Photobiomodulation therapy in dogs undergoing TPLO after cranial cruciate ligament rupture shows promise but no statistically significant difference in a randomized trial.

OBJECTIVE: Effect of photobiomodulation therapy (PBMT) in patients with CCLR after TPLO surgery by measuring C-reactive protein (CRP), percentage weight bearing, lameness using a short form of a composite measure pain scale, evaluated by the clinician and owners, and surgical site infection. SAMPLE: 54 client-owned dogs with CCLR undergoing unilateral TPLO surgery were enrolled in this study between April 5, 2021, through April 10, 2022. METHODS: The study population was randomly assigned to either a treatment group receiving PMBT (24 dogs) or a control group (30 dogs). PMBT was performed on the treatment group immediately after induction, and 6 hours, 24 hours, 48 hours, and 8 weeks postoperatively. The control group received sham PMBT (device turned off) at the same time. Evaluation of CRP, CMPS-SF, evidence of SSI, and %WB were evaluated for all dogs 24 hours preoperatively, and then 24 hours, 48 hours, and 8 weeks postoperatively. Owners completed CMPS-SF and subjective evaluations weekly for 8 weeks postoperatively. RESULTS: No statistically significant differences were found between treatment groups when evaluating CRP, %WB, and CMPS-SF by the clinician and weekly evaluation of the CMPS-SF by owners. Although no statistically significant differences were found in patients developing surgical site infections between treatment groups, SSI was only observed in patients in the control group (5/30, 16.6%). Most were minor/superficial infections (4/30 13.3%), and a single dog (1/30, 3.3%) had a major/deep surgical site infection. CLINICAL RELEVANCE: Although with promising but not statistically significant differences between groups, surgical site infections may be reduced after PBMT application.

Effect of potassium tellurite concentration in a chromogenic agar medium on isolation of tellurite-resistant "Top Seven" Shiga toxin-producing Escherichia coli from ground beef.

The effect of potassium tellurite concentration in a chromogenic agar medium on the detection of tellurite-resistant "top seven" Shiga toxin-producing Escherichia coli (STEC) in beef was evaluated. Samples of ground beef were inoculated with tellurite-resistant STEC O26, O45, O103, O111, O121, O145, or O157 strains at geometric mean (±standard error of the mean) levels of 0, 49 (±1), 490 (±1), or 4900 (±1) CFU/10 g and enriched 1:10 (90 mL) in EC broth (40°C for 6 h). Following enrichment, aliquots of broth culture were treated by immunomagnetic separation with one of three pools of beads against STEC serogroups; pool I: O26, O45, and O121; pool II: O103, O111, and O145; and pool III: O157. After immunomagnetic separation, 50 µL of washed bead suspensions in buffered peptone water was spiral plated onto a modified Possé medium containing 0.5, 1.0, or 1.5 mg/L potassium tellurite, and incubated at 37°C for 18 h. Up to four isolated colonies were picked from each spiral plate based on expected colony phenotypes for STEC, and isolate identity was confirmed with an 11-plex PCR assay targeting the O serogroups and virulence genes. Overall, across all inoculum levels and strains, modified Possé media containing 0.5, 1.0, or 1.5 mg/L potassium tellurite each had a positive predictive value of 100%, and medium containing 0.5 mg/L potassium tellurite had numerically the highest sensitivity (100%) and negative predictive value (100%), which was significantly different from 1.5 mg/L (92.9% and 40.0%, respectively; P < 0.05). Similarly, there was an inverse relationship between potassium tellurite concentration and analytical specificity (number of colonies tested that were STEC-positive): 0.5 (1463 of 1482; 98.7%), 1.0 (1356 of 1411; 96.1%), and 1.5 mg/L (1187 of 1278; 92.9%; P < 0.05). These results suggest that 0.5 mg/L gives better performance than 1.0 or 1.5 mg/L of potassium tellurite in Possé medium for isolation of tellurite-resistant "top seven" STEC from ground beef.

A clinically utilized intravenous continuous rate infusion of diltiazem does not significantly decrease systolic function in healthy dogs.

OBJECTIVE: To determine if left ventricular systolic function on echocardiography, systemic blood pressure, and electrocardiography change with a clinically accepted intravenous (IV) diltiazem constant rate infusion (CRI) compared to a control. ANIMALS: 10 healthy client-owned adult dogs. PROCEDURES: Prospective, masked, crossover study from May 27, 2021, to August 22, 2021. Dogs were randomized to receive diltiazem (loading dose of 240 µg/kg, IV followed by a CRI of 6 µg/kg/min for 300 minutes) or the same volume of 5% dextrose in water (D5W) administered IV followed by the opposite intervention after a 7-day washout. Blood pressure was monitored during each CRI, and echocardiographic and electrocardiographic studies were performed immediately before the CRI and during the last hour of the CRI. RESULTS: Postdiltiazem systolic time interval (STI) (median, 0.30; range, 0.16 to 0.34) was significantly lower than post-D5W STI (median, 0.32; range, 0.22 to 0.40; P = .046). All other echocardiographic parameters did not differ significantly between each of the groups after receiving diltiazem or D5W. Systemic blood pressure did not change significantly with either diltiazem (P = .450) or D5W (P = .940), and none of the dogs became hypotensive at any point in the study. Expectedly, negative dromotropy was observed with diltiazem. CLINICAL RELEVANCE: A significant decrease in left ventricular systolic function was not appreciated in healthy dogs receiving diltiazem at a clinically accepted intravenous infusion rate at this dosing regimen. Further studies are needed in dogs with cardiac disease.

The application, value, and impact of outcomes research in animal health and veterinary medicine.

Outcomes research is a relatively recent field of study in animal health and veterinary medicine despite being well-established in human medicine. As the field of animal health is broad-ranging in terms of animal species, objectives, research methodologies, design, analysis, values, and outcomes, there is inherent versatility in the application and impact of the discipline of outcomes research to a variety of stakeholders. The major themes of outcomes relevant to the animal health industry have been distilled down to include, but are not limited to, health, production, economics, and marketing. An outcomes research approach considers an element of value along with an outcome of interest, setting it apart from traditional research approaches. Elements of value are determined by the stakeholders' use of products and/or services that meet or exceed functional, emotional, life-changing, and/or societal needs. Stakeholder perception of value depends on many factors such as the purpose of the animal (e.g., companion vs. food production) and the stakeholder's role (e.g., veterinarian, client, pet-owner, producer, consumer, government official, industry representative, policy holder). Key areas of application of outcomes research principles include comparative medicine, veterinary product development, and post-licensure evaluation of veterinary pharmaceuticals and/or biologics. Topics currently trending in human healthcare outcomes research, such as drug pricing, precision medicine, or the use of real-world evidence, offer novel and interesting perspectives for addressing themes common to the animal health sector. An approach that evaluates the benefits of practices and interventions to veterinary patients and society while maximizing outcomes is paramount to combating many current and future scientific challenges where feeding the world, caring for our aging companion animals, and implementing novel technologies in companion animal medicine and in production animal agriculture are at the forefront of our industry goals.

Economic assessments from experimental research trials of feedlot cattle health and performance: a scoping review.

Animal husbandry decisions for feedlot cattle may be based on economic or financial impacts reported from livestock research trials comparing interventions such as health practices or performance technologies. Despite the importance of economic assessments to production management decisions, there are no consensus guidelines for their methods or reporting. Thus, we hypothesized that methods and reporting of economic assessments in the scientific literature are inconsistent. This scoping review describes the types of economic assessments used to evaluate the costs and benefits of interventions in feedlot trials, how measured health and performance outcomes are utilized in economic evaluations, and the completeness of reporting. A structured search was used to retrieve peer-reviewed articles (published in English) on experimental trials performed in Australia, North America, or South Africa, which reported feedlot cattle health, performance, or carcass characteristics and included an economic outcome. A total of 7,086 articles were screened for eligibility; 91 articles (comprising 113 trials) met the inclusion criteria. Trial characteristics, methods, and reporting data were extracted. A primary outcome was stated in only 36% (41/113) of the trials. Of these 41 trials, an economic outcome was reported as a primary outcome in 18 (44%). Methodology for the economic assessment was reported for 54 trials (48%), the type of economic assessment was explicitly stated for 21 trials (19%), and both the type of economic assessment and methodology used were reported for 29 trials (26%); neither were reported for nine trials (8%). Eight types of economic assessments were explicitly reported: cost-effectiveness, cost-benefit analysis, enterprise analysis, partial budget, break-even analysis, profitability, decision analysis, and economic advantage. From the trials that did not report an assessment type, three were identified: partial budget, enterprise analysis, and gross margin analysis. Overall, only 32 trials (28%) reported economics as an outcome of interest, the methodology used or the type of assessment, and values, sources, and dates for at least some of the price data used in the analysis. Given the variability in methods and inconsistent reporting for feedlot trials identified by this scoping review, a guideline to facilitate consistency on appropriate methods and reporting is warranted.

A systematic review and meta-analysis of published literature on prevalence of non-O157 Shiga toxin-producing Escherichia coli serogroups (O26, O45, O103, O111, O121, and O145) and virulence genes in feces, hides, and carcasses of pre- and peri-harvest cattle worldwide.

OBJECTIVE: The objective of this study was to summarize peer-reviewed literature on the prevalence and concentration of non-O157 STEC (O26, O45, O103, O111, O121, and O145) serogroups and virulence genes (stx and eae) in fecal, hide, and carcass samples in pre- and peri-harvest cattle worldwide, using a systematic review of the literature and meta-analyses. DATA SYNTHESIS: Seventy articles were eligible for meta-analysis inclusion; data from 65 articles were subjected to random-effects meta-analysis models to yield fecal prevalence estimates. Meta-regression models were built to explore variables contributing to the between-study heterogeneity. RESULTS: Worldwide pooled non-O157 serogroup, STEC, and EHEC fecal prevalence estimates (95% confidence interval) were 4.7% (3.4-6.3%), 0.7% (0.5-0.8%), and 1.0% (0.8-1.1%), respectively. Fecal prevalence estimates significantly differed by geographic region (P < 0.01) for each outcome classification. Meta-regression analyses identified region, cattle type, and specimen type as factors that contribute to heterogeneity for worldwide fecal prevalence estimates. CONCLUSIONS: The prevalence of these global foodborne pathogens in the cattle reservoir is widespread and highly variable by region. The scarcity of prevalence and concentration data for hide and carcass matrices identifies a large data gap in the literature as these are the closest proxies for potential beef contamination at harvest.

Association between Tulathromycin Treatment for Bovine Respiratory Disease and Antimicrobial Resistance Profiles among Gut Commensals and Foodborne Bacterial Pathogens Isolated from Feces of Beef Steers.

ABSTRACT: This study was conducted to evaluate the association between a therapeutic dose of tulathromycin for bovine respiratory disease in beef steers and the antimicrobial and multidrug resistance profiles of the gastrointestinal tract commensals Escherichia coli and Enterococcus spp. and the foodborne pathogens Salmonella enterica and Campylobacter spp. isolated from fecal samples. Individual fecal samples were collected on days 0, 14, and 28 from 70 beef steers that were housed in a single pen and had been treated or not treated with tulathromycin. Samples were cultured for bacterial isolation, and isolates were tested for antimicrobial susceptibility with the broth microdilution method to determine the MICs of clinically relevant antimicrobials used in both human and veterinary medicine. Generalized linear mixed effects models were fitted to estimate the prevalence of the bacterial species and the prevalence of resistant isolates over time and between treated and nontreated cattle and of multidrug-resistant isolates. Model-adjusted mean prevalences of E. coli, Enterococcus spp., S. enterica, and Campylobacter spp. were 99.5, 85.9, 1.5, and 17.7%, respectively. The prevalence of erythromycin-resistant Enterococcus spp. was significantly higher on day 14 (59.7%) than on day 28 (22.2%). A higher prevalence of erythromycin-resistant Enterococcus spp. was found in samples from treated (59.3%) than in samples from nontreated (27.6%) animals. Multidrug resistance (three or more antimicrobial classes) was observed in 8.4% of E. coli isolates and 62.7% of Enterococcus isolates. The administration of tulathromycin was significantly associated with an increased prevalence of erythromycin-resistant Enterococcus spp. isolates.

Assessment of bovine respiratory disease progression in calves challenged with bovine herpesvirus 1 and Mannheimia haemolytica using point-of-care and laboratory-based blood leukocyte differential assays.

Blood leukocyte differentials can be useful for understanding changes associated with bovine respiratory disease (BRD) progression. By improving turnaround time, point-of-care leukocyte differential assays (PCLD) may provide logistical advantages to laboratory-based assays. Our objective was to assess BRD progression in steers challenged with bovine herpesvirus 1 and Mannheimia haemolytica using point-of-care and laboratory-based blood leukocyte differentials. Thirty Holstein steers (average body weight of 211 kg + 2.4 kg) were inoculated intranasally on day 0 with bovine herpesvirus 1 and intrabronchially on day 6 with Mannheimia haemolytica. Blood leukocytes differentials were measured using both assays from study days 0 to 13. Linear mixed models were fitted to evaluate the associations between: (1) the type of assay (laboratory-based or PCLD) with respect to leukocyte, lymphocyte, and neutrophil concentrations; (2) study day with cell concentrations; and (3) cell concentrations with lung consolidation measured at necropsy. Point-of-care leukocyte, lymphocyte, and neutrophil concentrations were significantly associated (P < 0.05) with the respective cell concentrations obtained from the laboratory-based leukocyte differential. Cell concentrations reported by both assays differed significantly (P < 0.05) over time, indicating shifts from healthy to viral and bacterial disease states. Lymphocyte concentrations, lymphocyte/neutrophil ratios obtained from both assays, and band neutrophil concentrations from the laboratory-based assay were significantly associated (P < 0.05) with lung consolidation, enhancing assessments of disease severity. The PCLD may be a useful alternative to assess BRD progression when laboratory-based leukocyte differentials are impractical.

In Vitro Infection Dynamics of Japanese Encephalitis Virus in Established Porcine Cell Lines.

Japanese encephalitis virus (JEV) is a zoonotic mosquito-borne pathogen that regularly causes severe neurological disease in humans in Southeast Asia and the Western Pacific region. Pigs are one of the main amplifying hosts of JEV and play a central role in the virus transmission cycle. The objective of this study was to identify in vitro cell systems to investigate early effects of JEV infection including viral replication and host cell death. Here, we demonstrate the susceptibility of several porcine cell lines to the attenuated genotype III JEV strain SA14-14-2. Monolayers of porcine nasal turbinate (PT-K75), kidney (SK-RST), testis (ST), and monocyte-derived macrophage (CΔ2+) cells were infected with SA14-14-2 for up to five days at a multiplicity of infection (MOI) of 0.1. The hamster kidney cell line BHK-21, previously shown to be susceptible to SA14-14-2, was used as a positive control. Culture supernatants and cells were collected between 0 and 120 h post infection (hpi), and monolayers were observed for cytopathic effect (CPE) using brightfield microscopy. The number of infectious virus particles was quantified by plaque assay and cell viability was determined using trypan blue staining. An indirect immunofluorescence assay was used to detect the presence of JEV NS1 antigens in cells infected at 1 MOI. All four porcine cell lines demonstrated susceptibility to SA14-14-2 and produced infectious virus by 12 hpi. Virus titers peaked at 48 hpi in CΔ2+, BHK-21, and SK-RST cells, at 72 hpi in PT-K75, and at 120 hpi in ST cells. CPE was visible in infected CΔ2+ and BHK-21 cells, but not the other three cell lines. The proportion of viable cells, as measured by trypan blue exclusion, declined after 24 hpi in BHK-21 and 48 hpi in CΔ2+ cells, but did not substantially decline in SK-RST, PT-K75 or ST cells. At 48 hpi, JEV NS1 was detected in all infected cell lines by fluorescence microscopy. These findings demonstrate several porcine cell lines which have the potential to serve as useful research tools for investigating JEV infection dynamics and host cell mechanisms in a natural amplifying host species, such as pigs, in vitro.

Evaluation of specific immunoglobulin A in nasal secretions and neutralizing antibodies in serum collected at multiple time points from young beef calves following intranasal or subcutaneous administration of a modified-live bovine respiratory syncytial virus vaccine.

OBJECTIVE: To determine anti-bovine respiratory syncytial virus (BRSV) antibody titers for nasal secretions and serum from beef calves following administration of a modified-live (MLV) BRSV vaccine. ANIMALS: 60 healthy newborn purebred beef calves. PROCEDURES: Calves were randomly assigned to 1 of 3 groups: intranasal (IN)-SC (IN MLV BRSV vaccine within 24 hours of birth and SC MLV BRSV vaccine at 2 months of age), SC-IN (SC MLV BRSV vaccine within 24 hours of birth and IN MLV BRSV vaccine at 2 months of age), or NO-IN (no vaccine within 24 hours of birth and IN MLV BRSV vaccine at 2 months of age). Nasal secretion and serum samples were collected for determination of anti-BRSV antibodies within 24 hours of birth and 2 and 6 months of age. RESULTS: Titers of anti-BRSV IgA antibodies in nasal secretions and BRSV neutralizing antibodies in serum were similar among groups at each sampling time. Within 24 hours of birth, nasal anti-BRSV IgA titers were negligible. At 2 months, mean nasal anti-BRSV IgA titers for calves in IN-SC, SC-IN, and NO-IN groups were 192.84, 224.49, and 114.71, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Concentrations of anti-BRSV IgA antibodies in the nasal secretions and BRSV neutralizing antibodies in the serum of young beef calves following an MLV BRSV vaccine protocol that consisted of IN or SC vaccine within 24 hours of birth and vice versa at 2 months of age were not different from that following only an IN MLV BRSV vaccine at 2 months of age. However, the lack of any differences may have been attributed to other factors.

Weather conditions associated with death attributed to bovine respiratory disease complex in high-risk auction market-sourced male beef calves.

OBJECTIVE: To evaluate associations between weather conditions and management factors with the incidence of death attributable to bovine respiratory disease complex (BRDC) in high-risk auction-sourced beef calves. ANIMALS: Cohorts (n = 3,339) of male beef calves (545,866) purchased by 1 large cattle feeding operation from 216 locations and transported to 1 of 89 feeding locations (backgrounding location or feedlot) with similar management protocols. PROCEDURES: Associations between weather conditions and management factors on the day of purchase (day P) and during the first week at the feeding location and cumulative BRDC mortality incidence within the first 60 days on feed were estimated in a mixed-effects negative binomial regression model. RESULTS: Significant factors in the final model were weaning status; degree of commingling; body weight; transport distance; season; precipitation, mean wind speed, and maximum environmental temperature on day P; environmental temperature range in the first week after arrival at the feeding location; and interactions between distance and wind speed and between body weight and maximum environmental temperature. Precipitation and wind speed on day P were associated with lower cumulative BRDC mortality incidence, but wind speed was associated only among calves transported long distances (≥ 1,082.4 km). Higher mean maximum temperature on day P increased the incidence of cumulative mortality among calves with low body weights (< 275.5 kg). CONCLUSIONS AND CLINICAL RELEVANCE: Several weather conditions on day P and during the first week after arrival were associated with incidence of BRDC mortality. The results may have implications for health- and economic-risk management, especially for high-risk calves and calves that are transported long distances.

Detecting and quantifying marijuana metabolites in serum and urine of 19 dogs affected by marijuana toxicity.

Veterinarians diagnose marijuana toxicity based on clinical signs and history, or in conjunction with an over-the-counter (OTC) human urine drug screen. With the legalization of recreational marijuana use becoming more prevalent in the United States, a more accurate test to aid in the diagnosis of canine marijuana toxicity is needed. We collected urine and serum samples from 19 dogs with confirmed or suspected marijuana toxicosis from multiple veterinary hospitals and analyzed them with a novel UPLC-MS/MS method. Calibrations from 0.1 to 100 ng/mL and QC materials were prepared. Samples were extracted, purified, and eluted with solid-phase extraction. Urine samples were tested with an OTC human urine drug screen. The limit of detection (LOD) and lower limit of quantification (LLOQ) ranges for marijuana metabolites in serum were 0.05-0.25 ng/mL and 0.1-0.5 ng/mL, respectively. In urine, the LOD and LLOQ ranges for the metabolites were 0.05-0.1 ng/mL and 0.1-0.5 ng/mL, respectively. In serum, median and range of metabolite concentrations (ng/mL) detected included: THC, 65.0 (0.14-160); 11-OH-Δ9-THC, 4.78 (1.15-17.8); 11-nor-9-carboxy-Δ9-THC, 2.18 (0.71-7.79); CBD, 0.28 (0.11-82.5); and THC-glucuronide, 2.05 (0.72-18.3). In the 19 urine samples, metabolite: creatinine (ng: mg) values detected included: THC, 0.22 (0.05-0.74); 11-OH-Δ9-THC, 0; 11-nor-9-carboxy-Δ9-THC, 1.32 (0.16-11.2); CBD, 0.19 (0.12-0.26); THC-COOH-glucuronide, 0.08 (0.04-0.11); and THC-glucuronide, 0.98 (0.25-10.7). Twenty of 21 urine samples tested negative for THC on the urine drug screen. All 19 serum samples contained quantifiable concentrations of THC using our novel UPLC-MS/MS method. Utilizing a UPLC-MS/MS method can be a useful aid in the diagnosis of marijuana toxicosis in dogs, whereas using an OTC human urine drug test is not a useful test for confirming marijuana exposure in dogs because of the low concentration of THC-COOH in urine.

Identification, Shiga toxin subtypes and prevalence of minor serogroups of Shiga toxin-producing Escherichia coli in feedlot cattle feces.

Shiga toxin-producing Escherichia coli (STEC) are foodborne pathogens that cause illnesses in humans ranging from mild to hemorrhagic enteritis with complications of hemolytic uremic syndrome and even death. Cattle are a major reservoir of STEC, which reside in the hindgut and are shed in the feces, a major source of food and water contaminations. Seven serogroups, O26, O45, O103, O111, O121, O145 and O157, called 'top-7', are responsible for the majority of human STEC infections in North America. Additionally, 151 serogroups of E. coli are known to carry Shiga toxin genes (stx). Not much is known about fecal shedding and prevalence and virulence potential of STEC other than the top-7. Our primary objectives were to identify serogroups of STEC strains, other than the top-7, isolated from cattle feces and subtype stx genes to assess their virulence potential. Additional objective was to develop and validate a novel multiplex PCR assay to detect and determine prevalence of six serogroups, O2, O74, O109, O131, O168, and O171, in cattle feces. A total of 351 strains, positive for stx gene and negative for the top-7 serogroups, isolated from feedlot cattle feces were used in the study. Of the 351 strains, 291 belonged to 16 serogroups and 60 could not be serogrouped. Among the 351 strains, 63 (17.9%) carried stx1 gene and 300 (82.1%) carried stx2, including 12 strains positive for both. The majority of the stx1 and stx2 were of stx1a (47/63; 74.6%) and stx2a subtypes (234/300; 78%), respectively, which are often associated with human infections. A novel multiplex PCR assay developed and validated to detect six serogroups, O2, O74, O109, O131, O168, and O171, which accounted for 86.9% of the STEC strains identified, was utilized to determine their prevalence in fecal samples (n = 576) collected from a commercial feedlot. Four serogroups, O2, O109, O168, and O171 were identified as the dominant serogroups prevalent in cattle feces. In conclusion, cattle shed in the feces a number of STEC serogroups, other than the top-7, and the majority of the strains isolated possessed stx2, particularly of the subtype 2a, suggesting their potential risk to cause human infections.

Cantaloupe Facilitates Salmonella Typhimurium Survival Within and Transmission Among Adult House Flies (Musca domestica L.).

Salmonella enterica serovar Typhimurium is a pathogen harbored by livestock and shed in their feces, which serves as an acquisition source for adult house flies. This study used a green fluorescent protein (GFP) expressing strain of Salmonella Typhimurium to assess its acquisition by and survival within house flies, and transmission from and between flies in the presence or absence of cantaloupe. Female house flies were exposed to manure inoculated with either sterile phosphate-buffered saline or GFP-Salmonella Typhimurium for 12 h, then used in four experiments each performed over 24 h. Experiment 1 assessed the survival of GFP-Salmonella Typhimurium within inoculated flies. Experiment 2 determined transmission of GFP-Salmonella Typhimurium from inoculated flies to cantaloupe. Experiment 3 assessed fly acquisition of GFP-Salmonella Typhimurium from inoculated cantaloupe. Experiment 4 evaluated transmission of GFP-Salmonella Typhimurium between inoculated flies and uninoculated flies in the presence and absence of cantaloupe. GFP-Salmonella Typhimurium survived in inoculated flies but bacterial abundance decreased between 0 and 6 h without cantaloupe present and between 0 and 6 h and 6 and 24 h with cantaloupe present. Uninoculated flies acquired GFP-Salmonella Typhimurium from inoculated cantaloupe and bacterial abundance increased in cantaloupe and flies from 6 to 24 h. More uninoculated flies exposed to inoculated flies acquired GFP-Salmonella Typhimurium when cantaloupe was present than when absent. We infer that the presence of a shared food source facilitated the transfer of GFP-Salmonella Typhimurium from inoculated to uninoculated flies. Our study demonstrated that house flies acquired, harbored, and excreted viable GFP-Salmonella Typhimurium and transferred bacteria to food and each other. Understanding the dynamics of bacterial acquisition and transmission of bacteria between flies and food helps in assessing the risk flies pose to food safety and human health.

Effectiveness of a Direct-Fed Microbial Product Containing Lactobacillus acidophilus and Lactobacillus casei in Reducing Fecal Shedding of Escherichia coli O157:H7 in Commercial Feedlot Cattle.

The objective of this study was to evaluate the effectiveness of a direct-fed microbial (DFM) product in reducing fecal shedding of Escherichia coli O157:H7 in finishing commercial feedlot cattle in Kansas (KS) and Nebraska (NE). Utilizing a randomized complete block design within the feedlot (KS, n = 1; NE, n = 1), cattle were randomly allocated to 20 pens, grouped in blocks of two based on allocation date, and then, within the block, randomly assigned to a treatment group (DFM or negative control). The DFM product was included in the diet at a targeted daily dose of 1 × 109 colony-forming units (CFU) of the Lactobacillus acidophilus and Lactobacillus casei combination per animal for at least 60 d before sampling. Feedlots were sampled for four consecutive weeks; weekly sampling consisted of collecting 20 pen floor fecal samples per pen. Fecal samples were subjected to culture-based methods for detection and isolation of E. coli O157, and positive samples were quantified using real-time polymerase chain reaction. Primary outcomes of interest were fecal prevalence of E. coli O157:H7 and E. coli O157 supershedding (≥104 CFU/g of feces) prevalence. Data for each feedlot were analyzed at the pen level using mixed models accounting for the study design features. Model-adjusted mean E. coli O157:H7 fecal prevalence estimates (standard error of the mean [SEM]) for DFM and control groups were 8.2% (SEM = 2.2%) and 9.9% (SEM = 2.5%) in KS and 14.6% (SEM = 2.8%) versus 14.3% (SEM = 2.6%) in NE; prevalence did not differ significantly between treatment groups at either site (KS, p = 0.51; NE, p = 0.92). Mean E. coli O157 supershedding prevalence estimates for DFM and control groups were 2.2% (SEM = 0.7%) versus 1.8% (SEM = 0.7%) in KS (p = 0.66) and 6.7% (SEM = 1.5%) versus 3.2% (SEM = 1.0%) in NE (p = 0.04). In conclusion, administering the DFM product in the finishing diet of feedlot cattle did not significantly reduce E. coli O157:H7 fecal prevalence or supershedding prevalence in study pens at either commercial feedlot.

Performance of Chromogenic Agar Media for Isolation of Shiga Toxin-Producing Escherichia coli from Ground Beef.

ABSTRACT: The performance of three chromogenic agar media for detection of the "top seven" Shiga toxin-producing Escherichia coli (STEC) in beef was compared. Samples of retail ground beef were inoculated with STEC O26, O45, O103, O111, O121, O145, or O157 at geometric mean (±standard error of the mean) levels of 0, 48 (±1), 420 (±1), 4,100 (±1), or 45,000 (±1) CFU/10 g and enriched 1:10 (90 mL) in EC broth (40°C for 6 h). Following enrichment, aliquots of broth culture were treated by immunomagnetic separation with one of three pools of beads against the seven STEC serogroups: pool I, O26, O45, and O121; pool II, O103, O111, and O145; and pool III, O157. After immunomagnetic separation, 50 µL of washed bead suspensions in buffered peptone water were spiral plated onto modified Rainbow Agar O157 (mRBA), CHROMagar STEC (CS), or modified Possé differential medium (mPossé2) and incubated at 37°C for 18 h. Up to six isolated colonies were picked from each spiral plate based on expected colony phenotypes for STEC on the respective media, and isolate identity was confirmed with an 11-plex PCR assay targeting the O serogroups and virulence genes. Overall, mRBA had the highest sensitivity (99.2%), correctly detecting a significantly higher proportion of STEC serogroups than either CS (79.4%; P < 0.05) or mPossé2 (91.7%; P < 0.05). mRBA also had the highest negative predictive value (90.0%), correctly identifying a significantly higher proportion of true-negative samples compared with CS (25.7%; P < 0.05) and mPossé2 (46.2%; P < 0.05). However, mRBA also had the lowest analytical specificity of 83.2% (P < 0.05), yielding the lowest proportion of colonies tested that were STEC positive (3,548 of 4,263) compared with 97.7% (3,607 of 3,693) for mPossé2 and 98.0% (2,875 of 2,935) for CS. Reduced specificity results in more work and higher expense due to the increased number of colonies that must be tested. Further improvements in agar culture media for non-O157 STEC isolation are needed.

Evaluation of a 3-dimensional ultrasound device for noninvasive measurement of urinary bladder volume in dogs.

BACKGROUND: The BladderScan Prime Plus (BPP; Verathon, Bothell, Washington) is an application-specific, three-dimensional ultrasound device used for human, point-of-care volumetry of the urinary bladder. OBJECTIVE: To estimate the BPP's accuracy, repeatability, and optimized settings for assessing urinary bladder volumes in dogs, a variable utilized in assessing micturition disorders. ANIMALS: Twenty-four, client-owned, healthy, male dogs presenting for routine examination. METHODS: Prospective examinations were conducted by an experienced ultrasonographer and a novice, selecting the BPP's "man" or "child" setting, and were compared to urine volume obtained by catheterization. RESULTS: Mean urine volume significantly varied by operator (P = .05), device setting (P < .001), and weight (P = .01); the "man" setting produced mean volumes nearer to catheterized volumes. The mean difference between BPP's "man" setting and catheterized volume was 0.88 mL, with maximal positive and negative disagreement of +23.2 mL to -55.3 mL (SD 19.0). Percent disagreement between BPP and catheterized volumes demonstrated a mean of -4.5%, with maximal positive and negative disagreement of +58.1% to -74.1% (SD 34.9). The experienced operator recorded volumes significantly (P = .05) higher than the novice, with difference in means of 3.2 mL. In dogs weighing >5.5 kg (n = 18/24), mean difference between BPP's "man" setting and catheterized measurements, regardless of operator, was not significant. CONCLUSIONS: Although small magnitude interuser variability is present in BPP examinations, the device provides accurate, though imprecise quantification of bladder volume in canids weighing >5.5 kg.

Associations Between Season, Processing Plant, and Hide Cleanliness Scores with Prevalence and Concentration of Major Shiga Toxin-Producing Escherichia coli on Beef Cattle Hides.

The objectives of this study were (1) to estimate the prevalence and concentration of the seven major Shiga toxin-producing Escherichia coli (STEC) serogroups (O26, O45, O103, O111, O121, O145, and O157), collectively called STEC-7, on cattle hides collected in different seasons and beef processing plants; and (2) to determine associations of season, plant, and hide cleanliness scores with the prevalence and concentration of STEC-7. A total of 720 hide surface samples (240/season) were collected over three seasons (summer and fall 2015 and spring 2016) from beef cattle carcasses in four commercial processing plants in the United States. Samples were subjected to selective culture and spiral plating methods. Overall model-adjusted mean prevalence (95% confidence interval) was 0.3% (0.03-2.3%) for STEC O26; 0.05% (<0.01-8.5%) for STEC O45; 0.2% (0.02-1.9%) for STEC O103; 0.05% (<0.01-8.5%) for STEC O145; and 3.1% (0.6-15.2%) for STEC O157. Four percent of hide samples were enumerable for STEC O157; mean concentration (standard deviation) = 2.1 (0.7) log10 colony-forming units (CFUs)/100 cm2. No samples were enumerable for non-O157 STEC. Hide-on prevalence of STEC O157 and STEC non-O157 (specifically of STEC O103) was higher in summer and spring, respectively. Across seasons and plants, the most common STEC non-O157 serogroups in this study (O26 and O103) were associated with a higher prevalence of STEC O157. Season and plant played a role in prevalence and concentration of STEC in beef cattle hides, varying by serogroup. Tailoring mitigation strategies at the plant can be challenging and processors would benefit from supplementary preharvest interventions to reduce overall contamination pressure at the plant, especially in fall and spring months when hide-on prevalence of STEC non-O157 is higher.

Association between antimicrobial drug class selection for treatment and retreatment of bovine respiratory disease and health, performance, and carcass quality outcomes in feedlot cattle.

Treatment and control of bovine respiratory disease (BRD) is predicated on the use of two categories of antimicrobials, namely bacteriostatic drugs that inhibit bacterial growth and replication (STATIC), and bactericidal drugs that kill bacteria in in vitro culture systems (CIDAL). Recently, we reported that initial BRD treatment with a STATIC antimicrobial followed by retreatment with a CIDAL antimicrobial was associated with a higher frequency of multidrug-resistant bacteria isolated from field cases of BRD submitted to a veterinary diagnostic laboratory. The present study was conducted to test the hypothesis that calves administered the same class of antimicrobial for first and second BRD treatment (i.e., CIDAL-CIDAL or STATIC-STATIC) would have improved health and performance outcomes at the feedlot compared to calves that received a different antimicrobial class for retreatment (i.e., STATIC-CIDAL or CIDAL-STATIC). The association between antimicrobial treatments and health, performance, and carcass quality outcomes were determined by a retrospective analysis of 4,252 BRD treatment records from a commercial feedlot operation collected from 2001 to 2005. Data were compared using generalized linear mixed statistical models that included gender, season, and arrival weight as covariates. The mean (±SE) probability of BRD cases identified as requiring four or more treatments compared to three treatments was greater in calves that received STATIC-CIDAL (73.58 ± 2.38%) or STATIC-STATIC (71.32 ± 2.52%) first and second antimicrobial treatments compared to calves receiving CIDAL-CIDAL (50.35 ± 3.46%) first and second treatments (P < 0.001). Calves receiving CIDAL-CIDAL first and second treatments also had an increased average daily gain (1.11 ± 0.03 kg/d) compared to calves receiving STATIC-CIDAL (0.95 ± 0.03 kg/d) and STATIC-STATIC (0.84 ± 0.02 kg/d) treatments (P < 0.001). Furthermore, CIDAL-CIDAL-treated calves had a higher probability of a choice quality grade at slaughter (36.44 ± 4.80%) compared to STATIC-CIDAL calves (28.09 ± 3.88%) (P = 0.037). There was no effect of antimicrobial treatment combination on BRD mortality (P = 0.855) or yield grade (P = 0.240) outcomes. These observations suggest that consideration should be given to antimicrobial pharmacodynamics when selecting drugs for retreatment of BRD. These findings have implications for developing BRD treatment protocols that address both post-treatment production and antimicrobial stewardship concerns.