Infectious Bovine Respiratory Diseases in Adult Cattle: An Extensive Necropsic and Etiological Study

In young cattle, bovine respiratory disease (BRD) is a major cause of death and Mannheimia haemolytica is a frequent pathogen. Knowledge of fatal BRD in adult cattle is more limited. We assessed the importance of infectious BRD as a cause of death in adult cattle and determined the associated pathogens. We analyzed data from 737 adult cattle necropsies at the Pathology Unit for Large Animals at Oniris, Nantes, France over a 6 year period (2013–2019). Each carcass was subjected to a complete necropsy. Lungs showing macroscopic lesions were classified into three categories: infectious primary pulmonary (IPP) lesions, thromboembolic pneumonia (TEP) and others (aspiration pneumonia, verminous pneumonia, and local extension of an extra-pulmonary inflammatory process). Half of the lungs with IPP macroscopic lesions were sampled for histology and submitted for polymerase chain reaction. BRD was the second leading cause of death (15.7%) after digestive diseases (32.2%). A strong predominance of IPP lesions (42.3%) and TEP lesions (39.6%) was also demonstrated. In IPP macroscopic lesions, fibrinous, hemorrhagic and/or hecrotic (FHN) bronchopneumonia accounted for 77.6% of macroscopic lesions. Mannheimia haemolytica was significantly associated with FHN bronchopneumonia macroscopic lesions. This study suggests that Mannheimia haemolytica should be included in the differential diagnosis of BRD in adult cattle.

Evaluation of the PBMC Proliferation, Apoptosis and Cytokines Profiling in Cattle Infected with Mycoplasma bovis Strain 07801

Bovine respiratory diseases are widespread and too costly disease impacting contributes to economically essential diseases like mastitis and pneumonia worldwide. The present study aimed to explore the influence of M. bovis field strain 07801 on the status of PBMCs in the challenged cattle group and the immunized one. For this aim, the PBMCs proliferation, apoptosis, and cytokine profile changes were determined. In this study, M. bovis strain Mb 07801 and reference strain PG45 were used; ten calves (2-3 months old) were arranged into two groups (5 calve each): 1) PBS-challenged group (with M. bovis 07801 at about 1010 CFU/ml), and 2) immunized group (by Inactivated M. bovis 07801); both groups were treated nasally and intra-tracheal. Blood samples were obtained from both groups and examined for PBMCs proliferation and apoptosis, as well as serum cytokine profile, before infection and at day zero and days 7, 14, 21, and 36 post bacterial treatment. The results revealed that M. bovis strain (07801and PG45) antigen increased the proliferative response of the stimulated PBMC compared with the unstimulated cells and ConA-alone stimulated. The PBMC apoptosis showed a non-significant increase in both challenged and immunized groups compared to the negative and positive control (treated with apoptosis inducer) groups. Besides, the levels of cytokines profile showed a significant up-regulation in IFN-γ, IL-4, IL-2, IL-18, TNF-α, IFN-α, IL-6, IL-10 IL-1β, and IL-13, in both groups, except IL-4 and IL-18 those recorded a down-regulation in the immunized group at days 7,14, 21, and 36 post-M. bovis infection. In conclusion, immunization markedly ameliorated the immune deterioration induced by M. bovis strain 07801.

Minimizing the number of origins in batches of weaned calves to reduce their risks of developing bovine respiratory diseases

Bovine respiratory diseases (BRD) are a major concern for the beef cattle industry, as beef calves overwhelmingly develop BRD symptoms during the first weeks after their arrival at fattening units. These cases occur after weaned calves from various cow-calf producers are grouped into batches to be sold to fatteners. Cross-contaminations between calves from different origins (potentially carrying different pathogens), together with increased stress because of the process of batch creation, can increase their risks of developing BRD symptoms. This study investigated whether reducing the number of different origins per batch is a strategy to reduce the risk of BRD cases. We developed an algorithm aimed at creating batches with as few origins as possible, while respecting constraints on the number and breed of the calves. We tested this algorithm on a dataset of 137,726 weaned calves grouped into 9701 batches by a French organization. We also computed an index assessing the risks of developing BRD because of the batch composition by considering four pathogens involved in the BRD system. While increasing the heterogeneity of batches in calf bodyweight, which is not expected to strongly impact the performance, our algorithm successfully decreased the average number of origins in the same batch and their risk index. Both this algorithm and the risk index can be used as part of decision tool to assess and possibly minimize BRD risk at batch creation, but they are generic enough to assess health risk for other production animals, and optimize the homogeneity of selected characteristics.

In Silico Identification of Novel Immunogenic Secreted Proteins of Mycoplasma bovis from Secretome Data and Experimental Verification

Mycoplasma bovis is a major pathogen, responsible for bovine respiratory diseases worldwide. The present lack of effective control measures leaves cattle owners at considerable perpetual risk of M. bovis outbreaks. In this study, we identified M. bovis secreted immunogenic proteins in silico as potential candidates for novel diagnostic agents and vaccines. We used immunoinformatics to analyze 438 M. bovis proteins previously identified with a label-free proteomics analysis of virulent M. bovis HB0801 (P1) and its attenuated P150 strains. The subcellular localization of these proteins was preliminarily screened and 59 proteins were found to be secreted extracellular proteins. Twenty-seven of these proteins contained a large number of predictive T-cell epitopes presented by major histocompatibility complex (MHC) class I and II molecules. Twenty-two of these 27 proteins had a high number of conformational B-cell epitopes, predicted from the corresponding 3D structural templates, including one unique to P1, two unique to P150, and 19 common to both strains. Five proteins were selected for further validation, and two of these, MbovP274 and MbovP570, were successfully expressed and purified. Both were confirmed to be secretory and highly immunogenic proteins that induced a mouse antibody response, reacted with cattle serum positive for M. bovis infection, and significantly increased the production of interleukin 8 (IL-8), IL-12 and interferon γ (IFN-γ) during the secretion of these three cytokines by both M. bovis mutants of these genes. These results should be useful in the development of novel immunological agents against M. bovis infection.

Genetic and Antigenic Characterization and Retrospective Surveillance of Bovine Influenza D Viruses Identified in Hokkaido, Japan from 2018 to 2020

Influenza D virus (IDV), which is a new member of the Orthomyxoviridae family, is potentially involved in bovine respiratory diseases (BRDs). Bovine IDVs (BIDVs) from Japan have been distributed nationwide since 2010 and are genetically distinct from foreign IDVs. We isolated BIDVs from three BRD outbreaks, in Hokkaido during 2018–2020, to understand their genetic and antigenic characteristics. Retrospective surveillance was performed using sera collected throughout the last decade in Hokkaido to investigate BIDV existence. Three BIDVs were isolated using cell culture. Comparative and phylogenetic analyses using sequence data of the three BIDVs and IDVs from Japan and other countries available in GenBank demonstrated that Japanese BIDVs, including the three BIDV isolates, were genetically distinct from other IDVs. Genotype classifications based on the rotavirus genotype classification revealed multiple genotypes of RNA segments 1–7. Two BIDVs were of a new genotype, different from those of other Japanese BIDVs. Neutralization assays against two BIDVs with different genotypes using sera collected in acute and recovery phases of BRD revealed differences in cross-reactivity to heterogenous BIDVs. Retrospective surveillance suggested that BIDV existed in Hokkaido, in 2009. Our findings suggest that BIDVs of different genotypes and antigenicity are distributed and maintained in Hokkaido and provide new insights into molecular characteristics and the evolution of IDVs.

Monitoring Mycoplasma bovis Diversity and Antimicrobial Susceptibility in Calf Feedlots Undergoing a Respiratory Disease Outbreak

Bovine respiratory diseases (BRD) are widespread in veal calf feedlots. Several pathogens are implicated, both viruses and bacteria, one of which, Mycoplasma bovis, is under-researched. This worldwide-distributed bacterium has been shown to be highly resistant in vitro to the main antimicrobials used to treat BRD. Our objective was to monitor the relative prevalence of M. bovis during BRD episodes, its diversity, and its resistance phenotype in relation to antimicrobial use. For this purpose, a two-year longitudinal follow-up of 25 feedlots was organized and 537 nasal swabs were collected on 358 veal calves at their arrival in the lot, at the BRD peak and 4 weeks after collective antimicrobial treatments. The presence of M. bovis was assessed by real-time PCR and culture. The clones isolated were then subtyped (polC subtyping and PFGE analysis), and their susceptibility to five antimicrobials was determined. The course of the disease and the antimicrobials used had no influence on the genetic diversity of the M. bovis strains: The subtype distribution was the same throughout the BRD episode and similar to that already described in France, with a major narrowly-variable subtype circulating, st2. The same conclusion holds for antimicrobial resistance (AMR) phenotypes: All the clones were already multiresistant to the main antimicrobials used (except for fluoroquinolones) prior to any treatments. By contrast, changes of AMR phenotypes could be suspected for Pasteurellaceae in two cases in relation to the treatments registered.

KA-1002, a Novel Lysophosphatidic Acid Signaling Antagonist, Alleviates Bovine Tracheal Cell Disruption and Inflammation

The industrial livestock environment can cause stress and weakened immunity in cattle, leading to microbial infections which reduce productivity. As such, there is a need for an effective therapeutic agent that can alleviate uncontrolled destructive respiratory inflammation. We found that lysophosphatidic acid (LPA), a potent endogenous stress-induced inflammatory agent, causes respiratory tissue damage and triggers inflammation in bovine bronchial cells. LPA also inflames pulmonary bovine blood vessel cells to produce inflammatory cytokines. These findings strongly suggest that LPA is a highly important endogenous material exacerbating bovine respiratory diseases. We further identified a novel LPA-signaling antagonist, KA-1002, and showed that it alleviated LPA-mediated bovine tracheal cell disruption and inflammation. Therefore, KA-1002 could potentially serve as a novel therapeutic agent to maintain physiologically healthy and balanced conditions in bovine respiratory tracts.

Characterisation of the First Bovine Parainfluenza Virus 3 Isolate Detected in Cattle in Turkey

A respiratory disease outbreak on a cattle farm in northern Turkey produced respiratory tract symptoms and severe pneumonia symptoms in 20 calves. Eight calves died, and a lung specimen from one carcass was analysed for bacteria and for viruses of the Bovine respiratory diseases complex. Bacteriological analysis was negative, but antigen detection ELISA and RT-PCR results indicated the presence of Bovine parainfluenza virus (BPIV). Virus isolation succeeded on Madin-Darby Bovine Kidney cells, and subsequent whole genome sequencing and phylogenetic analysis identified BPIV-3c. This is the first report of BPIV-3c isolation from cattle in Turkey, indicating the need for more virological and epidemiological studies.