Preparation and Characterization of Monoclonal Antibodies against Mycoplasma bovis
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Monoclonal antibodies (mabs) against Mycoplasma (M.) bovis were prepared for use in diagnosis of bovine mastitis. From the original 32 hybridomas actively secreting mabs against M. bovis, 6 stable lines were cloned. Two of them, Mb 5D8 and Mb 4F6, recognized M. bovis antigens of estimated molecular weights of 33 and 26 kDa, respectively. They showed no cross-reaction to other bovine mycoplasmas, thus rendering them useful for specific detection of this pathogen. All mabs investigated cross-reacted with M. agalactiae which is known to be closely related to M. bovis, but does not occur in cattle. Two other mabs, Mb 5D4 and Mb 1F6, exhibited further cross-reactions to a number of bovine mycoplasma species. Finally, mabs Mb 5D5 and Mb 2G5 reacted with all mycoplasmas tested. The possibility that they recognized constituents of the broth culture medium is discussed.Keywords:
Bovine milk
Abstract Background Long‐term breastfeeding is beneficial for both mothers and infants and mastitis is associated with the premature interruption of breastfeeding. Mastitis can be infectious or noninfectious. However, the effect of noninfectious mastitis on milk microbiota is not well‐understood. In this study, we aimed to clarify the relationship between noninfectious mastitis and the microbiota by conducting breast milk culture tests. Methods We compared the milk microbiota between women with noninfectious mastitis and without mastitis. Bacterial cultures were compared in 143 milk samples from January to November 2022, and bacterial diversity was evaluated based on the total number of bacterial species and bacterial species found per specimen. Results Women with noninfectious mastitis provided samples at a significantly later stage postpartum ( p < 0.01). The total bacterial count was significantly lower in samples from participants with noninfectious mastitis ( p < 0.01). The bacterial diversity of milk from participants with noninfectious mastitis was lower than that without mastitis: nine bacterial species identified in the former and 21 in the latter. The number of Rothia spp. was significantly higher, whereas the number of Staphylococcus aureus , Staphylococcus epidermidis and Pseudomonas fluorescens was significantly lower in samples from women with mastitis. There was no correlation between postpartum week and the number of bacterial species or presence of Rothia spp. Conclusions Noninfectious mastitis is associated with a decrease in the diversity of human milk microbiota, indicating impaired immune, metabolic, and neuroendocrine development functions in infants. Rothia spp. may also be associated with noninfectious mastitis, suggesting a possible target for future research.
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The study in 209 mothers with lactational mastitis has employed an AGA-780M thermovisor. Thermograms underwent a qualitative and quantitative isotherm-based evaluation. Diagnostic sensitivity of telethermography in mastitis was 97.1%. It could not differentiate types of mastitis in most cases and thus had a low specificity. Telethermography allows early identification of mastitis, control of therapy and reduction of the incidence of suppurative mastitis. The opportunity of telethermographic differentiation of milk stasis and initial mastitis has important practical implications since it allows one to avoid inappropriate use of antibiotics.
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The ability of Mycobacterium bovis (M. bovis) to survive in bovine milk has emerged as a serious public health concern. The first objective of this study was to evaluate the diagnostic utility of IS1081-targeted real-time PCR for the detection of M. bovis DNA in different fractions of bovine milk. In a model study, bovine milk samples were spiked with serially diluted M. bovis BCG to investigate the detection limit of M. bovis DNA in whole milk and milk fractions (cream, pellet, and pellet + cream combined) using IS1081 real-time PCR. The assay was then used to detect M. bovis DNA in whole milk and milk fractions from naturally infected animals. The results showed that the IS1081 real-time PCR was more sensitive when detecting M. bovis DNA in the cream layer alone and cream + pellet combined compared to whole milk or the pellet alone. While PCR-based diagnostic assays for the detection of M. bovis in milk samples provide a quicker diagnostic tool for bovine tuberculosis, safe processing, and handling of M. bovis-infected milk samples remain a challenge and pose a human health risk. PrimeStore Molecular Transport Medium (MTM) has been shown to rapidly inactivate infected specimens while preserving nucleic acid for subsequent Molecular analysis. Therefore, the secondary objective of this study was to evaluate the ability of MTM to inactivate M. bovis BCG in spiked milk samples as well as its ability to preserve BCG DNA for the PCR assay. The results showed that MTM can successfully inactivate BCG alone or in spiked milk samples while preserving DNA for the PCR assay. The CT values of M. bovis BCG alone and spiked milk samples aliquoted in MTM and without MTM were similar at various dilutions. Taken together, our results indicate that using DNA extracted from the milk cream fraction alone or combined milk cream and pellet improved the recovery rate of M. bovis DNA in bovine milk samples. MTM has the potential to provide a safe and rapid sample processing tool for M. bovis inactivation in milk samples and preserve DNA for molecular diagnostics.
Bovine milk
Bovine tuberculosis
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The assay was aimed to investigate and analysis on main pathogen in cow mastitis and pathogen sensitivity to antibiotics in dairy farm of Jilin city.We collected aseptically cow mastitis milk samples,and conducted isolation,cultivation and drug sesitivity test.The results was indicated that hemolytic Streptococcus was main pathogenic bacteria to cause cow mastitis in dairy farm.Not only the pathogen had strong drug resistance,but also it could resist many antibiotics.In conclusion,the pathogenic bacteria had strong pathogenicity and drug resistance in dairy farm.
Isolation
Pathogenic bacteria
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Disease Control
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Immunopathological Responses to the Bovine Mastitis Associated with Staphylococcus Species Infection
Bovine mastitis is a disease that concerns animals' welfare and increases the economic production losses. Bacterial agents such as Staphylococcus species are the main causative agent of bovine mastitis. This bacterial agent expresses some inflammatory cytokines that might enhance the cell-mediated, which may promote the pathogenesis of mastitis. The objective of the current study was to investigate the bovine innate immune response circulating levels of pro-inflammatory and anti-inflammatory cytokines. A total of 10 mL of milk specimens were collected randomly from 100 clinically mastitic cows, and another 20 clinically healthy cows were considered as a control group for the California Mastitis test. The microbiological cultures of milk specimens were performed. The interleukins (ILs)that involved IL-4, IL-6, and IL-10 were detected using the ELISA test for the evaluation of the pro-inflammatory bovine mastitis pathophysiology. The results of this study showed that Staphylococcus aureus detection was in 31.2% of mastitic milk and 8.7% of non-mastitic milk specimens; and the coagulase-negative Staphylococcus was detected in 14.8% and 18.7% in the mastitic and non-mastitic milk specimens, respectively. The IL-6 level was shown significantly higher (P<0.05)in the specimens of mastitic milk (194±12.8 pg/mL) compared to the non-mastitic milk (31±2.9 pg/mL). In conclusion, the elevated level of expression of IL-6 cytokine in the milk of cows with mastitis suggested that IL-6 might be used as a potentially suitable biomarker for early bovine mastitis diagnosis
California mastitis test
Pathogenesis
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ABSTRACT Mycobacterium bovis is the causative agent of bovine tuberculosis, a disease that is increasing in incidence in United Kingdom cattle herds. In addition to increasing economic losses, the rise in bovine tuberculosis poses a human health risk. There is an urgent requirement for effective strategies for disease eradication; this will likely involve vaccination in conjunction with current test and slaughter policies. A policy involving vaccination would require an accurate diagnosis of M. bovis -infected animals and the potential to distinguish these animals from vaccinates. Currently used diagnostic tests, the skin test and gamma interferon (IFN-γ) blood test, have a sensitivity of up to 95%. A further complication is that M. bovis BCG-vaccinated animals are also scored positive by these tests. We tested the hypothesis that the quantification of IFN-γ-producing lymphocytes by flow cytometric analysis of intracellular IFN-γ expression would provide a more accurate discrimination of M. bovis -infected animals from BCG vaccinates. Significant numbers of IFN-γ-expressing CD4 + T cells were detected following culture of heparinized blood from M. bovis -infected animals, but not from BCG vaccinates, with purified protein derived from M. bovis (PPD-B) or live mycobacteria. Only 1 of 17 BCG-vaccinated animals had a significant number of CD4 + T lymphocytes expressing IFN-γ, compared with 21/22 M. bovis -infected animals. This assay could allow an accurate diagnosis of M. bovis and allow the discrimination of BCG-vaccinated cattle from infected cattle.
Bovine tuberculosis
BCG vaccine
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Staphylococcal mastitis is one of the commonest infection associated with dairy animals, mostly prevails in sub-clinical form and once left unidentified, usually flares up with clinical mastitis resulting in huge economic losses to the dairy owners. Therefore, the present study was undertaken to know the incidence of clinical and sub-clinical mastitis caused by Staphylococcus aureus in bovines in the Proddatur region of Andhra Pradesh by means of conventional and molecular methods. A total of 61 clinical mastitis milk samples were collected and 105 milk samples were screened for the presence of sub-clinical mastitis by California mastitis test (CMT). The clinical samples and the samples positive for sub-clinical infection were then subjected for specific culture and morphological identification by gram’s staining. The positive isolates were further confirmed for Staphylococcus genus and S. aureus by PCR targeting the 16S rRNA and nuc gene respectively. The overall incidence of Staphylococcus Sp and S. aureus in bovine mastitis was found to be 89% and 54% respectively in clinical mastitis and 71% and 50% in sub-clinical mastitis as confirmed by PCR. These findings clearly indicate the prevalence of S. aureus infection in bovine mastitis in this specific region and suggest the need for the routine screening of dairy herds for S. aureus associated mastitis infection, use of better management and animal husbandry practices for the prevention and early cure of bovine mastitis.
California mastitis test
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One of the most important problems in milk production, causing great economic loses is certainly mastitis. In order to minimize economic losses from mastitis dairy farms introduce different mastitis management programs. These programs include mastitis therapy and prevention. In mastitis control prevention is most important and when mastitis occurs cost of therapy and milk discharge is very important. In our study we examined cost of mastitis treatment and milk loss in different mastitis management programs. We concluded that most costly are mastitis caused by specific pathogens. Cost of milk loss is 2.4 times bigger than cost of drug consumption. Applying of tit-dipping has great importance in reduction of mastitis caused by specific pathogens and less importance for conditional saprophytes. In total, cost of mastitis treatment on whole farm was almost the same for all mastitis management programs, while the effect of the program on farm C was the most expensive in the cows with the finding of specific pathogens.
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