Prevalence of extended-spectrum cephalosporin-, carbapenem-, and fluoroquinolone-resistant members of the family Enterobacteriaceae isolated from the feces of horses and hospital surfaces at two equine specialty hospitals.

OBJECTIVE: To estimate the prevalence of extended-spectrum cephalosporin-, carbapenem-, and fluoroquinolone-resistant bacteria of the family Enterobacteriaceae in the feces of hospitalized horses and on hospital surfaces. SAMPLE: Fecal and environmental samples were collected from The Ohio State University Galbreath Equine Center (OSUGEC) and a private referral equine hospital in Kentucky (KYEH). Feces were sampled within 24 hours after hospital admission and after 48 hours and 3 to 7 days of hospitalization. PROCEDURES: Fecal and environmental samples were enriched, and then selective media were inoculated to support growth of Enterobacteriaceae bacteria that expressed resistance phenotypes to extended-spectrum cephalosporins, carbapenems, and fluoroquinolones. RESULTS: 358 fecal samples were obtained from 143 horses. More samples yielded growth of Enterobacteriaceae bacteria that expressed resistance phenotypes (AmpC ß-lactamase, OR = 4.2; extended-spectrum beta-lactamase, OR = 3.2; and fluoroquinolone resistance, OR = 4.0) after 48 hours of hospitalization, versus within 24 hours of hospital admission. Horses hospitalized at KYEH were at greater odds of having fluoroquinolone-resistant bacteria (OR = 2.2). At OSUGEC, 82%, 64%, 0%, and 55% of 164 surfaces had Enterobacteriaceae bacteria with AmpC ß-lactamase phenotype, extended-spectrum beta-lactamase phenotype, resistance to carbapenem, and resistance to fluoroquinolones, respectively; prevalences at KYEH were similarly distributed (52%, 32%, 1%, and 35% of 315 surfaces). CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that antimicrobial-resistant Enterobacteriaceae may be isolated from the feces of hospitalized horses and from the hospital environment. Hospitalization may lead to increased fecal carriage of clinically important antimicrobial-resistance genes.

Adoption of recommended hand hygiene practices to limit zoonotic disease transmission at agricultural fairs.

Rapid transmission and spread of infectious pathogens are enhanced by the agricultural fair environment, where large numbers of livestock and people from numerous backgrounds congregate for several days. The transmission of influenza A virus and zoonotic enteric pathogens to fairgoers is a considerable risk and has occurred several times over the past decade. In an effort to mitigate zoonotic disease transmission in these settings, public health guidelines and recommendations including hand sanitation stations have been implemented. While hand hygiene recommendations to prevent the spread of zoonotic disease are well communicated, it is hypothesized that the adoption of these recommendations by agricultural fairs and fairgoers is low. To test this hypothesis, hand hygiene data collected from 658 agricultural fairs between 2012 and 2019 was analyzed to determine frequency and function of hand sanitation stations at the fairs, as well as utilization of educational signage. In addition, an observational study was performed to calculate the proportion of fairgoers who use hand sanitation stations at the fair. Lastly, samples were taken from working hand sanitation stations present at the exits of livestock barns and tested for the presence of influenza A virus and antimicrobial resistant coliform bacteria. Hand sanitation stations were present at most fairs (77.4 %) as recommended, but only 142 out of 2021 (7.0 %) visitors were observed using the stations. Health risk signage was displayed at more than half of fairs while the proper wash procedure was displayed at less than half. No influenza A virus was detected on any of the hand sanitation stations, however antimicrobial resistant coliform bacteria were recovered from 75.5 % of the sampled hand sanitation stations. Fairs should employ educational material along with functional hand sanitation stations in order to promote hand hygiene at fairs. Stations should be maintained and cleaned often to ensure effectiveness, as hand hygiene continues to be recommended to fairgoers when exiting animal barns to reduce zoonotic disease transmission.

Pulsed electric field application reduces carbapenem- and colistin-resistant microbiota and blaKPC spread in urban wastewater.

Wastewater flows from metropolitan areas, especially those with healthcare inputs, can serve as transport reservoirs for the dissemination of clinically-relevant antimicrobial resistant bacteria (ARB) such as carbapenem- (CR) and colistin-resistant (CoR) strains. Pulsed electric field (PEF) is an emerging wastewater management tool for reducing bacterial loads without generating environmentally harmful byproducts, but it's ability to reduce ARB and their genetic determinants is not well reported. We collected 86, 10-L raw wastewater influent samples from a large metropolitan wastewater treatment plant in Columbus, Ohio and subjected them to low (34 kV cm-1 for 67 µsec) and high (36 kV cm-1 for 89 µsec) PEF treatment. We quantified the PEF effectiveness by measuring concentrations of total coliform bacteria, CR and CoR bacteria, and the epidemic carbapenemase gene, blaKPC, before and after PEF treatment. Utilizing marginal linear regression models with generalized estimating equations, we observed that low and high PEF treatment resulted in a 1.94 (95% CI 2.06-1.81; P < 0.001) and 2.32 (95% CI 2.46-2.18; P < 0.001) log reduction of total coliform bacteria concentrations, respectively. Low and high PEF treatment produced similar log reductions between CR E. coli (2.01 (95% CI 2.15-1.86; P < 0.001); 2.14 (95% CI: 5.30-4.61; P < 0.001)) and CR Enterobacteriaceae concentrations (1.55 (95% CI 1.70-1.41; P < 0.001); 1.86 (95% CI 2.05-1.68; P < 0.001)), and resulted in a 1.15 log (95% CI 1.38-0.93, P < 0.001) and 1.28 log (95% CI 1.54-1.03, P < 0.001) reduction of absolute blaKPC concentrations. Log CoR E. coli concentrations were reduced by 2.47 (95% CI 2.78-2.15; P < 0.001) and 2.52 (95% CI 2.91-2.15; P < 0.001) and CoR Enterobacteriaceae by 2.24 (95% CI 2.52-1.95; P < 0.001) and 2.50 (95% CI 2.89-2.11; P < 0.001) following low and high PEF application. PEF can be applied for wastewater management as an independent treatment method, particularly at critical control points, such as an on-site management of wastewater from hospitals or other healthcare facilities, or in series with other conventional methods to reduce total bacterial loads and concentrations of clinically-relevant ARB.

Effect of student transrectal palpation on early pregnancy loss in dairy cattle.

Transrectal palpation of the reproductive tract is the most common method for pregnancy determination in cattle and is considered a veterinary skill that new veterinary medicine (DVM) graduates should perform proficiently. However, using privately owned cattle to train students can be difficult because producers may believe that transrectal palpation by inexperienced students increases the risk of pregnancy wastage compared with examination by an experienced clinician. We used a randomized field trial of 1,216 healthy Holstein and Jersey cattle in 2 commercial dairy herds to estimate the effect of veterinary student transrectal palpation on early pregnancy loss. All cattle were determined to be pregnant using transrectal ultrasonography at approximately 37 d after artificial insemination. Cattle were then allocated into 2 groups based upon their ear tag number (study group = 598; control group = 618). Cattle in the study group were immediately palpated after ultrasonography by a fourth-year veterinary student, whereas control cattle were not subject to any additional pregnancy assessment. For analysis, the student palpators were divided into 2 groups: students who had previously had formal palpation training via an elective bovine palpation class (n = 30) and students who had not had palpation training (n = 134). All cattle were reevaluated using transrectal ultrasonography approximately 70 d after artificial insemination. A total of 53 (4.36%) animals lost their pregnancy between the first and second pregnancy assessments. Of the animals that lost their pregnancy, 26 (4.35%) were study group cows and 27 (4.37%) were control cows. Of the 26 cows documented to have had pregnancy loss within the study group, 20 out of 378 (5.3%) had been palpated by students who had not taken the palpation elective and 6 out of 220 (2.7%) had been palpated by students who had completed the elective. We found no difference in pregnancy loss between student-palpated and clinician-ultrasounded cattle, supporting the safety of using privately owned animals for student bovine palpation and pregnancy diagnosis training without affecting early pregnancy loss.

Carbapenemase-producing Enterobacteriaceae and Aeromonas spp. present in wastewater treatment plant effluent and nearby surface waters in the US.

Carbapenemase-producing bacteria (CPB) are rare, multidrug resistant organisms most commonly associated with hospitalized patients. Metropolitan wastewater treatment plants (WWTP) treat wastewater from large geographic areas which include hospitals and may serve as epidemiologic reservoirs for the maintenance or expansion of CPB that originate from hospitals and are ultimately discharged in treated effluent. However, little is known about the potential impact of these WWTP CPB on the local surface water and their risk to the public health. In addition, CPB that are present in surface water may ultimately disseminate to intensively-managed animal agriculture facilities where there is potential for amplification by extended-spectrum cephalosporins. To better understand the role of WWTPs in the dissemination of CPB in surface waters, we obtained samples of treated effluent, and both upstream and downstream nearby surface water from 50 WWTPs throughout the US. A total of 30 CPB with clinically-relevant genotypes were recovered from 15 WWTPs (30%) of which 13 (50%) serviced large metropolitan areas and 2 (8.3%) represented small rural populations (P < 0.05). Recovery of CPB was lowest among WWTPs that utilized ultraviolet radiation for primary disinfection (12%), and higher (P = 0.11) for WWTPs that used chlorination (42%) or that did not utilize disinfection (50%). We did not detect a difference in CPB recovery by sampling site, although fewer CPB were detected in upstream (8%) compared to effluent (20%) and downstream (18%) samples. Our results indicate that WWTP effluent and nearby surface waters in the US are routinely contaminated with CPB with clinically important genotypes including those producing Klebsiella pneumoniae carbapenemase (KPC) and New Delhi metallo-beta-lactamase (NDM). This is a concern for both public health and animal agriculture because introduction of CPB into intensively managed livestock populations could lead to their amplification and foodborne dissemination.

Enterobacter cloacae Complex Sequence Type 171 Isolates Expressing KPC-4 Carbapenemase Recovered from Canine Patients in Ohio.

Companion animals are likely relevant in the transmission of antimicrobial-resistant bacteria. Enterobacter xiangfangensis sequence type 171 (ST171), a clone that has been implicated in clusters of infections in humans, was isolated from two dogs with clinical disease in Ohio. The canine isolates contained IncHI2 plasmids encoding blaKPC-4 Whole-genome sequencing was used to put the canine isolates in phylogenetic context with available human ST171 sequences, as well as to characterize their blaKPC-4 plasmids.

ß-Lactam and Fluoroquinolone-Resistant Enterobacteriaceae Recovered from the Environment of Human and Veterinary Tertiary Care Hospitals.

The dissemination of Enterobacteriaceae expressing resistance to clinically important antibiotics such as extended-spectrum cephalosporins (ESC), carbapenems, and fluoroquinolones is of critical concern to both human and veterinary medicine. In healthcare facilities, the movement of patients, personnel, and equipment provides an opportunity for Enterobacteriaceae carrying antibiotic resistance genes to disseminate in the hospital environment where they pose a threat to patients, staff, and hospital visitors. We collected environmental samples using electrostatic cloths to estimate the frequency of resistant Enterobacteriaceae contamination on human and veterinary hospital surfaces. Samples were enriched in nutrient broth modified with antibiotics to provide specific selection pressure to select for bacteria expressing AmpC ß-lactamase (blaCMY), extended-spectrum ß-lactamase (ESBL, blaCTX-M), carbapenemase, quinolone, and fluoroquinolone-resistant phenotypes. A total of 31 contact surfaces were sampled at each of five visits to two human hospitals. In addition, 34 surfaces were sampled at each of three visits to a veterinary hospital serving both farm animal and companion animal patients. Isolates expressing the blaCMY phenotype were recovered from 0.6%, 100%, and 18.2% of human hospitals, veterinary farm animal, and veterinary companion animal hospital surfaces, respectively. Isolates expressing the blaCTX-M phenotype were recovered from 0.6%, 55%, and 16.7% of human hospitals, farm animal, and companion animal veterinary hospital surfaces, respectively. Carbapenemase-producing Enterobacteriaceae (CPE) were detected from 1.3% of human hospital surfaces, but none were recovered from the veterinary hospital. Fluoroquinolone resistance was detected in 0.6%, 5.0%, and 37.9% of human hospitals, farm animal, and companion animal veterinary hospital surfaces, respectively. Our results indicate that ESC and fluoroquinolone-resistant Enterobacteriaceae can contaminate surfaces in both human and veterinary medical settings, with higher prevalence observed in veterinary hospitals, although the recovery of CPE from human hospital environments is concerning. Hospital medical trolleys or carts may serve as fomites for the dissemination of clinically relevant resistant bacteria.

Risk of anesthesia-related complications in brachycephalic dogs.

OBJECTIVE To determine whether brachycephalic dogs were at greater risk of anesthesia-related complications than nonbrachycephalic dogs and identify other risk factors for such complications. DESIGN Retrospective cohort study. ANIMALS 223 client-owned brachycephalic dogs undergoing general anesthesia for routine surgery or diagnostic imaging during 2012 and 223 nonbrachycephalic client-owned dogs matched by surgical procedure and other characteristics. PROCEDURES Data were obtained from the medical records regarding dog signalment, clinical signs, anesthetic variables, surgery characteristics, and complications noted during or following anesthesia (prior to discharge from the hospital). Risk of complications was compared between brachycephalic and nonbrachycephalic dogs, controlling for other factors. RESULTS Perianesthetic (intra-anesthetic and postanesthetic) complications were recorded for 49.1% (n = 219) of all 446 dogs (49.8% [111/223] of brachycephalic and 48.4% [108/223] of nonbrachycephalic dogs), and postanesthetic complications were recorded for 8.7% (39/446; 13.9% [31/223] of brachycephalic and 3.6% [8/223] of nonbrachycephalic dogs). Factors associated with a higher perianesthetic complication rate included brachycephalic status and longer (vs shorter) duration of anesthesia; the risk of perianesthetic complications decreased with increasing body weight and with orthopedic or radiologic procedures (vs soft tissue procedures). Factors associated with a higher postanesthetic complication rate included brachycephalic status, increasing American Society of Anesthesiologists status, use of ketamine plus a benzodiazepine (vs propofol with or without lidocaine) for anesthetic induction, and invasive (vs noninvasive) procedures. CONCLUSIONS AND CLINICAL RELEVANCE Controlling for other factors, brachycephalic dogs undergoing routine surgery or imaging were at higher risk of peri- and postanesthetic complications than nonbrachycephalic dogs. Careful monitoring is recommended for brachycephalic dogs in the perianesthetic period.

Maintenance of Carbapenemase-Producing Enterobacteriaceae in a Farrow-to-Finish Swine Production System.

Carbapenemase-producing Enterobacteriaceae (CPE) threaten both agriculture and public health. While carbapenems are restricted in food-producing animals, other ß-lactams, such as ceftiofur, are frequently applied in livestock. While the relationship is not fully elucidated, ceftiofur use may provide selective pressure that promotes carbapenem resistance. Recently reported in U.S. livestock, plasmid-mediated CPE are also present in livestock in Europe and Asia. We previously reported the rare carbapenemase gene, blaIMP-64, in the environment of a large farrow-to-finish swine operation. To better understand CPE in this swine production system, in 2016 we followed a cohort of 350+ pigs over 5 months from late sow gestation to the final finishing phase. We screened both environmental and fecal samples for CPE using our selective enrichment protocol, with resulting phenotypic CPE isolates further characterized. Of 55 environmental and 109 sow fecal samples collected from a farrowing barn on our initial visit, 35 (64%) environmental and 15 (14%) sow fecal samples yielded isolates of multiple Enterobacteriaceae species carrying the metallo-ß-lactamase gene blaIMP-64 on an IncQ plasmid. The frequency of IMP-64-positive environmental (n = 32), sow fecal (n = 30), and piglet fecal swab (n = 120) samples was highest for all groups when the market pig cohort was between 1 and 10 days, with observed prevalence of 97%, 28%, and 18%, respectively. After weaning, blaIMP-64 was detected in a single environmental sample from a nursery pen, with no CPE recovered in the finishing phase. Used in U.S. swine production to treat and control disease, ceftiofur is administered to piglets on this farm at birth, with males receiving a second dose at castration (≈day 7). Once introduced into animal agriculture, the common use of ceftiofur may provide the selection pressure required for CPE dissemination throughout large, intensively managed food animal populations housed in animal-dense environments.

Arterial anomalies of the celiac trunk and median arcuate ligament compression in dogs and cats assessed by computed tomography angiography.

OBJECTIVE: To identify abnormalities of the celiac artery (CA) and major branches in dogs and cats by computed tomography angiography (CTA). STUDY DESIGN: Multi-institutional retrospective case series. ANIMALS: Two hundred fifty-four dogs and 13 cats. METHODS: Abdominal CTA images from 2009 to 2017 were reviewed. Logistic regression models were used to evaluate the relationship between CA abnormalities and sex, age, size of dog, concurrent venous anomaly, or presence of gastrointestinal signs. RESULTS: Abnormalities in the CA were observed in 32 animals (11.9%) including 9 with abnormal branching (3.4%) and 23 with CA compression (8.6%). A celiacomesenteric trunk was observed in 8 (2.9%; 6 dogs, 2 cats). The splenic artery originated from the cranial mesenteric artery in 1 dog; the hepatic arterial branches originated from the left gastric artery in another. Four out of 32 animals (12.5%) with an arterial anomaly had another vascular abnormality. Large breed dogs were more likely to have an arterial anomaly (OR 4.3, 95% CI: 1.18-15.5, P = .02) and 12 times more likely to have CA compression (OR 12.0, 95% CI: 1.4-97.7, P = .02) compared to small breed dogs. Dogs with CA compression were more likely to present for gastrointestinal signs (OR 3.6, 95% CI: 1.2-10.3, P = .01). CONCLUSION: Anomalies of the celiac trunk are apparent on CTA and may impact surgical or image-guided intervention. Compression at the origin of the CA was apparent on imaging, similar to the median arcuate ligament syndrome in people, although the significance of this finding in dogs is unknown.

Extended-Spectrum Cephalosporin-Resistant Enterobacteriaceae in Enteric Microflora of Wild Ducks.

We tested 772 cloacal swabs from wild ducks to estimate the prevalence of enteric bacteria resistant to extended-spectrum cephalosporins (ESC). We found a low prevalence of the important ESC resistance genotypes, blaCMY (5.7%) and blaCTX-M (0.3%). This suggests a minor role for wild ducks in the movement of resistant bacteria in the environment.

Carbapenemase-Producing Aeromonas veronii Disseminated in the Environment of an Equine Specialty Hospital.

The emergence of bacterial resistance to carbapenem antibiotics is an urgent public health threat. Carbapenem drugs are a last resort treatment option for life-threatening infections. The frequent use of broad-spectrum antibiotics to treat hospitalized patients provides significant selection pressure favoring the emergence and dissemination of resistant organisms, including carbapenem-resistant Enterobacteriaceae (CRE). CREs have been reported in animal populations, but only rarely in horses. Our objective was to determine the prevalence of CRE in the environment of a referral equine specialty hospital. Environmental samples were collected on seven different sampling dates. Four clonal carbapenemase-producing Aeromonas veronii were recovered from 315 sampled surfaces (1.3%). All four isolates harbored the carbapenemase-producing, metallo-ß-lactamase gene blacphA, although corresponding minimum inhibitory concentrations were within the susceptible range for imipenem and meropenem. All had an identical multilocus sequence type with a previously unreported allelic profile and contained multiple plasmids. To our knowledge, this recovery of blacphA-harboring A. veronii is the first report of carbapenemase-producing bacteria in the environment of an equine veterinary hospital. However, the low recovery rate suggests that environmental contamination is uncommon. Appropriate hospital cleaning and disinfection protocols are necessary to maintain a low risk of contamination for patients and personnel.

Enterobacteriaceae Harboring AmpC (blaCMY) and ESBL (blaCTX-M) in Migratory and Nonmigratory Wild Songbird Populations on Ohio Dairies.

Extended-spectrum ß-lactamases (ESBLs) confer bacterial resistance to critically important antimicrobials, including extended-spectrum cephalosporins (ESCs). Livestock are important reservoirs for the zoonotic food-borne transmission of ESC-resistant enteric bacteria. Our aim is to describe the potential role of migratory and resident wild birds in the epidemiology of ESBL-mediated bacterial resistance on dairy farms. Using mist nets, we sampled wild migratory and resident birds either immediately adjacent to or 600 ft away from free-stall barns on three Ohio dairy farms during the 2014 and 2015 spring migrations. Individual swabs were used to obtain both a cloacal and external surface swab from each bird. Samples were inoculated into MacConkey broth containing cefotaxime then inoculated onto MacConkey agar with cefoxitin, cefepime, or meropenem to identify the blaCMY, blaCTX-M, and carbapenemase phenotypes, respectively. Six hundred twenty-three birds were sampled, 19 (3.0%) of which harbored bacteria with blaCMY and 32 (5.1%) harbored bacteria with blaCTX-M from either their cloacal sample or from their external swab. There was no difference in the prevalence of either gene between migratory and resident birds. Prevalence of blaCMY and blaCTX-M was higher among birds sampled immediately outside the barns compared with those sampled 600 ft away. Our results suggest that wild birds can serve as mechanical and/or biological vectors for Enterobacteriaceae with resistance to ESCs. Birds live in close contact with dairy cows and their feed, therefore, transmission locally between farms is possible. Finding a similar prevalence in migratory and nonmigratory birds suggests the potential for regional and intercontinental movement of these resistance genes via birds.

Influenza A Virus Surveillance in Underrepresented Avian Species in Ohio, USA, in 2015.

We surveyed passerines and other terrestrial avian species for influenza A virus, resulting in molecular detection of virus from 1.5% of the 615 birds. However, no viral isolates were recovered. Little is known about the role that these undersurveilled avian species play in the ecology of influenza A virus.

Carbapenemase-Producing Enterobacteriaceae Recovered from the Environment of a Swine Farrow-to-Finish Operation in the United States.

Carbapenem-resistant Enterobacteriaceae (CRE) present an urgent threat to public health. While use of carbapenem antimicrobials is restricted for food-producing animals, other ß-lactams, such as ceftiofur, are used in livestock. This use may provide selection pressure favoring the amplification of carbapenem resistance, but this relationship has not been established. Previously unreported among U.S. livestock, plasmid-mediated CRE have been reported from livestock in Europe and Asia. In this study, environmental and fecal samples were collected from a 1,500-sow, U.S. farrow-to-finish operation during 4 visits over a 5-month period in 2015. Samples were screened using selective media for the presence of CRE, and the resulting carbapenemase-producing isolates were further characterized. Of 30 environmental samples collected from a nursery room on our initial visit, 2 (7%) samples yielded 3 isolates, 2 sequence type 218 (ST 218) Escherichia coli and 1 Proteus mirabilis, carrying the metallo-ß-lactamase gene blaIMP-27 on IncQ1 plasmids. We recovered on our third visit 15 IMP-27-bearing isolates of multiple Enterobacteriaceae species from 11 of 24 (46%) environmental samples from 2 farrowing rooms. These isolates each also carried blaIMP-27 on IncQ1 plasmids. No CRE isolates were recovered from fecal swabs or samples in this study. As is common in U.S. swine production, piglets on this farm receive ceftiofur at birth, with males receiving a second dose at castration (≈day 6). This selection pressure may favor the dissemination of blaIMP-27-bearing Enterobacteriaceae in this farrowing barn. The absence of this selection pressure in the nursery and finisher barns likely resulted in the loss of the ecological niche needed for maintenance of this carbapenem resistance gene.