OPHIDIOMYCES OPHIODIICOLA ON A CAPTIVE BLACK RACER (COLUBER CONSTRICTOR) AND A GARTER SNAKE (THAMNOPHIS SIRTALIS) IN PENNSYLVANIA.

Two snakes were presented to the Pennsylvania State University Animal Diagnostic Laboratory with one suffering from external lesions where the scales were raised and discolored, and the other with oral lesions and swelling extending to the left eye, which was opaque. Histopathological analysis revealed multifocal granulomas containing fungal hyphae. Morphological and DNA sequence analyses revealed both suffered from infection by Ophidiomyces ophiodiicola, an emerging pathogen of snakes. This is the first report of this disease in Pennsylvania.

A retrospective study of equine actinobacillosis cases: 1999-2011.

Several Actinobacillus spp. are common commensal bacteria of the oral cavity, gastrointestinal tract, and reproductive tract of horses and can cause disease in both foals and adults. The current retrospective study was designed to review Actinobacillus spp. isolated from clinical samples or necropsies of 99 horses during 1999-2011. The cases consisted of 43 foals (<6 months of age), 4 young adults (6 months-2 years), 39 adults (>2 years of age), 2 aborted fetuses, and 11 with unspecified ages. Clinical history, signs, bacterial species isolated, and associated lesions were documented. Actinobacillus spp. were isolated 111 times. The most common isolates were Actinobacillus equuli subsp. equuli (38.7%) and hemolytic Actinobacillus spp. (24.3%). Other isolates were Actinobacillus lignieresii (5.4%), Actinobacillus pleuropneumoniae (1.8%), and unclassified Actinobacillus spp. (28.8%). Actinobacillus equuli subsp. equuli was most commonly isolated from clinical and necropsy cases of septicemia and respiratory disease in both foals and adults. Embolic nephritis, the classical septicemic lesion of equine neonatal actinobacillosis, was also present in several adult septicemic actinobacillosis cases. Predisposing factors such as failure of passive transfer of colostral antibodies as well as concurrent pathogenic bacterial or viral infections were present in numerous actinobacillosis cases. There were many cases, however, for which a predisposing factor or concurrent infection was not documented or apparent, suggesting that Actinobacillus spp. can be primary pathogens under the right circumstances and in the right location.

Canine intestinal histoplasmosis containing hyphal forms.

A 12-year-old intact male Miniature Schnauzer dog with chronic diarrhea that was unresponsive to empirical treatment was presented to a referring veterinarian. A laparotomy was performed, and formalin-fixed biopsies of duodenum, jejunum, and colon were sent to Oklahoma Animal Disease Diagnostic Laboratory for evaluation. Histologic examination revealed a severe, diffuse, granulomatous enteritis and colitis with intralesional yeast and hyphal forms. Grocott methenamine silver stains revealed short, aseptate hyphae co-mingled with 2-8 µm, oval to round yeast organisms consistent with Histoplasma capsulatum. The atypical presentation of both yeast and hyphal forms prompted identification of the organism. Direct sequencing of a polymerase chain reaction product from paraffin-embedded intestinal samples confirmed the presence of Ajellomyces capsulatus with a homology over 99% to several sequences in GenBank. Ajellomyces capsulatus is the holomorphic name for H. capsulatum. Therefore, the mycelial form of a dimorphic fungus such as H. capsulatum can coexist with yeast cells within lesions of histoplasmosis. Following diagnosis, the dog was treated with itraconazole for 6 months and has improved.

Caudal vena caval thrombosis following treatment of deep digital sepsis.

A diagnosis of caudal vena caval thrombosis was made by ultrasonography of a Holstein cow presented for lethargy and poor milk production. Medical treatment was unsuccessful and the cow was euthanized. The diagnosis was confirmed at necropsy and Fusobacterium necrophorum was isolated from the thrombus. This paper discusses potential novel sources of caval thrombosis in this case.

Comparison of antemortem antimicrobial treatment regimens to antimicrobial susceptibility patterns of postmortem lung isolates from feedlot cattle with bronchopneumonia.

A retrospective study was performed to compare the treatment regimens in feedlot cattle that died with bovine respiratory disease (BRD) to the antimicrobial susceptibility patterns of the microorganisms isolated from lungs. Forty-three cattle submitted by the Willard Sparks Beef Research Center (WSBRC) to the Oklahoma Animal Disease Diagnostic Laboratory for postmortem examination during 2007 had bronchopneumonia (acute = 16, subacute = 5, or chronic = 22). Lungs from cattle were cultured aerobically (40 cattle) and for Mycoplasma spp. (34 cattle). Susceptibility panels were performed. At least 1 BRD pathogen (Mannheimia haemolytica, Pasteurella multocida, Histophilus somni, Mycoplasma bovis, or Arcanobacterium pyogenes) was isolated from 39 cattle, and 77% (30/39) had multiple organisms recovered. Mycoplasmal infections were common (25/34) and a major component of mixed infections (24/25). The majority (60%) of the M. haemolytica, P. multocida, and H. somni isolates were resistant to tetracycline. Most of the H. somni isolates (67%) were susceptible to tilmicosin (Ti), enrofloxacin (En), ceftiofur (Ce), and florfenicol, despite extensive treatment with Ti, En, and Ce (75% of isolates were from cattle that received each antimicrobial once). Most of the M. haemolytica (65%) and P. multocida (79%) isolates were susceptible to En and Ce, despite antemortem treatment of cattle with these antimicrobials. Hence, the current study reports a discrepancy between the antemortem treatment of clinical BRD and the susceptibility patterns of the bacteria isolated from lungs postmortem. Based on these findings, factors other than antimicrobial resistance are playing a role in the death of feedlot cattle with BRD.

Disseminated protothecosis diagnosed by evaluation of CSF in a dog.

A 5-year-old female spayed Shetland Sheepdog Mix dog was evaluated for a history of recent seizure activity, progressive hind limb ataxia, polyuria, and polydipsia and no history of gastrointestinal signs. Physical examination findings included conscious proprioceptive deficits, ataxia, and anterior uveitis along with a hypermature cataract in the right eye. Results of a CBC, serum biochemical profile, urinalysis, and computed tomography scan of the brain were unremarkable. Cerebrospinal fluid (CSF) analysis revealed marked eosinophilic pleocytosis and rare organisms consistent with Prototheca spp within neutrophils and macrophages. On postmortem histologic examination, mononuclear inflammation and numerous intralesional algal organisms, similar to those seen on the cytologic preparation of CSF, were found in the brain, eyes, kidneys, and heart. Abnormalities were not detected on gross and histologic examination of the gastrointestinal tract. Cultures of CSF and subdural/olfactory bulb, but not intestinal tract, yielded growth of Prototheca spp, and PCR analysis and DNA sequencing confirmed the organism as Prototheca zopfii genotype 2. We have reported a rare case of disseminated protothecosis that was diagnosed by evaluation of CSF in a dog presented with neurologic signs and no overt enteric disease. Protothecosis should be considered as a rare cause of seizures, even in the absence of obvious enteric signs, and should be included in the differential diagnosis of eosinophilic pleocytosis.

Genetic and phenotypic characterization of the bla(CMY) gene from Escherichia coli and Salmonella enterica isolated from food-producing animals, humans, the environment, and retail meat.

The bla(CMY-2) family of the ampC beta-lactamase genes confer broad-spectrum resistance to beta-lactam antimicrobials, including ceftriaxone and ceftiofur, as well as to beta-lactamase inhibitors, such as clavulanic acid. Organisms with the bla(CMY-2) phenotype have been recovered from the environment and from retail meat products, posing a potential public health risk. The objectives of this study were to sequence the bla(CMY-2) gene from Escherichia coli and Salmonella enterica from multiple sources that had a reduced susceptibility to ceftriaxone and to determine the effect of observed mutations in the bla(CMY-2) gene on the antimicrobial resistance phenotype (spectrum and minimum inhibitory concentration/susceptibility patterns) of the isolates. The bla(CMY-2) genes from 52 bacterial isolates were sequenced for this study. Sixty-two percent (32/52) were E. coli and 38% (20/52) were S. enterica. Of the 32 E. coli isolates, 30 were found to carry a beta-lactamase gene that was 100% homologous to bla(CMY-2). One of the E. coli isolates was found to contain a gene that was 90% homologous to bla(CMY-2). This isolate also had lower minimum inhibitory concentrations to tetracyclines, streptomycin, and the sulfonamide antimicrobials than are commonly expected for isolates containing the bla(CMY-2). Of the 20 genes obtained from Salmonella isolates, 8 (40%) were found to be homologous to bla(CMY-2), with no altered susceptibility phenotypes observed.

Pathogenesis of coxsackievirus-B5 acquired from intra-renal porcine islet cell xenografts in diabetic mice.

BACKGROUND: We previously demonstrated the ability of a human isolate of coxsackievirus-B5 (CVB5) to infect productively adult porcine islet cells (PICs) in vitro. PICs infected with CVB5 remain viable, and upon transplantation reversed diabetes in C56BL/6 mice for up to 5 days. METHODS: In the present work, we expanded this graft-to-host xenozoonosis model by examining the long-term functionality of CVB5-infected PIC xenografts in immunosuppressed mice. And, we characterized the pathogenesis of CVB5 infection in mice resulting from directional transmission of the virus from PIC xenografts to surrounding tissues in a mouse model for immunosuppressed human PIC xenograft recipients. RESULTS: Both acutely (12 h) and chronically (72 h) infected PIC xenografts functioned in vivo to reverse diabetes in mice. The efficacy of both infected and un-infected PICs was transient beyond 5 days post-inoculation and the long-term functionality of the grafts was compromised by host-to-graft rejection. CVB5-infected PIC xenografts transmitted infectious virus to immunosuppressed recipient mice resulting in extensive histopathologic changes. The virus replicated in the heart, liver, spleen, kidney, pancreas, brain and skeletal muscle in higher levels in severe-combined immunodeficient (SCID) mice that were directly inoculated with virus when compared to controls. In addition, infectious virus was recovered for up to 22 days after inoculation in SCID mice whereas it was only detected up to Day 4 PI in non-SCID mice. CONCLUSIONS: Immunosuppressed PIC xenograft recipients may be more susceptible to infection with CVB5 which could target the xenograft leading to disseminated infection in the host.

Parasite age-intensity relationships in red-spotted newts: does immune memory influence salamander disease dynamics?

Acquired immune memory in vertebrates influences transmission and persistence of infections, with consequences for parasite dynamics at both the individual and population levels. The potential impact of acquired immunity is of particular interest for salamanders, whose acquired immune systems are thought to be less effective than those of frogs and other tetrapods. One way to examine the importance of acquired immunity to parasite dynamics at the population level is by examining the relationship between host age and parasite infection intensity. Acquired immunity reduces infection rates in older animals, causing decreased parasite intensity in older age classes and leading to curvilinear age-intensity relationships for persistent parasites and convex age-intensity relationships for transient parasites. We used age-intensity relationships to look for the signature of acquired immunity for 12 parasite taxa of red-spotted newts (Notophthalmus viridescens), using data from a 2-year parasitological survey of six newt populations. We estimated ages from snout-vent length (SVL) based on the relationship between SVL and skeletochronologically-derived ages in a subset of newts. We found evidence of acquired immunity to two parasite taxa, bacterial pathogens and the protist Amphibiocystidium viridescens, whose convex age-intensity relationships could not be easily explained by alternative mechanisms. Our results suggest that the acquired immune response of newts is sufficient to influence the dynamics of at least some parasites.

Comparison of five culture methods for Salmonella isolation from swine fecal samples of known infection status.

The current study was conducted to evaluate 5 bacteriologic culture methods (methods 1, 2, 3, 4, and 5) for recovery of Salmonella enterica from swine feces, both for sensitivity of detection (ability to recover Salmonella from a positive sample) and for specificity (not to inadvertently identify an organism as a Salmonella species in a negative sample). Fifty-six negative samples and 46 positive samples were processed using each of the 5 methods, which differed primarily in the combinations of enrichment media used. All negative samples were negative for Salmonella when cultured by all 5 methods (100% specificity). Two of the methods (methods 1 and 4) resulted in the recovery of significantly less (P < 0.05) Salmonella when compared with the remaining 3 methods (methods 2, 3, and 5). No one method was successful in recovering Salmonella from all positive samples, although recovery with method 2 was statistically similar to the total number of positive samples analyzed (42 vs. 46 Salmonella-positive samples, P > 0.05). This study shows that culture methods differ significantly in their performance regarding the isolation of Salmonella from swine fecal samples.

Mortality in Pennsylvania captive elk (Cervus elaphus): 1998-2006.

The necropsy records of 65 elk ranging from 1 day to 13(1/2) years of age were reviewed to determine the primary cause of death of each animal. The animal carcasses were submitted from 22 separate Pennsylvania elk farms over an approximately 8-year period. The most common causes of mortality in animals in which the cause of death was determined were gastrointestinal parasitism (21 cases), pneumonia (7 cases), enterocolitis (5 cases), malnutrition (5 cases), and rumenitis/acidosis (5 cases). The cause of death was undetermined in 4 of the 65 elk. Nematode organisms (20 cases) were the primary parasites identified in cases of mortality owing to gastrointestinal parasitism. Arcanobacterium pyogenes (3 cases), Escherichia coli (3 cases), and Streptococcus sp. (2 cases) were the most commonly isolated bacteria from the lung in the cases of pneumonia. Fungal organisms, consistent with Aspergillus sp. were present histologically within the pulmonary lesions in 5 cases. Bacterial agents associated with enterocolitis included Clostridium perfringens (2 cases), E. coli (1 case), Salmonella Newport (1 case) and Salmonella Thompson (1 case). The majority (56.3%) of the death loss in elk of known ages occurred in animals 1 year of age or less, with 71.4% of the gastrointestinal parasitism, 60.0% of the malnutrition, 60.0% of the enterocolitis, and 57.1% of the pneumonia cases occurring in animals within this age range. In conclusion, gastrointestinal parasitism, pneumonia, malnutrition, enterocolitis and rumenitis/acidosis were considered the most common causes of death in captive elk in this study.

Antimicrobial-resistant enteric bacteria from dairy cattle.

A study was conducted to understand the descriptive and molecular epidemiology of antimicrobial-resistant gram-negative enteric bacteria in the feces of healthy lactating dairy cattle. Gram-negative enteric bacteria resistant to ampicillin, florfenicol, spectinomycin, and tetracycline were isolated from the feces of 35, 8, 5, and 42% of 213 lactating cattle on 74, 39, 9, 26, and 82% of 23 farms surveyed, respectively. Antimicrobial-resistant gram-negative bacteria accounted for 5 (florfenicol) to 14% (tetracycline) of total gram-negative enteric microflora. Nine bacterial species were isolated, of which Escherichia coli (87%) was the most predominant species. MICs showing reduced susceptibility to ampicillin, ceftiofur, chloramphenicol, florfenicol, spectinomycin, streptomycin, and tetracycline were observed in E. coli isolates. Isolates exhibited resistance to ampicillin (48%), ceftiofur (11%), chloramphenicol (20%), florfenicol (78%), spectinomycin (18%), and tetracycline (93%). Multidrug resistance (> or =3 to 6 antimicrobials) was seen in 40% of E. coli isolates from healthy lactating cattle. Of 113 tetracycline-resistant E. coli isolates, tet(B) was the predominant resistance determinant and was detected in 93% of isolates, while the remaining 7% isolates carried the tet(A) determinant. DNA-DNA hybridization assays revealed that tet determinants were located on the chromosome. Pulsed-field gel electrophoresis revealed that tetracycline-resistant E. coli isolates (n = 99 isolates) belonged to 60 subtypes, which is suggestive of a highly diverse population of tetracycline-resistant organisms. On most occasions, E. coli subtypes, although shared between cows within the herd, were confined mostly to a dairy herd. The findings of this study suggest that commensal enteric E. coli from healthy lactating cattle can be an important reservoir for tetracycline and perhaps other antimicrobial resistance determinants.

Dissemination of Salmonella enterica subsp. enterica Serovar Typhimurium var. Copenhagen clonal types through a contract heifer-raising operation.

Salmonella enterica subsp. enterica serovar Typhimurium var. Copenhagen isolates from a heifer-raising operation and from 11 dairy herds that had their calves contracted to the heifer-raising operation were examined for their phenotypic and genotypic characteristics. Results of the study showed that the heifer-raising operation could serve as a clearinghouse for Salmonella serovar Typhimurium var. Copenhagen and perhaps other Salmonella serotypes.

A retrospective study of mortality in Pennsylvania captive white-tailed deer (Odocoileus virginianus): 20000--2003.

The postmortem records of 160 white-tailed deer (Odocoileus virginianus) submitted for necropsy examination from 59 separate Pennsylvania captive deer farms over a 3.5-year period were reviewed to determine the primary cause of death of each animal. The most common causes of death were bronchopneumonia (39 cases), enterocolitis (30 cases), malnutrition (13 cases), and trauma (11 cases). Other causes of mortality included severe gastrointestinal parasitism (6 cases), cellulitis with septicemia (5 cases), degenerative myopathy (4 cases), ruminal acidosis (4 cases), and nephritis (4 cases). The cause of death was undetermined in 13 of the 160 animals. Arcanobacterium pyogenes (19 cases), Fusobacterium necrophorum (10 cases), Escherichia coli (7 cases), and Mannheimia haemolytica (4 cases) were the most commonly isolated bacteria from the pneumonic lungs. Bacterial agents associated with enterocolitis included Clostridium perfringens (15 cases), E. coli (12 cases), and Mycobacterium avium subsp. paratuberculosis (2 cases). The majority (52.2%) of the death loss in white-tailed deer of known ages occurred in animals 1 year of age or less, with 46.2% of the bronchopneumonia cases and 50.0% of the enterocolitis cases occurring during this time period. Cases of degenerative myopathy, myocardial degeneration, hepatic necrosis, meningoencephalitis, peritonitis, and urolithiasis considered severe enough to be the primary cause of death appeared early in life, affecting deer 6 months of age or less in all cases. In conclusion, bronchopneumonia, enterocolitis, malnutrition, and trauma were considered the most common causes of death in confined white-tailed deer in this study.

Prevalent human coxsackie B-5 virus infects porcine islet cells primarily using the coxsackie-adenovirus receptor.

BACKGROUND: We have previously demonstrated that transplanting porcine encephalomyocarditis virus (EMCV)-infected porcine islet cells (PICs) results in transmission of the virus to recipient mice, which is manifested by acute fatal infection within 5 to 8 days. Here, we determined PIC susceptibility to a related and highly prevalent human picornavirus, coxsackie B-5 virus (CVB-5). METHODS: PICs were inoculated with CVB-5 in vitro for up to 96 hours and infectivity, level of virus replication, and cellular function determined. Subsequently, monoclonal and polyclonal antibody blocking experiments were used to investigate the receptor CVB-5 uses to enter PICs, and the ability of CVB-5-infected islets to reverse diabetes analyzed in mice. RESULTS: Adult pig islets inoculated with CVB-5 in vitro showed a typical picornaviral replication cycle with a 2-h lag phase followed by a 4-h exponential phase during which the virus titer increased by 4 logs. However, CVB-5 was less cytolytic to PICs than EMCV, resulting in a persistent productive infection lasting for up to 96 h, with minimal evidence of cell lysis. Double immunostaining confirmed the presence of CVB-5 antigens in insulin-producing islets. Infection of PICs in the presence of antibodies against human coxsackie-adenovirus receptor (CAR) resulted in near complete blockage in production of infectious virus particles whereas blocking with anti-porcine decay-accelerating factor (DAF, also called CD55) or anti-porcine membrane cofactor protein (MCP, also called CD46) only slightly decreased the number of infectious CVB-5 particles produced. Immunofluoresence staining showed CAR and MCP expression on the islet surface, but not DAF. Transplanting CVB-5-infected PICs into diabetic C57BL/6 mice resulted in reversal of diabetes. CONCLUSION: Although PICs are susceptible to human CVB-5, the infection does not appear to affect xenograft function in vitro or in vivo in the short term.

Comparison of management practices between Ohio, USA dairy farms participating in whole-herd Johne's disease testing programs and those not participating.

The purpose of our mail survey was to compare the adoption of management practices recommended for Johne's disease (JD) control between herds involved in whole-herd testing programs versus those that do not routinely test the entire herd for JD. A questionnaire consisted of 38 closed-ended questions that inquired about: general herd characteristics; management practices related to JD control; changes that occurred within the last 5 years regarding management practices recommended for the control of JD; producer knowledge of JD; the perceived infection status of the herd by the producer; and herd JD-testing history. The questionnaire was mailed to 810 Ohio dairy producers in September 2002; 266 questionnaires were returned (32.8% response). We used univariable logistic-regression models to assess the relationship between whole-herd testing status (TESTING versus NON-TESTING) and each management practice, each change in management practice and producer knowledge about JD. Because it is conceivable that only producers who believe their herds to be infected would be motivated to adopt the management practices recommended for control of JD, the comparisons were repeated with models that controlled for producer-perceived infection status. Of the 20 management practices recommended for JD control that we evaluated, 7 differed between TESTING and NON-TESTING herds. Additionally, TESTING herds more-frequently reported adopting changes within the past 5 years relative to NON-TESTING herds with respect to 7 of 9 management practices evaluated. Producers with TESTING herds also reported greater familiarity with JD than those with NON-TESTING herds.

Prevalence of fecal shedding of Salmonella spp in dairy herds.

OBJECTIVE: To estimate prevalence of Salmonella spp in Ohio dairy farms and to identify potential risk factors for fecal shedding of salmonellae. DESIGN: Cross-sectional study. SAMPLE POPULATION: 105 Ohio dairy farms. PROCEDURE: Individual fecal samples from all mature cows in study herds were tested for Salmonella spp by use of standard bacteriologic culture procedures. Herds were identified as infected if at least 1 cow was shedding Salmonella spp. Information regarding herd characteristics, management practices, and health history were collected. Potential risk factors for herd-level Salmonella infection were identified. RESULTS: In 31% of the study herds (95% confidence interval, 22 to 40%), at least 1 cow was shedding Salmonella spp. Six percent of 7,776 fecal samples contained Salmonella organisms; prevalence within infected herds ranged from < 1 to 97%. Herd size, use of free stalls for lactating and nonlactating cows, and use of straw bedding in nonlactating cows were significantly associated with fecal shedding of Salmonella spp, as determined by use of univariate analysis. By use of multivariate analysis, large herds were more likely to be infected than smaller herds; however, no other factors were associated with Salmonella infection after adjustment for herd size. CONCLUSIONS AND CLINICAL RELEVANCE: Subclinical shedding of Salmonella spp is common in Ohio dairy herds, although we could not identify specific interventions that may influence the prevalence of Salmonella spp on dairy farms. It appears that large herd size and intensive management may provide an environment conducive to Salmonella shedding and chronic dairy herd infection.

Persistent fecal Salmonella shedding in five dairy herds.

OBJECTIVE: To monitor patterns of Salmonella fecal shedding in naturally infected dairy herds, determine the association between fecal shedding and individual animal production measures, and evaluate potential risk factors for shedding of Salmonella organisms among cattle in dairy herds. DESIGN: Longitudinal study. SAMPLE POPULATION: 5 Ohio dairy herds. PROCEDURE: For 3 herds, fecal samples were collected from all mature cows and unweaned calves 7 times during an 18-month period. For the remaining 2 herds, fecal samples were collected from 50 lactating cows 6 times during a 12-month period. Individual animal production records for 3 herds were used to examine associations between individual fecal Salmonella shedding status and 305-day mature-equivalent milk production, somatic cell count, milk fat content, and milk protein content. Multivariable logistic regression was used to test for associations between fecal shedding status and breed, lactation status, lactation number, and duration of lactation. RESULTS: None of the adult animals had clinical signs of salmonellosis, but prevalence of fecal Salmonella shedding at individual collection times ranged from 0 to 99% for cows and from 0 to 67% for unweaned calves. Mature cows were more likely to be shedding Salmonella organisms than were unweaned calves. Within herds, lactation status and duration of lactation for individual animals were associated with Salmonella shedding status. Salmonella fecal shedding status was not associated with individual cow production measures. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that subclinical fecal Salmonella shedding can persist in dairy herds for up to 18 months with no measurable effects on health or production of individual cows.