Sébastien Assié

Transmission dynamics of Mannheimia haemolytica in newly-received beef bulls at fattening operations.

The primary objective of this study was to determine, at the lung level, whether single or multiple clones of Mannheimia haemolytica are present within a pen during a bovine respiratory disease (BRD) episode. A secondary objective was to assess whether M. haemolytica isolates obtained from nasal swabs (NS) are identical to those isolated deeper within the respiratory tract. Sixteen BRD episodes that naturally occurred in 12 pens of eight to 12 bulls (n=112) newly-received at three fattening operations were investigated. One hundred and seventy five M. haemolytica isolates were collected from 239 pairs of trans-tracheal aspirations (TTA) and NS performed during these 16 BRD episodes. M. haemolytica isolates were characterized by pulsed-field gel electrophoresis (PFGE). PFGE types obtained from NS and TTA were then compared. M. haemolytica was isolated during 14 BRD episodes. Two to three different clones of M. haemolytica were recovered during 10 episodes whereas only one clone was recovered in four episodes. A moderate agreement (kappa=0.50) between NS and TTA for M. haemolytica isolation was observed. Identical PFGE types were only observed in 77% of matched NS-TTA pairs. The significant within-pen diversity of M. haemolytica during BRD episodes indicates that the disease is not primarily due to the spread of a single virulent clone among cattle and highlights the importance of predisposing factors that enable the resident flora to overcome the cattles immune system. The results also demonstrate that isolates recovered from NS are not always representative of the isolates present deeper within the respiratory tract.

Exposure to pathogens and incidence of respiratory disease in young bulls on their arrival at fattening operations in France

The incidence of clinical respiratory disease in 698 young beef bulls kept in 68 pens, and their exposure to respiratory pathogens after their arrival at 51 fattening operations in western France were assessed. Antibodies against bovine respiratory syncytial virus (BRSV), bovine herpesvirus type 1 (BHV-1), Mannheimia haemolytica and Mycoplasma bovis were measured by ELISA. The incidence risk of respiratory disease was 18.5 per cent during the first six weeks. Cases occurred in 37 of the 68 pens, and in these pens 30.9 per cent of the bulls were affected. Their exposure to BHV-1 was very limited. When they arrived a high proportion of the bulls were seropositive to M haemolytica and a high proportion seroconverted to BRSV, M haemolytica and M bovis within the first six weeks. The risk of incidence of respiratory disease was lower in the pens in which the bulls had been vaccinated against M haemolytica. Higher proportions of the bulls were affected in pens in which small proportions of the bulls were seropositive to M haemolytica or BRSV on arrival, and in pens in which high proportions of the bulls were exposed to M haemolytica or BRSV during the first six weeks.

Disruption of Bcchs4, Bcchs6 or Bcchs7 chitin synthase genes in Botrytis cinerea and the essential role of class VI chitin synthase (Bcchs6)

Chitin synthases play critical roles in hyphal development and fungal pathogenicity. Previous studies on Botrytis cinerea, a model organism for necrotrophic pathogens, have shown that disruption of Bcchs1 and more particularly Bcchs3a genes have a drastic impact on virulence (Soulié et al., 2003, 2006). In this work, we investigate the role of other CHS including BcCHS4, BcCHS6 and BcCHS7 during the life cycle of B. cinerea. Single deletions of corresponding genes were carried out. Phenotypic analysis indicates that: (i) BcCHS4 enzyme is not essential for development and pathogenicity of the fungus; (ii) BcCHS7 is required for pathogenicity in a host dependant manner. For Bcchs6 gene disruption, we obtained only heterokaryotic strains. Indeed, sexual or asexual purification assays were unsuccessful. We concluded that class VI chitin synthase could be essential for B. cinerea and therefore BcCHS6 represents a valuable antifungal target.

Management- and housing-related risk factors of respiratory disorders in non-weaned French Charolais calves

Our aim was to determine at calf-batch level the management- and housing-related risk factors of respiratory disorders in non-weaned Charolais calves. Farmers recorded cases according to the definition provided i.e. the association of at least one respiratory sign and, in the same calf or another calf of the same batch, at least one general sign on the same day or the day before. During farm visits, quality of farmers records was checked and questionnaires were applied to gather farm and herd characteristics and to describe farming practices and housing facilities. Data were suitable for analysis for 172 batches where no metaphylactic treatment was implemented. Batches had great disparity in incidence of respiratory disorders. In the 120 batches with at least one case, the quartiles of incidence rate were, respectively, 0.95, 2.15 and 3.59 cases per 1000 calf-days at-risk. For risk-factor analysis, the statistical unit was the calf-batch located in a given batch. We used a ZINB model because (i) there was significant overdispersion of incidence rates of respiratory disorders (overdispersion test statistic of O=1049 (P<0.001)), (ii) probability was high that a two-group modelling process existed (Vuong statistic of V=2.44 (P=0.0073)) and (iii) the ZINB model fit significantly better than the ZIP model (likelihood-ratio statistic of 340.14 (P<0.001)). The risk was higher in open-fronted barns or in open barns compared to closed ones (incidence-rate ratios (IRRs): 1.9 and 1.8), in barns where no annual disinfection was implemented (IRR: 1.5) and in part slope with straw/part scraped and in part straw-bedded/part scraped barns compared to completely straw-bedded barns (IRRs: 1.9 and 1.6). Larger calf-batches were more at-risk than smaller batches (OR for the effect of an increase of 10 calves on the probability of a 0 count: 0.5).

Visually undetected fever episodes in newly received beef bulls at a fattening operation: Occurrence, duration, and impact on performance

Monitoring body temperature of newly received cattle allows for identification of fever episodes not visually detected by feedlot personnel (FENO). Information concerning the occurrence, duration, and impact on performance of these FENO is not available in the literature. Such information is crucial to assess the potential benefit of the identification and treatment of FENO. Therefore, the objectives of this study were to describe the occurrence and duration of FENO and to evaluate their impact on ADG. One hundred twelve beef bulls (initial BW = 346 ± 36 kg) were studied for 40 d after arrival at 3 French fattening operations. At d 1, each animal was administered orally a reticulo-rumen bolus, which allowed continuous measurement and recording of reticulo-rumen temperature. Animals were weighed on d 1 and 40. Bulls were observed twice daily by personnel for visual signs of apparent disease. Bulls that appeared ill, had a rectal temperature ≥39.7°C, and demonstrated symptoms consistent with bovine respiratory disease (BRD) were treated with antibiotics. After d 40, data obtained from the boluses were retrospectively analyzed using a cumulative sum test to detect significant increases in reticulo-rumen temperature considered as fever episodes. Numerous fever episodes (n = 449) were retrospectively detected in 110 bulls. Of these 449 fever episodes, 74% were not associated with any visually detected clinical signs of disease and thus were identified as FENO. These FENO were often transitory (75% lasted less than 47 h). However, 25% lasted from 47 to 263 h. Of the 112 bulls, 88 were treated for BRD with 20 and 7 animals treated, respectively, 2 and 3 times. In treated animals, fever episodes began 4 to 177 h (mean = 50 h) before BRD treatment. The duration of FENO was associated (P = 0.002) with a lesser ADG (d 1 to 40): -33 g/d for daily FENO duration. Our results demonstrated that FENO occurred frequently in bulls during the first weeks after entrance into a fattening operation and can last up to 11 d. The impact of FENO observed on ADG in this study indicated a potential benefit of treating affected animals, specifically those with FENO of long duration. However, further research is needed to determine the medical and economic relevance of such treatment.

Transmission dynamics of Mycoplasma bovis in newly received beef bulls at fattening operations

Mycoplasma bovis is an important cause of bovine respiratory disease (BRD) in newly received cattle at fattening operations. However, little information on its within-pen transmission dynamics during a BRD outbreak is available. Such information is nevertheless crucial to adapt control measures during M. bovis–associated BRD outbreaks. The objective of the current study was to determine whether single or multiple clones of M. bovis are present within a pen during a BRD outbreak that occurs early in the feeding period. Sixteen BRD outbreaks that naturally occurred in 12 pens of 8–12 bulls each (n = 112) newly received at 3 fattening operations were investigated. Two hundred and thirty-nine transtracheal aspirations (TTA) were performed during the outbreaks, and the M. bovis isolates obtained were characterized by pulsed-field gel electrophoresis (PFGE). Mycoplasma bovis isolates were recovered from TTA in 8 of the 16 BRD outbreaks that occurred. The within-pen prevalence of bulls positive for M. bovis during these outbreaks ranged from 8% to 100%. The PFGE analysis revealed that, even though bulls came from multiple origins, a single clone of M. bovis was present within a pen during BRD outbreaks with a high prevalence of M. bovis infection. The study therefore indicates that, even if M. bovis can recrudesce from carriers after stressful events such as transportation and commingling, the increased prevalence of M. bovis pulmonary infection observed during BRD outbreaks that are early occurring in the feeding period seems primarily due to the horizontal transmission of only 1 clone among cattle.

Detection by real-time RT-PCR of a bovine respiratory syncytial virus vaccine in calves vaccinated intranasally

Seventeen four- to five-week-old calves that were not shedding bovine respiratory syncytial virus (BRSV) were vaccinated intranasally against the disease and sampled by nasal swabbing on 16 different days for up to 20 days after vaccination. BRSV vaccine virus was detected in 15 of the 17 calves. Five of the calves were PCR positive on only one swab, eight were PCR positive on two to five swabs and two were PCR positive on more than five swabs. Twelve of the calves were positive only before day 14 and three were positive after day 14. The nasal shedding of BRSV vaccine virus was very variable.

Impact of Timing and Dosage of a Fluoroquinolone Treatment on the Microbiological, Pathological, and Clinical Outcomes of Calves Challenged with Mannheimia haemolytica

The efficacy of an early and low inoculum-adjusted marbofloxacin treatment was evaluated on microbiological and clinical outcomes in calves infected with 4.107 CFU of Mannheimia haemolytica A1. Twenty-two calves were included based on their rectal temperature rise in the 10 h after challenge and allocated in four groups, receiving a single intramuscular injection of saline (CON), 2 mg/kg marbofloxacin 2–4 h after inclusion (early treatment, E2), 2 or 10 mg/kg marbofloxacin 35–39 h after inclusion (late treatments, L2, L10). In CON calves, M. haemolytica DNA loads in bronchoalveolar lavages continuously increased from inclusion to day 4, and were associated with persistent respiratory clinical signs and lung lesions. At times of early and late treatments, M. haemolytica loads ranged within 3.5–4 and 5.5–6 log10 DNA copies/mL, respectively. Early 2 mg/kg marbofloxacin treatment led to rapid and total elimination of bacteria in all calves. The late treatments induced a reduction of bacterial loads, but 3 of 6 L2 and 1 of 6 L10 calves were still positive for M. haemolytica at day 4. Except for CON calves, all animals exhibited clinical improvement within 24 h after treatment. However, early 2 mg/kg treatment was more efficacious to prevent pulmonary lesions, as indicated by the reduction of the extension and severity of gross lesions and by the histopathological scores. These results demonstrated for the first time that a reduced antibiotic regimen given at an early stage of the disease and targeting a low bacterial load could be efficacious in a natural bovine model of pneumonia.

Evaluation of a Commercial Real-Time Reverse Transcription Polymerase Chain Reaction Kit for the Diagnosis of Bovine Respiratory Syncytial Virus Infection

Recently a commercial real-time reverse transcription polymerase chain reaction (RT-PCR) kit has been marketed for the detection of Bovine respiratory syncytial virus (BRSV). However, diagnostic interpretation of the results of this kit requires its comparison to commonly used methods. Therefore, the objective of this study was to evaluate the performance of this kit in comparison with the conventional direct fluorescent antibody test (FAT). Twenty BRSV strains and 14 heterologous bovine viruses were used to check the kits sensitivity and specificity. The efficiency and detection limit of the kit were determined by testing dilution series of a BRSV strain. The comparison between real-time RT-PCR kit and FAT was performed with 94 clinical samples from calves with clinical signs of respiratory disease including lung tissues (n = 55), transtracheal aspiration samples (n = 20), and nasal swab samples (n = 19). All of the BRSV strains tested were detected by real-time RT-PCR. No cross-reaction was shown with the 14 heterologous bovine viruses. The real-time RT-PCR was 99.3% efficient with a detection limit of 0.1 TCID50 (50% tissue culture infective dose). The results of real-time RT-PCR and FAT were concordant for 65 of the 94 clinical samples tested. The remaining 29 clinical samples were positive by real-time RT-PCR and negative by FAT, demonstrating the higher sensitivity of real-time RT-PCR. In conclusion, the kit evaluated in this study was sensitive, specific, and had a low threshold of detection. Furthermore, the use of this kit instead of FAT allows an improvement of the sensitivity for the detection of BRSV in clinical samples.

Implementing Precision Antimicrobial Therapy for the Treatment of Bovine Respiratory Disease: Current Limitations and Perspectives

The therapeutic efficacy of an early treatment protocol with an infection-stage adjusted fluoroquinolone regimen was evaluated in a field study on young bulls (YBs) presenting signs of bovine respiratory disease (BRD). A total of 195 YB (Charolais, Limousin, and Rouge-des-Prés breeds) from 6 farms implementing or not prophylactic antimicrobial treatments (PROPHY or absence) were randomly assigned to 1 of 2 experiment groups based on time of detection of BRD and first-line marbofloxacin regimen, early adjusted dose [Early 2 (E2)] or late standard dose [Late 10 (L10)]. Each YB was administered orally a reticulo-rumen bolus, allowing continuous monitoring of ruminal temperature. In the E2 group, YB presenting early signs of BRD, i.e., an increase in ruminal temperature over 40.2°C and persisting more than 12u2009h, confirmed by a clinical examination showing no or mild signs of BRD, were given 2u2009mg/kg of marbofloxacin. In the L10 group, YBs presenting moderate or severe signs of BRD at visual inspection, confirmed at clinical examination, were given 10u2009mg/kg of marbofloxacin. If needed, YBs were given a relapse treatment. The YBs were followed for 30u2009days. The proportions of first and relapse treatments were calculated, as well as the therapeutic efficacy at day 10. In the E2 group, the first-line treatments’ proportion was significantly higher (Pu2009<u20090.05), while the relapse treatments’ proportion tended to be higher (Pu2009=u20090.08), than in the L10 group. Evolution of clinical scores (CSs) of diseased YB was followed for 10u2009days. In both groups, CS and rectal temperature decreased significantly 24u2009h after treatment (Pu2009<u20090.05). Treatment incidences (TI) representing antimicrobial consumption assessed on used daily doses (UDD) were calculated. Antimicrobial consumption of marbofloxacin and relapse treatments were not significantly different between the groups. These values were strongly influenced by the recourse to a prophylactic antimicrobial treatment, accounting for more than 90% of the antimicrobial amount in the herds implementing prophylaxis. The higher number of treatments in the groups treated on the basis of ruminal temperature monitoring, the accuracy of the detection method, and the necessary conditions to implement precision antimicrobial therapy in the field are discussed in this article.