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The objectives of this study were to investigate the effects of group housing (three calves per group) on bovine respiratory disease (BRD), diarrhea and antimicrobial resistance (AMR) to fecal commensal Escherichia coli (EC) and enterococci/streptococci (ES). Our study comprised two arms, one experimental and one observational. In the experimental arm, preweaned calves on a California dairy were randomized to either individual (IND; n = 21) or group (GRP; n = 21) housing, using a modified California-style wooden hutch. The study period lasted from birth to 56 days of age, during which calves were health scored daily. Cumulative incidence and hazard ratios were estimated for disease. Antimicrobial resistance outcomes were assessed using a prospective cohort design; feces were collected from each calf three times per week and EC and ES were evaluated for AMR using the broth microdilution method against a panel of 19 antimicrobial drugs (AMD). Analysis of treatment records was used to select calves that had been exposed (EXP) to an AMD-treated calf. In GRP, exposure occurred when a calf was a hutchmate with an AMD-treated calf. In IND, exposure occurred when a calf was a neighbor with an AMD-treated calf (TRT). Age-matched unexposed calves (UNEXP) were then selected for comparison. Proportions of AMR in fecal commensals among EXP, UNEXP, and TRT calves were compared between GRP and IND. Accelerated failure time survival regression models were specified to compare differences in minimum inhibitory concentration (MIC) of fecal commensals between EXP and UNEXP calves within each of GRP and IND calves separately. Group calves had a BRD hazard 1.94 times greater that of IND calves (p = 0.03), using BRD treatment records as the outcome. For AMR in EC isolates, higher resistance to enrofloxacin was detected in enrofloxacin-EXP GRP isolates compared with enrofloxacin-EXP IND isolates, and UNEXP GRP calves had lower resistance to ceftiofur compared with enrofloxacin-EXP and enrofloxacin-TRT calves. A significant housing-by-time interaction was detected for EC ceftiofur MIC in EXP GRP calves at 4–14 days post exposure to enrofloxacin (MIC EXP-UNEXP: µg/mL (95% CI): 10.62 (1.17, 20.07)), compared with UNEXP calves. The findings of this study show an increase in BRD hazard for group-housed calves and an increase in ceftiofur resistance in group-housed calves exposed to an enrofloxacin-treated calf.

Pseudogymnoascus destructans is the etiological agent of white-nose syndrome (WNS), a fungal skin infection of hibernating bats. Pathophysiology of the disease involves disruption of bat metabolism and hibernation patterns, which subsequently causes premature emergence and mortality. However, information on the mechanism(s) and virulence factors of P. destructans infection is minimally known. Typically, fungal adherence to host cells and extracellular matrix (ECM) is the critical first step of the infection. It allows pathogenic fungi to establish colonization and provides an entry for invasion in host tissues. In this study, we characterized P. destructans conidial adherence to laminin and fibronectin. We found that P. destructans conidia adhered to laminin and fibronectin in a dose-dependent, time-dependent and saturable manner. We also observed changes in the gene expression of secreted proteases, in response to ECM exposure. However, the interaction between fungal conidia and ECM was not specific, nor was it facilitated by enzymatic activity of secreted proteases. We therefore further investigated other P. destructans proteins that recognized ECM and found glyceraldehyde-3-phosphate dehydrogenase and elongation factor 1-alpha among the candidate proteins. Our results demonstrate that P. destructans may use conidial surface proteins to recognize laminin and fibronectin and facilitate conidial adhesion to ECM. In addition, other non-specific interactions may contribute to the conidial adherence to ECM. However, the ECM binding protein candidates identified in this study highlight additional potential fungal virulence factors worth investigating in the P. destructans mechanism of infection in future studies.

OBJECTIVE To determine the ability of 0.2% polyhexamethylene biguanide (PHMB)-impregnated gauze to inhibit the growth of bacteria isolated from equine infected sites. STUDY DESIGN In vitro study. METHODS Nine bacterial isolates were obtained from cultures submitted from equine patients presenting with penetrating injuries of the hoof (n = 4), septic osteitis (n = 1), synovial sepsis (n = 1), wounds (n = 2), and incisional infection following laparotomy (n = 1). Two standardized strains were also included. A standard inoculum of each isolate was placed on 12 Muller-Hinton agar plates. Squares (2.5 cm × 2.5 cm) of 0.2% PHMB-impregnated (n = 6) and nonimpregnated control gauze (n = 6) were placed on inoculated agar plates. Bacterial growth under each gauze square was assessed after a 24-h incubation period and areas of inhibition were measured to a standardized scale, using image-processing software. Mean ± SD growth inhibition (%) using 0.2% PHMB-impregnated gauze was compared to the nonimpregnated gauze for each isolate using Student's t test (p

Infection with Veronaea botryosa can result in rare cutaneous or disseminated, granulomatous to pyogranulomatous phaeohyphomycosis in humans, although disease due to the fungus has also been reported in non-mammalian vertebrates. This report documents disease due to V. botryosa in captive, juvenile to subadult or young adult white sturgeon (Acipenser transmontanus Richardson) from California USA and complements a previous report of the disease in captive Siberian sturgeon (Acipenser baerii) from Florida USA. Pathological examinations revealed granulomatous to pyogranulomatous inflammation of multiple organs. Isolates of the fungal agent were phenotypically consistent with V. botryosa, and molecular analyses of the D1/D2 region of the fungal 28S rRNA gene and the internal transcribed spacer (ITS) region located between the fungal 18S and 28S rRNA genes confirmed the aetiologic agent as V. botryosa. The disease in captive sturgeon results in a considerable economic encumbrance to the producer due to the loss of the cumulative financial resources invested in the production of older subadult to young adult sturgeon.

OBJECTIVE To determine the pharmacokinetics of danofloxacin following IM administration of a single dose (10 mg/kg) in koi (Cyprinus carpio). ANIMALS 69 healthy adult koi housed in a 980-L flow-through-system tank. PROCEDURES 3 fish were kept as untreated controls, and the remaining 66 fish were assigned to 11 treatment groups with 6 fish/group. Fish in the treatment groups were given a single dose of danofloxacin (10 mg/kg) IM in the left epaxial musculature. Fifteen, 30, and 45 minutes and 1, 4, 12, 24, 72, 96, 120, and 144 hours after administration of danofloxacin, fish in each treatment group were euthanized, and blood samples and samples of liver, spleen, gill, anterior kidney, posterior kidney, skin and muscle, and scales were collected. Plasma and tissue drug concentrations were determined by liquid chromatography–tandem mass spectrometry, and noncompartmental pharmacokinetic analyses were performed. Tissues from the untreated control fish and fish euthanized 144 hours after danofloxacin administration were examined histologically. RESULTS Maximum plasma concentration (mean, 8,315.7 ng/mL) was reached approximately 45 minutes after danofloxacin administration; plasma elimination half-life was 15 hours. Danofloxacin was detected in all examined tissues from all 6 fish euthanized 15 minutes after drug administration and was detected in some tissues from 3 of the 6 fish euthanized 144 hours after drug administration. For all tissues, results of histologic examination were unremarkable. CONCLUSIONS AND CLINICAL RELEVANCE IM administration of a single dose (10 mg/kg) of danofloxacin in koi resulted in rapid absorption, with maximum plasma concentration reached approximately 45 minutes after drug administration; the drug could still be detected in some tissues 144 hours after administration.

Objectives The aim of this study was to evaluate the susceptibility pattern of the incisional site Staphylococcus spp. to three commonly used perioperative antimicrobials in dogs. Materials and Methods A cohort of client-owned dogs was included in this prospective observational study. After induction of general anaesthesia and hair clipping, culture samples were collected from the incisional site of healthy dogs before orthopaedic surgery. The isolated colonies were identified. The susceptibility pattern of staphylococcal isolates was determined using Clinical and Laboratory Standard Institute standards. The staphylococcal susceptibilities to amoxicillin/clavulanic acid and cefpodoxime were compared with cefazolin using two-sided equality of proportions test. Results A total of 391 bacterial isolates were identified in 115 dogs. Bacillus spp. (n = 200) and Staphylococcus spp. (n = 95) were the most common bacteria isolated. Coagulase- positive and -negative staphylococci were isolated in 44 and 38% of dogs respectively. Cefazolin, amoxicillin/clavulanic acid and cefpodoxime staphylococcal susceptibilities were 68, 77 and 76% respectively. There were no significant differences in Staphylococcus spp. susceptibility between amoxicillin/clavulanic acid or cefpodoxime and cefazolin (p = 0.19, p = 0.25). Thirty-six per cent of coagulase-negative staphylococci and 6% of coagulase-positive staphylococci isolated were methicillin resistant. Conclusion Cefazolin, amoxicillin/clavulanic acid and cefpodoxime had similar susceptibilities for staphylococcal isolates. Routine use of amoxicillin/clavulanic acid or cefpodoxime in replace of cefazolin should be practiced with caution.

Outbreaks of an infectious disease affecting cultured white sturgeon (Acipenser transmontanus) were investigated. Clinical signs included erratic swimming, arching of the back and mortality. Necropsy findings included poorly demarcated yellow to dark-red and friable lesions in the epaxial muscle, ulcerative skin lesions and haemorrhages in the swim bladder and coelomic wall. Histological evaluation revealed areas of necrotizing and heterophilic myositis with aggregates of bacterial cocci. The lumen of blood vessels in the dermis, under ulcerated areas, and in the posterior kidney, was occluded by fibrin thrombi. Aggregates of Gram-positive cocci were observed in the muscle lesions and within the fibrin thrombi in the dermis and kidney. Genetically homogeneous Streptococcus iniae strains were recovered from affected fish from different outbreaks. The isolates shared high degree of similarity at gene locus (gyrB) with previously characterized S. iniae from cultured fish in California, confirming the emergence of this particular strain of S. iniae in US aquaculture.

Background Intravenous catheter (IVC) use in hospitalized ruminants is a common procedure. Limited information is available describing complications associated with IVCs. Hypotheses Prevalence of IVC infections in hospitalized ruminants is >50%. Intravenous catheters maintained for >5 days are more likely to be infected than those maintained for 4 days had a higher likelihood of being infected than those maintained for 4 days to reduce IVC infection.

CASE DESCRIPTION An 8-year-old sexually intact female eclectus parrot (Eclectus roratus) with a 4-day history of hyporexia and lethargy and a 1-day history of tenesmus was examined. CLINICAL FINDINGS Severe leukocytosis characterized by severe heterophilia and moderate monocytosis was present. Marked dilation of the proventriculus and ventriculus and ascites were identified by means of radiography, coelomic ultrasonography, and contrast-enhanced CT, with no clinically relevant motility noted on ultrasonography. Results of coelomic fluid analysis were consistent with pyogranulomatous effusion. Endoscopy of the upper gastrointestinal tract following proventricular and ventricular lavage showed a thick caseous plaque occupying 30% of the caudal proventricular mucosa. Abundant yeast organisms were evident during cytologic examination of a proventricular and ventricular wash sample, and fecal culture yielded Candida glabrata. TREATMENT AND OUTCOME The bird was treated with SC fluids, assisted feedings, nystatin, fluconazole, amoxicillin–clavulanic acid, enrofloxacin, gastroprotectants, maropitant, and analgesics and slowly improved during hospitalization. A marked decrease in proventricular dilation was evident on serial radiographs obtained over a 12-month period. One year after diagnosis, the bird was presented with a 1-week history of hyporexia and lethargy, and fecal culture grew C glabrata. Antifungal treatment was resumed for 3 months. The bird had no clinical signs of infection 16 months after this recurrence, and subsequent fecal cultures were negative for fungal growth. CLINICAL RELEVANCE Findings illustrate the importance of upper gastrointestinal endoscopy in diagnosing proventricular and ventricular dilation in birds and emphasize the need for long-term antifungal treatment and monitoring in birds with fungal infections.

Enterococci have been increasing in prevalence in foal sepsis over the past three decades. There are no published studies in the peer-reviewed literature documenting common sites of infection, antimicrobial susceptibility, or outcome specifically associated with enterococcal infections in foals. Our objectives were to evaluate the sites of origin, antimicrobial susceptibility, and survival outcome to discharge in foals with enterococcal infections compared with foals with sepsis of another bacterial etiology. Seventy-five foals 0-30 days of age with cultures positive for Enterococcus and 170 control foals 0-30 days of age with cultures positive for other bacteria were included. Enterococcus was 2.67 times (95% confidence interval [CI], 1.49-4.80; P = 0.0012) more likely to be isolated from the lower urogenital tract of foals than were other bacteria. Enterococci were less likely to be isolated from blood cultures than other bacteria, with an odds ratio (OR) of 0.17 (95% CI 0.09-0.35; P 0.0001). For Enterococcus isolates, 48% (n = 29/61) had a multiple antimicrobial resistance (MAR) index of ≥30% and 46% (n = 28/61) had a multiple drug resistance (MDR) index of ≥30%. Foals with enterococcal infections were less likely to survive to discharge (49.9% vs. 63.5%; P = 0.03). Enterococcus is commonly isolated from the lower urogenital tract of foals, is often multidrug resistant, and foals with enterococcal infections were less likely to survive. Multidrug resistance is common among enterococcal isolates, and therefore antimicrobial susceptibility testing of cultured isolates is warranted.

Southern sea otters (SSO: Enhydra lutris nereis) are a federally-listed threatened subspecies found almost exclusively in California, USA. Despite their zoonotic potential and lack of host specificity, K. pneumoniae and Klebsiella spp. have largely unknown epizootiology in SSOs. Klebsiella pneumoniae is occasionally isolated at necropsy, but not from live SSOs. Hypermucoviscous (HMV) K. pneumoniae strains are confirmed pathogens of Pacific Basin pinnipeds, but have not been previously isolated from SSOs. We characterized the virulence profiles of K. pneumoniae isolates from necropsied SSOs, evaluated killing of marine mammal K. pneumoniae following in vitro exposure to California sea lion (CSL: Zalophanus californianus) whole blood and serum, and characterized lesion patterns associated with Klebsiella spp. infection in SSOs. Four of 15 SSO K. pneumoniae isolates were HMV and all were recovered from SSOs that stranded during 2005. Many K. pneumoniae infections were associated with moderate to severe pathology as a cause of death or sequela. All HMV infections were assessed as a primary cause of death or as a direct result of the primary cause of death. Klebsiella-infected SSOs exhibited bronchopneumonia, tracheobronchitis and/or pleuritis, enteritis, Profilicollis sp. acanthocephalan peritonitis, septic peritonitis, and septicemia. All SSO HMV isolates were capsular type K2, the serotype most associated with HMV infections in CSLs. Multiplex PCR revealed two distinct virulence gene profiles within HMV isolates and two within non-HMV isolates. In vitro experiments investigating CSL whole blood and serum killing of K. pneumoniae suggest that HMV isolates are more resistant to serum killing than non-HMV isolates.

The objectives of this study were to evaluate the prevalence of extended spectrum β-lactamase (ESBL) genes, AmpC-type β-lactamase (ACBL) genes, and plasmid mediated quinolone resistance (PMQR) genes in Salmonella isolated at a Veterinary Medical Teaching Hospital microbiology laboratory, examine trends in presence of these resistance genes, and to explore the correlation between phenotypic resistance and presence of specific genes. The presence of ESBL, ACBL, and PMQR genes were detected using a single, novel multiplex qPCR. Only the genes blaCMY–2 and blaTEM were detected in the 110 Salmonella isolates tested. PMQR genes were not detected in isolates screened. Of 94 third-generation cephalosporin resistant isolates, representing eight serotypes, 48% (n = 45) were positive for blaCMY–2 only and 50% (n = 47) were simultaneously positive for blaCMY–2 and blaTEM. Two third-generation cephalosporin resistant isolates were tested negative for all β-lactamase genes in our qPCR assay and likely house ESBL genes not screened for by our qPCR assay. A logistic regression model revealed that for serotype Dublin isolates (n = 38) the odds ratio for testing positive for blaTEM when compared to all other serotypes was 51.6 (95% CI: 4.01–664.03, p = 0.0029). For serotype Typhimurium (n = 9) the odds ratio for testing positive for blaTEM when compared to all other serotypes was 43.3 (95% CI: 1.76–1000, p = 0.0216). Overall, our results suggest that the prevalence of resistance to cephalosporins and fluoroquinolones due to ESBLs, ACBLs, and PMQR genes present in bovine nontyphoidal Salmonella enterica isolates has remained relatively constant in the isolates screened over a 14-year period.

Most of the world’s sea otters reside in Alaska, but there has never been an assessment of long-term mortality patterns for this keystone predator. We examined data collected from 780 northern sea otter (Enhydra lutris kenyoni)carcasses recovered in Alaska from 2002 to 2012 to evaluate the causes of mortality and risk factors associated with death. A smaller group (n= 144, 18%) of fresh non-frozen carcasses were included in a more detailed mortality analysis. Forty-four% of the fresh dead otters were determined to have died from infectious endocarditis, meningoencephalitis, and/or septicemia due to systemic streptococcosis (“Strepsyndrome”).Streptococcus lutetiensis, a member of theStreptococcus bovis/equinus(SB/E) group was most commonly isolated, although other members of the SB/E group were identified. There were fewer cases whereS. phocaeand other streptococci were isolated. A regression analysis revealed age and location risk factors forStrepsyndrome. Subadults were the highest risk age group, and otters recovered from the Kachemak Bay region were 3.6 times (95% CI: 2.2–5.9) more likely to die fromStrepsyndrome than otters recovered elsewhere. Diagnosis of thisStrepsyndrome had not been reported in other marine mammals in Alaska. Sporadic cases of septicemia and infectious endocarditis due toS. lutetiensishave been seen in the southern sea otter population in California. Other causes of death for the fresh otters included neurologic diseases (10%), trauma (8%), nutritional diseases (7%), cardiovascular diseases (7%), gastrointestinal diseases /parasites (6%), undetermined (5%), septicemia (3%), and neoplasia (3%). All other causes of death (oiling, hepatobiliary, fungal, marine biotoxins, pulmonary) were at or below 1%. Twenty percent of fresh animals were positive for phocine distemper virus (PDV) by polymerase chain reaction (PCR, 11/55), 18% seropositive for PDV (7/38), and paramyxovirus-like particles were demonstrated by electron microscopy within inclusion bodies from one animal. Low concentrations of the harmful algal bloom toxins domoic acid and saxitoxin were also detected in 26% and 22% of fresh animals, respectively. Protozoal disease was rare. These patterns of disease differ from sea otters in other regions and possible reasons are discussed.

Background Understanding the effects of herd management practices on the prevalence of multidrug-resistant pathogenic Salmonella and commensals Enterococcus spp. and Escherichia coli in dairy cattle is key in reducing antibacterial resistant infections in humans originating from food animals. Our objective was to explore the herd and cow level features associated with the multi-drug resistant, and resistance phenotypes shared between Salmonella, E. coli and Enterococcus spp. using machine learning algorithms. Methods Randomly collected fecal samples from cull dairy cows from six dairy farms in central California were tested for multi-drug resistance phenotypes of Salmonella, E. coli and Enterococcus spp. Using data on herd management practices collected from a questionnaire, we built three machine learning algorithms (decision tree classifier, random forest, and gradient boosting decision trees) to predict the cows shedding multidrug-resistant Salmonella and commensal bacteria. Results The decision tree classifier identified rolling herd average milk production as an important feature for predicting fecal shedding of multi-drug resistance in Salmonella or commensal bacteria. The number of culled animals, monthly culling frequency and percentage, herd size, and proportion of Holstein cows in the herd were found to be influential herd characteristics predicting fecal shedding of multidrug-resistant phenotypes based on random forest models for Salmonella and commensal bacteria. Gradient boosting models showed that higher culling frequency and monthly culling percentages were associated with fecal shedding of multidrug resistant Salmonella or commensal bacteria. In contrast, an overall increase in the number of culled animals on a culling day showed a negative trend with classifying a cow as shedding multidrug-resistant bacteria. Increasing rolling herd average milk production and spring season were positively associated with fecal shedding of multidrug- resistant Salmonella. Only six individual cows were detected sharing tetracycline resistance phenotypes between Salmonella and either of the commensal bacteria. Discussion Percent culled and culling rate reflect the increase in culling over time adjusting for herd size and were associated with shedding multidrug resistant bacteria. In contrast, number culled was negatively associated with shedding multidrug resistant bacteria which may reflect producer decisions to prioritize the culling of otherwise healthy but low-producing cows based on milk or beef prices (with respect to dairy beef), amongst other factors. Using a data-driven suite of machine learning algorithms we identified generalizable and distant associations between antimicrobial resistance in Salmonella and fecal commensal bacteria, that can help develop a producer-friendly and data-informed risk assessment tool to reduce shedding of multidrug-resistant bacteria in cull dairy cows.

To determine the pharmacokinetics of danofloxacin following IM administration of a single dose (10 mg/kg) in koi (69 healthy adult koi housed in a 980-L flow-through-system tank.3 fish were kept as untreated controls, and the remaining 66 fish were assigned to 11 treatment groups with 6 fish/group. Fish in the treatment groups were given a single dose of danofloxacin (10 mg/kg) IM in the left epaxial musculature. Fifteen, 30, and 45 minutes and 1, 4, 12, 24, 72, 96, 120, and 144 hours after administration of danofloxacin, fish in each treatment group were euthanized, and blood samples and samples of liver, spleen, gill, anterior kidney, posterior kidney, skin and muscle, and scales were collected. Plasma and tissue drug concentrations were determined by liquid chromatography-tandem mass spectrometry, and noncompartmental pharmacokinetic analyses were performed. Tissues from the untreated control fish and fish euthanized 144 hours after danofloxacin administration were examined histologically.Maximum plasma concentration (mean, 8,315.7 ng/mL) was reached approximately 45 minutes after danofloxacin administration; plasma elimination half-life was 15 hours. Danofloxacin was detected in all examined tissues from all 6 fish euthanized 15 minutes after drug administration and was detected in some tissues from 3 of the 6 fish euthanized 144 hours after drug administration. For all tissues, results of histologic examination were unremarkable.IM administration of a single dose (10 mg/kg) of danofloxacin in koi resulted in rapid absorption, with maximum plasma concentration reached approximately 45 minutes after drug administration; the drug could still be detected in some tissues 144 hours after administration.

Polymerase chain reaction (PCR)–based detection assays for Streptococcus equi subspecies equi often overestimate the prevalence of samples containing viable organisms. The objective of this study was to determine if viability could be determined using genome quantitation and detection of messenger RNA (mRNA) transcripts for the SeM gene of S. equi in pre- and post-cultured samples. Nasal secretions collected from 42 horses with suspected strangles were tested by culture and by quantitative PCR (qPCR) before and 24 hours after a culture step. Viable S. equi was determined based on the detection of S. equi via culture, the detection of mRNA transcripts for the SeM gene of S. equi by qPCR, and/or an increase in absolute number of SeM target genes of S. equi between pre- and post-cultured samples. Viability was determined in 28/42 samples based on isolation of S. equi (11 samples), the presence of mRNA transcripts for the SeM gene of S. equi (25), and/or an increase in absolute quantitation of the SeM gene of S. equi between pre- and post-culture (17). The overall agreement between culture alone and the three criteria to determine viability was 59%. The overall agreement for the detection of mRNA transcripts and increase in absolute target genes was 88% and 74%, respectively. The combination of mRNA transcripts and increase in absolute target genes was able to determine the viability status in all 42 samples. In the absence of a culture-positive result for S. equi, the determination of viability can be achieved by using molecular strategies applied to samples undergoing a 24-hour culture step.

Interactions with livestock in public settings such as county and state fairs can expose people and other livestock to faecal material capable of spreading zoonotic enteric pathogens. The goal of this study was to understand these risks by screening livestock faeces (n = 245) and livestock bedding (n = 155) for common zoonotic pathogens (Giardia, Cryptosporidium, Salmonella and Campylobacter spp.) and by measuring faecal indicator, Escherichia coli, concentrations in drinking water (n = 153), feed containers (n = 124) and bedding material (n = 157) in four livestock species (cattle, sheep, goats and swine) from county fairs in California, USA. Results indicated that sheep were most likely to have pathogens detected in faeces and that Giardia was the most frequently detected pathogen in both faeces (11%) and bedding (21%) across all livestock species. Additionally, increasing the number of animals in a holding pen at fairs, increasing the stocking density of animals in transport trailers to fairs, and having access to water in transport trailers significantly increased the odds of detecting pathogens in livestock faeces of any animal species. Observing solid material in water, stale feed and soiled bedding was associated with detecting higher E. coli concentrations. These findings provide evidence of faecal pathogens present at county fairs and suggest that site observations can aid in assessing levels of faecal exposure. The findings also indicate that the use of biosecurity measures such as (a) routine changing of livestock drinking water, feed and bedding, (b) not overstocking animals in holding pens and trailers and (c) keeping species in separate holding areas may reduce the risk of humans and livestock being exposed to faecal pathogens.

The objectives of this study were to evaluate the prevalence of extended spectrum β-lactamase (ESBL) genes, AmpC-type β-lactamase (ACBL) genes, and plasmid mediated quinolone resistance (PMQR) genes in Salmonella isolated at a Veterinary Medical Teaching Hospital microbiology laboratory, examine trends in presence of these resistance genes, and to explore the correlation between phenotypic resistance and presence of specific genes. The presence of ESBL, ACBL, and PMQR genes were detected using a single, novel multiplex qPCR. Only the genes blaCMY–2 and blaTEM were detected in the 110 Salmonella isolates tested. PMQR genes were not detected in isolates screened. Of 94 third-generation cephalosporin resistant isolates, representing eight serotypes, 48% (n = 45) were positive for blaCMY–2 only and 50% (n = 47) were simultaneously positive for blaCMY–2 and blaTEM. Two third-generation cephalosporin resistant isolates were tested negative for all β-lactamase genes in our qPCR assay and likely house ESBL genes not screened for by our qPCR assay. A logistic regression model revealed that for serotype Dublin isolates (n = 38) the odds ratio for testing positive for blaTEM when compared to all other serotypes was 51.6 (95% CI: 4.01–664.03, p = 0.0029). For serotype Typhimurium (n = 9) the odds ratio for testing positive for blaTEM when compared to all other serotypes was 43.3 (95% CI: 1.76–1000, p = 0.0216). Overall, our results suggest that the prevalence of resistance to cephalosporins and fluoroquinolones due to ESBLs, ACBLs, and PMQR genes present in bovine nontyphoidal Salmonella enterica isolates has remained relatively constant in the isolates screened over a 14-year period.

Background Mycobacterial infections in cats are challenging to treat and incompletely described. Hypothesis/objectives To describe the features of mycobacterial infections in cats from northern California. Animals Nineteen cats, all with nontuberculous mycobacterial (NTM) infections; 4 with Mycobacterium avium infection, 15 with rapid‐growing mycobacterial (RGM) infection. Methods Retrospective study. Cases with positive mycobacterial culture, species identification, and susceptibility testing were included. Descriptive statistics were used. Fisher's exact test and Mann‐Whitney U test were used for comparisons between M avium and RGM infections (P ≤ .05). Results Rapid‐growing mycobacterial cases included Mycobacterium smegmatis (9), Mycobacterium fortuitum (4), Mycobacterium abscessus (1), and Mycobacterium thermoresistibile (1). Mycobacterium avium infections were more likely than RGM infections to be disseminated (3/4 vs 0/15; P = .004). Disease of the skin/subcutis (15/15 vs 0/4; P

Background The presence of Escherichia coli O157:H7 (E. coli O157:H7) super-shedding cattle in feedlots has the potential to increase the overall number (bio-burden) of E. coli O157:H7 in the environment. It is important to identify factors to reduce the bio-burden of E. coli O157 in feedlots by clarifying practices associated with the occurrence of super-shedders in feedlot cattle. Methods The objective of this study is to (1) identify host, pathogen, and management risk factors associated with naturally infected feedlot cattle excreting high concentrations of E. coli O157:H7 in their feces and (2) to determine whether the ingested dose or the specific strain of E. coli O157:H7 influences a super-shedder infection within experimentally inoculated feedlot cattle. To address this, (1) pen floor fecal samples and herd parameters were collected from four feedlots over a 9-month period, then (2) 6 strains of E. coli O157:H7, 3 strains isolated from normal shedder steers and 3 strains isolated from super-shedder steers, were inoculated into 30 one-year-old feedlot steers. Five steers were assigned to each E. coli O157:H7 strain group and inoculated with targeted numbers of 102, 104, 106, 108, and 1010 CFU of bacteria respectively. Results In the feedlots, prevalence of infection with E. coli O157:H7 for the 890 fecal samples collected was 22.4%, with individual pen prevalence ranging from 0% to 90% and individual feedlot prevalence ranging from 8.4% to 30.2%. Three samples had E. coli O157:H7 levels greater than 104 MPN/g feces, thereby meeting the definition of super-shedder. Lower body weight at entry to the feedlot and higher daily maximum ambient temperature were associated with increased odds of a sample testing positive for E. coli O157:H7. In the experimental inoculation trial, the duration and total environmental shedding load of E. coli O157:H7 suggests that the time post-inoculation and the dose of inoculated E. coli O157:H7 are important while the E. coli O157:H7 strain and shedding characteristic (normal or super-shedder) are not. Discussion Under the conditions of this experiment, super-shedding appears to be the result of cattle ingesting a high dose of any strain of E. coli O157:H7. Therefore strategies that minimize exposure to large numbers of E. coli O157:H7 should be beneficial against the super-shedding of E. coli O157:H7 in feedlots.

Tularemia is a rare zoonotic disease found worldwide. The agent responsible for disease, Francisella tularensis, is one of the most highly infectious pathogens known, one that is capable of causing life-threatening illness with inhalation of

Emergent hypermucoviscous (HMV) strains of Klebsiella pneumoniae have been reported in multiple marine mammal species; however, there is limited information regarding the epidemiology and pathogenesis of this infection in these species. We determined the prevalence of HMV K. pneumoniae in wild-caught and stranded marine mammal populations on the US Pacific Coast. Samples were collected from 270 free-ranging California sea lions (CSLs; Zalophus californianus) captured at three discrete sampling sites and from 336 stranded marine mammals of various species. We recovered HMV K. pneumoniae only from CSLs, with a prevalence of 1.5% (4 of 275) in stranded animals, compared with 1.1% (3 of 270) in wild-caught animals. We assessed the phenotypic and genotypic variability of recovered HMV K. pneumoniae isolates recovered from CSLs ( n=11) and of archival HMV and non-HMV isolates from stranded marine mammals ( n=19). All but two HMV isolates were of the K2 serotype, whereas none of the non-HMV isolates belonged to this serotype. Of the HMV isolates, 96% (24 of 25) were PCR positive for the HMV-associated gene p- rmpA, whereas 92% (23 of 25) were PCR positive for p- rmpA2. Genetic fingerprinting by repetitive extragenic palindromic PCR showed four discrete clusters, demonstrating genotypic variability that loosely correlated with phenotype. Antimicrobial susceptibility testing revealed all isolates from stranded CSLs were susceptible to ceftiofur, indicating this antimicrobial agent is an appropriate choice for treatment of HMV K. pneumoniae infections in stranded CSLs. Our culture assay could reliably detect HMV K. pneumoniae from concentrations as low as 10

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was compared to conventional biochemical testing methods and nucleic acid analyses (16S rDNA sequencing, hippurate hydrolysis gene testing, whole genome sequencing [WGS]) for species identification of Campylobacter isolates obtained from chickens ( Gallus gallus domesticus, n = 8), American crows ( Corvus brachyrhynchos, n = 17), a mallard duck ( Anas platyrhynchos, n = 1), and a western scrub-jay ( Aphelocoma californica, n = 1). The test results for all 27 isolates were in 100% agreement between MALDI-TOF MS, the combined results of 16S rDNA sequencing, and the hippurate hydrolysis gene PCR ( p = 0.0027, kappa = 1). Likewise, the identifications derived from WGS from a subset of 14 isolates were in 100% agreement with the MALDI-TOF MS identification. In contrast, biochemical testing misclassified 5 isolates of C. jejuni as C. coli, and 16S rDNA sequencing alone was not able to differentiate between C. coli and C. jejuni for 11 sequences ( p = 0.1573, kappa = 0.0857) when compared to MALDI-TOF MS and WGS. No agreement was observed between MALDI-TOF MS dendrograms and the phylogenetic relationships revealed by rDNA sequencing or WGS. Our results confirm that MALDI-TOF MS is a fast and reliable method for identifying Campylobacter isolates to the species level from wild birds and chickens, but not for elucidating phylogenetic relationships among Campylobacter isolates.