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Frontiers in Veterinary Science 2023Bovine respiratory disease (BRD) is a multifactorial disease complex in which bacteria in the upper respiratory tract play an important role in disease development....
INTRODUCTION
Bovine respiratory disease (BRD) is a multifactorial disease complex in which bacteria in the upper respiratory tract play an important role in disease development. Previous studies have related the presence of four BRD-pathobionts (, , , and ) in the upper respiratory tract to BRD incidence and mortalities in the dairy and beef cattle industry, but these studies typically only use one time point to compare the abundance of BRD-pathobionts between apparently healthy and BRD-affected cattle. The objective of this study was to characterize the longitudinal development of the nasopharyngeal (NP) microbiome from apparently healthy calves, and in calves with clinical signs of BRD, the microbiota dynamics from disease diagnosis to recovery.
METHODS
Deep nasopharyngeal swabs were taken from all calves immediately after transport (day 0). If a calf was diagnosed with BRD ( = 10), it was sampled, treated with florfenicol or tulathromycin, and sampled again 1, 5, and 10 days after antibiotic administration. Otherwise, healthy calves ( = 20) were sampled again on days 7 and 14. Bacterial community analysis was performed through 16S rRNA gene amplicon sequencing.
RESULTS
The NP microbiome of the healthy animals remained consistent throughout the study, regardless of time. The NP microbiota beta diversity and community composition was affected by tulathromycin or florfenicol administration. Even though BRD-pathobionts were identified by 16S rRNA gene sequencing in BRD-affected animals, no difference was observed in their relative abundance between the BRD-affected and apparently healthy animals. The abundance of BRD-pathobionts was not predictive of disease development while the relative abundance of BRD pathobionts was unique to each BRD-affected calf. Interestingly, at the end of the study period, the genera was the most abundant genus in the healthy group, while was the most abundant genus in the animals that recovered from BRD.
DISCUSSION
This study highlights that injected antibiotics seem to improve the NP microbiome composition (higher abundance of and lower abundance of ), and that the relative abundance of BRD-pathobionts differs between individual calves but is not strongly predictive of BRD clinical signs, indicating that additional factors are likely important in the clinical progression of BRD.
PubMed: 38033643
DOI: 10.3389/fvets.2023.1297158 -
Journal of Advanced Veterinary and... Sep 2023This research aimed to assess the prevalence of caprine pasteurellosis, isolate and identify pasteurellosis ( and ) in pneumonic goats, and discover the main bacterial... (Review)
Review
OBJECTIVE
This research aimed to assess the prevalence of caprine pasteurellosis, isolate and identify pasteurellosis ( and ) in pneumonic goats, and discover the main bacterial cause of pneumonia.
MATERIALS AND METHODS
One hundred and five samples (94 nasal swabs and 11 lung tissues) from goats suspected of having pneumonia were taken and transferred aseptically to the laboratory. Following the processing of the collected samples, spp. was isolated with the aid of plate culture methods. Biochemical characteristics were used to identify all bacterial isolates, which were then verified by polymerase chain reaction (PCR). Antimicrobial susceptibility testing was also carried out to evaluate the sensitivity profiles of various antibiotics. The serotype-specific antigen (PHSSA) gene was used to identify isolates of , and the KMT1 gene was used to identify isolates of .
RESULTS
From the 105 clinically suspicious samples, 51 (48.57%) were identified to be through bacteriological testing and also by PCR targeting the gene. Of these, 47.87% (45/94) were nasal swabs, and 54.55% (6/11) were lung tissues. Among confirmed samples, 70.59% (36/51) were identified as and 29.41% (15/51) were identified as . Resistance to tetracycline, streptomycin, oxytetracycline, gentamicin, and ceftriaxone was found in 50%-83% of the isolates. In addition, PCR identified the PHSSA and KMT1 genes from isolates of and , respectively.
CONCLUSION
The present study revealed that and primarily caused pasteurellosis in pneumonic goats in Bangladesh. However, when treating these animals, the proper choice of antimicrobials should be made to control this disease.
PubMed: 37969786
DOI: 10.5455/javar.2023.j707 -
Brazilian Journal of Microbiology :... Dec 2023Bovine respiratory disease (BRD) is a multifactorial and predominantly multietiological disease that affects dairy cattle herds worldwide, being more frequent in young...
Bovine respiratory disease (BRD) is a multifactorial and predominantly multietiological disease that affects dairy cattle herds worldwide, being more frequent in young animals. The occurrence of BRD was investigated in lactating cows from two high-yielding dairy herds in southern Brazil. To determine the etiology of the clinical cases of acute respiratory disease, nasal swab samples were collected from cows with clinical signs of BRD and evaluated using PCR and RT-PCR for nucleic acid detection of the main BRD etiological agents, including Mycoplasma bovis, Mannheimia haemolytica, Pasteurella multocida, Histophilus somni, bovine respiratory syncytial virus, bovine coronavirus, bovine viral diarrhea virus, bovine alphaherpesvirus 1, and bovine parainfluenza virus 3. Only three microorganisms (M. bovis, H. somni, and P. multocida) were identified in both single and mixed infections. We concluded that 40.0% of the cows were infected with M. bovis and 75.0% with H. somni in herd A. Considering both single and mixed infections, the analyses performed in herd B showed that 87.5%, 25.0%, and 50.0% of the cows were infected with M. bovis, H. somni, and P. multocida, respectively. M. bovis and H. somni are considered fastidious bacteria and laboratory diagnosis is neglected. Subsequently, most clinical cases of mycoplasmosis and histophilosis in cattle remain undiagnosed. This study demonstrates the importance of M. bovis and H. somni infections in adult cows with BRD. These results highlight the importance of including these bacteria in the group of etiological agents responsible for the occurrence of BRD in cattle, especially in adult cows with unfavorable immunological conditions, such as recent calving and peak lactation.
Topics: Animals; Female; Cattle; Coinfection; Lactation; Cattle Diseases; Bacterial Infections; Bacteria; Pasteurella multocida
PubMed: 37917227
DOI: 10.1007/s42770-023-01165-1 -
International Journal of Systematic and... Oct 2023Forty-one isolates of Bisgaard taxon 6 obtained from guinea pigs, pandas, pigs and muskrat and isolates of taxon 10 from horses and horse bites in humans were subjected...
Forty-one isolates of Bisgaard taxon 6 obtained from guinea pigs, pandas, pigs and muskrat and isolates of taxon 10 from horses and horse bites in humans were subjected phenotypic characterization. Production of acid from (-)-d-mannitol, (-)-d-sorbitol and (+)-d-glycogen separated taxon 10 (positive) from taxon 6 (negative), while from two to 11 phenotypic characteristics separated taxa 6 and 10 from the 32 genera of reported so far. Forty-four strains were genetically characterized. Sequencing of 16S rRNA genes documented a monophyletic relationship at the species level and the highest 16S rRNA gene sequence similarity of 95.6 % to other species was found between strain CCUG 15568 and the type strain of (CCUG 38457). Digital DNA-DNA hybridization (dDDH) values predicted from whole genomic sequences between CCUG 15568 and other characterized strains of taxa 6 and 10 were 69.3-99.9 %. The average nucleotide identity values were higher than 95 % for all strains. The highest dDDH value of 29 % outside the taxa 6 and 10 group was obtained with the genome of the type strain of [] , indicating a separate taxonomic status at species level to taxa 6 and 10. The phylogenetic comparison of concatenated conserved protein sequences showed the unique position of the taxa investigated in the current study which qualified for the status of a new genus since the highest identity was found with with 79 %, well below the upper threshold between genera of 85 %. Based upon the low genetic similarity to other genera of the family and a unique phenotype, we suggest that Bisgaard taxa 6 and 10 should be classified as gen. nov., sp. nov. The G+C of the type strain of , 8.5 (=CCUG 15568=DSM 115565), is 46.2 mol%, calculated from the whole genome.
Topics: Humans; Animals; Guinea Pigs; Horses; Phylogeny; RNA, Ribosomal, 16S; Sequence Analysis, DNA; DNA, Bacterial; Bacterial Typing Techniques; Base Composition; Fatty Acids; Pasteurellaceae
PubMed: 37882672
DOI: 10.1099/ijsem.0.006092 -
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 -
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