Search tips
Search criteria

Results 1-25 (64)

Clipboard (0)

Select a Filter Below

Year of Publication
1.  Staphylococcus warneri, a resident skin commensal of rainbow trout (Oncorhynchus mykiss) with pathobiont characteristics 
Veterinary microbiology  2013;169(0):80-88.
Commensal microorganisms live in association with the mucosal surfaces of all vertebrates. The skin of teleost fish is known to harbor commensals. In this study we report for the first time the presence of an intracellular Gram positive bacteria, Staphylococcus warneri that resides in the skin epidermis of rainbow trout (Oncorhynchus mykiss). S. warneri was isolated from healthy hatchery trout skin epithelial cells. In situ hybridization confirmed the intracellular nature of the bacterium. Skin explants exposed in vitro to S. warneri or the extracellular pathogen Vibrio anguillarum show that S. warneri is able to induce an anti-inflammatory cytokine status via TGF-β 1b compared to the pro-inflammatory responses (IL-1β , IL-6 and TNF-α) elicited by V. anguillarum. In vivo experiments showed that S. warneri is not pathogenic to rainbow trout when injected intraperitoneally at high concentrations. However, S. warneri is able to stimulate V. anguillarum growth and biofilm formation on rainbow trout scales. Our results demonstrate that rainbow trout skin commensals such as S. warneri have the potential to become indirect pathobionts by enhancing growth and biofilm formation of pathogens such as V. anguillarum. These results show that fish farming practices (i.e. handling and other manipulations) can alter the skin microbiota and compromise the skin health of rainbow trout.
PMCID: PMC3967797  PMID: 24438987
rainbow trout; skin; Staphylococcus warneri; Vibrio anguillarum; pathobiont
2.  Microarray-based detection of viruses causing vesicular or vesicular-like lesions in livestock animals 
Veterinary microbiology  2008;133(0):145-153.
Definitive diagnosis of vesicular or vesicular-like lesions in livestock animals presents challenges both for veterinary clinicians and diagnostic laboratories. It is often impossible to diagnose the causative disease agent on a clinical basis alone and difficult to collect ample vesicular epithelium samples. Due to restrictions of time and sample size, once laboratory tests have ruled out foot-and-mouth disease, vesicular stomatitis and swine vesicular disease a definitive diagnosis may remain elusive. With the ability to test a small quantity of sample for a large number of pathogens simultaneously, DNA microarrays represent a potential solution to this problem. This study describes the application of a long oligonucleotide microarray assay to the identification of viruses known to cause vesicular or vesicular-like lesions in livestock animals. Eighteen virus isolates from cell culture were successfully identified to genus level, including representatives of each foot-and-mouth disease virus serotype, two species of vesicular stomatitis virus, swine vesicular disease virus, vesicular exanthema of swine virus, bovine herpesvirus 1, orf virus, pseudocowpox virus, bluetongue virus serotype 1 and bovine viral diarrhoea virus 1. Vesicular stomatitis virus and vesicular exanthema of swine virus were also identified in vesicular epithelium samples, with varying levels of sensitivity. The results indicate that with further development this microarray assay could be a valuable tool for the diagnosis of vesicular and vesicular-like diseases.
PMCID: PMC4310687  PMID: 18621489
3.  Short Communication: New Recognition Of Enterovirus Infections In Bottlenose Dolphins (Tursiops Truncatus) 
Veterinary microbiology  2009;139(0):170-175.
An enterovirus was cultured from an erosive tongue lesion of a bottlenose dolphin (Tursiops truncatus). The morphology of virions on negative staining electron microscopy was consistent with those of enteroviruses. Analysis of 2613 bp of the polyprotein gene identified the isolate as a novel enterovirus strain, tentatively named bottlenose dolphin enterovirus (BDEV), that nests within the species Bovine enterovirus. Serologic evidence of exposure to enteroviruses was common in both free ranging and managed collection dolphins. Managed collection dolphins were more likely to have high antibody levels, although the highest levels were reported in free ranging populations. Associations between enterovirus antibody levels, and age, sex, complete blood counts, and clinical serum biochemistries were explored. Dolphins with higher antibody levels were more likely to be hyperproteinemic and hyperglobulinemic.
PMCID: PMC4310689  PMID: 19581059
4.  Pathogenesis of GIII.2 bovine norovirus, CV186-OH/00/US strain in gnotobiotic calves 
Veterinary microbiology  2013;168(1):202-207.
The pathogenesis of GIII.2 bovine norovirus (BoNoV) is not well understood. Our study demonstrated persisting diarrhea and prolonged fecal shedding, but with a lack of significant intestinal lesions in gnotobiotic (Gn) calves infected with GIII.2 BoNoV, CV186-OH/00/US strain. Nine 4 to 7-day-old Angus/Jersey crossbred Gn calves were orally inoculated with 10.0-11.9 log10 genomic equivalents (GE)/calf of CV186-OH (n=7) or mock (n=2). Calves were euthanized at post-inoculation day (PID) 1 (n=1) when moderate to severe lethargy was observed and at PIDs 2-6 (n=4) after lethargy had subsided. Two calves were kept longer term (until PID 30) for monitoring fecal shedding patterns by TaqMan real-time RT-PCR (qRT-PCR). Most infected calves exhibited two clinical signs: i) acute but persisting diarrhea and ii) acute moderate to severe lethargy. The two infected calves, followed longer-term, had prolonged fecal viral RNA shedding [peak average titer of 11.8 (± 0.2) log10 GE/ml] at least until PID 20. By qRT-PCR, 5 infected calves had low viral RNA titers in serum, ranging from 4.0 to 5.8 log10 GE/ml, at PIDs 1-5, but not (<2.7 log10 GE/ml) at PIDs 6-30. The latter observation coincided with the presence of serum IgG antibody to BoNoV at PIDs 8-30. Collectively, the GIII.2 BoNoV strain CV186-OH induced only mild enteropathogenicity, evident by the lack of significant intestinal lesions, but it led to persisting mild diarrhea and prolonged fecal virus shedding in Gn calves. The prolonged fecal shedding of GIII.2 BoNoV might partially explain how this virus is maintained as endemic infections in cattle.
PMCID: PMC3905316  PMID: 24309403
Norovirus; Pathogenesis; Cattle; Prolonged virus shedding
5.  Putative Human and Avian Risk Factors for Avian Influenza Virus Infections in Backyard Poultry in Egypt 
Veterinary microbiology  2013;168(1):208-213.
Highly pathogenic influenza A virus subtype H5N1 causes significant poultry mortality in the six countries where it is endemic and can also infect humans. Egypt has reported the third highest number of poultry outbreaks (n=1,084) globally. The objective of this cross-sectional study was to identify putative risk factors for H5N1 infections in backyard poultry in 16 villages in Damietta, El Gharbia, Fayoum, and Menofia governorates from 2010–2012. Cloacal and tracheal swabs and serum samples from domestic (n=1242)and wild birds (n=807) were tested for H5N1 via RT-PCR and hemagglutination inhibition, respectively. We measured poultry rearing practices with questionnaires (n=306 households) and contact rates among domestic and wild bird species with scan sampling. Domestic birds (chickens, ducks, and geese, n = 51) in three governorates tested positive for H5N1 by PCR or serology. A regression model identified a significant correlation between H5N1 in poultry and the practice of disposing of dead poultry and poultry feces in the garbage (F = 15.7, p< 0.0001). In addition, contact between domestic and wild birds was more frequent in villages where we detected H5N1 in backyard flocks (F= 29.5, p< 0.0001).
PMCID: PMC3890107  PMID: 24315038
Disease reservoirs; Influenza in birds; Influenza A virus – H5N1 subtype; Poultry diseases; Zoonoses
6.  Pathogenicity study in sheep using reverse-genetics-based reassortant bluetongue viruses 
Veterinary Microbiology  2014;174(1-2):139-147.
•Use of reverse genetics to generate reassortant BTV viruses for testing in animals.•Two structural and one non-structural proteins are involved in pathogenicity.•Molecular basis of bluetongue disease appears to be highly complex.
Bluetongue (BT) disease, caused by the non-enveloped bluetongue virus (BTV) belonging to the Reoviridae family, is an economically important disease that affects a wide range of wild and domestic ruminants. Currently, 26 different serotypes of BTV are recognized in the world, of which BTV-8 has been found to exhibit one of the most virulent manifestations of BT disease in livestock. In recent years incursions of BTV-8 in Europe have resulted in significant morbidity and mortality not only in sheep but also in cattle. The molecular and genetic basis of BTV-8 pathogenesis is not known. To understand the genetic basis of BTV-8 pathogenicity, we generated reassortant viruses by replacing the 3 most variable genes, S2, S6 and S10 of a recent isolate of BTV-8, in different combinations into the backbone of an attenuated strain of BTV-1. The growth profiles of these reassortant viruses were then analyzed in two different ovine cell lines derived from different organs, kidney and thymus. Distinct patterns for each reassortant virus in these two cell lines were observed. To determine the pathogenicity of these reassortant viruses, groups of BTV-susceptible sheep were infected with each of these viruses. The data suggested that the clinical manifestations of these two different serotypes, BTV-1 and BTV-8, were slightly distinct and BTV-1, when comprising all 3 genome segments of BTV-8, behaved differently to BTV-1. Our results also suggested that the molecular basis of BT disease is highly complex.
PMCID: PMC4220015  PMID: 25307940
Bluetongue virus; Serotype 8; Reassortment; Non-structural protein NS3; Pathogenicity in sheep
7.  Characterization of Recombinant H9N2 Influenza Viruses Isolated from Wild Ducks in China 
Veterinary microbiology  2013;166(0):327-336.
Wild birds are considered to be the natural reservoirs for avian influenza A viruses (AIV). During active influenza surveillance in Poyang Lake of southeast China, we isolated and characterized eleven H9N2 viruses from two species of wild ducks. Phylogenetic analysis showed that the eleven isolates were almost identical with 99.3% to 100% nucleotide homology in their entire genome, and they all closely related in whole eight genes (95.6-99.4% homology) to human H9N2 isolates (HK/33982/2009) and clustered in the same sublineage. The isolates belonged to triple reassortant H9N2 genotype viruses containing Ck/Bei-like NA genes, Y439-like PA genes and six other G1-like genes. We also found that the subtype of virus replicated efficiently in the lungs and tracheas of BALB/c mice and caused mortality in 20-40% of infected groups after 3-6 days, which indicates that the subtype of virus is capable of establishing lethal mammalian infections. However, whether or not the virus has features transmittable from wild ducks to humans is not known. This study showed that the subtype of virus was detected for the first time in wild birds, and also suggested that wild birds may carry the virus for a long time and spread it over long distances along migratory routes, so more attention should be paid to the continued surveillance of wild birds.
PMCID: PMC3773574  PMID: 23830774
H9N2; human infection; triple reassortant; wild ducks
8.  Inactivated vaccine with adjuvants consisting of pattern recognition receptor agonists confers protection against avian influenza viruses in chickens 
Veterinary microbiology  2014;172(0):120-128.
Use of adjuvant containing pathogen pattern recognition receptor agonists is one of the effective strategies to enhance the efficacy of licensed vaccines. In this study, we investigated the efficacy of avian influenza vaccines containing an adjuvant (CVCVA5) which was composed of polyriboinosinic polyribocytidylic, resiquimod, imiquimod, muramyl dipeptide and levomisole. Avian influenza vaccines adjuvanted with CVCVA5 were found to induce significantly higher titers of hemagglutiniton inhibition antibodies (P ≤ 0.01) than those of commercial vaccines at 2-, 3- and 4-week post vaccination in both specific pathogen free (SPF) chickens and field application. Furthermore, virus shedding was reduced in SPF chickens immunized with H9-CVCVA5 vaccine after H9 subtype heterologous virus challenge. The ratios of both CD3+CD4+ and CD3+CD8+ lymphocytes were slowly elevated in chickens immunized with H9-CVCVA5 vaccine. Lymphocytes adoptive transfer study indicates that CD8+ T lymphocyte subpopulation might have contributed to improved protection against heterologous virus challenge. Results of this study suggest that the adjuvant CVCVA5 was capable of enhancing the potency of existing avian influenza vaccines by increasing humoral and cellular immune response.
PMCID: PMC4208718  PMID: 24894132
Avian influenza virus; Inactivated vaccine; Adjuvants; Pattern recognition receptor; Agonist; Toll-like receptor
9.  Transcriptome analysis of CNS immediately before and after the detection of PrPSc in SSBP/1 sheep scrapie 
Veterinary Microbiology  2014;173(3-4):201-207.
•Arrays and DGE-tags quantified gene expression in the CNS during sheep scrapie.•Neurological receptors were increased with disease progression.•Clues to basis of psychiatric changes.•Step changes to gene expression after the detection of PrPSc in CNS.
Sheep scrapie is a transmissible spongiform encephalopathy (TSE), progressive and fatal neurodegenerative diseases of the central nervous system (CNS) linked to the accumulation of misfolded prion protein, PrPSc. New Zealand Cheviot sheep, homozygous for the VRQ genotype of the PRNP gene are most susceptible with an incubation period of 193 days with SSBP/1 scrapie. However, the earliest time point that PrPSc can be detected in the CNS is 125 days (D125). The aim of this study was to quantify changes to the transcriptome of the thalamus and obex (medulla) at times immediately before (D75) and after (D125) PrPSc was detected. Affymetrix gene arrays were used to quantify gene expression in the thalamus and Illumina DGE-tag profiling for obex. Ingenuity Pathway Analysis was used to help describe the biological processes of scrapie pathology.
Neurological disease and Cancer were common Bio Functions in each tissue at D75; inflammation and cell death were major processes at D125. Several neurological receptors were significantly increased at D75 (e.g. CHRNA6, GRM1, HCN2), which might be clues to the molecular basis of psychiatric changes associated with TSEs. No genes were significantly differentially expressed at both D75 and D125 and there was no progression of events from earlier to later time points. This implies that there is no simple linear progression of pathological or molecular events. There seems to be a step-change between D75 and D125, correlating with the detection of PrPSc, resulting in the involvement of different pathological processes in later TSE disease.
PMCID: PMC4206282  PMID: 25183238
Scrapie; Prion; Gene expression; Thalamus; Medulla; CNS+
10.  Genetic diversity of porcine group A rotavirus strains in the UK 
Veterinary Microbiology  2014;173(1-2):27-37.
•This is the first study of rotavirus genotypes circulating in UK pigs.•Rotavirus transmission between pigs and humans is not thought to be common in the UK.•Human rotavirus genotype P[8] found in a UK pig.•The uncommon rotavirus genotype P[32] is widespread in UK pig herds.
Rotavirus is endemic in pig farms where it causes a loss in production. This study is the first to characterise porcine rotavirus circulating in UK pigs. Samples from diarrheic pigs with rotavirus enteritis obtained between 2010 and 2012 were genotyped in order to determine the diversity of group A rotavirus (GARV) in UK pigs. A wide range of rotavirus genotypes were identified in UK pigs: six G types (VP7); G2, G3, G4, G5, G9 and G11 and six P types (VP4); P[6], P[7], P[8], P[13], P[23], and P[32]. With the exception of a single P[8] isolate, there was less than 95% nucleotide identity between sequences from this study and any available rotavirus sequences.
The G9 and P[6] genotypes are capable of infecting both humans and pigs, but showed no species cross-over within the UK as they were shown to be genetically distinct, which suggested zoonotic transmission is rare within the UK. We identified the P[8] genotype in one isolate, this genotype is almost exclusively found in humans. The P[8] was linked to a human Irish rotavirus isolate in the same year. The discovery of human genotype P[8] rotavirus in a UK pig confirms this common human genotype can infect pigs and also highlights the necessity of surveillance of porcine rotavirus genotypes to safeguard human as well as porcine health.
PMCID: PMC4158422  PMID: 25123085
Rotavirus; Porcine; Phylogenetic; Zoonosis
11.  Prevalence and genetic heterogeneity of porcine group C rotaviruses in nursing and weaned piglets in Ohio, USA and identification of a potential new VP4 genotype 
Veterinary microbiology  2013;164(0):27-38.
Swine fecal samples collected from seven farms were screened for group C rotaviruses (RVCs) using a reverse transcription-polymerase chain reaction assay. A total of 380 samples were tested and 19.5% were positive. Of the 128 samples collected in 2012, 23.5% from nursing piglets and 8.5% from weaned piglets were RVC positive, with a higher RVC frequency in diarrheic (28.4%) than in non-diarrheic (6.6%) piglets. Two strains (RVC/Pig-wt/USA/RV0104/2011/G3PX and RVC/Pig-wt/USA/RV0143/2012/G6Px) from two different farms were characterized genetically to gain information on virus diversity based on full length sequences of the inner capsid VP6, enterotoxin NSP4 and the outer capsid VP7 and VP4 (partial for RV0104) genes. The VP6 gene of the two strains showed high (99%) nucleotide identity to one another, 84–91% identity to other porcine RVCstrains and 81–82% identity to human and bovine RVC strains. The NSP4 gene analysis revealed that RVC/Pig-wt/USA/RV0104/2011/G3PX and RVC/Pig-wt/USA/RV0143/2012/G6Px strains were not closely related to each other (87% identity), but shared higher identity with prototype RVC/Pig-wt/USA/Cowden/1980/G1Px strain (93% and 89%, respectively) and were more distantly related to human strains (72–76% identity). The VP7 gene analysis indicated that the two strains were distantly related to one another (72% identity). RVC/Pig-wt/USA/RV0143/2012/G6Px was most closely related to porcine RVC G6 strains (82–86% identity), whereas RVC/Pig-wt/USA/RV0104/2011/G3PX was most closely related to porcine HF (G3) strain (94% identity). Analysis of the full length nucleotide sequence of the VP4 gene revealed that RVC/Pig-wt/USA/RV0143/2012/G6Px was distantly related to porcine (75%), bovine (74%) and human (70%) strains. The deduced amino acid identities (69.5–75.6%) of VP4 between RVC/Pig-wt/USA/RV0143/2012/G6Px and other RVCs were low; hence, we propose that this strain comprises a new VP4 genotype. Our results indicate high genetic heterogeneity in RVCs genes and the concurrent co-circulation of different genotypes at the same time. Our findings are useful for the development of more accurate diagnostic tools, for basic research to understand gene function and to provide information for RVC diversity germane to vaccine development.
PMCID: PMC4094028  PMID: 23428382
Group C rotavirus; Prevalence; Genetic diversity; Swine
12.  Immune response to ORF5a protein immunization is not protective against Porcine Reproductive and Respiratory Syndrome Virus infection 
Veterinary microbiology  2013;164(0):281-285.
Porcine reproductive and respiratory syndrome virus (PRRSV) is an enveloped RNA virus responsible for PRRS in swine, a disease with globally significant animal welfare and economic concerns. There is no specific treatment and variably effective immune protection. Molecular mechanisms responsible for virulence, pathogenesis and protective immune response remain poorly understood. These factors limit progress towards development of effective measures for prevention and treatment of PRRS. A novel PRRSV ORF5a protein, encoded in an open reading frame (ORF) that overlaps the major envelope glycoprotein GP5 ORF, was recently identified. Because ORF5a is highly conserved in diverse PRRSV isolates, is a structural protein in the virion, and elicits a specific antibody response in infected pigs, we investigated its potential role in immune protection against PRRSV infection. Pigs immunized with ORF5a protein had robust serologic responses. However, the antibodies did not neutralize virus, and immunity did not protect against challenge infection. We conclude from these findings that the ORF5a antibody response is neither neutralizing nor protective.
PMCID: PMC3684206  PMID: 23578707
Swine; immunology; arterivirus; PRRSV; neutralizing antibody
13.  The prevalence and genetic diversity of group A rotaviruses on pig farms in the Mekong Delta region of Vietnam 
Veterinary Microbiology  2014;170(3-4):258-265.
Group A rotaviruses (ARoVs) are a common cause of severe diarrhea among children worldwide and the cause of approximately 45% of pediatric hospitalizations for acute diarrhea in Vietnam. ARoVs are known to cause significant economic losses to livestock producers by reducing growth performance and production efficiencies, however little is known about the implications of asymptomatic endemic circulation of ARoV. We aimed to determine the prevalence and predominant circulating genotypes of ARoVs on pig farms in a southern province of Vietnam. We found overall animal-level and farm-level prevalence of 32.7% (239/730) and 74% (77/104), respectively, and identified six different G types and 4 P types in various combinations (G2, G3, G4, G5, G9, G11 and P[6], P[13], P[23], and P[34]). There was no significant association between ARoV infection and clinical disease in pigs, suggesting that endemic asymptomatic circulation of ARoV may complicate rotavirus disease attribution during outbreaks of diarrhea in swine. Sequence analysis of the detected ARoVs suggested homology to recent human clinical cases and extensive genetic diversity. The epidemiological relevance of these findings for veterinary practitioners and to ongoing pediatric ARoV vaccine initiatives in Vietnam merits further study.
PMCID: PMC4003349  PMID: 24679960
Rotaviruses; Pigs; Vietnam
14.  The enterohemorrhagic Escherichia coli effector protein NleF binds mammalian Tmp21 
Veterinary microbiology  2013;164(0):164-170.
The human pathogens enterohemorrhagic and enteropathogenic Escherichia coli (EHEC and EPEC), as well as the mouse pathogen Citrobacter rodentium encode type III secretion system (T3SS) effector proteins to promote their survival in the infected host. The mechanisms of action and the host targets of T3SS effectors are under active investigation because of their importance to bacterial virulence. The non-locus of enterocyte effacement (LEE)-encoded protein F, NleF, contributes to E. coli and C. rodentium colonization of piglets and mice, respectively. Here we sought to characterize the host binding partners of NleF. Using a yeast two-hybrid screen, we identified Tmp21, a type-I integral membrane protein and COPI-vesicle receptor involved in trans-Golgi network function, as an NleF-binding partner. We confirmed this interaction using immunoprecipitation and bimolecular fluorescence complementation (BiFC). We expressed a temperature-sensitive vesicular stomatitis virus glycoprotein (tsVSVG) to monitor protein trafficking and determined that NleF slows the intracellular trafficking of tsVSVG from the endoplasmic reticulum to the Golgi.
PMCID: PMC3640833  PMID: 23434013
Effector; Enterohemorrhagic E. coli; NleF; Tmp21; Type III secretion
15.  Enteropathogenic Escherichia coli Prevalence in Laboratory Rabbits 
Veterinary microbiology  2013;163(3-4):395-398.
Rabbit-origin enteropathogenic E. coli (EPEC) causes substantial diarrhea-associated morbidity and has zoonotic potential. A culture-based survey was undertaken to ascertain its prevalence. EPEC was isolated from 6/141 (4.3%) commercially-acquired laboratory rabbits. Three of these did not have diarrhea or EPEC-typical intestinal lesions; they instead had background plasmacytic intestinal inflammation. Asymptomatically infected rabbits may function as EPEC reservoirs.
PMCID: PMC3594591  PMID: 23391439
Escherichia coli infections; laboratory animals; rabbits; diarrhea; zoonoses; inflammation
16.  Characterization of a Moraxella species that causes epistaxis in macaques 
Veterinary microbiology  2010;147(0):367-375.
Bacteria of the genus Moraxella have been isolated from a variety of mammalian hosts. In a prior survey of bacteria that colonize the rhesus macaque nasopharynx, performed at the Tulane National Primate Research Center, organisms of the Moraxella genus were isolated from animals with epistaxis, or “bloody nose syndrome.” They were biochemically identified as Moraxella catarrhalis, and cryopreserved. Another isolate was obtained from an epistatic cynomolgus macaque at the U.S. Army Medical Research Institute of Infectious Diseases. Based on differences in colony and cell morphologies between rhesus and human M. catarrhalis isolates, we hypothesized that the nonhuman primate Moraxella might instead be a different species. Despite morphological differences, the rhesus isolates, by several biochemical tests, were indistinguishable from M. catarrhalis. Analysis of the cynomolgus isolate by Vitek 2 Compact indicated that it belonged to a Moraxella group, but could not differentiate among species. However, sequencing of the 16S ribosomal RNA gene from four representative rhesus isolates and the cynomolgus isolate showed closest homology to Moraxella lincolnii, a human respiratory tract inhabitant, with 90.16% identity. To examine rhesus macaques as potential hosts for M. catarrhalis, eight animals were inoculated with human M. catarrhalis isolates. Only one of the animals was colonized and showed disease, whereas four of four macaques became epistatic after inoculation with the rhesus Moraxella isolate. The nasopharyngeal isolates in this study appear uniquely adapted to a macaque host and, though they share many of the phenotypic characteristics of M. catarrhalis, appear to form a genotypically distinct species.
PMCID: PMC3971920  PMID: 20667430
Moraxella; Epistaxis; Nonhuman primate; Macaca
17.  Rabies virus glycoprotein is an important determinant for the induction of innate immune responses and the pathogenic mechanisms 
Veterinary microbiology  2012;162(2-4):601-613.
Our previous studies have suggested that street and fixed rabies viruses (RABV) induce diseases in the mouse model via different mechanisms. In the present study, attempts were made to determine if it is the glycoprotein (G) that is responsible for the observed differences in the pathogenic mechanisms. To this end, an infectious clone from fixed virus B2c was established and used as a backbone for exchange of the G from street viruses. The rate of viral replication, expression of viral proteins, and the induction of innate immune responses were compared in cells or in mice infected with each of the viruses. Furthermore, the infiltration of inflammatory cells into the CNS and the enhancement of blood-brain-barrier (BBB) permeability were also compared. It was found that fixed viruses induced stronger innate immune responses (expression of chemokines, infiltration of inflammatory cells, and enhancement of BBB permeability) than street RABV or recombinant viruses expressing the G from street RABVs. Fixed viruses induce disease via an immune-mediated pathogenic mechanism while street viruses or recombinant viruses expressing the G from street RABVs induce diseases via a mechanism other than immune-mediated pathogenesis. Therefore, RABV G is an important determinant for the induction of innate immune responses and consequently the pathogenic mechanisms.
PMCID: PMC3568536  PMID: 23265241
Rabies virus; innate immune response; inflammation; blood brain barrier
18.  Multidrug-resistant Escherichia coli from canine urinary tract infections tend to have commensal phylotypes, lower prevalence of virulence determinants and ampC-replicons☆ 
Veterinary Microbiology  2014;169(3-4):171-178.
Multidrug-resistant Escherichia coli is an emerging clinical challenge in domestic species. Treatment options in many cases are limited. This study characterized MDR E. coli isolates from urinary tract infections in dogs, collected between 2002 and 2011. Isolates were evaluated in terms of β-lactamase production, phylogenetic group, ST type, replicon type and virulence marker profile. Comparisons were made with antibiotic susceptible isolates also collected from dogs with urinary tract infections. AmpC β-lactamase was produced in 67% of the MDR isolates (12/18). Of these, 8 could be specifically attributed to the CMY-2 gene. None of the isolates tested in either group expressed ESBLs. Phylo-group distribution was as expected in the susceptible isolates, with an over representation of the pathogenic B2 phylo-group (67%). In contrast, the phylogenetic background for the MDR group was mixed, with representation of commensal phylo-groups A and B1. The B2 phylo-group represented the smallest proportion (A, B1, B2 or D was 28%, 22%, 11% and 33%, respectively). Virulence marker profiles, evaluated using Identibac® microarray, discriminated between the two groups. Marker sequences for a core panel of virulence determinants were identified in most of the susceptible isolates, but not in most of the MDR isolates. These findings indicate that for MDR isolates, plasmid-mediated AmpC is an important resistance mechanism, and while still capable of causing clinical disease, there is evidence for a shift towards phylogenetic groups of reduced inferred virulence potential. There was no evidence of zoonotic potential in either the susceptible or MDR urinary tract isolates in this study.
PMCID: PMC3969583  PMID: 24485933
Escherichia coli; Plasmid-mediated AmpC; Urinary tract infection; Dog; Multidrug-resistant; β-Lactamase
19.  Correlation of cell surface marker expression with African swine fever virus infection☆ 
Veterinary Microbiology  2014;168(2-4):413-419.
The expression of surface markers on African swine fever virus (ASFV) infected cells was evaluated to assess their involvement in infection. Previous findings indicated CD163 expression was correlated with ASFV susceptibility. However, in this study the expression of porcine CD163 on cell lines did not increase the infection rate of these cells indicating other factors are likely to be important in determining susceptibility to infection. On adherent porcine bone marrow (pBM) cells the expression of CD45 was strongly correlated with infection. CD163 and CD203a expression correlated at intermediate levels with infection, indicating cells expressing these markers could become infected but were not preferentially infected by the virus. Most of the cells expressing MHCII were infected, indicating that they may be preferentially infected although expression of MHCII was not essential for infection and a large percentage of the infected cells were MHCII negative. CD16 showed a marked decrease in expression following infection and significantly lower levels of infected cells were shown to express CD16. Altogether these results suggest CD163 may be involved in ASFV infection but it may not be essential; the results also highlight the importance of other cell markers which requiring further investigation.
PMCID: PMC3969584  PMID: 24398227
African swine fever virus; Surface markers; CD163; MHCII; CD203a; CD45
20.  Gene expression analysis of Salmonella enterica SPI in macrophages indicates differences between serovars that induce systemic disease from those normally causing enteritis☆ 
Veterinary Microbiology  2013;167(3-4):675-679.
Global gene expression of the invasive Salmonella serovars S. Enteritidis and S. Typhimurium, and the less-invasive S. Infantis and S. Hadar was studied during infection of a chicken macrophage cell line. Major functional gene groups responsible for intracellular physiological changes were regulated similarly in all four serovars. However, SPI1 and SPI4 genes of S. Enteritidis and S. Typhimurium were strongly repressed in the macrophages whereas S. Infantis, S. Hadar and other similar serovars maintained up-regulation of these gene sets. This phenomenon may explain some of the biological differences between invasive and non-invasive Salmonella serovars.
PMCID: PMC3878769  PMID: 24080352
Salmonella; Macrophage; Gene expression; Microarray; SPI
21.  Molecular detection of murine noroviruses in laboratory and wild mice 
Veterinary microbiology  2012;160(3-4):463-467.
Fecal specimens collected from 121 laboratory mice, 30 striped field mice (Apodemus agrarius), 70 yellow-necked mice (Apodemus flavicollis), and 3 bank voles (Myodes glareolus) were tested in sample pools for the presence of murine noroviruses (MNV). Ten of 41 laboratory mice and 2 of 3 striped field mice pooled samples were positive for MNV. All laboratory mouse MNVs were closely related to previously described MNVs. The complete ORF2 (VP1) of both striped field mouse MNVs identified in this study was 1623 nt (541 aa) long and differed at 12% nt (8% aa) positions from each other, at 22–24% nt (15–18% aa) positions from the laboratory mouse MNVs and at 20–22% nt (13–14% aa) positions from the recently described wood mouse (Apodemus sylvaticus) MNVs. This study provides further evidence for the circulation of novel, genetically diverse MNVs in wild mice.
PMCID: PMC3469783  PMID: 22748629
mouse; calicivirus; murine norovirus
22.  Co-infection of porcine dendritic cells with porcine circovirus type 2a (PCV2a) and genotype II porcine reproductive and respiratory syndrome virus (PRRSV) induces CD4+CD25+FoxP3+ T cells in vitro 
Veterinary microbiology  2012;160(1-2):233-239.
Porcine circovirus associated disease (PCVAD) is currently one of the most economically important diseases in the global swine industry. Porcine circovirus type 2 (PCV2) is the primary causative agent, however co-infection with other swine pathogens such as porcine reproductive and respiratory syndrome virus (PRRSV) is often required to induce the full spectrum of clinical PCVAD. While the specific mechanisms of viral co-infection that lead to clinical disease are not fully understood, immune modulation by the co-infecting viruses likely plays a critical role. We evaluated the ability of dendritic cells (DC) infected with PRRSV, PCV2 or both to induce regulatory T cells (Tregs) in vitro. DCs infected with PCV2 significantly increased CD4+CD25+FoxP3+ Tregs (p<0.05) and DCs co-infected with PRRSV and PCV2 induced significantly higher numbers of Tregs than with PCV2 alone (p<0.05). Cytokine analysis indicated that the induction of Tregs by co-infected DCs may be dependent on TGF-β and not IL-10. Our data support the immunomodulatory role of PCV2/PRRSV co-infection in the pathogenesis of PCVAD, specifically via Treg mediated immunosuppression.
PMCID: PMC3443269  PMID: 22633482
porcine reproductive and respiratory syndrome virus (PRRSV); porcine circovirus type 2 (PCV2); dendritic cells; porcine circovirus associated disease (PCVAD); regulatory T cell; co-infection
23.  Bordetella avium Antibiotic Resistance, Novel Enrichment Culture, and Antigenic Characterization 
Veterinary microbiology  2012;160(1-2):189-196.
Bordetella avium continues to be an economic issue in the turkey industry as the causative agent of bordetellosis, which often leads to serious secondary infections. This study presents a broad characterization of the antibiotic resistance patterns in this diverse collection of B. avium strains collected over the past thirty years. In addition, the plasmid basis for the antibiotic resistance was characterized. The antibiotic resistance pattern allowed the development of a novel enrichment culture method that was subsequently employed to gather new isolates from diseased turkeys and a healthy sawhet owl. While a healthy turkey flock was shown to seroconvert by four weeks-of-age, attempts to culture B. avium from healthy turkey poults were unsuccessful. Western blot of B. avium strains using pooled serum from diseased and healthy commercial turkey flocks revealed both antigenic similarities and differences between strains. In sum, the work documents the continued exposure of commercial turkey flocks to B. avium and the need for development of an effective, inexpensive vaccine to control spread of the disease.
PMCID: PMC3469198  PMID: 22721730
Bordetellosis incidence; serology; antibiotic resistance; Bordetella avium; poultry
24.  Initial sequence characterization of the rhabdoviruses of squamate reptiles, including a novel rhabdovirus from a caiman lizard (Dracaena guianensis) 
Veterinary Microbiology  2012;158(3-4):274-279.
Rhabdoviruses infect a variety of hosts, including non-avian reptiles. Consensus PCR techniques were used to obtain partial RNA-dependent RNA polymerase gene sequence from five rhabdoviruses of South American lizards; Marco, Chaco, Timbo, Sena Madureira, and a rhabdovirus from a caiman lizard (Dracaena guianensis). The caiman lizard rhabdovirus formed inclusions in erythrocytes, which may be a route for infecting hematophagous insects. This is the first information on behavior of a rhabdovirus in squamates. We also obtained sequence from two rhabdoviruses of Australian lizards, confirming previous Charleville virus sequence and finding that, unlike a previous sequence report but in agreement with serologic reports, Almpiwar virus is clearly distinct from Charleville virus. Bayesian and maximum likelihood phylogenetic analysis revealed that most known rhabdoviruses of squamates cluster in the Almpiwar subgroup. The exception is Marco virus, which is found in the Hart Park group.
PMCID: PMC3371314  PMID: 22397930
Rhabdoviridae; phylogeny; Dracaena guianensis; Marco virus; Chaco virus; Timbo virus; Sena Madureira virus
25.  Detection of classical and newly described staphylococcal superantigen genes in coagulase-negative staphylococci isolated from bovine intramammary infections 
Veterinary microbiology  2010;147(0):149-154.
The coagulase negative staphylococci (CNS) are the most prevalent mastitis pathogen group yet their virulence characteristics have not been well described. We investigated the presence of 19 classical and newly described staphylococcal superantigen (SAg) genes in CNS isolates from bovine intramammary infections (IMI). A total of 263 CNS representing 11 different Staphylococcus spp. were examined, and 31.2% (n = 82) of CNS isolates had one or more SAg genes; there were 21 different SAg gene combinations. The most prevalent combination of SAg genes (seb, seln, and selq; n = 45) was found in S. chromogenes, S. xylosus, S. haemolyticus, S. sciuri subsp. carnaticus, S. simulans and S. succinus. The genes for SAgs appear to be widely distributed amongst CNS isolated from bovine IMI.
PMCID: PMC3689430  PMID: 20667668
coagulase-negative staphylococci; staphylococcal superantigens; multiplex PCR

Results 1-25 (64)