The potential risk associated with foodborne pathogens in a watershed: Escherichia coli O157:H7 in dairy cattle.

We carried out an investigation to determine the occurrence of Escherichia coli O157:H7 in dairy herds in the Delaware County watershed and to identify the factors that play a role in the likelihood of presence of this organism among animals on these farms. The pathogen poses risk of environmental degradation and health to the inhabitants. A total of 2162 fecal samples were collected per rectum from a representative sample of cattle on 27 dairy farms. The samples were investigated for the presence of E. coli O157:H by initially enriching using a bacteriological media and detection of the pathogen using real-time polymerase chain reaction technique. Escherichia coli O157:H7 was detected in 74% of the herds in the target population and in 3.7% of samples collected. An additional 54 animals were identified that were infected with O157 non-H7 strains of E. coli among 15 farms. Several putative risk factors were associated with the detection of the pathogen on the enrolled farms included age, housing calves indoors, group housing for calves, housing calves in the calf barn, presence of dogs on the farm, and housing post-weaned calves in a cow barn or heifer barn versus a greenhouse. In conclusion, E. coli O157:H7 was present on the dairy farms of Delaware County and may pose a threat to the people that live and work there. The risk associated with the detection of this pathogen could be reduced by adjusting management factors that were identified in this study.

Geography Shapes the Population Genomics of Salmonella enterica Dublin.

Salmonella enterica serotype Dublin (S. Dublin) is a bovine-adapted serotype that can cause serious systemic infections in humans. Despite the increasing prevalence of human infections and the negative impact on agricultural processes, little is known about the population structure of the serotype. To this end, we compiled a manually curated data set comprising of 880 S. Dublin genomes. Core genome phylogeny and ancestral state reconstruction revealed that region-specific clades dominate the global population structure of S. Dublin. Strains of S. Dublin in the UK are genomically distinct from US, Brazilian, and African strains. The geographical partitioning impacts the composition of the core genome as well as the ancillary genome. Antibiotic resistance genes are almost exclusively found in US genomes and are mediated by an IncA/C2 plasmid. Phage content and the S. Dublin virulence plasmid were strongly conserved in the serotype. Comparison of S. Dublin to a closely related serotype, S. enterica serotype Enteritidis, revealed that S. Dublin contains 82 serotype specific genes that are not found in S. Enteritidis. Said genes encode metabolic functions involved in the uptake and catabolism of carbohydrates and virulence genes associated with type VI secretion systems and fimbria assembly respectively.

Detection of Salmonella spp. in veterinary samples by combining selective enrichment and real-time PCR.

Rapid screening for enteric bacterial pathogens in clinical environments is essential for biosecurity. Salmonella found in veterinary hospitals, particularly Salmonella enterica serovar Dublin, can pose unique challenges for culture and testing because of its poor growth. Multiple Salmonella serovars including Dublin are emerging threats to public health given increasing prevalence and antimicrobial resistance. We adapted an automated food testing method to veterinary samples and evaluated the performance of the method in a variety of matrices including environmental samples ( n = 81), tissues ( n = 52), feces ( n = 148), and feed ( n = 29). A commercial kit was chosen as the basis for this approach in view of extensive performance characterizations published by multiple independent organizations. A workflow was established for efficiently and accurately testing veterinary matrices and environmental samples by use of real-time PCR after selective enrichment in Rappaport-Vassiliadis soya (RVS) medium. Using this method, the detection limit for S. Dublin improved by 100-fold over subculture on selective agars (eosin-methylene blue, brilliant green, and xylose-lysine-deoxycholate). Overall, the procedure was effective in detecting Salmonella spp. and provided next-day results.

Infectious crystalline keratopathy in dogs and cats: clinical, in vivo confocal microscopic, histopathologic, and microbiologic features of eight cases.

OBJECTIVE: To describe clinical, in vivo confocal microscopic, histopathologic, and microbiologic features of canine and feline cases of infectious crystalline keratopathy (ICK). ANIMALS STUDIED: Six dogs and two cats with naturally acquired ICK. PROCEDURES: Medical records of dogs and cats with a clinical diagnosis of ICK were reviewed. Signalment, medical history, clinical findings, and diagnostic evaluations were retrieved, including corneal cytology, histopathology, in vivo confocal microscopy, and microbiology results. RESULTS: All animals presented with fine, needle-like, and branching white crystalline anterior stromal opacities emanating from corneal facets or corneal epithelial defects. Mild conjunctival hyperemia and anterior uveitis were frequently present. Concurrent ocular and systemic diseases were common, including keratoconjunctivitis sicca, corneal sequestrum, diabetes mellitus, hyperadrenocorticism, and malignant neoplasia. Bacteria, with minimal or absent leukocytes, were identified by cytology and histopathology. Histopathologically, the crystalline corneal opacities corresponded with dense accumulations of bacteria present in the interlamellar stromal spaces and forming cord-like projections within the stroma. In vivo confocal microscopy demonstrated deposits of reflective crystalline or amorphous structures within the stroma with a paucity of associated inflammatory changes. The most frequently cultured bacteria were alpha-hemolytic Streptococcus and Staphylococcus species. Resolution of clinical lesions was achieved in most cases with long-term medical or surgical therapy; however, the initiation of medical treatment was associated with an acute, dramatic onset of severe keratitis and anterior uveitis in some animals. CONCLUSIONS: Infectious crystalline keratopathy in dogs and cats shares many features with this condition in human patients. Prolonged medical therapy, or surgical intervention, is required for resolution.

The risk of carriage of Salmonella spp. and Listeria monocytogenes in food animals in dynamic populations.

Salmonella spp. and Listeria monocytogenes are foodborne pathogens of global importance. We assessed their risks and associated factors in a highly dynamic population of animals. Animal and environmental samples were collected from dairy cattle, sheep, camel and chickens at either the farms or the abattoirs. The pathogens were detected using a combination of bacterial enrichment culture and real-time polymerase chain reaction (PCR). Data on putative risk factors were collect and analysed for their significance of association with these pathogens. Salmonella spp. were detected at higher proportions in sheep faeces and sheep carcasses in comparison to cattle faeces (odds ratio = 2.4 and 2.2, respectively). This pathogen was less common in milk or carcasses samples from cattle or chickens. Sheep and camel carcass samples were highly contaminated with Salmonella spp. Faecal samples from cattle had the most diverse serovars of Salmonella enterica including S. Newport, S. Haifa, S. Kedougou, S. Kentucky, S. Mbandaka and S. Goettingen. Exotic serovars in sheep included S. Eastbourne, S. Chester and S. Kottnus. Serovars that were shed in camel faeces included S. Newport, S. Bovismorbificans and S. Infantis. In all sampled populations, detection of Salmonella spp. was more likely during warmer months than cold months. Listeria monocytogenes was not common in the targeted populations and was detected at a rate of 2.4%, mainly from sheep carcasses. The study highlights the role of food animals as reservoirs of pathogens across boundaries since all feed are imported in that population from different parts of the world.

Recombinant antigens rLipL21, rLoa22, rLipL32 and rLigACon4-8 for serological diagnosis of leptospirosis by enzyme-linked immunosorbent assays in dogs.

Animal leptospirosis is one of the most common zoonotic diseases in the United States and around the world. In a previous study, we applied four recombinant antigens, rLipL21, rLoa22, rLipL32 and rLigACon4-8 of Leptospira interrogans (L. interrogans) for the serological diagnosis of equine leptospirosis (Ye et al, Serodiagnosis of equine leptospirosis by ELISA using four recombinant protein markers, Clin. Vaccine. Immunol. 21:478-483). In this study, the same four recombinant antigens were evaluated for their potential to diagnose canine leptospirosis by ELISA. A total of 305 canine sera that were Leptospira microscopic agglutination test (MAT)-negative (n = 102) and MAT-positive (n = 203) to 5 serovars (Pomona, Grippotyphosa, Icterohaemorrhagiae, Canicola and Hardjo) were tested. When individual recombinant antigens were used, the sensitivity and specificity of ELISA were 97.5% and 84.3% for rLigACon4-8; 89.7% and 81.4% for rLoa22; 92.6% and 84.3% for rLipL32 and 99.5% and 84.3% for rLipL21, respectively compared to the MAT. The sensitivity and specificity of ELISA were, 92.6% and 91.2% for rLigACon4-8 and rLipL32, 97.5% and 84.3% for rLigACon4-8 and rLipL21, 89.7% and 87.3% for rLigACon4-8 and rLoa22, 89.7% and 87.3% to rLipL21 and rLoa22, 92.6% and 91.2% for rLipL21 and rLipL32 and 89.2% and 94.1% for rLoa22 and rLipL32 when one of the two antigens was test positive. The use of all four antigens in the ELISA assay was found to be sensitive and specific, easy to perform, and agreed with the results of the standard Leptospira Microscopic Agglutination test (MAT) for the diagnosis of canine leptospirosis.

Salmonella enterica serovar Oranienburg outbreak in a veterinary medical teaching hospital with evidence of nosocomial and on-farm transmission.

Nosocomial salmonellosis continues to pose an important threat to veterinary medical teaching hospitals. The objectives of this study were to describe an outbreak of salmonellosis caused by Salmonella enterica serovar Oranienburg within our hospital and to highlight its unique features, which can be used to help mitigate or prevent nosocomial outbreaks in the future. We retrospectively analyzed data from patients that were fecal culture-positive for Salmonella Oranienburg between January 1, 2006, and June 1, 2011, including historical, clinical, and pulsed-field gel electrophoresis (PFGE) data. Salmonella Oranienburg was identified in 20 horses, five alpacas, and three cows during this time frame, with dates of admission spanning the period from August, 2006, through January, 2008. We consider most of these patients to have become infected through either nosocomial or on-farm transmission, as evidenced by molecular subtyping results and supportive epidemiologic data. Interpretation of PFGE results in this outbreak was challenging because of the identification of several closely related Salmonella Oranienburg subtypes. Furthermore, a high percentage of cases were fecal culture-positive for Salmonella Oranienburg within 24 h of admission. These patients initially appeared to represent new introductions of Salmonella into the hospital, but closer inspection of their medical records revealed epidemiologic links to the hospital following the index case. Cessation of this outbreak was observed following efforts to further heighten biosecurity efforts, with no known cases or positive environmental samples after January, 2008. This study demonstrates that a Salmonella-positive culture result within 24 h of admission does not exclude the hospital as the source of infection, and it underscores the important role played by veterinary medical teaching hospitals as nodes of Salmonella infection that can promote transmission outside of the hospital setting.

Serodiagnosis of equine leptospirosis by enzyme-linked immunosorbent assay using four recombinant protein markers.

Leptospirosis, caused by Leptospira spp., is one of the most common zoonotic diseases in the world. We tested four recombinant proteins of Leptospira interrogans, namely, rLipL21, rLoa22, rLipL32, and rLigACon4-8, to evaluate their potential for use as antigens for the diagnosis of equine leptospirosis. We employed equine sera (n = 130) that were microscopic agglutination test (MAT) negative and sera (n = 176) that were MAT positive for the 5 serovars that most commonly cause equine leptospirosis. The sensitivity and specificity of ELISA compared to MAT were 82.39% and 86.15%, respectively, for LigACon4-8, 77.84% and 92.31%, respectively, for Loa22, 77.84% and 86.15%, respectively, for LipL32, and 84.66% and 83.85%, respectively, for LipL21. When one of the two antigens was test positive, the sensitivity and specificity of ELISA were 93.75% and 78.46%, respectively, for rLigACon4-8 and LipL32, 93.18% and 76.15%, respectively, for rLigACon4-8 and LipL21, 89.77% and 80.77%, respectively, for rLigACon4-8 and Loa22, 91.48% and 78.46%, respectively, for LipL21 and Loa22, 93.75% and 76.92%, respectively, for LipL21 and LipL32, and 90.34% and 80.77%, respectively, for Loa22 and LipL32. In conclusion, we have developed an indirect ELISA utilizing rLigACon4-8, rLoa22, rLipL32, and rLipL21 as diagnostic antigens for equine leptospirosis. The use of four antigens in the ELISA was found to be sensitive and specific, the assay was easy to perform, and the results concurred with the results of the standard Leptospira MAT.

Molecular detection of Salmonella species in bovine fecal samples.

A study was performed to assess the validity of the BAX automated polymerase chain reaction system (DuPont Nutrition & Health, Wilmington, Delaware) to detect the shedding of Salmonella species in bovine fecal samples. A total of 133 bovine fecal samples that were submitted to the Cornell University Animal Health Diagnostic Laboratory for Salmonella culture were also tested in the BAX system with a modified version of the manufacturer's enrichment protocol. Using culture as the gold standard test, the BAX system was found to have a sensitivity of 85.7% and a specificity of 90.5%. There was excellent agreement (κ = 0.71, standard error = 0.072) and no significant differences between the 2 methods (McNemar χ(2) = 0.180).

Multidrug resistance is common in Escherichia coli associated with ileal Crohn's disease.

BACKGROUND: Escherichia coli is increasingly implicated in the pathogenesis of ileal Crohn's disease (ICD), offering a potential therapeutic target for disease management. Empirical antimicrobial targeting of ileal E. coli has advantages of economy and speed of implementation, but relies on uniform susceptibility of E. coli to routinely selected antimicrobials to avoid apparent treatment failure. Therefore, we examined the susceptibility of ileal E. coli to such antimicrobials. METHODS: E. coli from 32 patients with ICD and 28 with normal ileum (NI) were characterized by phylogroup, pathotype, antimicrobial susceptibility, and presence of antimicrobial resistance genes. RESULTS: In all, 17/32 ICD and 12/28 NI patients harbored ≥ 1 E. coli strain; 10/24 E. coli strains from ICD and 2/14 from NI were nonsuscepti-ble to ≥ 1 antimicrobial in ≥ 3 categories (multidrug-resistant). Resistance to amoxicillin/clavulanic-acid, cefoxitin, chloramphenicol, ciprofloxa-cin, gentamicin, and rifaximin was restricted to ICD, with 10/24 strains from 8/17 patients resistant to ciprofloxacin or rifaximin (P < 0.01). Adherent-invasive E. coli (AIEC) were isolated from 8/32 ICD and 5/28 NI, and accounted for 54% and 43% of E. coli strains in these groups. In all, 8/13 AIEC strains from ICD (6/8 patients) versus 2/6 NI (2/5 patients) showed resistance to the macrophage-penetrating antimicrobials ciprofloxacin, clarithromycin, rifampicin, tetracycline, and trimethoprim/sulfamethoxazole. Resistance was associated with tetA, tetB, tetC, bla-(TEM), bla(oxa)-1, sulI, sulII, dhfrI, dhfrVII, ant(3″)-Ia, and catI genes and prior use of rifaximin (P < 0.01). CONCLUSIONS: ICD-associated E. coli frequently manifest resistance to commonly used antimicrobials. Clinical trials of antimicrobials against E. coli in ICD that are informed by susceptibility testing, rather than empirical selection, are more likely to demonstrate valid outcomes of such therapy.

The effect of clinical outbreaks of salmonellosis on the prevalence of fecal Salmonella shedding among dairy cattle in New York.

The objective of this study was to determine if the within-herd prevalence of fecal Salmonella shedding is higher in dairy herds with clinical outbreaks of disease, as compared to herds with subclinical infections only. Data were collected prospectively from dairy herds throughout New York that had at least 150 lactating cows and that received clinical service from participating veterinarians. After enrollment, Salmonella surveillance consisted of both environmental screening and disease monitoring within the herd. Herds positive by either environmental or fecal culture were sampled during three visits to estimate the within-herd prevalence of Salmonella. We characterized isolates by serovar and antimicrobial resistance pattern. Among 57 enrolled herds, 44 (77%) yielded Salmonella-positive samples during the study period; 27 (61%) of the positive herds had Salmonella isolated from environmental samples only, and 17 (39%) had one or more laboratory-confirmed clinical cases. The within-herd prevalence of fecal Salmonella shedding ranged from 0 to 53%. Salmonella Cerro was the predominant serovar, accounting for 56% of all isolates. Antimicrobial resistance ranged from zero to nine drugs, and 14 (32%) of the positive farms generated multidrug-resistant isolates. Herds with laboratory-confirmed clinical cases had a higher prevalence of fecal Salmonella shedding than herds that only generated positive environmental samples, as estimated by a Poisson regression model (prevalence ratio, 2.7; p = 0.01). An association between dairy herd outbreaks of salmonellosis and a higher prevalence of asymptomatic shedding should help guide strategies for reducing the public health threat of Salmonella, as the ability to recognize high-risk herds by clinical laboratory submissions presents an obvious opportunity to maximize food safety at the preharvest level. This is in contrast with other foodborne zoonotic pathogens, such as Campylobacter jejuni and Escherichia coli O157:H7, which occur widely in adult cattle without accompanying clinical disease.

Molecular identification of veterinary yeast isolates by use of sequence-based analysis of the D1/D2 region of the large ribosomal subunit.

Conventional methods of yeast identification are often time-consuming and difficult; however, recent studies of sequence-based identification methods have shown promise. Additionally, little is known about the diversity of yeasts identified from various animal species in veterinary diagnostic laboratories. Therefore, in this study, we examined three methods of identification by using 109 yeast samples isolated during a 1-year period from veterinary clinical samples. Comparison of the three methods-traditional substrate assimilation, fatty acid profile analysis, and sequence-based analysis of the region spanning the D1 and D2 regions (D1/D2) of the large ribosomal subunit-showed that sequence analysis provided the highest percent identification among the three. Sequence analysis identified 87% of isolates to the species level, whereas substrate assimilation and fatty acid profile analysis identified only 54% and 47%, respectively. Less-stringent criteria for identification increased the percentage of isolates identified to 98% for sequence analysis, 62% for substrate assimilation, and 55% for fatty acid profile analysis. We also found that sequence analysis of the internal transcribed spacer 2 (ITS2) region provided further identification for 36% of yeast not identified to the species level by D1/D2 sequence analysis. Additionally, we identified a large variety of yeast from animal sources, with at least 30 different species among the isolates tested, and with the majority not belonging to the common Candida spp., such as C. albicans, C. glabrata, C. tropicalis, and the C. parapsilosis group. Thus, we determined that sequence analysis of the D1/D2 region was the best method for identification of the variety of yeasts found in a veterinary population.

Pulsed-field gel electrophoresis diversity of human and bovine clinical Salmonella isolates.

Pulsed-field gel electrophoresis (PFGE) characterization of 335 temporally and spatially matched clinical, bovine, and human Salmonella enterica subsp. enterica isolates revealed 167 XbaI PFGE patterns. These isolates were previously classified into 51 serotypes and 73 sequence types, as determined by multilocus sequence typing. Discriminatory power of PFGE (Simpson's index, D = 0.991) was considerably higher than that of multilocus sequence typing (D = 0.920) or serotyping (D = 0.913). Although 128 PFGE types each only represented a single isolate, 8 PFGE types represented >4 isolates, including (i) three serotype Enteritidis and Heidelberg patterns that were only identified among human isolates, (ii) two PFGE patterns (each representing serotypes Bardo and Newport) that were significantly more common among bovine isolates as compared with human isolates; (iii) two PFGE types that each includes two serotypes (4,5,12:i:- and Typhimurium; Thompson and 1,7:-:1,5); and (iv) one PFGE type that includes eight Typhimurium isolates from humans and cattle. Characterization of isolates collected over multiple farm visits indicated that given specific PFGE types persisted over time on 11 farms. On an additional seven farms, isolates with a given sequence type represented multiple PFGE type, which typically only differed by <3 bands, suggesting PFGE type diversification during strain persistence. Sixteen PFGE types were isolated from 2 or more farms, including two widely distributed serotype Newport-associated PFGE types each found on 10 farms. In six instances two or three human isolates collected in the same county in the same or consecutive months represented the same subtypes, suggesting small human case clusters. PFGE-based characterization and surveillance of human and animal isolates can provide improved understanding of Salmonella diversity and epidemiology, including identification of possible host-associated and common, widely distributed PFGE types.

Temporal clusters of bovine Salmonella cases at a veterinary medical teaching hospital, 1996-2007.

The objectives of this study were to identify and characterize temporal clusters of bovine Salmonella cases at a veterinary medical teaching hospital and to determine which clusters were likely to have involved nosocomial transmission. Data on fecal Salmonella shedding status, serotype, and antimicrobial resistance were collected retrospectively for all cattle admitted to the Cornell University Equine and Farm Animal Hospital between January 1, 1996, and June 1, 2007. Pulsed-field gel electrophoresis (PFGE) was performed on all available isolates. Cluster analysis was used to identify temporal clusters of cases. A total of 5398 cattle were admitted during the study period; the prevalence of fecal Salmonella shedding among clinical suspects was 6.5%, whereas that among nonsuspects tested through routine surveillance was 2.5%. Eight temporal clusters (including 57 cattle) were investigated as possible outbreaks involving nosocomial transmission, ranging in size from 4 to 10 cases. All but one cluster were centered over the month of August or September. A total of 15 Salmonella serotypes were represented, with the most common being Typhimurium (33%), Newport (23%), and Agona (12%). Among the isolates available for PFGE analysis, there were 19 PFGE types represented. The majority of temporal clusters during the study period were not nosocomial in origin. However, two of the clusters were outbreaks directly resulting from nosocomial Salmonella transmission, based on case histories, serotype data, antimicrobial resistance patterns, and PFGE analysis. The clear seasonal pattern exhibited by these clusters underscores the need for heightened Salmonella vigilance during the late summer and early fall. The combination of statistical methods, routine bacteriologic data, and PFGE analysis is an effective means of conducting surveillance and outbreak investigations in a hospital setting.

Fecal shedding of, antimicrobial resistance in, and serologic response to Salmonella Typhimurium in dairy calves.

OBJECTIVE: To determine the duration of fecal shedding of and serologic response to Salmonella spp after natural infection in dairy calves and characterize Salmonella organisms recovered from these herds. DESIGN: Longitudinal study. ANIMALS: Calves from 2 dairy herds (A and B) in the northeast United States that were identified at the beginning of a Salmonella outbreak. PROCEDURES: Fecal samples were collected twice per week (herd A) or once per week (herd B); blood samples were collected for serologic testing once per week in both herds. Bacteriologic culture of fecal samples was performed, and Salmonella isolates were characterized by serotype, pulsed-field gel electrophoresis (PFGE) pattern, and antimicrobial resistance profile. RESULTS: All Salmonella isolates from herd A were serovar Typhimurium var Copenhagen, had the same PFGE pattern, and were resistant to at least 9 antimicrobials. All isolates from herd B were Salmonella Typhimurium, represented 2 PFGE patterns, and were susceptible to all antimicrobials evaluated. The estimated duration of fecal shedding was 14 days in herd A and 9 days in herd B. Few calves were seropositive for antibody against Salmonella lipopolysaccharide within the first week after birth (0 of 20 in herd A and 13 of 79 in herd B) or seroconverted (6 in herd A and 4 in herd B). Fecal shedding was more common in calves that seroconverted, but overall, there was not a strong association between seropositivity and fecal shedding of Salmonella organisms. CONCLUSIONS AND CLINICAL RELEVANCE: Although the herds differed in serologic response and Salmonella subtype, the duration of fecal shedding among calves was similar between herds.

Fecal shedding of Salmonella spp among cattle admitted to a veterinary medical teaching hospital.

OBJECTIVE- To estimate the prevalence of fecal shedding of Salmonella spp among bovine patients at a veterinary teaching hospital, to identify risk factors for fecal shedding of Salmonella organisms, and to characterize the serotypes. DESIGN- Retrospective cohort study. SAMPLE POPULATION- 5,398 hospitalized cattle. PROCEDURES- Data were collected for all cattle admitted during an 11-year period. Fecal shedding of Salmonella spp was determined by means of standard bacteriologic culture. Multivariable logistic regression models were used to identify risk factors for shedding of Salmonella spp among patients. RESULTS- The prevalence of Salmonella shedding among clinical suspects was 6.5% (50/768), whereas that among nonsuspects tested through routine surveillance was 2.5% (50/2,020). Among clinical suspect calves, fecal shedding of Salmonella spp was more likely for those admitted in the fall (odds ratio [OR], 5.9), those with septicemia (OR, 3.3), or those with an umbilical hernia (OR, 8.6). Among clinical suspect adult cattle, those with enteritis (OR, 9.9) or metritis (OR, 5.2) were more likely to be shedding Salmonella spp. Among nonsuspect cattle, none of the variables were significant predictors of shedding status. Twenty-one serotypes were detected during the study period, with the most common being Salmonella enterica serotypes Typhimurium (33%), Newport (23%), and Agona (12%). CONCLUSIONS AND CLINICAL RELEVANCE- Seasonal and disease risk factors for fecal shedding of Salmonella spp were evident among clinical suspect cattle admitted to a veterinary teaching hospital. In contrast, lack of significant associations among nonsuspect cattle would suggest that targeted screening within this population is not warranted.

Yersinia pseudotuberculosis in a closed colony of Egyptian fruit bats (Rousettus aegyptiacus).

An outbreak of Yersinia pseudotuberculosis (Yptb) occurred in a closed colony of Egyptian fruit bats (Rousettus aegyptiacus) and resulted in the death of seven bats over a 6-week period. An initial survey of the remaining bats revealed visceral abscessation characteristic of pseudotuberculosis in five of the 12 bats examined (41.7%), inciting depopulation of the colony. At necropsy, 70% of the 115 bats in the colony exhibited gross evidence suggestive of Yptb infection, including mesenteric lymphadenopathy (ML), hepatic abscessation (HA), and/or splenomegaly (SPM). Thirty of these bats (13 females and 17 males of various ages) were chosen at random and their tissues submitted for bacterial culture and histopathologic examination. Twenty-three of these 30 bats had one or more gross lesions considered consistent with Yptb, including ML, HA, and SPM. On histopathology, four of the 30 bats had necrotizing lesions containing Gram-negative bacteria in multiple organs, while 18 others exhibited mild mesenteric lymphadenitis and hepatitis. Four of the 30 bats had positive cultures for Yptb. Bats with gross evidence of mesenteric lymphadenopathy, splenomegaly, or histopathologic presence of demodicosis or bacteria in tissues were more likely (P < 0.05) to have a positive Yersinia culture. Examination of the correlation between population density and mortality rates of the colony revealed that the mortality rate of subadult bats increased dramatically at the time of the outbreak, when the population density was at its highest. It is suspected that stress, primarily from severe overcrowding, predisposed the bat colony to morbidity and mortality from this organism, which likely originated from a rodent reservoir.

Multilocus variable-number tandem-repeat method for typing Salmonella enterica serovar Newport.

In recent years, the proportion of Salmonella enterica infections represented by S. enterica serovar Newport has increased markedly among humans and animals. Multilocus variable-number tandem-repeat analysis (MLVA) has proven to be useful in discriminating other highly clonal Salmonella serovars. Here, we report on the development of a highly discriminatory MLVA for Salmonella serovar Newport.

Identification of potential on-farm sources of Listeria monocytogenes in herds of dairy cattle.

OBJECTIVE: To elucidate the ecology of Listeria monocytogenes on dairy cattle farms by determining the prevalence of the organism in various samples. SAMPLE POPULATION: Dairy cattle operations in central New York State. PROCEDURES: A repeated cross-sectional study design was used. Various samples were obtained from cattle (feces, composite udder milk, and udders), their environment (silage, feed bunks, water troughs, and floor bedding), inline milk filters, and bulk tank milk from 50 dairy farms. Samples were tested for L monocytogenes by use of a PCR assay with 2 steps of bacterial enrichment. Data were analyzed with mixed-effect logistic regression to control for the potential clustering of L monocytogenes on particular farms. RESULTS: L monocytogenes was detected in composite milk, udder swab samples, and fecal samples at prevalences of 13%, 19%, and 43%, respectively. There was no significant clustering of the pathogen by farm. Listeria monocytogenes was more common in samples obtained from cattle and the environment during winter and summer versus the fall. The prevalence of L monocytogenes was twice as high in samples obtained from feed bunks, water troughs, and bedding, compared with that in samples obtained from silage (65%, 66%, 55%, and 30%, respectively). CONCLUSIONS AND CLINICAL RELEVANCE: L monocytogenes was more prevalent in samples obtained from dairy cattle and their environment than in milk samples. Strategies to control the pathogen in dairy operations should focus on cow hygiene and sanitary milk harvesting on the farm.

Adherent and invasive Escherichia coli is associated with granulomatous colitis in boxer dogs.

The mucosa-associated microflora is increasingly considered to play a pivotal role in the pathogenesis of inflammatory bowel disease. This study explored the possibility that an abnormal mucosal flora is involved in the etiopathogenesis of granulomatous colitis of Boxer dogs (GCB). Colonic biopsy samples from affected dogs (n = 13) and controls (n = 38) were examined by fluorescent in situ hybridization (FISH) with a eubacterial 16S rRNA probe. Culture, 16S ribosomal DNA sequencing, and histochemistry were used to guide subsequent FISH. GCB-associated Escherichia coli isolates were evaluated for their ability to invade and persist in cultured epithelial cells and macrophages as well as for serotype, phylogenetic group, genome size, overall genotype, and presence of virulence genes. Intramucosal gram-negative coccobacilli were present in 100% of GCB samples but not controls. Invasive bacteria hybridized with FISH probes to E. coli. Three of four GCB-associated E. coli isolates adhered to, invaded, and replicated within cultured epithelial cells. Invasion triggered a "splash"-type response, was decreased by cytochalasin D, genistein, colchicine, and wortmannin, and paralleled the behavior of the Crohn's disease-associated strain E. coli LF 82. GCB E. coli and LF 82 were diverse in serotype and overall genotype but similar in phylogeny (B2 and D), in virulence gene profiles (fyuA, irp1, irp2, chuA, fepC, ibeA, kpsMII, iss), in having a larger genome size than commensal E. coli, and in the presence of novel multilocus sequence types. We conclude that GCB is associated with selective intramucosal colonization by E. coli. E. coli strains associated with GCB and Crohn's disease have an adherent and invasive phenotype and novel multilocus sequence types and resemble E. coli associated with extraintestinal disease in phylogeny and virulence gene profile.