Prevalence, Risk Factors, and Antimicrobial Resistance Profile of Respiratory Pathogens Isolated From Suckling Beef Calves to Reprocessing at the Feedlot: A Longitudinal Study.

Here, we investigated the prevalence and risk factors for the presence of Histophilus somni, Mannheimia haemolytica, Mycoplasma bovis, and Pasteurella multocida in the respiratory tract of calves from the spring processing to the reprocessing at feedlots. Additionally, we characterized, phenotypically and genotypically, the antimicrobial resistance (AMR) profile of the four species. Calves from 22 cow-calf operations were enrolled in the study (n = 30 calves per operation) and sampled by deep nasopharyngeal swabs at three time points: spring processing, weaning, or induction into feedlots, and at reprocessing at the feedlot. Isolates were tested for susceptibility using the minimum inhibitory concentration (MIC) test against commonly administered antimicrobials. Additionally, a subset of isolates underwent whole-genome sequencing to infer presence of AMR genes and resistance determinants. Among studied pathogens, P. multocida was the most prevalent species, regardless of time point, followed by M. haemolytica, M. bovis, and H. somni. For M. bovis, a sharp increase in prevalence was detected at the reprocessing sampling, whereas for P. multocida, an increase in prevalence was observed at the weaning/induction sampling. Comingling and co-location of feedlots were not associated with prevalence of any respiratory pathogen. In terms of AMR, resistance against macrolides was prevalent in M. bovis, with most isolates resistant against tildipirosin, tilmicosin, and tylosin. In general, there was limited evidence to support an increase in resistance rates of respiratory bacteria from the spring processing to reprocessing at feedlots, with the exception of florfenicol resistance in M. bovis, which increased at reprocessing. Metaphylactic administration of tetracyclines at feedlot induction was not associated with the MIC of tetracyclines in any respiratory bacteria. Conversely, there were clear associations between the parenteral use of macrolides as metaphylaxis at the feedlot induction, and increased MIC against macrolides in P. multocida, M. haemolytica, and H. somni. Overall, the AMR phenotypes were corroborated by presence of AMR genes. We hypothesize that the administration of macrolides such as tulathromycin at feedlot induction contributes to historical changes in macrolides MIC data of respiratory bacteria of beef cattle.

Prevalence and Risk Factors Associated With Antimicrobial Resistance in Bacteria Related to Bovine Respiratory Disease-A Broad Cross-Sectional Study of Beef Cattle at Entry Into Canadian Feedlots.

A broad, cross-sectional study of beef cattle at entry into Canadian feedlots investigated the prevalence and epidemiology of antimicrobial resistance (AMR) in Mannheimia haemolytica, Pasteurella multocida, Histophilus somni, and Mycoplasma bovis, bacterial members of the bovine respiratory disease (BRD) complex. Upon feedlot arrival and before antimicrobials were administered at the feedlot, deep nasopharyngeal swabs were collected from 2,824 feedlot cattle in southern and central Alberta, Canada. Data on the date of feedlot arrival, cattle type (beef, dairy), sex (heifer, bull, steer), weight (kg), age class (calf, yearling), source (ranch direct, auction barn, backgrounding operations), risk of developing BRD (high, low), and weather conditions at arrival (temperature, precipitation, and estimated wind speed) were obtained. Mannheimia haemolytica, P. multocida, and H. somni isolates with multidrug-resistant (MDR) profiles associated with the presence of integrative and conjugative elements were isolated more often from dairy-type than from beef-type cattle. Our results showed that beef-type cattle from backgrounding operations presented higher odds of AMR bacteria as compared to auction-derived calves. Oxytetracycline resistance was the most frequently observed resistance across all Pasteurellaceae species and cattle types. Mycoplasma bovis exhibited high macrolide minimum inhibitory concentrations in both cattle types. Whether these MDR isolates establish and persist within the feedlot environment, requires further evaluation.

Impact of meloxicam on respiratory virus titers and health outcomes when administered concurrently with a modified live respiratory vaccine in abruptly weaned beef steers.

Abruptly weaned crossbred steer calves (N = 271) were used in a randomized, blinded 2-arm clinical trial to assess the impact of a long-acting non-steroidal anti-inflammatory drug on bovine herpesvirus type 1, bovine respiratory syncytial virus, parainfluenza virus type 3, and coronavirus titers and health outcomes when administered concurrently with a modified live respiratory vaccine upon arrival at a feedlot. Treatment groups included a control (saline; n = 135) and an experimental group (injectable meloxicam; n = 136). Viral antibody titers and body weight were measured on arrival, day 7, and day 21, along with a final weight on day 45. Body weight and antibody titers for all viruses increased over time (P < 0.001); however, there were no differences by treatment group or a significant group × time interaction when evaluated using repeated measures analysis of variance. Interestingly, the use of meloxicam was associated with increased treatment risk (P < 0.05). In conclusion, the administration of meloxicam may adversely affect health; however, a decreased vaccine response is likely not a contributing factor.

Des bouvillons croisés sevrés rapidement (N = 271) ont été utilisés dans un essai clinique randomisé en aveugle à deux bras pour évaluer l'impact d'un anti-inflammatoire non stéroïdien à action prolongée sur les titres du virus de la rhinotrachéite infectieuse bovine, du virus respiratoire syncytial bovin, du virus parainfluenza 3 et du coronavirus, et les résultats pour la santé lorsqu'administré en même temps qu'un vaccin vivant modifié respiratoire à l'arrivée dans un parc d'engraissement. Les groupes de traitement comprenaient un témoin (solution saline; n = 135) et un groupe expérimental (méloxicam injectable; n = 136). Les titres d'anticorps viraux et le poids corporel ont été mesurés à l'arrivée, au jour 7 et au jour 21, ainsi qu'un poids final au jour 45. Le poids corporel et les titres d'anticorps pour tous les virus ont augmenté avec le temps (P < 0,001); cependant, il n'y avait aucune différence selon le groupe de traitement ou une interaction groupe × temps significative lors de l'évaluation à l'aide de mesures répétées d'analyse de la variance. Fait intéressant, l'utilisation du méloxicam était associée à un risque de traitement accru (P < 0,05). En conclusion, l'administration de méloxicam peut nuire à la santé; cependant, une réponse vaccinale réduite n'est probablement pas un facteur contributif.(Traduit par Docteur Serge Messier).

Clinical impacts of administering a nonsteroidal anti-inflammatory drug to beef calves after assisted calving on pain and inflammation, passive immunity, health, and growth.

Assisted calves are often born weak, injured, or oxygen deprived and have a higher risk of morbidity and mortality. The objective was to investigate the impact of using pain mitigation at birth in assisted beef calves on physiological indicators of pain and inflammation, passive immunity, health, and growth. Thirty-three primiparous cows and their calves requiring assistance at birth on two ranches located in southern Alberta were enrolled. Data collected at birth include date and time of calving, calf sex, meconium staining, presentation of calf, and calving difficulty (easy assist: one person manually delivered the calf; difficult assist: delivery by two or more people, or mechanical assistance). Within 10 min of birth, calves were stratified by calving difficulty, randomized to a medication group, and received a subcutaneous dose of meloxicam (0.5 mg/kg BW) or an equivalent volume of placebo. Cow-calf pairs were then placed in individual box stalls for observation and sampling. At birth, 1, 4, and 24 h after birth, heart rate, respiratory rate, and rectal temperature were assessed and blood samples collected to measure indicators of pain and inflammation (cortisol, corticosterone, substance P, and haptoglobin). Serum IgG concentration and failed transfer of passive immunity (serum IgG concentration <24 g/L) were assessed in the 24-h blood samples. Preweaning treatment for disease and mortality information was collected and calves were weighed at 7 to 10 d of age and at weaning. Of the 33 calves enrolled, 17 calves received meloxicam and 16 calves received a placebo. Meloxicam-medicated calves had significantly greater ADG to 7 to 10 d of age (P = 0.05) (mean = 0.9 kg/d; SE = 0.10) compared with placebo-medicated calves (mean = 0.6 kg/d; SE = 0.12). There was no significant effect of meloxicam on physiological indicators of pain and inflammation, standing or nursing by 1 h, passive immunity, health outcomes, or ADG to weaning (P > 0.1). Although this was a small sample population, meloxicam given to assisted calves at birth improved ADG in the first week of life, which may indicate an important production management tool for improving well-being in assisted calves.

Prevalence and lameness-associated risk factors in Alberta feedlot cattle.

Lameness in cattle is a health and welfare concern; however, limited information is available on risk factors and the relationship between lameness and common diseases like bovine respiratory disease (BRD). Therefore, the objectives of this study were to: 1) identify prevalence of lameness in feedlot cattle and related risk factors of cattle diagnosed as lame; and 2) determine associations between BRD occurrence and lameness. Feedlot cattle health records were available from 28 feedlots for 10 yr. The data set consisted of 663,838 cattle records, with 13.9% (92,156) diagnosed with a disease, including 32.3%, 46.0%, and 22.0% with lameness, BRD, and other diagnoses, respectively. Lameness was classified into four categories: foot rot (FR), joint infections (JI), lame with no visible swelling (LNVS), and injuries (INJ), with a prevalence of 74.5%, 16.1%, 6.1%, and 3.1%, respectively. Lameness was compared across cattle types (arrival date and weight) as well as age classification (calf vs. yearling), gender (steer vs. heifer), and season of placement in the feedlot (spring, summer, fall, and winter). Within the disease-diagnosed population, lameness represented 28.5% of treated fall-placed calves, 38.5% of winter-placed calves, and 40.8% of treated yearlings. Foot rot was the most common diagnosis with 74.5% of all lameness diagnoses, with winter- and fall-placed calves more likely to be diagnosed with FR compared to yearlings (OR: 1.19, 95% CI: 1.10-1.30 and OR: 1.46, 95% CI: 1.38-1.55, respectively). Joint infections were the second most common diagnosis (16.1%). Compared to yearlings, fall-placed calves had a higher odds (OR: 3.64, 95% CI: 3.12-4.24) for JI. Injuries and LNVS were the least common but again fall-placed calves had higher odds of this diagnosis compared to yearlings (OR: 2.26, 95% CI: 1.70-2.99 and OR: 9.10, 95% CI: 6.26-13.2, respectively). Gender was significantly different for JI as steers were less likely affected compared to heifers (OR: 0.687, 95% CI: 0.545-0.867), and more likely affected by LNVS (OR: 2.46, 95% CI: 1.57-3.84). Of all lameness-associated deaths, JI accounted for almost 50%. Finally, cattle diagnosed with BRD were subsequently more likely to be diagnosed with INJ, JI, or LNVS (P < 0.001 for all comparisons). In conclusion, animal type and gender were associated with type of lameness diagnoses, allowing feedlots to allocate resources to groups at highest risk and focus on early intervention strategies.

Cattle Immunized with a Recombinant Subunit Vaccine Formulation Exhibits a Trend towards Protection against Histophilus somni Bacterial Challenge.

Histophilosis, a mucosal and septicemic infection of cattle is caused by the Gram negative pathogen Histophilus somni (H. somni). As existing vaccines against H. somni infection have shown to be of limited efficacy, we used a reverse vaccinology approach to identify new vaccine candidates. Three groups (B, C, D) of cattle were immunized with subunit vaccines and a control group (group A) was vaccinated with adjuvant alone. All four groups were challenged with H. somni. The results demonstrate that there was no significant difference in clinical signs, joint lesions, weight change or rectal temperature between any of the vaccinated groups (B,C,D) vs the control group A. However, the trend to protection was greatest for group C vaccinates. The group C vaccine was a pool of six recombinant proteins. Serum antibody responses determined using ELISA showed significantly higher titers for group C, with P values ranging from < 0.0148 to < 0.0002, than group A. Even though serum antibody titers in group B (5 out of 6 antigens) and group D were significantly higher compared to group A, they exerted less of a trend towards protection. In conclusion, the vaccine used in group C exhibits a trend towards protective immunity in cattle and would be a good candidate for further analysis to determine which proteins were responsible for the trend towards protection.

Reverse vaccinology as an approach for developing Histophilus somni vaccine candidates.

Histophilosis of cattle is caused by the Gram negative bacterial pathogen Histophilus somni (H. somni) which is also associated with the bovine respiratory disease (BRD) complex. Existing vaccines for H. somni include either killed cells or bacteria-free outer membrane proteins from the organism which have proven to be moderately successful. In this study, reverse vaccinology was used to predict potential H. somni vaccine candidates from genome sequences. In turn, these may protect animals against new strains circulating in the field. Whole genome sequencing of six recent clinical H. somni isolates was performed using an Illumina MiSeq and compared to six genomes from the 1980's. De novo assembly of crude whole genomes was completed using Geneious 6.1.7. Protein coding regions was predicted using Glimmer3. Scores from multiple web-based programs were utilized to evaluate the antigenicity of these predicted proteins which were finally ranked based on their surface exposure scores. A single new strain was selected for future vaccine development based on conservation of the protein candidates among all 12 isolates. A positive signal with convalescent serum for these antigens in western blots indicates in vivo recognition. In order to test the protective capacity of these antigens bovine animal trials are ongoing.

Single nucleotide polymorphisms in the bovine Histophilus somni genome; a comparison of new and old isolates.

Histophilus somni, a causative agent of the bovine respiratory disease complex, can also cause a variety of systemic disorders, including bronchopneumonia, myocarditis, pericarditis, arthritis, pleuritis, and infectious thrombotic meningoencephalitis. The purpose of this study was to determine if currently circulating strains differ from those of the 1980s by identifying genomic changes. Single nucleotide polymorphisms (SNPs) and insertion and deletion (INDEL) sites were examined by whole-genome sequencing in 12 samples, 6 old and 6 new. The 31 028 SNP/INDELs recorded were compared against the reference genome sequence of the pathogenic H. somni strain 2336. The distribution of about 75% of these SNPs within a specified gene differed between old and new isolates and did not follow any particular pattern. The other 25% clustered into 2 groups containing the same SNPs in various genes: group I included 5 old isolates and 1 new isolate; group II included 5 new isolates and 1 old isolate. For putative virulence genes there were more SNPs in group I compared with strain 2336, itself an older isolate, than in group II. Although only 25% of all the SNPs formed 2 clusters, the results suggest some genetic difference in various genes between old and new strains.

Histophilus somni est l'un des agents majeurs du complexe respiratoire bovin (CRB), qui peut aussi causer diverses pathologies dont de la bronchopneumonie, myocardite, péricardite, arthrite, pleurésie et de la méningo-encéphalite thrombotique. L'objectif général de l'étude était de comparer les souches actuellement en circulation avec les souches isolées dans les années 80. Plus spécifiquement les changements génétiques survenus entre des isolats récents et des isolats collectés il y a une trentaine d'années ont été analysés. Les polymorphismes d'un seul nucléotide (single nucleotide polymorphism, SNP) ont été examinés en utilisant une approche de séquençage global de tout le génome pour 12 échantillons, six anciens et six nouveaux. Un total de 31 028 SNPs a été identifié et une analyse comparative de ces SNPs avec la séquence génomique de référence de la souche pathogène 2336 de H. somni a été effectuée. La distribution génique d'environ 75 % de ces SNPs entre les souches anciennes et récentes est différente et ne suit pas de tendance particulière. Toutefois, 25 % des SNPs se répartissent rapidement en deux groupes distincts. Le groupe I inclut cinq isolats anciens et un récent alors que le groupe II comprend cinq isolats récents et un isolat ancien qui se regroupent ensemble pour de mêmes SNPs dans plusieurs gènes. La présence des SNPs dans des gènes potentiellement liés à la virulence est plus manifeste dans le groupe I, comparé à l'ancien isolat 2336, que dans le groupe II. Bien que seulement 25 % des SNPs totaux se répartissent en deux groupes, les résultats suggèrent des variations génétiques significatives entre souches anciennes et récentes dans les séquences de nombreux gènes.(Traduit par Docteur François Meurens).

Evaluating the cost implications of a radio frequency identification feeding system for early detection of bovine respiratory disease in feedlot cattle.

New technologies to identify diseased feedlot cattle in early stages of illness have been developed to reduce costs and welfare impacts associated with bovine respiratory disease (BRD). However, the economic value of early BRD detection has never been assessed. The objective was to simulate cost differences between two BRD detection methods during the first 61 d on feed (DOF) applied in moderate- to large-sized feedlots using an automated recording system (ARS) for feeding behavior and the current industry standard, pen-checking (visual appraisal confirmed by rectal temperature). Economic impact was assessed with a cost analysis in a simple decision model. Scenarios for Canadian and US feedlots with high- and low-risk cattle were modeled, and uncertainty was estimated using extensive sensitivity analyses. Input costs and probabilities were mainly extracted from publicly accessible market observations and a large-scale US feedlot study. In the baseline scenario, we modeled high-risk cattle with a treatment rate of 20% within the first 61 DOF in a feedlot of >8000 cattle in Canada. Early BRD detection was estimated to result in a relative risk of 0.60 in retreatment and 0.66 in mortality compared to pen-checking (based on previously published estimates). The additional cost of monitoring health with ARS in Canadian dollar (CAD) was 13.68 per steer. Scenario analysis for similar sized US feedlots and low-risk cattle with a treatment rate of 8% were included to account for variability in costs and probabilities in various cattle populations. Considering the cost of monitoring, all relevant treatment costs and sale price, ARS was more costly than visual appraisal during the first 61 DOF by CAD 9.61 and CAD 9.69 per steer in Canada and the US, respectively. This cost difference increased in low-risk cattle in Canada to CAD 12.45. Early BRD detection with ARS became less expensive if the costs for the system decreased to less than CAD 4.06/steer, or if the underlying true BRD incidence (not treatment rate) within the first 61 DOF exceeded 47%. The model was robust to variability in the remaining input variables. Some of the assumptions in the baseline analyses were conservative and may have underestimated the real value of early BRD detection. Systems such as ARS may reduce treatment costs in some scenarios, but the investment costs are currently too high to be cost-effective when used solely for BRD detection compared to pen-checking.