Antimicrobial Activities of α-Helix and ß-Sheet Peptides against the Major Bovine Respiratory Disease Agent, Mannheimia haemolytica.

Bovine respiratory disease (BRD) is the leading cause of morbidity and mortality in cattle raised in North America. At the feedlot, cattle are subject to metaphylactic treatment with macrolides to prevent BRD, a practice that may promote antimicrobial resistance and has resulted in an urgent need for novel strategies. Mannheimia haemolytica is one of the major bacterial agents of BRD. The inhibitory effects of two amphipathic, α-helical (PRW4, WRL3) and one ß-sheet (WK2) antimicrobial peptides were evaluated against multidrug-resistant (MDR) M. haemolytica isolated from Alberta feedlots. WK2 was not cytotoxic against bovine turbinate (BT) cells by the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. All three peptides inhibited M. haemolytica, with WK2 being the most efficacious against multiple isolates. At 8-16 µg/mL, WK2 was bactericidal against Mh 330 in broth, and at 32 µg/mL in the presence of BT cells, it reduced the population by 3 logs CFU/mL without causing cytotoxic effects. The membrane integrity of Mh 330 was examined using NPN (1-N-phenylnaphthylamine) and ONPG (o-Nitrophenyl ß-D-galactopyranoside), with both the inner and outer membranes being compromised. Thus, WK2 may be a viable alternative to the use of macrolides as part of BRD prevention and treatment strategies.

Modelling the effects of antimicrobial metaphylaxis and pen size on bovine respiratory disease in high and low risk fattening cattle.

Bovine respiratory disease (BRD) dramatically affects young calves, especially in fattening facilities, and is difficult to understand, anticipate and control due to the multiplicity of factors involved in the onset and impact of this disease. In this study we aimed to compare the impact of farming practices on BRD severity and on antimicrobial usage. We designed a stochastic individual-based mechanistic BRD model which incorporates not only the infectious process, but also clinical signs, detection methods and treatment protocols. We investigated twelve contrasted scenarios which reflect farming practices in various fattening systems, based on pen sizes, risk level, and individual treatment vs. collective treatment (metaphylaxis) before or during fattening. We calibrated model parameters from existing observation data or literature and compared scenario outputs regarding disease dynamics, severity and mortality. The comparison of the trade-off between cumulative BRD duration and number of antimicrobial doses highlighted the added value of risk reduction at pen formation even in small pens, and acknowledges the interest of collective treatments for high-risk pens, with a better efficacy of treatments triggered during fattening based on the number of detected cases.

Antimicrobial resistance in bovine respiratory disease: Auction market- and ranch-raised calves.

This study compared changes in prevalence and antimicrobial susceptibility of Mannheimia haemolytica, Pasteurella multocida, and Histophilus somni in feedlot calves derived from the auction market (AUCT; n = 299) and from a single-ranch source (RANCH; n = 300). In the AUCT calves, the prevalence of Mannheimia haemolytica decreased, whereas Histophilus somni increased over the feeding period. The AUCT calves showed an increase in isolates not susceptible to tulathromycin for all bovine respiratory disease (BRD) pathogens, an increase in Pasteurella multocida and Histophilus somni isolates not susceptible to oxytetracycline, and an increase in Pasteurella multocida isolates not susceptible to florfenicol. In the RANCH calves, the prevalence of all 3 BRD pathogens was high at feedlot entry and decreased significantly during the study period. In RANCH calves, there was a significant increase in Pasteurella multocida isolates not susceptible to oxytetracycline, tulathromycin, and florfenicol. Surprisingly, there was a significant decrease in Mannheimia haemolytica isolates that were not susceptible to oxytetracycline, tilmicosin, and tulathromycin.

Résistance aux antimicrobiens lors de maladies respiratoires bovines : veaux provenant de marché aux enchères et ceux élevés en ranch. Cette étude a comparé les changements dans la prévalence et la sensibilité aux antimicrobiens de Mannheimia haemolytica, Pasteurella multocida et Histophilus somni isolés de veaux en parc d'engraissement provenant du marché aux enchères (AUCT; n = 299) et d'un seul ranch (RANCH; n = 300). Chez les veaux AUCT, la prévalence de M. haemolytica a diminué, tandis que celle d'H. somni a augmenté au cours de la période d'alimentation. Les veaux AUCT ont montré une augmentation des isolats non sensibles à la tulathromycine pour tous les agents pathogènes des maladies respiratoires bovines (BRD), une augmentation des isolats de P. multocida et H. somni non sensibles à l'oxytétracycline, et une augmentation des isolats de P. multocida non sensibles au florfénicol. Chez les veaux du RANCH, la prévalence des 3 agents pathogènes BRD était élevée à l'entrée du parc d'engraissement et a diminué de manière significative au cours de la période d'étude. Chez les veaux RANCH, il y a eu une augmentation significative des isolats de P. multocida non sensibles à l'oxytétracycline, à la tulathromycine et au florfénicol. Étonnamment, il y a eu une diminution significative des isolats de M. haemolytica qui n'étaient pas sensibles à l'oxytétracycline, à la tilmicosine et à la tulathromycine.(Traduit par Dr Serge Messier).

Models of antimicrobial pressure on intestinal bacteria of the treated host populations.

Antimicrobial drugs are used to treat pathogenic bacterial infections in animals and humans. The by-stander enteric bacteria of the treated host's intestine can become exposed to the drug or its metabolites reaching the intestine in antimicrobially active form. We consider which processes and variables need to be accounted for to project the antimicrobial concentrations in the host's intestine. Those include: the drug's fraction (inclusive of any active metabolites) excreted in bile; the drug's fractions and intestinal segments of excretion via other mechanisms; the rates and intestinal segments of the drug's absorption and re-absorption; the rates and intestinal segments of the drug's abiotic and biotic degradation in the intestine; the digesta passage time through the intestinal segments; the rates, mechanisms, and reversibility of the drug's sorption to the digesta and enteric microbiome; and the volume of luminal contents in the intestinal segments. For certain antimicrobials, the antimicrobial activity can further depend on the aeration and chemical conditions in the intestine. Model forms that incorporate the inter-individual variation in those relevant variables can support projections of the intestinal antimicrobial concentrations in populations of treated host, such as food animals. To illustrate the proposed modeling framework, we develop two examples of treatments of bovine respiratory disease in beef steers by oral chlortetracycline and injectable third-generation cephalosporin ceftiofur. The host's diet influences the digesta passage time, volume, and digesta and microbiome composition, and may influence the antimicrobial loss due to degradation and sorption in the intestine. We consider two diet compositions in the illustrative simulations. The examples highlight the extent of current ignorance and need for empirical data on the variables influencing the selective pressures imposed by antimicrobial treatments on the host's intestinal bacteria.

Assessment of nebulisation of sodium ceftiofur in the treatment of calves naturally infected with bovine respiratory disease.

Twelve screened cases of bovine respiratory disease (BRD) in calves were enrolled. Six of the calves were treated intramuscularly with sodium ceftiofur (1 mg/kg), and six were treated with nebulised sodium ceftiofur (1 mg/kg). Comparative evaluation of the two therapeutic modalities was based on repetitive analysis of hematological profile of calves on days 0, 5, and 10 post-therapy. The mortality rate in the group of calves treated with the nebulised sodium ceftiofur was significantly (p < 0.001) lower, and their clinical and hematological parameters returned to normal significantly (p < 0.001) faster than in calves treated intramuscularly. Nebulisation of sodium ceftiofur is the most effective treatment in calves with BRD under field conditions. Nasal lavage fluid analysis indicating a high rise of neutrophil count and macrophages may be used as an alternative method to detect pulmonary inflammation in BRD-affected calves.

Pharmacokinetics and pharmacodynamics of gamithromycin in pulmonary epithelial lining fluid in naturally occurring bovine respiratory disease in multisource commingled feedlot cattle.

The objectives of this study were to determine (i) whether an association exists between individual pharmacokinetic parameters and treatment outcome when feeder cattle were diagnosed with bovine respiratory disease (BRD) and treated with gamithromycin (Zactran(®) ) at the label dose and (ii) whether there was a stronger association between treatment outcome and gamithromycin concentration in plasma or in the pulmonary epithelial lining fluid (PELF) effect compartment. The study design was a prospective, blinded, randomized clinical trial utilizing three groups of 60 (362-592 lb) steers/bulls randomly allocated within origin to sham injection or gamithromycin mass medication. Cattle were evaluated daily for signs of BRD by a veterinarian blinded to treatment. Animals meeting the BRD case definition were enrolled and allocated to a sample collection scheme consisting of samples for bacterial isolation (bronchoalveolar lavage fluid and nasopharyngeal swabs) and gamithromycin concentration determination (PELF and plasma). Gamithromycin susceptibility of M. haemolytica (n = 287) and P. multocida (n = 257) were determined using broth microdilution with frozen panels containing gamithromycin at concentrations from 0.03 to 16 µg/mL. A two-compartment plasma pharmacokinetic model with an additional compartment for gamithromycin in PELF was developed using rich data sets from published and unpublished studies. The sparse data from our study were then fit to this model using nonlinear mixed effects modeling to estimate individual parameter values. The resulting parameter estimates were used to simulate full time-concentration profiles for each animal in this study. These profiles were analyzed using noncompartmental methods so that PK/PD indices (AUC24 /MIC, AUC∞ /MIC, CMAX /MIC) could be calculated for plasma and PELF (also T>MIC) for each individual. The calculated PK/PD indices were indicative that for both M. haemolytica and P. multocida a higher drug exposure in terms of concentration, and duration of exposure relative to the MIC of the target pathogen, was favorable to a successful case outcome. A significant association was found between treatment success and PELF AUC0-24 /MIC for P. multocida. The calves in this study demonstrated an increased clearance and volume of distribution in plasma as compared to the healthy calves in two previously published reports. Ultimately, the findings from this study indicate that higher PK/PD indices were predictive of positive treatment outcomes.

The effect of antimicrobial treatment and preventive strategies on bovine respiratory disease and genetic relatedness and antimicrobial resistance of Mycoplasma bovis isolates in a western Canadian feedlot.

Feedlot calves (n = 3784) were systematically randomized and allocated in a 2 × 2 factorial study to receive metaphylactic oxytetracycline (OTC) on arrival or no antimicrobial, as well as florfenicol once subcutaneously or twice intramuscularly (48 h apart) if diagnosed with bovine respiratory disease (BRD). Calves of different treatment groups were comingled and followed from placement to re-implantation (~100 days). Animals receiving OTC had a reduced risk of BRD, an increased risk of arthritis, and no significant differences in average daily gain, BRD relapse, overall mortality, or BRD mortality. There were no significant differences between treatment protocols. Deep nasal swabs (n = 233) taken at arrival (n = 122), treatment (n = 77), and swabs from lungs and joints at postmortem (n = 34) were cultured for Mycoplasma bovis from 61 animals ill or dying of chronic pneumonia and arthritis and from 61 healthy calves. There was significant variation in diversity among isolates (n = 51) between study years and different cattle. Metaphylaxis or antimicrobial treatment did not affect the diversity of isolates. Except for tilmicosin, isolates were largely susceptible to tested antimicrobials.

Effet du traitement antimicrobien et des stratégies préventives sur le complexe respiratoire bovin ainsi que la relation génétique et l'antibiorésistance des isolats deMycoplasma bovisdans un parc d'engraissement de l'Ouest canadien. Les veaux d'un parc d'engraissement (n = 3784) ont été systématiquement randomisés et répartis dans une étude factorielle 2 × 2 pour recevoir de l'oxytétracycline métaphylactique (OTC) à l'arrivée ou pas d'antimicrobien, ainsi qu'une injection sous-cutanée ou deux injections intramusculaires (à intervalle de 48 h) de florfénicol s'ils étaient diagnostiqués avec le complexe respiratoire bovin (CRB). Les veaux de différents groupes de traitement ont été regroupés pêle-mêle et suivis du placement à la réimplantation (~100 jours). Les animaux recevant l'OTC avaient un risque réduit de CRB, un risque accru d'arthrite et ne présentaient pas de différences significatives pour le gain de poids quotidien moyen, la rechute du CRB, la mortalité globale ou la mortalité associée au CRB. Il n'y avait aucune différence significative entre les protocoles de traitement. Des écouvillonnages nasaux profonds (n = 233) prélevés à l'arrivée (n = 122), au traitement (n = 77) et des écouvillonnages des poumons et des articulations post mortem (n = 34) ont été cultivés pour Mycoplasma bovis à partir de 61 animaux malades ou mourants de pneumonie chronique et d'arthrite et de 61 veaux en santé. Il n'y avait aucune variation significative dans la diversité des isolats (n = 51) entre les années d'étude et les différents bovins. La métaphylaxie ou le traitement antimicrobien n'a pas affecté la diversité des isolats. Sauf pour la tilmicosine, les isolats étaient largement sensibles aux antimicrobiens testés.(Traduit par Isabelle Vallières).

Evaluation of predictive models to determine final outcome for feedlot cattle based on information available at first treatment for bovine respiratory disease.

OBJECTIVE: To evaluate predictive model ability to determine whether an animal finished the feeding period using data known at first treatment for bovine respiratory disease (BRD). Additional comparisons evaluated the potential benefits of predictions by adding weather data, utilizing balancing techniques, and creating models for individual feedyards. ANIMALS: This retrospective study included animal, pen, and feedyard data from 12 US feedyards from 2016 to 2021. The final dataset consisted of 96,382 BRD cases of which 14.2% did not finish the feeding phase. PROCEDURES: Five predictive models were trained and underwent threshold probability adjustment to maximize F1 score. Model performance was evaluated using accuracy, sensitivity specificity, positive and negative predictive values, and area under the receiver operating characteristics curve (AUC). RESULTS: Overall, model performance was low with a median AUC value of 0.675. The addition of weather data had little effect on AUC but resulted in more variation in sensitivity and specificity. Resampling the dataset had a limited effect on performance. Individual feedlot models had higher AUC values than others with the decision tree typically performing best in most feedyards. CLINICAL RELEVANCE: Results indicated some utility of predictive models evaluating BRD cases to predict cattle that did not finish the feeding phase. These models could be valuable in assisting health providers making decisions on individual cases.

Calves severely affected by bovine respiratory disease have reduced protection against histone toxicity and exhibit lower complement activity.

Bovine respiratory disease (BRD) remains the greatest challenge facing the beef industry. Calves affected by BRD can manifest illness ranging from subclinical infection to acute death. In pathologies similar to BRD, extracellular histones have been implicated as major contributors to lung tissue damage. Histones are basic proteins responsible for DNA organization in cell nuclei, however when released extracellularly during cell injury or via neutrophil activation they become cytotoxic. Cattle suffering severe cases of BRD demonstrate reduced capacity to protect against the cytotoxic effects of histones, however, the protective mechanism(s) of serum remain(s) unknown. Therefore, the objective was to identify components within serum that contribute to protection against histone toxicity. Serum proteins from animals considered protective (P; N = 4) and nonprotective (NP; N = 4) against the toxic effects of histones were precipitated by the addition and incubation of exogenous histones. Proteins that interact with histones from both groups were isolated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and identified via label free "shotgun" proteomics. Sixteen candidate proteins increased by ≥2-fold change in P vs. NP animals were identified, with several associated with the complement system. A subsequent study was conducted to evaluate complement system activity and serum protective capacity against exogenous histones in feedlot heifers. Serum samples were collected from 118 heifer calves (BW at arrival = 229 ±â€…2.4 kg) at feedlot arrival. Animals were retrospectively assigned to groups consisting of: calves not requiring treatment with antibiotics for BRD (CONT; N = 80), calves treated once (1TRT; N = 21), calves treated twice (2TRT; N = 5), calves treated thrice (3TRT; N = 3), or calves that died from BRD within 1 wk of entering the feedlot (DA; N = 9). Serum from DA animals was less protective than CONT (P = 0.0005) animals against histone toxicity. Complement activity of DA animals was reduced compared to CONT (P = 0.0044) animals. Additionally, the use of both assays as a ratio resulted in increased ability to detect DA animals. Results suggest that cattle predisposed to severe cases of respiratory disease may have impaired complement activity presumably contributing to reduced protective capacity against histone toxicity.

Bovine respiratory disease (BRD) remains the leading cause of feedlot calf sickness and death. In respiratory disease affecting humans and mice, major tissue damage is caused by release of histones. Histones are proteins found in the nucleus of cells that condense DNA, however, cells that become damaged release histones extracellularly. Research has shown that calves with severe cases of BRD are less able to protect against the toxic effects of histones residing outside of the cell. It is speculated that components within the blood may interact with histones and confer protection from histone toxicity. This study evaluated serum from protective and nonprotective cattle against histone toxicity and identified 16 proteins that were elevated in protective animals. Several proteins were associated with the complement system of the innate immune system. To evaluate immune complement activity and protective capacity against histone toxicity, serum was collected from heifers at feedlot arrival. Calves suffering from a severe case of BRD demonstrated reduced capacity to protect against histone toxicity. Complement activity of calves severely affected with BRD was reduced as well. Results suggest that cattle susceptible to severe cases of BRD may have impaired complement activity likely contributing to reduced protective capacity against histone toxicity.

Hematological and immunological responses to naturally occurring bovine respiratory disease in newly received beef calves in a commercial stocker farm.

The objective was to determine temporal changes in hematological and immune parameters in response to naturally occurring bovine respiratory disease (BRD) in commercially managed stocker calves. Forty newly weaned beef steers purchased from auction markets were housed at a commercial stocker operation in Crossville, TN. Blood samples, rectal temperature, and thoracic ultrasonography (TUS; 1: normal to 3: severe) were collected on days 0, 7, 14, and 21. Castration status (FC: freshly castrated; PC: previously castrated) was determined on arrival based on presence of a fresh castration site at the scrotum. Calves received antibiotics for BRD based on clinical severity scoring (CSS; 0: moribund, 4: moribund) and rectal temperature. Complete blood counts (CBC) were performed. Calves were categorized based on the number of treatments (NumTrt) received (0x, 1x, and 2x). Temporal variations in CBC and immune parameters were analyzed using mixed model repeated measure ANOVA (Proc GLIMMIX; SAS 9.4). Variation of CBCs and immune parameters based on TUS was determined using mixed model ANOVA. There was a NumTrt by day interaction effect on the responses of white blood cells (WBC) (P = 0.04) and haptoglobin (HPT) (P = 0.04). On day 21, WBC were greater in the 2x NumTrt group than other groups, but there were no differences in WBC between NumTrt levels on other days. Haptoglobin was greater in the 2x group on days 14 and 21 than 0x or 1x. Red blood cells (RBC) (P = 0.02) and WBC (P = 0.04) differed between FC and PC groups, and lower RBC and WBC were observed in the FC group. A castration status × day effect for mean corpuscular volume (MCV; P = 0.04) was observed where FC group had higher MCV at days 14 and 21 than the PC group. Tumor necrosis factor-α differed based on NumTrt (P = 0.03) and higher concentrations were found in 2x group. We observed a day effect for IL-1ß (P = 0.009) and TNF-α (P = 0.001). Significant effect of TUS on HPT at day 14 (P = 0.0004) and day 21 (P = 0.002) was observed. Combining HPT and platelet explained 15% of the variability in treatment status on a given day, whereas HPT and hemoglobin explained 10% of the variability in lung consolidation status. Although hematological and immunological parameters varied largely in our study, the potential of combining HPT with hematological variables should be studied further. Results from this study would help in understanding temporal changes in CBC and immune parameters in newly received stocker cattle.

Blood and immune parameters are altered during bovine respiratory disease (BRD) progression and can be used for predicting disease status. We aimed looking at the dynamics of hematology and immunology in newly received stocker cattle in naturally occurring BRD. Forty newly received stocker cattle were managed by a local producer and monitored for BRD occurrence for 21 d after receiving during the high-risk period. Newly weaned calves were monitored as they experience several stress factors and become prone to BRD. Additionally, there are limited data related to immunological changes that occur in high-risk stocker cattle. Since there is no perfect diagnostic test for BRD, the diagnosis of BRD is likely missed when only visual signs are used. We observed that haptoglobin (HPT) was the most important parameter to differentiate BRD severity. The combination of HPT with blood parameters (hemoglobin and platelets) was useful to predict treatment and lung infection status. Therefore, measuring hematological and immunological parameters might be helpful to determine BRD status and facilitate treatment decisions in newly received stocker cattle.

A systematic review of the utility of biomarkers as aids in the early diagnosis and outcome prediction of bovine respiratory disease complex in feedlot cattle.

Bovine respiratory disease complex (BRDC) is a common, serious problem in feedlot cattle worldwide. Early diagnosis and outcome prediction are critical for making decisions to prevent economic loss and to limit antimicrobial use. Diagnosing BRDC is commonly based on visual signs and behavioral changes; both assessments are considered to have low diagnostic accuracy. Biomarkers are important for supporting the diagnosis of BRDC, determining the necessity and potential outcomes of treatment, and assisting in research in which differentiating diseased animals is required. There are few reviews summarizing the biomarkers available and utilized. We systematically evaluated the detection and prognostic potential of biomarkers from the literature published between January 1990 and December 2020. We performed a descriptive analysis of 5 biomarker categories: acute-phase proteins, stress-related hormones, other blood biomarkers, omics biomarkers, and non-blood biomarkers. The retrieved articles consisted of studies or trials that assessed the detection value and treatment and/or outcome prediction efficacy of biomarkers for BRDC in feedlot cattle; 23 manuscripts for review and analysis satisfied the selection criteria. Based on our review, we cannot recommend a specific biomarker as the sole method for the early detection or outcome prediction for BRDC, given that the application and efficacy of biomarkers varies in different situations. Our systematic review may serve as a reference for clinical and research investigations of early detection and outcome prediction of BRDC.

Limitations of bacterial culture, viral PCR, and tulathromycin susceptibility from upper respiratory tract samples in predicting clinical outcome of tulathromycin control or treatment of bovine respiratory disease in high-risk feeder heifers.

A cross-sectional prospective cohort study including 1026 heifers administered tulathromycin due to high risk of clinical signs of bovine respiratory disease (BRD), measured poor association between BRD clinical outcomes and results of bacterial culture and tulathromycin susceptibility from BRD isolates of deep nasopharyngeal swabs (DNS) and adequate association with viral polymerase chain reaction (PCR) results from nasal swabs. Isolation rates from DNS collected on day-0 and at 1st BRD-treatment respectively were: Mannheimia haemolytica (10.9% & 34.1%); Pasteurella multocida (10.4% & 7.4%); Mycoplasma bovis (1.0% & 36.6%); and Histophilus somni (0.7% & 6.3%). Prevalence of BRD viral nucleic acid on nasal swabs collected exclusively at 1st BRD-treatment were: bovine parainfluenza virus type-3 (bPIV-3) 34.1%; bovine viral diarrhea virus (BVDV) 26.3%; bovine herpes virus type-1 (BHV-1) 10.8%; and bovine respiratory syncytial virus (BRSV) 54.1%. Increased relative risk, at 95% confidence intervals, of 1st BRD-treatment failure was associated with positive viral PCR results: BVDV 1.39 (1.17-1.66), bPIV-3 1.26 (1.06-1.51), BHV-1 1.52 (1.25-1.83), and BRSV 1.35 (1.11-1.63) from nasal swabs collected at 1st BRD-treatment and culture of M. haemolytica 1.23 (1.00-1.51) from DNS collected at day-0. However, in this population of high-risk feeder heifers, the predictive values of susceptible and resistant isolates had inadequate association with BRD clinical outcome. These results indicate, that using tulathromycin susceptibility testing of isolates of M. haemolytica or P. multocida from DNS collected on arrival or at 1st BRD-treatment to evaluate tulathromycin clinical efficacy, is unreliable.

Comprehensive at-arrival transcriptomic analysis of post-weaned beef cattle uncovers type I interferon and antiviral mechanisms associated with bovine respiratory disease mortality.

BACKGROUND: Despite decades of extensive research, bovine respiratory disease (BRD) remains the most devastating disease in beef cattle production. Establishing a clinical diagnosis often relies upon visual detection of non-specific signs, leading to low diagnostic accuracy. Thus, post-weaned beef cattle are often metaphylactically administered antimicrobials at facility arrival, which poses concerns regarding antimicrobial stewardship and resistance. Additionally, there is a lack of high-quality research that addresses the gene-by-environment interactions that underlie why some cattle that develop BRD die while others survive. Therefore, it is necessary to decipher the underlying host genomic factors associated with BRD mortality versus survival to help determine BRD risk and severity. Using transcriptomic analysis of at-arrival whole blood samples from cattle that died of BRD, as compared to those that developed signs of BRD but lived (n = 3 DEAD, n = 3 ALIVE), we identified differentially expressed genes (DEGs) and associated pathways in cattle that died of BRD. Additionally, we evaluated unmapped reads, which are often overlooked within transcriptomic experiments. RESULTS: 69 DEGs (FDR<0.10) were identified between ALIVE and DEAD cohorts. Several DEGs possess immunological and proinflammatory function and associations with TLR4 and IL6. Biological processes, pathways, and disease phenotype associations related to type-I interferon production and antiviral defense were enriched in DEAD cattle at arrival. Unmapped reads aligned primarily to various ungulate assemblies, but failed to align to viral assemblies. CONCLUSION: This study further revealed increased proinflammatory immunological mechanisms in cattle that develop BRD. DEGs upregulated in DEAD cattle were predominantly involved in innate immune pathways typically associated with antiviral defense, although no viral genes were identified within unmapped reads. Our findings provide genomic targets for further analysis in cattle at highest risk of BRD, suggesting that mechanisms related to type I interferons and antiviral defense may be indicative of viral respiratory disease at arrival and contribute to eventual BRD mortality.

Ivermectin also inhibits the replication of bovine respiratory viruses (BRSV, BPIV-3, BoHV-1, BCoV and BVDV) in vitro.

Bovine respiratory disease (BRD) complex is an important viral infection that causes huge economic losses in cattle herds worldwide. However, there is no directly effective antiviral drug application against respiratory viral pathogens; generally, the metaphylactic antibacterial drug applications are used for BRD. Ivermectin (IVM) is currently used as a broad-spectrum anti-parasitic agent both for veterinary and human medicine on some occasions. Moreover, since it is identified as an inhibitor for importin α/ß-mediated nuclear localization signal (NLS), IVM is also reported to have antiviral potential against several RNA and DNA viruses. Since therapeutic use of IVM in COVID-19 cases has recently been postulated, the potential antiviral activity of IVM against bovine respiratory viruses including BRSV, BPIV-3, BoHV-1, BCoV and BVDV are evaluated in this study. For these purposes, virus titration assay was used to evaluate titers in viral harvest from infected cells treated with non-cytotoxic IVM concentrations (1, 2.5 and 5 µM) and compared to titers from non-treated infected cells. This study indicated that IVM inhibits the replication of BCoV, BVDV, BRSV, BPIV-3 and BoHV-1 in a dose-dependent manner in vitro as well as number of extracellular infectious virions. In addition, it was demonstrated that IVM has no clear effect on the attachment and penetration steps of the replication of the studied viruses. Finally, this study shows for the first time that IVM can inhibit infection of BRD-related viral agents namely BCoV, BPIV-3, BVDV, BRSV and BoHV-1 at the concentrations of 2.5 and 5 µM. Consequently, IVM, which is licensed for antiparasitic indications, also deserves to be evaluated as a broad-spectrum antiviral in BRD cases caused by viral pathogens.

In-vitro antibiotic resistance phenotypes of respiratory and enteric bacterial isolates from weaned dairy heifers in California.

Antimicrobial drug (AMD) use for bovine respiratory disease (BRD) continues to be concerning for development of antimicrobial resistance (AMR) in respiratory and enteric bacteria of cattle. This study aimed to provide data regarding AMR in respiratory isolates, and identify relationships between respiratory and enteric AMD susceptibility, in weaned dairy heifers. A cross-sectional study was performed between June of 2019 and February 2020, on 6 calf rearing facilities in California. Deep nasopharyngeal and rectal swabs were collected from 341 weaned heifers and submitted for selective bacterial culture and AMR testing. Mannheimia haemolytica, Pasteurella multocida, and Histophilus somni were selectively isolated from respiratory samples; Escherichia coli and Enterococcus spp. were selectively isolated from rectal swabs. Minimum inhibitory concentrations (MIC) were determined for selected isolates against 19 AMD. The proportion of resistant isolates was calculated using Clinical Laboratory Standards Institute (respiratory) or USDA NARMS (enteric) breakpoints; when no applicable breakpoint was available, the distribution of MIC was described and compared. Association between AMR in a calf's respiratory isolate and a higher or lower MIC of the matched enteric isolates was determined. More than 50% of P. multocida isolates were resistant to each of 7 AMD commonly used to treat BRD (florfenicol, gamithromycin, tildipirosin, tilmicosin, danofloxacin, enrofloxacin and tetracycline). Resistance in respiratory isolates was only associated with higher matched enteric MIC for gamithromycin and tulathromycin. Multidrug resistance was reported in >70% of P. multocida and M. haemolytica isolates. Antimicrobial resistance, including multidrug resistance, in respiratory isolates appears to be widespread in weaned dairy heifers; this finding has not previously been reported and raises concern for the future efficacy of AMD used to treat respiratory diseases in weaned dairy heifers. Enteric bacterial MIC appear to have limited direct association with respiratory isolate AMR classification.

Pharmacokinetics of gamithromycin in cattle with comparison of plasma and lung tissue concentrations and plasma antibacterial activity.

The pharmacokinetics (PK) and dose proportionality of gamithromycin (ZACTRAN), a novel azalide, after a single intravenous (i.v.) dose of 3 mg/kg or subcutaneous (s.c.) injection at 3, 6 and 9 mg/kg body weight were studied in 13 male castrate and 13 female Angus cattle. Following i.v. administration, the mean area under the curve extrapolated to infinity (AUC(inf)) was 4.28 +/- 0.536 microgxh/mL, and mean elimination half-life (t(1/2)) was 44.9 +/- 4.67 h, with a large volume of distribution (V(ss)) of 24.9 +/- 2.99 L/kg and a high clearance rate (Cl(obs)) of 712 +/- 95.7 mL/h/kg. For cattle treated with s.c. injection of 3, 6 or 9 mg/kg, mean AUC(inf) values were 4.55 +/- 0.690, 9.42 +/- 1.11 and 12.2 +/- 1.13 microgxh/mL, respectively, and the mean elimination half-lives (t(1/2)) were 51.2 +/- 6.10, 50.8 +/- 3.80 and 58.5 +/- 5.50 h. Gamithromycin was well absorbed and fully bioavailable (97.6-112%) after s.c. administration. No statistically significant (alpha = 0.05) gender differences in the AUC(Inf) or elimination half-life values were observed. Dose proportionality was established based on AUC(Inf) over the range of 0.5 to 1.5 times of the recommended dosage of 6 mg/kg of body weight. Further investigations were conducted to assess plasma PK, lung/plasma concentration ratios and plasma antibacterial activity using 36 cattle. The average maximum gamithromycin concentration measured in whole lung homogenate was 18 500 ng/g at first sampling time of 1 day ( approximately 24 h) after treatment. The ratios of lung to plasma concentration were 265, 410, 329 and 247 at 1, 5, 10 and 15 days postdose. The lung AUC(inf) was 194 times higher than the corresponding plasma AUC(inf). The apparent elimination half-life for gamithromycin in lung was 90.4 h ( approximately 4 days). Antibacterial activity was observed with plasma collected at 6 h postdose with a corresponding average gamithromycin plasma concentration of 261 ng/mL. In vitro plasma protein binding in bovine plasma was determined to be 26.0 +/- 0.60% bound over a range of 0.1-3.0 microg/mL of gamithromycin. The dose proportionality of AUC, high bioavailability, rapid and extensive distribution to lung tissue and low level of plasma protein binding are beneficial PK parameters for an antimicrobial drug used for the treatment and prevention of bovine respiratory disease.

Pharmacological considerations of antibiotic failures in bovine respiratory disease cases.

Bovine respiratory disease (BRD) is one of the most common indications for antimicrobial therapy in beef cattle production and research trials demonstrate that antibiotic therapy greatly improves clinical outcome for BRD. These trials also show that BRD treatment success rates are less than 100% and that there are opportunities to optimize antimicrobial prescribing and improve clinical outcomes if the underlying cause(s) of BRD treatment failures can be identified and addressed. As the etiology of BRD in an individual animal is frequently multi-factorial in nature; it is likely that BRD treatment failures also result from complex interactions between the drug, drug administrator, animal host, pathogens, and the environment. This review will focus specifically on the pharmacological aspects, specifically the interactions between the host and the drug and the drug and the drug administrator, of BRD treatment failures and the actions that veterinary practitioners can take to investigate and mitigate therapeutic failures in future cases.

Bovine Respiratory Disease Treatment Failure: Impact and Potential Causes.

Bovine respiratory disease (BRD) is often attributed to complex interactions between the host, pathogen, and the environment. Likewise, many BRD treatment failures result from interactions between the host, pathogen, environment, drug, and drug administrator. Investigating and addressing the underlying causes of BRD treatment failures can improve clinical outcomes and animal welfare of future cases, improve morale of employees, reduce direct costs of dealing with BRD treatment failures, refine antimicrobial prescribing practices, and advance antimicrobial stewardship. This article discusses these interactions and provides guidance to veterinary practitioners on evaluating the success of treatment protocols.

Bovine respiratory disease treatment failure: definition and impact.

Bovine respiratory disease (BRD) treatment failure occurs when animals receiving a treatment regimen for BRD fail to directly return to health, resulting in chronic illness and a requirement for repeated treatments, sale for salvage slaughter, euthanasia or death. BRD treatment failure has both direct and indirect impacts. Direct impacts include costs to manage chronically ill animals, including those associated with BRD relapse treatment; reduced returns from animals sent for salvage slaughter, loss of the initial investment to purchase the animal and feed and other accumulated expenses to death, and costs associated with carcass disposal. Indirect impacts include costs of infrastructure requirements, and negative effects on animal welfare and employee morale.

Antimicrobial resistance in Mannheimia haemolytica: prevalence and impact.

Bovine respiratory disease (BRD) is the most common cause of morbidity and mortality in North American beef cattle. In recent years, isolation of strains of Mannheimia haemolytica that are resistant to multiple different classes of antimicrobials has become commonplace. New research would suggest that the routine use of antimicrobials by some cattle operations might be driving emerging resistance patterns, with the majority of the spread observed due to propagation of strains of M. haemolytica that have acquired integrative conjugative elements. To date, there is little information evaluating the impact of antimicrobial resistance on clinical outcome in cattle with BRD.