted marker vaccines as well as inactivated-virus vaccines may provide a method for control of IBR.
Evaluation of intraocular pressure in conscious Hermann's tortoises (Testudo hermanni) by means of rebound tonometry
Authors: Selleri Paolo , Di Girolamo Nicola , Andreani Valentina , Guandalini Adolfo , D'Anna Nunzio
Published: October 2012

Objective—To determine intraocular pressure (IOP) in healthy Hermann's tortoises (Testudo hermanni).

Animals—26 outdoor-housed Hermann's tortoises (13 males and 13 females); body weight ranged from 255 to 2,310 g, and age ranged from 4 to > 50 years.

Procedures—After a preliminary ophthalmic evaluation was performed, IOP was measured by means of a rebound tonometer in both eyes of each tortoise. Three measurements were obtained for each eye; successive measurements were obtained from alternate eyes. Each measurement was based on the mean of 6 values automatically provided by the rebound tonometer. Statistical analysis was used to evaluate correlations between variables and to identify sex- or size-related IOP variations, and changes in IOP over multiple measurements.

Results—Mean ± SEM IOP of the 52 eyes was 15.74 ± 0.20 mm Hg (range, 9 to 22 mm Hg). Results for t tests did not reveal significant differences in IOP between the right and left eyes or between males and females. A significant moderate negative correlation (r = −0.41; r2 = 0.169) between IOP and body weight was detected. Results of repeated-measures ANOVA revealed a significant increase in IOP over multiple measurements.

Conclusions and Clinical Relevance—Rebound tonometry was a practical and rapid means of determining IOP in small- to medium-sized tortoises that required minimal manual restraint of the animals. Establishing IOP values in healthy Hermann's tortoises will provide a reference frame for use during complete ophthalmic examinations, thus allowing clinicians to diagnose a broader spectrum of ocular pathological conditions in tortoises.
Evaluation of a novel suture material for closure of intestinal anastomoses in canine cadavers
Authors: Hansen Lane A. , Monnet Eric L.
Published: October 2012

Objective—To compare leakage and maximum intraluminal pressures for a novel suture material with pressures for comparable suture material when used in closure of intestinal anastomoses in canine cadavers.

Sample—Healthy intestines from cadavers of dogs euthanized for reasons unrelated to the study.

Procedures—18 anastomoses were performed on intestinal sections within 72 hours after dogs were euthanized and intestinal samples collected. Anastomoses were performed with a simple continuous suture pattern. Leakage and maximum intraluminal pressures were measured and recorded for 6 control segments and 18 anastomosed sections. A barbed glycomer 631 suture (size 4–0 United States Pharmacopeia [USP]) was compared with glycomer 631 sutures (sizes 3–0 and 4–0 USP). Results for leakage and maximum intraluminal pressures were compared via an ANOVA.

Results—The barbed glycomer 631 suture material leaked at a significantly higher pressure than did the comparable glycomer 631 suture materials. Maximum intraluminal pressures were not significantly different among the suture materials.

Conclusions and Clinical Relevance—Barbed glycomer 631 4–0 USP suture material was as effective as glycomer 631 suture materials and may be a safe alternative for use in closure of enterectomies in dogs.