Effects of dietary zinc on the gut microbiome and resistome of the gestating cow and neonatal calf.

Zinc is an essential trace element required in the diet of all species. While the effects of zinc have been studied in growing calves, little is known about the effect of zinc on the microbiota of the gestating cow or her neonatal calf. Understanding factors that shape the gut health of neonatal animals and evaluating the effect of dietary supplements in adult gestating animals is important in promoting animal health and informing feeding practices. The aims of this study were to determine the effect of dietary zinc on the microbiota and resistome of the gestating cow and calf. Gestating cows received standard (40 ppm) or high (205 ppm) dietary zinc levels from dry off to calving. Fecal samples were collected from cows upon enrollment and at calving and from neonatal calves. Fecal samples underwent 16S rRNA sequencing and a subset also underwent shotgun metagenomic sequencing. The effect of zinc supplementation on the diversity and composition of the cow and calf microbiome and resistome was assessed. Alpha and beta diversity and composition of the microbiota were significantly altered over time but not by treatment in the cows, with alpha diversity decreasing and 14 genera found at significantly higher relative abundances at calving compared to enrollment. Levels of 27 antimicrobial resistance genes significantly increased over time. Only a small number of taxa were differentially expressed at calving in treatment and control groups, including Faecalibacterium, Bacteroides, Turicibacter, and Bifidobacterium pseudolongum. No effect of the dam's treatment group was observed on the diversity or composition of the neonatal calf microbiota. The calf resistome, which was relatively rich and diverse compared to the cow, was also unaffected by the dam's treatment group. The impact of high levels of dietary zinc thus appeared to be minimal, with no observed changes in alpha or beta diversity, and few changes in the relative abundance of a small number of taxa and antimicrobial resistance genes.

The potential role of veterinary technicians in promoting antimicrobial stewardship.

BACKGROUND: A core principle of antimicrobial stewardship (AMS) in veterinary settings is the need for engagement of all stakeholders; however, no studies have addressed the role of veterinary technicians in AMS specifically. The objective of this study was to qualitatively assess knowledge, opinions, and practices related to AMS among technicians. Semi-structured interviews were conducted with 33 veterinary technicians with varied backgrounds, experience and roles. Interviews centered on participants work experience and interactions with their employer, perceptions of antimicrobial resistance and overuse in veterinary medicine, observed application of AMS principles, opinions on potential opportunities for technicians to contribute to AMS and concomitant potential barriers to these opportunities. Transcripts of interviews were coded thematically by two authors, then organized into a hierarchical framework, and the characterization of codes was compared across different categories of respondents. RESULTS: Most veterinary technicians were knowledgeable about antimicrobial drugs but could not provide a complete definition of antimicrobial resistance or AMS. Most veterinary technicians could identify examples of antimicrobial misuse. Participants identified areas of client education and discussion with veterinarians as potential areas to contribute to AMS. Barriers identified included hierarchical structures of veterinary practices and time-constraints. Most participants expressed a personal interest in participating in AMS. CONCLUSIONS: There is a possible appetite among some veterinary technicians to participate in AMS and they already play applicable roles in practices. Barriers such as educational needs, hierarchical structures of veterinary practices and time constraints will need to be addressed if technicians are included in AMS efforts.

Factors Influencing Equine Affiliate Opinion on Reporting of Equine Strangles in the United States.

Although equine strangles is reportable in all states, synchronous reporting of this disease does not occur across the country. States have variable regulations on reporting (actionable, notifiable, and monitored) and no mandatory comprehensive databases exist for tracking prevalence. In this study, which is a companion to our recent publication on veterinarians' opinions on this topic, we solicited equine affiliates' (including horse owners, horse business owners/trainers, breeders, and barn managers) opinions on reporting of strangles and factors influencing their opinion. A total of 518 equine affiliates/stakeholders living within the United States. A structured survey was administered online. Eighty-one participants (17.8%) believed that strangles should continue to be nationally monitored and that individual states should have jurisdiction over laboratory confirmed positive cases; 124 (27.2%) believed strangles should become nationally monitored with mandatory notification of positive cases to a central forum; 77 participants (16.9%) thought strangles should become notifiable nationally; and 157 (34.4%) thought strangles should become notifiable and actionable. Participants who ranked strangles as "important" or "very important" compared to other infectious disease were more likely to want increased reporting (OR = 3.62, P = .054), and participants who were more familiar with the disease were more likely to rank it as important: for every 1-point increase in correct total score on the 5-question basic knowledge exam included, there was a 49% increase in odds of ranking strangles as important or very important (P = .003). Equine stakeholders who perceived their horses to be at medium or high risk of acquiring the disease were 2.76 times more likely to rank strangles as important or very important (P = .014). Of the sampled equine stakeholder community, the majority of respondents (78.5%) favored increased reporting. Information obtained in this study regarding current views of those involved in the equine industry on strangles reporting could inform future policy regarding the disease.

Infrequent intrahousehold transmission of Clostridioides difficile between pet owners and their pets.

Companion animals have been shown to carry Clostridioides difficile strains that are similar or identical to strains found in people, and a small number of studies have shown that pets carry genetically identical C. difficile isolates as their owners, suggesting inter-species transmission. However, the directionality of transmission is ultimately unknown, and the frequency with which animals acquire C. difficile following their owners' infection is unclear. The goal of this study was to assess how often pets belonging to people with C. difficile infection carry genetically related C. difficile isolates. We enrolled pet owners from two medical institutions (University of Pennsylvania Health System (UPHS) and The Ohio State University Wexner Medical Center (OSUWMC)) who had diarrhoea with or without positive C. difficile assays and tested their faeces and their pets' faeces for C. difficile using both anaerobic culture and PCR assays. When microorganisms were obtained from both the owner and pet and had the same toxin profile or ribotype, isolates underwent genomic sequencing. Faecal samples were obtained from a total of 59 humans, 72 dogs and 9 cats, representing 47 complete households (i.e. where a sample was available from the owner and at least one pet). Of these, C. difficile was detected in 30 humans, 10 dogs and 0 cats. There were only two households where C. difficile was detected in both the owner and pet. In one of these households, the C. difficile isolates were of different toxin profiles/ribotypes (A+/B+ / RT 499 from the owner, A-/B- / RT PR22386 from the dog). In the other household, the isolates were genetically identical (one SNP difference). Interestingly, the dog from this household had recently received a course of antibiotics (cefpodoxime and metronidazole). Our findings suggest that inter-species transmission of C. difficile occurs infrequently in households with human C. difficile infections.

Evaluation of canine raw food products for the presence of extended-spectrum beta-lactamase- and carbapenemase-producing bacteria of the order Enterobacterales.

OBJECTIVE: To assess the potential contamination of commercial raw dog food products with bacteria of the Enterobacterales order that produce extended spectrum beta-lactamase (ESBL) and carbapenemase enzymes, determine risk factors for contamination, and understand isolate genetic diversity. SAMPLES: A total of 200 canine raw food products. METHODS: Products were cultured on selective chromogenic agar following enrichment steps. Whole-genome sequencing was performed for isolates that were confirmed to produce an ESBL. Isolates were characterized by antimicrobial resistance genes, and multilocus sequences typing, and compared to other isolates in the NCBI database for clonality. Preservation method and protein sources were assessed as potential risk factors for contamination with ESBL and carbapenemase-producing bacteria of the Enterobacterales order. RESULTS: No carbapenemase-producing Enterobacterales (CPE) were identified, but ESBL-producing Enterobacterales bacteria were isolated from 20/200 products (10.0%; 95% CI, 7.3 to 16.5%), all of which were frozen. Pork-derived protein source products were 8.1 times (P = .001; 95% CI, 2.53 to 26.2) more likely to carry ESBL-producing Enterobacterales bacteria than other protein sources. WGS analysis confirmed the presence of ESBL genes in a total of 25 distinct isolates (19 Escherichia coli, 5 Klebsiella pneumoniae, and 1 Citrobacter braakii). Genes encoding CTX-M type ESBL enzymes were the most common (24/25 isolates, 96.0%) with blaCTX-M-27 being the most common allele (8/25, 32.0%). CLINICAL RELEVANCE: Frozen, raw food products may serve as a route of transmission of ESBL-producing Enterobacterales bacteria to companion animals. Veterinarians should advise owners about the risks of raw food diets, including potential exposure to antimicrobial-resistant bacteria.

The Importance of Evaluating Positive Welfare Characteristics and Temperament in Working Therapy Dogs.

To date, investigations of the welfare of therapy dogs have focused largely on examining physiological and behavioral measures that could indicate if the animal is experiencing stress or distress. However, this approach does not fully address the definition of welfare which is often described as existing on a continuum from negative (or stressful) to positive. With therapy dogs, it would be worth addressing if they experience positive emotional affect while working since the quality and efficacy of animal-assisted interventions for the human recipient is likely to be influenced by the animal's emotional state during the interaction. The purpose of this review is to articulate how objective measurements of the HPA axis and measurements of behavioral observations and standardized questions can be used to evaluate positive welfare in therapy dogs. A potentially relevant indicator of positive welfare is the peripheral concentration of the neurohormone oxytocin, which has been found to increase in systemic circulation within a variety of species during positive social and affiliative contexts, including during human-dog interaction. Oxytocin is also a negative-feedback regulator of the Hypothalamic-Pituitary-Adrenal (HPA) axis, which culminates with the production of the stress hormone cortisol. Cortisol is widely used as a physiological indicator to assess negative welfare states in animals, including therapy dogs. Observable behavior during interactions with humans that may convey enjoyment could provide indicators of positive welfare in dogs such as engagement in play, or human-directed affiliative behaviors including leaning against, nudging, or licking the patient. However, in assessing positive welfare, it is also critical to consider that all animal behavioral displays and physiological responses are dependent on the dog's individual (and breed) temperament. Temperament directly drives how the animal copes and responds to its current physical and social environment, including during stressful situations such as when therapy dogs interact with unfamiliar humans in novel healthcare settings. Coupled with both positive and negative physiological and behavioral welfare indicators, questionnaire data can provide further context to, and enhance interpretations of, therapy dog welfare assessment results. Overall, to date, no studies have measured all of these factors to assess therapy dog welfare.

Effects of regional limb perfusion technique on concentrations of antibiotic achieved at the target site: A meta-analysis.

Intravenous regional limb perfusions (RLP) are widely used in equine medicine to treat distal limb infections, including synovial sepsis. RLPs are generally deemed successful if the peak antibiotic concentration (Cmax) in the sampled synovial structure is at least 8-10 times the minimum inhibitory concentration (MIC) for the bacteria of interest. Despite extensive experimentation and widespread clinical use, the optimal technique for performing a successful perfusion remains unclear. The objective of this meta-analysis was to examine the effect of technique on synovial concentrations of antibiotic and to assess under which conditions Cmax:MIC ≥ 10. A literature search including the terms "horse", "equine", and "regional limb perfusion" between 1990 and 2021 was performed. Cmax (µg/ml) and measures of dispersion were extracted from studies and Cmax:MIC was calculated for sensitive and resistant bacteria. Variables included in the analysis included synovial structure sampled, antibiotic dose, tourniquet location, tourniquet duration, general anesthesia versus standing sedation, perfusate volume, tourniquet type, and the concurrent use of local analgesia. Mixed effects meta-regression was performed, and variables significantly associated with the outcome on univariable analysis were added to a multivariable meta-regression model in a step-wise manner. Sensitivity analyses were performed to assess the robustness of our findings. Thirty-six studies with 123 arms (permutations of dose, route, location and timing) were included. Cmax:MIC ranged from 1 to 348 for sensitive bacteria and 0.25 to 87 for resistant bacteria, with mean (SD) time to peak concentration (Tmax) of 29.0 (8.8) minutes. Meta-analyses generated summary values (θ) of 42.8 x MIC and 10.7 x MIC for susceptible and resistant bacteria, respectively, though because of high heterogeneity among studies (I2 = 98.8), these summary variables were not considered reliable. Meta-regression showed that the only variables for which there were statistically significant differences in outcome were the type of tourniquet and the concurrent use of local analgesia: perfusions performed with a wide rubber tourniquet and perfusions performed with the addition of local analgesia achieved significantly greater concentrations of antibiotic. The majority of arms achieved Cmax:MIC ≥ 10 for sensitive bacteria but not resistant bacteria. Our results suggest that wide rubber tourniquets and concurrent local analgesia should be strongly considered for use in RLP and that adequate therapeutic concentrations (Cmax:MIC ≥ 10) are often achieved across a variety of techniques for susceptible but not resistant pathogens.

Factors Influencing Veterinarian Opinion on Reporting of Equine Strangles in the United States.

Although equine strangles is reportable in all states, synchronous reporting of this disease does not occur across the country. States have variable regulations on reporting (actionable, notifiable, and monitored) and no mandatory comprehensive databases exist for tracking prevalence. In this study, we solicited veterinarians' opinions on reporting of strangles and factors influencing their opinion. Two hundred and fifty veterinarians practicing within the United States A structured survey was administered online. A total of 250 veterinarians participated: 84 participants (34%) believed that strangles should continue to be nationally monitored and that individual states should have jurisdiction over laboratory-confirmed positive cases; 58 (23.2%) believed strangles should become nationally monitored with mandatory notification of positive cases to a central forum; 24 participants (9.6%) thought strangles should become notifiable nationally; and 44 (17.6%) thought strangles should become notifiable and actionable. Veterinarians who were currently in the habit of reporting strangles were also more likely to want increased reporting (Or=1.87), though this association was not quite statistically significant  (P = 0.054), as did veterinarians who ranked strangles as "very important" or "important" relative to other infectious disease (OR 3.77, P = .037). Veterinarians practicing in the Southwest (P = .01) and West (P = .04) were significantly less likely than northeast practitioners to rank strangles of higher importance. Opinions on equine strangles and desire for increased reporting were varied in the sampled veterinary community. Information obtained in this study regarding veterinarians' current views on strangles reporting could inform future policy regarding the disease.

Critically important antimicrobials are frequently used on equine racetracks.

OBJECTIVE: To characterize antimicrobial use on four racetracks in the eastern US during the peak racing 2017-2018 seasons. PROCEDURES: Handwritten daily treatment sheets provided by attending veterinarians who listed treatments administered to horses stabled at the racetrack were obtained. Information contained in the treatment sheets included the date, name of the horse and its trainer, type of treatment, and a brief (usually 1-word) indication for treatment. The handwritten data listed on the racetrack treatment sheets were manually transcribed and analyzed. RESULTS: A total of 2,684 antimicrobial prescriptions were recorded, representing 6.8% of all drug treatments. The most frequently prescribed antimicrobials were enrofloxacin, with 854 prescriptions (31.8% of antimicrobial treatments), followed by gentamicin (570 [21.2%] prescriptions), ceftiofur (388 [14.5%] prescriptions,), and penicillin (220 [8.2%] prescriptions). The relative frequencies of antimicrobial class and indication for treatment varied significantly by racetrack and by prescribing veterinarian. Limitations associated with the data precluded ascertainment of the proportion of horses treated or exact indications for treatment. CLINICAL RELEVANCE: Antimicrobials appeared to be prescribed relatively infrequently at racetracks relative to other drugs, but highly or critically important antimicrobials were most often used. The appropriateness of use of these drugs remains unknown.

Trends in the number of patients linked to potential vet-shopping behavior in the United States (2014-2019).

OBJECTIVE: To estimate the number of patients linked to vet-shopping behavior (the solicitation of controlled substance prescriptions from multiple veterinarians for misuse) in the United States using 2014-2019 data and characterize mandates for veterinarians to examine prescription drug monitoring programs (PDMPs) before prescribing controlled substances as of April 2021. SAMPLE: National database reporting prescription dispensing from 92% of US pharmacies from 2014 through 2019. PROCEDURES: The annual number of patients with dispensed prescriptions for opioid analgesics, opioid cough-and-cold medications, or benzodiazepines from ≥ 4 veterinarians was calculated. State veterinary medical associations were contacted for information on veterinarian PDMP use mandates. RESULTS: From 2014 through 2019, the number of patients with prescriptions for any class of controlled substances from ≥ 4 veterinarians tripled from 935 to 2,875 (+207.5%). The number of patients with opioid cough-and-cold medication prescriptions from ≥ 4 veterinarians rose from 150 to 1,348 (+798.9%). The corresponding number for benzodiazepines rose from 185 to 440 (+137.8%). The corresponding number for opioid analgesics peaked at 868 in 2016 before decreasing to 733 in 2019. In April 2021, 10 states mandated veterinarians to examine PDMP records of owners or animals before prescribing controlled substances; 3 mandates excluded benzodiazepines. CLINICAL RELEVANCE: Vet shopping in the US may be increasingly common. Mandates for veterinarians to examine PDMPs before prescribing controlled substances might facilitate detection of this behavior. However, benefits of mandates should be weighed against their potential burden on veterinarians.

Gut microbiota features associated with Clostridioides difficile colonization in dairy calves.

Diarrheal disease, a major cause of morbidity and mortality in dairy calves, is strongly associated with the health and composition of the gut microbiota. Clostridioides difficile is an opportunistic pathogen that proliferates and can produce enterotoxins when the host experiences gut dysbiosis. However, even asymptomatic colonization with C. difficile can be associated with differing degrees of microbiota disruption in a range of species, including people, swine, and dogs. Little is known about the interaction between C. difficile and the gut microbiota in dairy calves. In this study, we sought to define microbial features associated with C. difficile colonization in pre-weaned dairy calves less than 2 weeks of age. We characterized the fecal microbiota of 80 calves from 23 different farms using 16S rRNA sequencing and compared the microbiota of C. difficile-positive (n = 24) and C. difficile-negative calves (n = 56). Farm appeared to be the greatest source of variability in the gut microbiota. When controlling for calf age, diet, and farm location, there was no significant difference in Shannon alpha diversity (P = 0.50) or in weighted UniFrac beta diversity (P = 0.19) between C. difficile-positive and-negative calves. However, there was a significant difference in beta diversity as assessed using Bray-Curtiss diversity (P = 0.0077), and C. difficile-positive calves had significantly increased levels of Ruminococcus (gnavus group) (Adj. P = 0.052), Lachnoclostridium (Adj. P = 0.060), Butyricicoccus (Adj. P = 0.060), and Clostridium sensu stricto 2 compared to C. difficile-negative calves. Additionally, C. difficile-positive calves had fewer microbial co-occurrences than C. difficile-negative calves, indicating reduced bacterial synergies. Thus, while C. difficile colonization alone is not associated with dysbiosis and is therefore unlikely to result in an increased likelihood of diarrhea in dairy calves, it may be associated with a more disrupted microbiota.

Educational interventions to address misconceptions about antibiotic residues in milk can alter consumer perceptions and may affect purchasing habits.

The industrialization of the agri-food industry and resultant decrease in the number of people employed on farms has contributed to a knowledge gap among consumers about food production processes. A commonly reported concern of dairy consumers is the use of antibiotics in dairy animals, even though these drugs are an important tool for promoting animal health and welfare and food safety. The extent to which consumers are aware of antibiotic residue avoidance practices in dairy production is unknown, and it is unclear whether acquisition of such knowledge could affect purchasing behavior and perceptions of dairy farming. The objectives of this study were to assess consumers' perceptions about the quality and production of dairy products in the United States and determine whether educational materials on processes that limit the occurrence of antibiotic residues in milk can change consumers' perceptions of dairy products and purchasing behaviors. We surveyed 804 consumers and assigned them to 1 of 3 interventions: (1) a control arm (reading the content of the Dairy page of the USDA's myplate.gov website); (2) an educational brochure on the processes that prevent antibiotic residues in milk; and (3) a video on the same processes. We found that a majority (86.1%) of participants believe that the quality of dairy products in the United States is high, although many had concerns about the treatment of dairy animals and chemicals (pesticides, antibiotics, hormones) in dairy products. Compared with the control intervention, the brochure was associated with a significant decrease in the level of concern consumers had about chemicals in their milk [-0.20 points on a Likert scale, 95% confidence interval (CI), -0.32 to -0.08] and a significantly increased comfort in purchasing conventional dairy products (odds ratio 2.43, 95% CI, 1.62 to 3.66). The video was associated with even stronger effects: a 0.29-unit decrease in the level of concern about chemicals in milk (95% CI, -0.42 to -0.016) and 2.94 times greater odds of purchasing conventional dairy products (95%, CI 1.92 to 4.49). Although consumer food decision making is complex and driven by multiple factors, it appears that education about the processes that promote food safety can reassure consumers about their concerns and potentially affect purchasing habits.

Performance of commercial PCR assays to detect toxigenic Clostridioides difficile in the feces of puppies.

Clostridioides difficile is an important enteric pathogen that causes significant morbidity and mortality in humans. With community-acquired infections on the rise, it is important to identify reservoirs of the pathogen. Companion animals can be asymptomatic carriers of C. difficile and may therefore represent a reservoir, but epidemiological studies of C. difficile within the pet-owner unit are needed, along with validated methods to detect C. difficile in both people and animals. The goal of this study was to assess the performance of commercial qPCR assays and a multiplex PCR for C. difficile compared to toxigenic culture. These assays were tested on up to 103 fecal samples from puppies, a population in which the prevalence of C. difficile is the highest. The sensitivities, specificities, positive predictive values and negative predictive values were respectively 84.2%, 87.7%, 61.5%, and 95.9% for the Cepheid GeneXpert; 66.7%, 66.7%, 29.6%, and 90.9% for the DiaSorin Simplexa; and 94.4%, 85.0%, 65.4%, and 98.1%, for the multiplex qPCR. The agreement was highest between the GeneXpert and the multiplex PCR (90.1% agreement, with a kappa statistic of 0.77). For diagnostic purposes, the positive predictive values of the assays were low. However, the high sensitivities of the assays could render them useful for epidemiologic purposes.

Microbial Sharing between Pediatric Patients and Therapy Dogs during Hospital Animal-Assisted Intervention Programs.

Microbial sharing between humans and animals has been demonstrated in a variety of settings. However, the extent of microbial sharing that occurs within the healthcare setting during animal-assisted intervention programs is unknown. Understanding microbial transmission between patients and therapy dogs can provide important insights into potential health benefits for patients, in addition to addressing concerns regarding potential pathogen transmission that limits program utilization. This study evaluated for potential microbial sharing between pediatric patients and therapy dogs and tested whether patient-dog contact level and a dog decolonization protocol modified this sharing. Patients, therapy dogs, and the hospital environment were sampled before and after every group therapy session and samples underwent 16S rRNA sequencing to characterize microbial communities. Both patients and dogs experienced changes in the relative abundance and overall diversity of their nasal microbiome, suggesting that the exchange of microorganisms had occurred. Increased contact was associated with greater sharing between patients and therapy dogs, as well as between patients. A topical chlorhexidine-based dog decolonization was associated with decreased microbial sharing between therapy dogs and patients but did not significantly affect sharing between patients. These data suggest that the therapy dog is both a potential source of and a vehicle for the transfer of microorganisms to patients but not necessarily the only source. The relative contribution of other potential sources (e.g., other patients, the hospital environment) should be further explored to determine their relative importance.

Antimicrobial prescribing patterns in equine ambulatory practice.

Antimicrobial resistance is one of the most important public health risks facing our world today. Antimicrobials are commonly prescribed in equine veterinary medicine, but limited information exists documenting their use in practice. The goal of this study was to investigate antimicrobial prescription patterns in regards to prescription frequency, duration, drug class, clinician and affected body system in an equine ambulatory setting via retrospective analysis of billing and electronic medical records. Risk factors associated with antimicrobial prescription including the nature of the visit, submission of a culture, body system affected and clinician were assessed using multivariable regression. We found that antimicrobials were prescribed in 8.5 % of visits with a median number of 3.5 (IQR 0.8-12.2) animal-defined daily doses (ADD), defined as the number of daily doses of all antimicrobials prescribed to a patient at a single visit. Aminoglycosides were the most common class of antimicrobials prescribed and trimethoprim sulfamethoxazole was the most common drug prescribed overall. Amikacin was primarily used for patients presenting with musculoskeletal signs, and the median number of ADDs for visits where amikacin was prescribed was 1 (IQR 0.9-1.9), while the median number of antimicrobial ADDs for all other visits was 4.4 (IQR 0-14.1). Statistically significant differences in antimicrobial use patterns existed across clinicians, months, years and affected body systems. Horses presenting with ocular (OR 1199; 95 % CI 204-7,037; p < 0.001) and integumentary (OR 365; 95 % CI 87.2-1532; p < 0.001) signs were most likely to be prescribed an antimicrobial. Emergency visits (OR 5.61; 95 % CI 3.19-9.89; p < 0.001) and submission of a bacterial culture (OR 3.58; 95 % CI 2.11-6.09; p < 0.001) were associated with an increased likelihood of an antimicrobial prescription. This observational study was the first to quantitatively characterize antimicrobial use patterns in equine ambulatory practice in the United States, which is an important step needed to determine appropriateness of use and develop and evaluate antimicrobial stewardship guidelines.

Short communication: Comparison of the fecal bacterial communities in diarrheic and nondiarrheic dairy calves from multiple farms in southeastern Pennsylvania.

Diarrhea is a major cause of illness and death in preweaned calves and causes significant economic losses to producers. A better understanding of the fecal microbiota in diarrheic and nondiarrheic calves could lead to improved treatment and prevention strategies. The purpose of this study was to compare the fecal microbiota of diarrheic and nondiarrheic calves to improve our understanding of what constitutes a healthy fecal microbiota in preweaned calves. At each of 7 farms, fecal samples were obtained from 1 to 3 diarrheic Holstein dairy calves (2 to 17 d old at sampling time) and age-matched (within 5 d) nondiarrheic controls for a total of 20 samples. Calves were fed either acidified bulk milk, pasteurized or unpasteurized waste milk, or milk replacer depending on farm. Fecal samples were extracted for genomic DNA, PCR-amplified for the V1-V2 region of the 16S rRNA bacterial gene, sequenced on the Illumina MiSeq (Illumina Inc., San Diego, CA) platform, and analyzed using QIIME2. Firmicutes and Bacteroidetes were the most abundant phyla in both groups; Fusobacteria was numerically more abundant in the diarrheic group, whereas Proteobacteria and Actinobacteria were numerically more abundant in the nondiarrheic group. At the genus level, Bacteroides was the most abundant genus in both groups and was numerically more abundant in the nondiarrheic group. Results from the mixed-effects regression model showed that Faecalibacterium and Butyricimonas were more abundant in the nondiarrheic calves, whereas Clostridium and Peptostreptococcus were more abundant in the diarrheic calves. Our results indicate that commensal bacteria acquired in the neonatal period may have been replaced with potential pathogens in diarrheic calves, which may have contributed to the incidence of diarrhea either directly or indirectly.

Clostridioides difficile on dairy farms and potential risk to dairy farm workers.

Clostridioides difficile causes severe colitis in people and is a significant enteric pathogen in many species of animals, including swine, horses, and potentially cattle. C. difficile is shed in feces, and transmission occurs horizontally via the fecal-oral route. Livestock has been suggested as a potential reservoir for C. difficile, and while studies have shown that swine and farm workers can be colonized with identical clones of C. difficile, the zoonotic transmission of C. difficile from livestock to people has not been definitively demonstrated. The goal of this study was to determine whether dairy calves and dairy farm workers harbored genetically similar isolates of C. difficile. First, we validated a glove juice protocol for detecting C. difficile on farm workers' hands. We then visited 23 farms and collected 1) fecal samples from 92 dairy calves, 2) hand rinsates from 38 dairy farm workers, and 3) fecal samples from five of the dairy farm workers who were willing to submit them. All samples underwent anaerobic culture and qPCR to detect C. difficile. C. difficile was detected on 15 of the farms (65.2%, 95% confidence interval (CI) 42.7%-83.6%) and in 28 calves (30.4%, 95% CI 21.2-40.9%) but in none of the hand rinsates or human fecal samples. Thus, the zoonotic transmission of C. difficile on dairy farms could not be demonstrated, and dairy farmers did not appear to be at increased risk of acquiring C. difficile via the fecal-oral route.

Small and Large Animal Veterinarian Perceptions of Antimicrobial Use Metrics for Hospital-Based Stewardship in the United States.

Background: Robust measurement and tracking of antimicrobial use (AMU) is a fundamental component of stewardship interventions. Feeding back AMU metrics to individual clinicians is a common approach to changing prescribing behavior. Metrics must be meaningful and comprehensible to clinicians. Little is known about how veterinary clinicians working in the United States (US) hospital setting think about AMU metrics for antimicrobial stewardship. Objective: To identify hospital-based veterinary clinicians' attitudes toward audit and feedback of AMU metrics, their perceptions of different AMU metrics, and their response to receiving an individualized prescribing report. Methods: Semi-structured interviews were conducted with veterinarians working at two hospitals in the Eastern US. Interviews elicited perceptions of antimicrobial stewardship in veterinary medicine. Respondents were shown a personalized AMU Report characterizing their prescribing patterns relative to their peers and were asked to respond. Interviews were recorded, transcribed, and analyzed using the framework method with matrices. Results: Semi-structured interviews were conducted with 34 veterinary clinicians (22 small animal and 12 large animal). Respondents generally felt positive about the reports and were interested in seeing how their prescribing compared to that of their peers. Many respondents expressed doubt that the reports accurately captured the complexities of their prescribing decisions and found metrics associated with animal daily doses (ADDs) confusing. Only 13 (38.2%) respondents felt the reports would change how they used antimicrobials. When asked how the impact of the reports could be optimized, respondents recommended providing a more detailed explanation of how the AMU metrics were derived, education prior to report roll-out, guidance on how to interpret the metrics, and development of meaningful benchmarks for goal-setting. Conclusions: These findings provide important insight that can be used to design veterinary-specific AMU metrics as part of a stewardship intervention that are meaningful to clinicians and more likely to promote judicious prescribing.

The Effects of Frequency and Duration of Handling on the Development of Feline Upper Respiratory Infections in a Shelter Setting.

Reducing stress is important to maintaining the health of shelter cats and decreasing the risk of upper respiratory disease (URD). The aim of this study was to determine if the frequency and/or duration of daily routine handling of shelter cats affects the likelihood of URD development. At a closed admission shelter, each cat free of URD on intake was given a cage card for recording handling data. These data included: date and times when the cat was handled, duration of handling, if and when the cat developed signs of URD, and the handler identity. Cox regression was used to determine the relationship between these factors and URD development. We found cats that did not develop URD were handled significantly more than cats that did (1.1 times per day vs. 0.7 times per day, p < 0.001). Increased frequency of handling had a borderline significant effect on the hazard of developing URD (HR 0.37; CI: 0.13-1.1; p = 0.066). No other parameters were significantly associated with the development of URD; however, small sample size may be responsible for this finding. A larger study is needed to elucidate the relationship between handling and URD development.

Modification of empirical antimicrobial regimens in large animal medicine.

BACKGROUND: Empirical antimicrobial regimens can be modified following new diagnostic information or when empirical treatment fails. Little is known about the frequency or clinical context in which these modifications occur. We characterised these modifications in a large animal hospital to identify when antimicrobial use could be optimised. METHODS: Chart reviews were performed for all inpatients and outpatients administered antimicrobials at a large animal veterinary referral and teaching hospital in 2017-2018 (n=1163 visits) to determine when and why empirical regimens were modified. Multinomial logistic regression was performed to identify factors associated with reasons for modification. RESULTS: Empirical antimicrobial regimens were modified in 17.3 per cent of visits. The main reasons were parenteral-oral conversions in horses and failure of disease prevention or treatment in ruminants. Empirical therapy for disease prevention was more likely to be modified because of complications in ruminants and in animals on the emergency/critical care service. Empirical therapy for disease treatment was more often modified for reasons other than de-escalation in ruminants and in animals with longer lengths of stay. CONCLUSIONS: Empirical antimicrobial regimens were modified infrequently and mostly for purposes of parenteral-oral conversion in horses and lack of response in ruminants. De-escalation of antimicrobials administered for disease treatment, when guided by diagnostics, is a major tenet of judicious antimicrobial use. However, more research is needed to determine when and how antimicrobial regimens administered for disease prevention should be modified.