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PloS One 2023Respiratory disease is responsible for dramatic population declines in bighorn sheep (Ovis canadensis), and respiratory pathogen diagnostics contribute to the management...
Assessing shared respiratory pathogens between domestic (Ovis aries) and bighorn (Ovis canadensis) sheep; methods for multiplex PCR, amplicon sequencing, and bioinformatics to characterize respiratory flora.
Respiratory disease is responsible for dramatic population declines in bighorn sheep (Ovis canadensis), and respiratory pathogen diagnostics contribute to the management of bighorn populations. To create a comprehensive and consistent approach to bighorn sheep respiratory diagnostics, we created a culture-independent assay to detect and strain type Mannheimia haemolytica, Bibersteinia trehalosi, Pasteurella multocida, and Mycoplasma ovipneumoniae. The assay also detects and characterizes the Pasteurellaceae leukotoxin A gene, and broadly assesses the bacterial composition of each sample based on 16S rRNA sequences. The assay is based on a three-step approach: 1) Multiplex PCR to amplify targets including eight loci for each bacterial species, the Pasteurellaceae lktA gene, and the 16S rRNA gene 2) Library preparation, barcoding, and short-read Illumina sequencing to determine the genetic sequences of each target, and 3) Bioinformatics in the form of automated software to analyze genetic sequences. The assay was designed to assess shared pathogens between domestic and bighorn sheep, but could be useful for many applications in bighorn sheep respiratory disease research and management.
Topics: Animals; Sheep; Sheep, Bighorn; Sheep, Domestic; Multiplex Polymerase Chain Reaction; RNA, Ribosomal, 16S; Sheep Diseases; Mannheimia haemolytica; Respiratory Tract Diseases; Computational Biology
PubMed: 37856492
DOI: 10.1371/journal.pone.0293062 -
Microbiology Spectrum Dec 2023The Gram-negative coccobacillus is a natural inhabitant of the upper respiratory tract in ruminants and the most common bacterial agent involved in bovine respiratory...
The Gram-negative coccobacillus is a natural inhabitant of the upper respiratory tract in ruminants and the most common bacterial agent involved in bovine respiratory disease complex development. Key virulence factors harbored by are leukotoxin, lipopolysaccharide, capsule, adhesins, and neuraminidase which are involved in evading innate and adaptive immune responses. In this study, we have shown that CMP-sialic acid synthetase () is necessary for the incorporation of sialic acid onto the membrane, and inactivation of results in increased phagocytosis and complement-mediated killing of thus demonstrating that sialylation contributes to the virulence of .
Topics: Cattle; Animals; Mannheimia haemolytica; N-Acylneuraminate Cytidylyltransferase; Serogroup; Gene Deletion; Phagocytosis
PubMed: 37850751
DOI: 10.1128/spectrum.02944-23 -
Frontiers in Microbiology 2023Bovine respiratory disease (BRD) is a significant health problem in beef cattle production, resulting in considerable economic losses due to mortalities, cost of...
BACKGROUND
Bovine respiratory disease (BRD) is a significant health problem in beef cattle production, resulting in considerable economic losses due to mortalities, cost of treatment, and reduced feed efficiency. The onset of BRD is multifactorial, with numerous stressors being implicated, including transportation from farms to feedlots. In relation to animal welfare, regulations or practices may require mandatory rest times during transportation. Despite this, there is limited information on how transportation and rest stops affect the respiratory microbiota.
RESULTS
This study evaluated the effect of cattle source (ranch-direct or auction market-derived) and rest stop duration (0 or 8 h of rest) on the upper respiratory tract microbiota and its relationship to stress response indicators (blood cortisol and haptoglobin) of recently weaned cattle transported for 36 h. The community structure of bacteria was altered by feedlot placement. When cattle were off-loaded for a rest, several key bacterial genera associated with BRD (, , ) were increased for most sampling times after feedlot placement for the ranch-direct cattle group, compared to animals given no rest stop. Similarly, more sampling time points had elevated levels of BRD-associated genera when auction market cattle were compared to ranch-direct. When evaluated across time and treatments several genera including were positively correlated with blood cortisol concentrations.
CONCLUSION
This is the first study to assess the effect of rest during transportation and cattle source on the respiratory microbiota in weaned beef calves. The results suggest that rest stops and auction market placement may be risk factors for BRD, based solely on increased abundance of BRD-associated genera in the upper respiratory tract. However, it was not possible to link these microbiota to disease outcome, due to low incidence of BRD in the study populations. Larger scale studies are needed to further define how transportation variables impact cattle health.
PubMed: 37808284
DOI: 10.3389/fmicb.2023.1192763 -
Frontiers in Plant Science 2023Bovine respiratory disease (BRD) affects feedlot cattle across North America, resulting in economic losses due to animal treatment and reduced performance. In an effort...
Bovine respiratory disease (BRD) affects feedlot cattle across North America, resulting in economic losses due to animal treatment and reduced performance. In an effort to develop a vaccine candidate targeting a primary bacterial agent contributing to BRD, we produced a tripartite antigen consisting of segments of the virulence factor Leukotoxin A (LktA) and lipoprotein PlpE from , fused to a cholera toxin mucosal adjuvant (CTB). This recombinant subunit vaccine candidate was expressed in the leaves of plants, with accumulation tested in five subcellular compartments. The recombinant protein was found to accumulate highest in the endoplasmic reticulum, but targeting to the chloroplast was employed for scaling up production due the absence of post-translational modification while still producing feasible levels. Leaves were freeze dried, then orally administered to mice to determine its immunogenicity. Sera from mice immunized with leaf tissue expressing the recombinant antigen contained IgG antibodies, specifically recognizing both LktA and PlpE. These mice also had a mucosal immune response to the CTB+LktA+PlpE protein as measured by the presence of LktA- and PlpE-specific IgA antibodies in lung and fecal material. Moreover, the antigen remained stable at room temperature with limited deterioration for up to one year when stored as lyophilized plant material. This study demonstrated that a recombinant antigen expressed in plant tissue elicited both humoral and mucosal immune responses when fed to mice, and warrants evaluation in cattle.
PubMed: 37790785
DOI: 10.3389/fpls.2023.1251046 -
Journal of Veterinary Research Sep 2023Early vaccination of cattle with an inactivated commercial bacterial vaccine against bovine respiratory disease has been reported to increase antibody production and can...
INTRODUCTION
Early vaccination of cattle with an inactivated commercial bacterial vaccine against bovine respiratory disease has been reported to increase antibody production and can alleviate the disease. However, its dosage has been little investigated in young Holstein calves. This study addresses the need to establish guide values for vaccine dosage in these animals.
MATERIAL AND METHODS
Healthy calves received an inactivated vaccine for , and intramuscularly at the ages of 1 and 4 weeks. Administered vaccine doses were 1.0 mL for the primary and booster vaccinations (1.0 + 1.0 group), 0.5 mL for the primary and 1.0 mL for the booster vaccination (0.5 + 1.0 group), or 0.5 mL for both vaccinations (0.5 + 0.5 group).
RESULTS
Differences in the vaccine responses between the 1.0 + 1.0 group and 0.5 + 1.0 group were minor. However, the number of calves with a positive vaccine response to in the 0.5 + 0.5 group was less than half of that in the 1.0 + 1.0 and 0.5 + 1.0 groups. In logistic regression analysis, although the booster vaccination dose was positively correlated with seropositivity for , the primary vaccination dose was not correlated with vaccine response. The number of calves with positive vaccine responses to was low even after booster vaccination regardless of the dose.
CONCLUSION
The dose of 0.5 mL can be used for primary vaccinations in newborn Holstein calves, but 1.0 mL may be required for booster vaccinations.
PubMed: 37786851
DOI: 10.2478/jvetres-2023-0037 -
Frontiers in Cellular and Infection... 2023Bovine respiratory disease (BRD) is the most devastating disease affecting beef and dairy cattle producers in North America. An emerging area of interest is the... (Meta-Analysis)
Meta-Analysis
BACKGROUND
Bovine respiratory disease (BRD) is the most devastating disease affecting beef and dairy cattle producers in North America. An emerging area of interest is the respiratory microbiome's relationship with BRD. However, results regarding the effect of BRD on respiratory microbiome diversity are conflicting.
RESULTS
To examine the effect of BRD on the alpha diversity of the respiratory microbiome, a meta-analysis analyzing the relationship between the standardized mean difference (SMD) of three alpha diversity metrics (Shannon's Diversity Index (Shannon), Chao1, and Observed features (OTUs, ASVs, species, and reads) and BRD was conducted. Our multi-level model found no difference in Chao1 and Observed features SMDs between calves with BRD and controls. The Shannon SMD was significantly greater in controls compared to that in calves with BRD. Furthermore, we re-analyzed 16S amplicon sequencing data from four previously published datasets to investigate BRD's effect on individual taxa abundances. Additionally, based on Bray Curtis and Jaccard distances, health status, sampling location, and dataset were all significant sources of variation. Using a consensus approach based on RandomForest, DESeq2, and ANCOM-BC2, we identified three differentially abundant amplicon sequence variants (ASVs) within the nasal cavity, ASV5_, ASV19_, and ASV37_ However, no ASVs were differentially abundant in the other sampling locations. Moreover, based on SECOM analysis, ASV37_ had a negative relationship with ASV1_, ASV5_, and ASV4_. ASV19_ had negative relationships with ASV1_, ASV4_, ASV54_, ASV7_, and ASV8_.
CONCLUSIONS
Our results confirm a relationship between bovine respiratory disease and respiratory microbiome diversity and composition, which provide additional insight into microbial community dynamics during BRD development. Furthermore, as sampling location and sample processing (dataset) can also affect results, consideration should be taken when comparing results across studies.
Topics: Cattle; Animals; Respiratory Tract Diseases; Cattle Diseases; Clostridiales; Microbiota; Mycoplasma hyorhinis
PubMed: 37743862
DOI: 10.3389/fcimb.2023.1223090 -
Animals : An Open Access Journal From... Aug 2023Hu sheep, a locally bred species in China known for its high productivity, is currently suffering from pneumonia. Here, we combine high-throughput 16SrRNA gene...
Hu sheep, a locally bred species in China known for its high productivity, is currently suffering from pneumonia. Here, we combine high-throughput 16SrRNA gene sequencing and bacterial culturing to examine the bacterial community in pneumonic Hu Sheep lungs ( < 0.05). The results showed that the abundance and diversity of lung bacteria in healthy sheep were significantly higher than those in pneumonia sheep ( = 0.139), while there was no significant difference between moderate and severe pneumonia. Furthermore, the composition of the lung microbiota community underwent significant alterations between different levels of pneumonia severity. The application of LEfSe analysis revealed a notable enrichment of within the lungs of sheep afflicted with moderate pneumonia ( < 0.01), surpassing the levels observed in their healthy counterparts. Additionally, emerged as the prevailing bacterial group within the lungs of sheep suffering from severe pneumonia. Integrating the results of bacterial isolation and identification, we conclusively determined that was the primary pathogenic bacterium within the lungs of sheep afflicted with moderate pneumonia. Furthermore, the exacerbation of pneumonia may be attributed to the synergistic interplay between spp. and other bacterial species. Our results provide new insights for guiding preventive and therapeutic measures for pneumonia of different severities in sheep.
PubMed: 37685027
DOI: 10.3390/ani13172763 -
Journal of Dairy Science Dec 2023The high degree of commingling and accumulation of stressors during and after transport makes prevention of bovine respiratory disease (BRD) extremely challenging in the...
The high degree of commingling and accumulation of stressors during and after transport makes prevention of bovine respiratory disease (BRD) extremely challenging in the veal and dairy beef industry. Upon arrival, vaccination for agents involved in BRD is practically most achievable, but its efficacy under such conditions in dairy veal calves is unknown. Given the high prevalence of subclinical pneumonia in these settings, the primary objective of the present study was to determine the effect of 2 vaccination protocols administered upon arrival against bovine respiratory syncytial virus (BRSV), bovine parainfluenza type 3 virus (BPI-3), and Mannheimia haemolytica on clinical BRD and lung ultrasonographic findings in dairy veal calves. In addition, the effects of vaccination on average daily live weight gain and cold carcass weight were determined. In this randomized clinical trial, 443 male dairy calves were assigned to one of 3 groups: a negative, placebo-controlled group (n = 151), a vaccination group with 2 subcutaneous injections 4 wk apart with an inactivated vaccine containing BRSV, BPI-3, and M. haemolytica (parenteral [PE] group; n = 149) and a second vaccination group receiving an intranasal live-attenuated vaccine containing BRSV and BPI-3 and 2 subcutaneous vaccinations with the same inactivated vaccine as the PE vaccination group (intranasal-parenteral [IN-PE] group; n = 143). Clinical scoring and quick thoracic ultrasonography (qTUS) were performed on all calves on arrival (wk 0), at the peak of respiratory disease (outbreak; wk 1), at the end of the first antimicrobial group treatment (wk 3), and at a long-term evaluation point (wk 10). Culture and nanopore sequencing on nonendoscopic bronchoalveolar lavage (nBAL) samples were used to identify pathogens involved in the outbreak. Upon arrival, 15.1% of the calves had lung consolidation ≥1cm and incidence quickly rose to 42.8% during the outbreak. In both the PE and IN-PE group, the odds of pneumonia in wk 10 were reduced by 62% (odds ratio [OR] = 0.38; 95% confidence interval [CI] = 0.23-0.64) and 41% (OR = 0.59; 95% CI = 0.37-0.96), respectively. Short-term cure rate (50.3%), as determined immediately after the first group antimicrobial treatment, was not influenced by vaccination. In contrast, long-term cure rate, determined at wk 10, was affected by vaccination with higher cure in the PE group compared with the control group (69.4% vs. 51.2%; OR = 2.2; 95% CI = 1.1-5.0). Average daily gain in the first 10 wk of production was not affected by vaccination. Vaccination resulted in an increase in cold carcass weight of 3.5 and 4.3 kg in the PE (95% CI = -0.9-7.9) and IN-PE group (95% CI = -0.17-8.7), respectively. In conclusion, under the conditions of the present study, vaccination upon arrival resulted in a reduced prevalence of pneumonia at wk 10 of production, likely caused both by an improved cure rate of secondary infections and a reduced incidence of new cases between outbreak and long-term evaluation. The present protocol, using qTUS for pneumonia detection and nBAL diagnostics for pathogen identification adds a new dimension to randomized clinical trials on respiratory disease in calves.
Topics: Animals; Cattle; Male; Cattle Diseases; Vaccination; Pneumonia; Anti-Infective Agents; Ultrasonography; Vaccines, Inactivated; Respiratory Syncytial Virus, Bovine
PubMed: 37641351
DOI: 10.3168/jds.2023-23438 -
Scientific Reports Aug 2023Bovine respiratory disease (BRD) is a significant health issue in the North American feedlot industry, causing substantial financial losses due to morbidity and...
Bovine respiratory disease (BRD) is a significant health issue in the North American feedlot industry, causing substantial financial losses due to morbidity and mortality. A lack of effective vaccines against BRD pathogens has resulted in antibiotics primarily being used for BRD prevention. The aim of this study was to develop a mucosal vaccine against the BRD pathogen, Mannheimia haemolytica, using Bacillus subtilis spores as an adjuvant. A chimeric protein (MhCP) containing a tandem repeat of neutralizing epitopes from M. haemolytica leukotoxin A (NLKT) and outer membrane protein PlpE was expressed to produce antigen for adsorption to B. subtilis spores. Adsorption was optimized by comparing varying amounts of antigen and spores, as well as different buffer pH and reaction temperatures. Using the optimal adsorption parameters, spore-bound antigen (Spore-MhCP) was prepared and administered to mice via two mucosal routes (intranasal and intragastric), while intramuscular administration of free MhCP and unvaccinated mice were used as positive and negative control treatments, respectively. Intramuscular administration of MhCP elicited the strongest serum IgG response. However, intranasal immunization of Spore-MhCP generated the best secretory IgA-specific response against both PlpE and NLKT in all samples evaluated (bronchoalveolar lavage, saliva, and feces). Since proliferation of M. haemolytica in the respiratory tract is a prerequisite to lung infection, this spore-based vaccine may offer protection in cattle by limiting colonization and subsequent infection, and Spore-MhCP warrants further evaluation in cattle as a mucosal vaccine against M. haemolytica.
Topics: Cattle; Animals; Mice; Mannheimia haemolytica; Spores, Bacterial; Respiratory System; Bacterial Vaccines; Cattle Diseases
PubMed: 37563163
DOI: 10.1038/s41598-023-29732-4 -
American Journal of Veterinary Research Nov 2023Evaluate agreement among the antimicrobial susceptibility profiles of Mannheimia haemolytica or Pasteurella multocida obtained by transtracheal wash, nasal swab,...
Agreement of antimicrobial susceptibility testing of Pasteurella multocida and Mannheimia haemolytica isolates from preweaned dairy calves with bovine respiratory disease.
OBJECTIVE
Evaluate agreement among the antimicrobial susceptibility profiles of Mannheimia haemolytica or Pasteurella multocida obtained by transtracheal wash, nasal swab, nasopharyngeal swab, and bronchoalveolar lavage.
ANIMALS
100 Holstein and Holstein-cross bull calves with bovine respiratory disease.
METHODS
Calves > 30 days old with naturally occurring bovine respiratory disease were sampled sequentially by nasal swab, nasopharyngeal swab, transtracheal wash, and then bronchoalveolar lavage. Samples were cultured, and for each antimicrobial, the MIC of 50% and 90% of isolates was calculated, and isolates were categorized as susceptible or not. Categorical discrepancies were recorded. Percent positive agreement and kappa values were calculated between isolates for each of the sampling methods.
RESULTS
Antimicrobial susceptibility varied by pathogen and resistance to enrofloxacin, florfenicol, tilmicosin, and spectinomycin was detected. Minor discrepancies were seen in up to 29% of classifications, with enrofloxacin, penicillin, and florfenicol more frequently represented than other drugs. Very major and major discrepancies were seen when comparing florfenicol (1.9%) and tulathromycin (3.8 to 4.9%) across sampling methods. Some variability was seen in agreement for enrofloxacin for several comparisons (8.3 to 18.4%).
CLINICAL RELEVANCE
Susceptibility testing of isolates from 1 location of the respiratory tract can reliably represent susceptibility in other locations. Nevertheless, the potential for imperfect agreement between sampling methods does exist. The level of restraint available, the skill level of the person performing the sampling, the age and size of the animal, disease status, and treatment history all must be factored into which test is most appropriate for a given situation.
Topics: Humans; Cattle; Animals; Male; Pasteurella multocida; Mannheimia haemolytica; Enrofloxacin; Anti-Bacterial Agents; Cattle Diseases; Respiratory Tract Diseases; Microbial Sensitivity Tests
PubMed: 37558231
DOI: 10.2460/ajvr.23.06.0140