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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 -
Carbohydrate Research Dec 2023N-Glycosyltransferase (NGT) is an inverting glycosyltransferase for an unusual pathway of N-linked protein glycosylation and glycosylates polypeptides in the consensus...
N-Glycosyltransferase (NGT) is an inverting glycosyltransferase for an unusual pathway of N-linked protein glycosylation and glycosylates polypeptides in the consensus sequon (N-(X≠P)-T/S) with hexose monosaccharides. Here, we expressed and characterized a novel N-glycosyltransferase from Mannheimia haemolytica (named MhNGT). RP-HPLC and Mass Spectrometry were used to assay and quantify glycopeptide formation by MhNGT and determine its substrate specificities. MhNGT could utilize a variety of nucleotide-activated sugar donors, including UDP-Glc, UDP-Gal and UDP-Xyl, to glycosylate the tested peptides DANYTK, GGNWTT and PAVGNCSSALR with higher efficiency than ApNGT which was comprehensive studied. The optimum temperature of MhNGT was about 30 °C and the optimum pH was 7.5-8.0 in PBS-NaOH buffer. MhNGT exhibited a different position-specific residue preference of substrate peptides from the NGT of Actinobacillus pleuropneumoniae (ApNGT). The effective glycosylation of common short peptides by MhNGT demonstrated the enzyme's potential to alter therapeutically significant mammalian N-glycoproteins.
Topics: Animals; Mannheimia haemolytica; Glycosyltransferases; Peptides; Protein Isoforms; Uridine Diphosphate; Mammals
PubMed: 37783053
DOI: 10.1016/j.carres.2023.108947 -
Preventive Veterinary Medicine Oct 2023Bovine Respiratory Disease (BRD) affects young bulls, causing animal welfare and health concerns as well as economical costs. BRD is caused by an array of viruses and...
Bovine Respiratory Disease (BRD) affects young bulls, causing animal welfare and health concerns as well as economical costs. BRD is caused by an array of viruses and bacteria and also by environmental and abiotic factors. How farming practices influence the spread of these causal pathogens remains unclear. Our goal was to assess the impact of zootechnical practices on the spread of three causal agents of BRD, namely the bovine respiratory syncytial virus (BRSV), Mannheimia haemolytica and Mycoplasma bovis. In that extent, we used an individual based stochastic mechanistic model monitoring risk factors, infectious processes, detection and treatment in a farm possibly featuring several batches simultaneously. The model was calibrated with three sets of parameters relative to each of the three pathogens using data extracted from literature. Separated batches were found to be more effective than a unique large one for reducing the spread of pathogens, especially for BRSV and M.bovis. Moreover, it was found that allocating high risk and low risk individuals into separated batches participated in reducing cumulative incidence, epidemic peaks and antimicrobial usage, especially for M. bovis. Theses findings rise interrogations on the optimal farming practices in order to limit BRD occurrence and pave the way to models featuring coinfections and collective treatments p { line-height: 115%; margin-bottom: 0.25 cm; background: transparent}a:link { color: #000080; text-decoration: underline}a.cjk:link { so-language: zxx}a.ctl:link { solanguage: zxx}.
Topics: Animals; Cattle; Male; Farms; Respiratory Tract Diseases; Cattle Diseases; Mannheimia haemolytica; Agriculture; Bovine Respiratory Disease Complex
PubMed: 37688889
DOI: 10.1016/j.prevetmed.2023.106009 -
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 -
Veterinary Research Communications Feb 2024Exposure to gamma rays from cobalt 60 (Co) can induce a complete inactivation of Mannheimia haemolytica. The inactivated bacterial pathogen is a potential vaccine...
Exposure to gamma rays from cobalt 60 (Co) can induce a complete inactivation of Mannheimia haemolytica. The inactivated bacterial pathogen is a potential vaccine candidate for immunization of ruminants such as sheep. The subcutaneous administration of irradiated vaccine in a two-dose regimen (4.0 × 10 colony forming unit (CFU) per dose) results in no mortality in any of the vaccinated sheep during immunization and after subsequent challenge of the live bacteria of the same strain of M. haemolytica. A significant rise in serum IgG titer, detected through ELISA, is observed after the passage of two weeks from the inoculation of the first dose whereas, the peak of the mean serum antibody titer occurred after two weeks of booster dose. The vaccination does not bring significant change to the IFN-γ levels in serum. The bacterial challenge of the vaccinated sheep does not induce a further seroconversion relative to serum antibody titer. In conclusion, the vaccinated sheep are protected by the elevated IgG titer and increased levels of IL-4 (Th-2 response) compared to the non-vaccinated sheep. Radiation technology can provide the opportunity for mass production of immunologically safe vaccines against animal and zoonotic diseases. Ethics Approval by the National Research Center Ethics Committee (Trial Registration Number (TRN) no 13,602,023, 13/5/2023) was obtained.
Topics: Animals; Sheep; Mannheimia haemolytica; Gamma Rays; Bacterial Vaccines; Vaccination; Immunoglobulin G; Sheep Diseases
PubMed: 37642819
DOI: 10.1007/s11259-023-10207-w -
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 -
Tropical Animal Health and Production Aug 2023The lipopolysaccharide (LPS) endotoxin and outer membrane protein (OMP) are among the virulence factors of Gram-negative bacteria responsible for inducing pathogenicity...
The lipopolysaccharide (LPS) endotoxin and outer membrane protein (OMP) are among the virulence factors of Gram-negative bacteria responsible for inducing pathogenicity in the infected host. OMP and LPS occur on the outer membrane of M. haemolytica A2, the primary aetiological agent of pneumonic mannheimiosis in small ruminants. While the LPS is known to mediate Gram-negative bacterial infection by activating downstream inflammatory pathways, the potential role of OMP during inflammatory responses remained unclear. Hence, this study determined the effect of the OMP of M. haemolytica A2 on the serum concentration of pro-inflammatory cytokines and the male reproductive hormones (testosterone and Luteinizing Hormone). We randomly assigned twelve bucks to three groups (n = 4 bucks each): Group 1 was challenged with 2 mL PBS buffer (pH 7.0) intranasally; Group 2 received 2 mL of 1.2 X 10 CFU/mL whole M. haemolytica A2 intranasally; and Group 3 received 2 mL of OMP extract obtained from 1.2 X 10 CFU/mL M. haemolytica A2 intramuscularly. Serum samples collected at pre-determined intervals were used for the quantitative determination of the pro-inflammatory cytokines (IL-1β, IL-6, and TNFα) and reproductive hormones (testosterone and LH) using commercial sandwich enzyme-linked immunosorbent assay (ELISA). The serum concentration of IL1β was initially increased within the first-hour post-challenge in Groups 2 and 3, followed by a significant decrease in concentration at 21d and 35d (p < 0.05) in Group 3. Only mild fluctuations in IL-6 occurred in group 2, as opposed to the 1.7-fold rapid increase in TNFα within 2 h post-challenge before decreasing at 6 h. An increase in pro-inflammatory cytokines was accompanied by an acute febrile response of 39.5 ± 0.38 °C (p < 0.05) at 2 h and 40.1 ± 0.29 °C (p < 0.05) at 4 h in Group 2 and Group 3, respectively. Serum testosterone decreased significantly (p < 0.05) in both treatment groups but remained significantly (p > 0.05) lower than in Group 1 throughout the study. There was a moderate negative association between testosterone and IL1β (r = -0.473; p > 0.05) or TNFα (r = -0.527; p < 0.05) in Group 2. Serum LH also showed moderate negative associations with TNFα in Group 2 (r = -0.63; p < 0.05) and Group 3 (r = -0.54; p > 0.05). The results of this study demonstrated that M. haemolytica A2 and its OMP produced marked alterations in serum levels of pro-inflammatory cytokines and male reproductive hormones. The negative correlations between serum testosterone and inflammatory cytokines would suggest the potential role of OMP in causing male infertility by mediating innate inflammatory responses to suppress testosterone production in bucks.
Topics: Male; Animals; Membrane Proteins; Cytokines; Tumor Necrosis Factor-alpha; Mannheimia haemolytica; Interleukin-6; Lipopolysaccharides; Testosterone
PubMed: 37589856
DOI: 10.1007/s11250-023-03706-0 -
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