Bovine Respiratory Disease Research Articles

Influenza D virus infection in China, 2022–2023

ABSTRACT Influenza D virus (IDV) plays an important role in the bovine respiratory disease (BRD) complex. Its potential for the zoonotic transmission is of particular concern. In China, IDV has previously been identified in agricultural animals by molecular surveys with no live virus isolates reported. In this study, live IDVs were successfully isolated from cattle in China, which prompted us to further investigate the national prevalence, antigenic property, and infection biology of the virus. IDV RNA was detected in 11.1% (51/460) of cattle throughout the country in 2022–2023. Moreover, we conducted the first IDV serosurveillance in China, revealing a high seroprevalence (91.4%, 393/430) of IDV in cattle during the 2022–2023 winter season. Notably, all the 16 provinces from which cattle originated possessed seropositive animals, and 3 of them displayed the 100% IDV-seropositivity rate. In contrast, a very low seroprevalence of IDV was observed in pigs (3%, 3/100) and goats (1%, 1/100) during the same period of investigation. Furthermore, besides D/Yama2019 lineage-like IDVs, we discovered the D/660 lineage-like IDV in Chinese cattle, which has not been detected to date in Asia. Finally, the Chinese IDVs replicated robustly in diverse cell lines but less efficiently in the swine cell line. Considering the nationwide distribution, high seroprevalence, and appreciably genetic diversity, further studies are required to fully evaluate the risk of Chinese IDVs for both animal and human health in China, which can be evidently facilitated by IDV isolates reported in this study.

Influence of autolyzed yeast on the health of immunologically immature calves

Autolyzed yeast is a supplement option for calves due to its ability to stimulate the immune system and performance; however, there are few studies about the effect of different yeast derivatives. Autolyzed yeast is rich in immunomodulatory substances, such as mannan oligosaccharides and β-glucans. These substances enhance the innate and humoral immunity of calves, resulting in greater intestinal and respiratory health. Thus, the objective of the study was to evaluate whether supplementation with autolyzed yeast improves the health of calves, in the challenge of naturally acquired Eimeriosis and bovine respiratory disease (BRD). Twenty Holstein calves aged 15 days, which had already suckled colostrum, were studied for 36 days. At 15 days of life all animals had naturally acquired Eimeriosis. They were challenged with an intranasal BRD vaccine at 30 days of age. The Supplemented group (n=10) received autolyzed yeast (10 g animal -1 day -1), once a day in milk for 36 days, and the Control group (n=10) was not supplemented throughout the experiment. Blood neutrophil: lymphocyte ratio, oxidative metabolism of neutrophils (OM), serum immunoglobulins and haptoglobin, occurrence of diarrhea, BRD, and weight gain were measured. After vaccination, all animals increased the blood neutrophil: lymphocyte ratio, but only the control group showed a 20% reduction in OM three days after the vaccine. The supplement affected the immune response, preserving OM, increasing serum IgA levels by 10% (P=0.006), and reducing haptoglobin serum levels (P=0.05). In addition, the supplement attenuated diarrhea and BRD, which allowed for greater weekly weight gain (difference of 2 kg between treatments, P<0.05), concluding that the autolyzed yeast showed some evidence that it can increase the health of calves challenged with Eimeriosis and the BRD vaccine.

Use of Immunostimulators in the Prevention of Coronavirus Disease in Calves

Vaccination and passive immunization, disinfection and biosecurity, feeding and feeding, care, hygiene and immune status of animals are essential to prevent BCoV diseases, limit the use of antibacterial drugs and reduce the subsequent emergence of microbial resistance, improve the quality of animal products and animals. Multivalent vaccines are now available to limit the spread of the virus and achieve hyperimmune herd passive immunization in calves, which may protect or reduce the severity of BCoV diarrhea in calves. The importance of passive immunization is noted. An oral live vaccine has also been developed to prevent diarrhea in calves. Studies have also been conducted on the use of intranasal BCoV vaccination in calves against bovine respiratory disease and winter dysentery.

Evaluation of bovine colostrum replacer supplementation to improve weaning transition in Holstein dairy calves

The objective of this randomized controlled trial was to evaluate the efficacy of supplementing bovine colostrum replacer during weaning to reduce intestinal permeability and improve gain. For this experiment, 65 calves were enrolled and housed individually until 70 d of age. Calves were fed milk replacer (150 g/L) 3 times daily with 9 L/d, 10.5 L/d, 11.25 L/d, and 12 L/d offered from d 1-7, d 8-14, d 15-21, d 22-56, respectively. Calves were weaned over 8 d from d 57-64, receiving a total of 7.8 L in 2 meals/d from d 57-60 and 3.8 L/d in one feeding from d 61-64. At d 57, calves were blocked by birth weight and randomly assigned to one of 2 treatments, equal in metabolizable energy, which were fed once daily during weaning from d 57-64: control (CON; n = 31 calves): 3.8 L milk replacer (150 g/L) fed by nipple bottle, or colostrum supplementation (COL; n = 34): a mixture of 1 L bovine colostrum replacer (125 g/L) and 3 L milk replacer (150 g/L) with 3.8 L of the mixture fed by nipple bottle. Serum IgG was measured within 48 h of birth and body weight was taken at d 0, 57, 60, 64, 70, 77, and 84. Starter intake and bovine respiratory disease (BRD) score were measured daily from d 50-70 and fecal consistency was examined daily from d 56-70. Serum β-hydroxybutyrate (BHB) and lung consolidation were evaluated at d 57, 64, and 70 and intestinal permeability was assessed by recovery of chromium-EDTA, lactulose, and d-mannitol from plasma after oral administration at d 56 and 65. There was no difference in body weight between treatment groups at the start of weaning, but COL were 2.79 kg (95% CI: 0.90-4.68) and 2.76 kg (95% CI: 0.86-4.65) heavier than CON at d 77 and 84, respectively. Additionally, COL tended to gain 100.00 g/d more than CON calves (95% CI: -10.41-207.13) from d 57-84. There were no differences in any of the other variables measured. Supplementation of bovine colostrum replacer during weaning may improve weight gain, but the mechanism of action is not clear.

The Canadian Cow-Calf Surveillance Network - productivity and health summary 2018 to 2022.

Cow-calf surveillance data provide critical information about changing herd demographics, productivity, health, and management as well as the opportunity to investigate the impact of differing management practices. A national cow-calf network was established to collect baseline information on herd health, production performance measures, and management. Questionnaires were used to collect information on herd attributes as well as data for the calving season, breeding, pregnancy testing, and weaning for herds from across Canada. From 2018 to 2022, a total of 565 calving record questionnaires, representing 110,658 calving female records from 171 herds were returned, along with 543 herd breeding to weaning questionnaires. Suggested benchmark values based on the 25th percentiles from Western and Eastern Canada were determined to be <5% for non-pregnancy, <2% for calf death from birth to 24 h, and < 2% for calf death from 24 h to weaning. Herds from Eastern Canada were more likely to report any assistance at calving (p < 0.001) than herds from Western Canada. Participating herds from the east had longer breeding (p < 0.001) and calving (p < 0.001) seasons than those from the west and were also more likely to use artificial insemination or embryo transfer (p < 0.001). Timing of calving season and use of large pastures for calving were not associated with mortality at birth. Herds that started calving before April were more likely to have calves die before 30 d than those that started calving later; however, this difference was not observed between 30 d and weaning. Herds that started to calve earlier were also more likely to report treating more calves for bovine respiratory disease, diarrhea, and navel or joint infections as well as to calf deaths from respiratory disease. Calves from herds in the east were more likely to be treated or die from diarrhea than from the west. Females from herds that started calving later were less likely to be pregnant. The unique longitudinal productivity and health data resulting from this network established a national baseline to address region-specific needs for knowledge translation and solutions to enhance productivity and support sustainability.

Antimicrobial Activities of α-Helix and β-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 β-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 β-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.

Dogs' ability to detect an inflammatory immune response in cattle via olfaction.

Canine olfaction is a potential means for detection of respiratory disease in beef cattle. In a prior study, two dogs were trained to discriminate between nasal swabs from healthy cattle and cattle that developed Bovine Respiratory Disease. Dogs had some ability to identify samples from BRD-affected cattle, but results were ambiguous. The purpose of this study was to evaluate more dogs using better-controlled training and testing procedures. Nasal and saliva swabs were collected from 96 cattle before and after administering a vaccine to induce an inflammatory immune response. Samples were stored at -80°C for up to 11 months before use, and samples from animals with an elevated body temperature at baseline were omitted. An automated olfactometer apparatus was constructed to improve blinding procedures and reduce opportunities for odor contamination. Four dogs were trained to distinguish between swabs from healthy and sickness-model cattle, including the two dogs from the previous study ("Runnels" and "Cheaps") and two inexperienced dogs ("Molokai" and "Amy"). During a seven-month training period, dogs were exposed to samples from 28 animals. Dogs were tested on 59 sets of unfamiliar samples. Performance varied among dogs (χ2 = 10.48, p = 0.02). Molokai's performance was above chance (0.73 ± 0.06, p = 0.0006), while Amy (0.44 ± 0.06, p = 0.43), Cheaps (0.53 ± 0.07, p = 0.79), and Runnels (0.56 ± 0.06, p = 0.43) did not respond correctly at a rate different from chance. Accuracy did not differ between nasal swabs (0.63 ± 0.08) and saliva swabs (0.53 ± 0.08, χ2 = 0.81, p = 0.37). The results of this study indicate that canine olfaction may be an effective means of detecting illness in beef cattle. However, individual dogs' aptitude for this detection task varies.

Identifying and addressing barriers and opportunities for bovine respiratory disease complex vaccination: a consensus paper on practical recommendations for best practise vaccination.

Many questions are raised, and challenges faced in the new era of (intranasal) bovine respiratory disease complex vaccination. An increase in vaccination rate is expected, due to its positive impact on cattle health, reduction of antimicrobial use and economic factors. However, engagement of farmers and veterinarians with regard to vaccination is often affected by limitations, resulting in the development of barriers to vaccination, but also opportunities to overcome these. The objective of the report is to provide practical recommendations and a consensus on best practises for BRDC vaccination, addressing barriers faced by veterinarians and farmers. The report combines an evidence review with expert opinions and includes discussions on different vaccination approaches, such as intranasal and systemic protocols. As result of the discussions, several barriers to BRDC vaccination were identified, including a lack of knowledge or visibility of the disease's impact, the preference for blanket antibiotic use over vaccination, resistance to change, the need for visible success, uncertainty about the best time to vaccinate, and concerns about adverse reactions and vaccine efficacy in the presence of maternal antibodies. While these barriers seem substantial, they provide opportunities for the veterinary sector. Indeed, veterinarians are encouraged to use the argumentation presented, along with local case studies and diagnostic testing to highlight the impact of disease, while conducting calf health audits, ensuring expectations are managed to achieve visible success. Overall, this consensus paper aims to provide practical recommendations and support for veterinarians and farmers to overcome barriers and increase BRDC vaccination rates in cattle.

Investigation of the safety and protective efficacy of an attenuated and marker M. bovis-BoHV-1 combined vaccine in bovines.

Bovine respiratory disease (BRD) is one of the most common diseases in the cattle industry worldwide; it is caused by multiple bacterial or viral coinfections, of which Mycoplasma bovis (M. bovis) and bovine herpesvirus type 1 (BoHV-1) are the most notable pathogens. Although live vaccines have demonstrated better efficacy against BRD induced by both pathogens, there are no combined live and marker vaccines. Therefore, we developed an attenuated and marker M. bovis-BoHV-1 combined vaccine based on the M. bovis HB150 and BoHV-1 gG-/tk- strain previously constructed in our lab and evaluated in rabbits. This study aimed to further evaluate its safety and protective efficacy in cattle using different antigen ratios. After immunization, all vaccinated cattle had a normal rectal temperature and mental status without respiratory symptoms. CD4+, CD8+, and CD19+ cells significantly increased in immunized cattle and induced higher humoral and cellular immune responses, and the expression of key cytokines such as IL-4, IL-12, TNF-α, and IFN-γ can be promoted after vaccination. The 1.0× 108 CFU of M. bovis HB150 and 1.0 × 106 TCID50 BoHV-1 gG-/tk- combined strain elicited the most antibodies while significantly increasing IgG and cellular immunity after challenge. In conclusion, the M. bovis HB150 and BoHV-1 gG-/tk- combined strain was clinically safe and protective in calves; the mix of 1.0 × 108 CFU of M. bovis HB150 and 1.0 × 106 TCID50 BoHV-1 gG-/tk- strain was most promising due to its low amount of shedding and highest humoral and cellular immune responses compared with others. This study introduces an M. bovis-BoHV-1 combined vaccine for application in the cattle industry.

Antimicrobial Resistance Genes in Respiratory Bacteria from Weaned Dairy Heifers.

Bovine respiratory disease (BRD) is the leading cause of mortality and antimicrobial drug (AMD) use in weaned dairy heifers. Limited information is available regarding antimicrobial resistance (AMR) in respiratory bacteria in this population. This study determined AMR gene presence in 326 respiratory isolates (Pasteurella multocida, Mannheimia haemolytica, and Histophilus somni) from weaned dairy heifers using whole genome sequencing. Concordance between AMR genotype and phenotype was determined. Twenty-six AMR genes for 8 broad classes of AMD were identified. The most prevalent, medically important AMD classes used in calf rearing, to which these genes predict AMR among study isolates were tetracycline (95%), aminoglycoside (94%), sulfonamide (94%), beta-lactam (77%), phenicol (50%), and macrolide (44%). The co-occurrence of AMR genes within an isolate was common; the largest cluster of gene co-occurrence encodes AMR to phenicol, macrolide, elfamycin, β-lactam (cephalosporin, penam cephamycin), aminoglycoside, tetracycline, and sulfonamide class AMD. Concordance between genotype and phenotype varied (Matthew's Correlation Coefficient ranged from -0.57 to 1) by bacterial species, gene, and AMD tested, and was particularly poor for fluoroquinolones (no AMR genes detected) and ceftiofur (no phenotypic AMR classified while AMR genes present). These findings suggest a high genetic potential for AMR in weaned dairy heifers; preventing BRD and decreasing AMD reliance may be important in this population.

Variation in Pen-Level Prevalence of BRD Bacterial Pathogens and Antimicrobial Resistance Following Feedlot Arrival in Beef Calves.

Antimicrobials are crucial for treating bovine respiratory disease (BRD) in beef feedlots. Evidence is needed to support antimicrobial use (AMU) decisions, particularly in the early part of the feeding period when BRD risk is highest. The study objective was to describe changes in prevalence and antimicrobial susceptibility of BRD bacterial pathogens at feedlot processing (1 day on feed (1DOF)), 12 days later (13DOF), and for a subset at 36DOF following metaphylactic antimicrobial treatment. Mixed-origin steer calves (n = 1599) from Western Canada were managed as 16 pens of 100 calves, receiving either tulathromycin (n = 1199) or oxytetracycline (n = 400) at arrival. Deep nasopharyngeal swabs collected at all time points underwent culture and antimicrobial susceptibility testing (AST). Variability in the pen-level prevalence of bacteria and antimicrobial susceptibility profiles were observed over time, between years, and metaphylaxis options. Susceptibility to most antimicrobials was high, but resistance increased from 1DOF to 13DOF, especially for tetracyclines and macrolides. Simulation results suggested that sampling 20 to 30 calves per pen of 200 reflected the relative pen-level prevalence of the culture and AST outcomes of interest. Pen-level assessment of antimicrobial resistance early in the feeding period can inform the evaluation of AMU protocols and surveillance efforts and support antimicrobial stewardship in animal agriculture.

Risk Assessment Tools from the One Health Perspective: A Narrative Review.

61% of the infections around the world that have emerged to date are zoonotic. Evidence warns that the threat posed by zoonoses is on the rise, and the risk of a new pandemic is higher now than ever. Early identification of risk, populations at risk, and risk of transmission are essential steps towards a prevention, preparation and response to outbreaks. This review aims to look at the tools available for identifying and estimating risks and threats from one health perspective and finally propose a list of indicators which could assess the risk of transmission of disease at the humans, animals and the environment intersection. The databases like PubMed, google scholar, Embase and Scopus were used to extract the relevant articles. A search was carried out using a keyword. A total of 1311 articles were listed initially after the search and reviewed. Out of 1311, only 26 tools which assessed the risk of diseases mainly infectious or were relevant to risk of transmission of any infectious diseases were included in the review. The tools included in this review involve risk assessment at the environmental, animal and human dimensions. The tools are used to evaluate the contamination of the environment due to chemicals or toxins or the risk of transmission of infection due to environmental factors like air contamination, to identify the animal diseases like bovine respiratory disease and foot and mouth disease and to estimate the human health risk at the community or individual levels. Risk assessment tools are an essential part of the prevention of pandemics. These tools are helpful in assessing the risk of transmission of infections either from human to human, between human and animals, between animals and animals and so on. Thus this review gives us an insight into the existing risk assessment tools and the need for a One Health risk assessment tools to prevent outbreaks in future. It also provides a list of factors that can be included in a one health risk assessment tool.

Development of a detection chip for major pathogenic drug-resistant genes and drug targets in bovine respiratory system diseases.

Bovine respiratory disease (BRD) is a significant veterinary challenge, often exacerbated by pathogen resistance, hindering effective treatment. Traditional testing methods for primary pathogens - Mycoplasma bovis, Pasteurella multocida, and Mannheimia haemolytica - are notably time-consuming and lack the rapidity required for effective clinical decision-making. This study introduces a TaqMan MGB probe detection chip, utilizing fluorescent quantitative PCR, targeting key BRD pathogens and associated drug-resistant genes and sites. We developed 94 specific probes and primers, embedded into a detection chip, demonstrating notable specificity, repeatability, and sensitivity, reducing testing time to under 1 h. Additionally, we formulated probes to detect mutations in the quinolone resistance-determining region, associated with fluoroquinolone resistance in BRD pathogens. The chip exhibited robust sensitivity and specificity, enabling rapid detection of drug-resistant mutations in clinical samples. This methodology significantly expedites the diagnostic process for BRD and sensitive drug screening, presenting a practical advancement in the field.

Influenza D virus in cattle (review)

The influenza D virus was first detected and identified in 2011. The overall amino acid sequence of influenza D virus shares approximately 50% identity with that of influenza C virus, suggesting that both viruses had a common ancestor. Cattle is considered to be the primary natural reservoir for influenza D virus. The involvement of this virus into the bovine respiratory disease complex has been confirmed. The virus causes mild to moderate disease in calves and replicates in both the upper and lower respiratory tracts, promoting bronchopneumonia. The influenza D virus can be transmitted by contact or aerosol over short distances, has a high transmission rate and can potentiate the effects of other respiratory pathogens. There are currently no vaccines or specific treatment for influenza D virus. This virus can replicate and be transmitted by direct contact in ferrets and guinea pigs, which are surrogate models of human influenza infection, as well as in well-differentiated human airway epithelial cells (hAECs). Currently five distinctive lineages of influenza D virus have been identified, co-circulating in worldwide bovine and pig populations that may facilitate genetic re-assortment between different viral strains. The virus has a zoonotic potential, and if its pathogenicity for humans changes, its importance for public health will be great. Very high seropositivity rates among persons working with cattle in the USA and Italy have been reported. There is no data in the available literature on the circulation of the influenza D virus in the Russian Federation. Research is needed to study this new virus, as well as monitoring of the virus spread and circulation in our country to understand its role in bovine respiratory disease complex and its zoonotic potential.

Association of passive immunity and genetic composition, health, and performance of tropical dairy calves

This retrospective cohort study aimed to identify risk factors for the failure of transfer of passive immunity (FPI) in preweaned dairy calves, explore its associations with morbidity, mortality, genetics, and determine a standardized cut-off point for FPI. Analyzing data from 6011 calves, factors such as sire predicted transmitted ability for milk (PTA), birth season, retained placenta (RP), total serum protein concentration (TSP), morbidity (neonatal calf diarrhea - NCD, bovine respiratory disease - BRD, tick-borne disease - TBD), mortality, and average daily weight gain (ADG) were considered. The calves were categorized into predominantly Gyr (PG) and predominantly Holstein (PH) genetic composition groups. Multivariate mixed logistic regression revealed optimal TSP cut-off points for predicting morbidity (7.6 g/dL) and mortality (6.9 g/dL). PH calves exhibited 1.35 times higher odds of FPI and 1.48 times greater odds of disease. Calves from multiparous cows and those born to dams with RP had increased FPI odds. Disease prevalence was 53%, with 41% NCD, 18% BRD, and 10% TBD. Season, parity, PTA, and birth weight were associated with disease odds, though FPI was not a reliable predictor. The mortality percentage was 6%, with PH calves and those with a positive PTA having higher odds. The ADG was 0.64 kg, and FPI-affected calves gained less weight. The study compared and identified various risk factors that potentially impact calf immunity. However, the use of a standardized cut-off point for FPI assessment was not effective in predicting morbidity and mortality at this specific farm.

Construction and characterization of a reverse genetics system of bovine parainfluenza virus type 3c as a tool for rapid screening of antivirals in vitro.

Bovine parainfluenza virus type 3 (BPIV3) is a key pathogen associated with bovine respiratory disease complex (BRDC). However, its specific pathogenesis mechanisms have not been fully elucidated. Reverse genetics provides a useful method for understanding the pathogenic mechanism of BPIV3. To ensure the functionality of the rescue platforms, we first constructed a minigenome (MG) system of BPIV3 utilizing a 5-plasmid system in this investigation. Then, a full-length infection clone of BPIV3 was obtained from the SX-2021 strain, and different methods were employed to identify the rescued virus. Additionally, we recovered a recombinant BPIV3 using the reverse genetics system that could express enhanced green fluorescence protein (eGFP). Through the growth curve assays, the replicate capability of rBPIV3-SX-EGFP was found to be similar to that of the parental virus. Subsequently, the rBPIV3-SX-EGFP was used to determine the antiviral activity of ribavirin. The results showed that ribavirin had an anti-BPIV3 effect in MDBK cells. In conclusion, the successful development of a reverse genetic system for the SX-2021 strain establishes a foundation for future studies on BPIV3, including investigations into its pathogenic mechanism, gene function, and antiviral screening properties.

Prevalence, Molecular Characteristics and Virulence Identification of Bovine Parainfluenza Virus Type 3 in China.

Bovine parainfluenza virus type 3 (BPIV-3) is one of the major pathogens of the bovine respiratory disease complex (BRDC). BPIV-3 surveillance in China has been quite limited. In this study, we used PCR to test 302 cattle in China, and found that the positive rate was 4.64% and the herd-level positive rate was 13.16%. Six BPIV-3C strains were isolated and confirmed by electron microscopy, and their titers were determined. Three were sequenced by next-generation sequencing (NGS). Phylogenetic analyses showed that all isolates were most closely related to strain NX49 from Ningxia; the genetic diversity of genotype C strains was lower than strains of genotypes A and B; the HN, P, and N genes were more suitable for genotyping and evolutionary analyses of BPIV-3. Protein variation analyses showed that all isolates had mutations at amino acid sites in the proteins HN, M, F, and L. Genetic recombination analyses provided evidence for homologous recombination of BPIV-3 of bovine origin. The virulence experiment indicated that strain Hubei-03 had the highest pathogenicity and could be used as a vaccine candidate. These findings apply an important basis for the precise control of BPIV-3 in China.

148 Supplementing Bovacillus to newly weaned beef steers during a 90-d grazing period

Abstract This experiment compared performance, health, and physiological responses of high-risk stocker cattle receiving a Bacillus subtilis derivative during a 90-d grazing period. Crossbred steers (n = 120) were obtained from an auction yard and transported to the experimental facility. Steer shrunk body weight (BW) was recorded at arrival (d -1). This value was averaged with BW recorded on d 0 to represent initial BW (236 ± 2 kg). On d 0, steers were ranked by BW and allocated to 1 of 12 pastures (4-ha pastures; 10 steers/pasture). Pastures were randomly assigned to receive daily supplementation with dried distillers’ grains at 1% of BW containing either: 1) Bacillus licheniformis and B. subtilis ingredient (BOV; 2 g/steer daily of Bovacillus, Chr Hansen A/S, Horsholm, Denmark) or 2) no additive (CON). Cattle received treatments from d 0 to 90 while housed in pastures containing stockpiled grass forage, as well as free-choice access to water and a commercial mineral + vitamin mix without ionophores. Steers were assessed for bovine respiratory disease (BRD) signs daily, and shrunk BW recorded on d 91. Blood samples were collected, and BW recorded on d 0, 14, 28, 56, and 90. No treatment differences were detected (P ≥ 0.19) for glucose, non-esterified fatty acids, β-hydroxybutyrate, haptoglobin, or insulin-like growth factor 1, whereas a treatment × day interaction indicated increased cortisol in BOV cattle on d 56 (P &amp;lt; 0.01). A treatment × day interaction was detected for serum concentrations of parainfluenza type 3 antibodies, which was greater in BOV steers on d 90 (P &amp;lt; 0.01). No treatment differences were detected (P ≥ 0.23) for serum concentrations of antibodies against other BRD pathogen. Average daily gain (ADG) tended to be increased (P = 0.09) in BOV compared with CON steers from d 56 to d 90 (0.858 and 0.760 kg/d, respectively, although did not impact overall ADG (P = 0.48). No treatment differences were detected for steer final BW (P = 0.43). Total BW change per pasture was greater in BOV compared with CON (790 vs. 550 kg BW change; P = 0.02). No treatment differences were detected (P = 1.00) for BRD incidence, although steers removed from the experiment for health reasons tended to be increased in CON compared with BOV (P = 0.08). Despite lacking differences in health or physiological responses, B. licheniformis and B. subtilis supplementation increased overall production of BW in high-risk stocker steers during a 90-d grazing period.

88 Impacts of trace mineral source and ancillary drench on steer performance during a 60-day backgrounding phase

Abstract This experiment evaluated the impacts of supplementing organic complexed Cu, Co, Mn, and Zn on productive and health responses of high-risk beef cattle during a 60-d backgrounding phase. Crossbred steers (n = 120) were purchased at auction and transported to the experimental facility. Steer BW was recorded at arrival (d -1; initial shrunk BW = 227.7 ± 1.3 kg). On d 0, steers were ranked by BW and allocated to 1 of 8 groups (8 steers/group) and housed in drylot pens (6 m x 12 m) equipped with GrowSafe automated feeding systems (Model 8000; 2 bunks/pen; d 0 to 60). Groups were randomly assigned to receive a total mixed ration (TMR; 47% corn) containing: 1) sulfate sources of Cu, Co, Mn, and Zn (INR; n = 40); 2) organic complexed sources of Cu, Mn, Co, and Zn (AAC; Zinpro Availa 4; Zinpro Corp., Eden Prairie, MN; n = 40); or 3) AAC and an organic complexed trace mineral drench (APF; 30 mL/hd; Zinpro ProFusion, Zinpro Corp.) on d 0 and ancillary to morbidity treatment (n = 40). Diets provided the same daily amount of energy, protein, macro minerals and trace minerals based on 7 g/steer daily of Zinpro Availa 4. Steers were assessed for bovine respiratory disease (BRD) signs daily. Liver biopsies were performed on day 0, 28 and 60. Final BW was recorded on d 60 and 61. Blood samples were collected on d 0, 2, 6, 10, 13, 21, 28 and 45. No treatment differences were detected (P ≥ 0.23) for TMR intake, final BW, ADG, feed efficiency, or BRD incidence. Mean liver Co concentrations were greater (P = 0.02) in AAC and APF compared with INR steers. Mean liver Cu was greater (P = 0.02) in APF compared with AAC steers. Liver Zn tended to be greater (P = 0.10) on d 28 but less (P = 0.05) on d 60 for INR compared with AAC and APF steers. Plasma cortisol was least (P = 0.05) for steers supplemented with AAC on d 6, whereas AAC steers tended to have greater (P = 0.09) plasma cortisol on d 13 compared with APF. Plasma haptoglobin tended to be greater (P ≤ 0.10) for INR steers on d 28 and 45 compared with AAC and APF. While supplementing cattle with AAC or INR results in similar animal performance and clinical disease, AAC and APF reduces stress and acute phase protein responses.

Macrolide resistance in Mannheimia haemolytica isolates associated with bovine respiratory disease from the German national resistance monitoring program GERM-Vet 2009 to 2020.

Data collected from the German national resistance monitoring program GERM-Vet showed slowly increasing prevalence of macrolide resistance among bovine respiratory disease (BRD)-associated Pasteurellacae from cattle over the last decade. The focus of this study was to analyze the genetic basis of antimicrobial resistance (AMR) and the prevalence of multidrug-resistance (MDR)-mediating integrative and conjugative elements (ICEs) in 13 German BRD-associated Mannheimia haemolytica isolates collected between 2009 and 2020 via whole-genome sequencing. Antimicrobial susceptibility testing (AST) was performed via broth microdilution according to the recommendations of the Clinical and Laboratory Standards Institute for the macrolides erythromycin, tilmicosin, tulathromycin, gamithromycin, tildipirosin, and tylosin as well as 25 other antimicrobial agents. All isolates either had elevated MICs or were resistant to at least one of the macrolides tested. Analysis of whole-genome sequences obtained by hybrid assembly of Illumina MiSeq and Oxford Nanopore MinION reads revealed the presence of seven novel Tn7406-like ICEs, designated Tn7694, and Tn7724- Tn7729. These ICEs harbored the antimicrobial resistance genes erm(T), mef (C), mph(G), floR, catA3, aad(3")(9), aph(3')-Ia, aac(3)-IIa, strA, strB, tet(Y), and sul2 in different combinations. In addition, mutational changes conferring resistance to macrolides, nalidixic acid or streptomycin, respectively, were detected among the M. haemolytica isolates. In addition, four isolates carried a 4,613-bp plasmid with the β-lactamase gene blaROB - 1. The detection of the macrolide resistance genes erm(T), mef (C), and mph(G) together with other resistance genes on MDR-mediating ICEs in bovine M. haemolytica may explain the occurrence of therapeutic failure when treating BRD with regularly used antimicrobial agents, such as phenicols, penicillins, tetracyclines, or macrolides. Finally, pathogen identification and subsequent AST is essential to ensure the efficacy of the antimicrobial agents applied to control BRD in cattle.