The prevalence of mastitis, milk quality and health risks associated with milk consumption were investigated on 96 randomly selected traditional herds in Dodoma rural and Mvomero districts of Tanzania. Mastitis was investigated based on clinical signs, microbiology and California mastitis test (CMT), while milk quality was evaluated using total viable count (TVC)and total coliform count (TCC). Animals were tested for tuberculosis using a single comparative intradermal tuberculin test. The prevalence of subclinical mastitis based on CMT was low (8.3%). The major isolates were Staphylococcus aureus (35.3%), other staphylococci (20.8%), coliforms (27.7%), microcci (5.8%) and streptococci (9.8%). The average TVC of milk in Dodoma rural district (1.0 x10(7)+/-3.4 x10(7))was significantly higher than that in Mvomero district (8.9x10(5) 3.5x10(6)) (p<0.001)and the proportion of TCC-positive samples in Dodoma (70.7%)were significantly higher (p<0.001) than that of Mvomero sample(20.8%). Whereas no tuberculin reactor animal was detected in the study animals, atypical mycobacteria were isolated from milk and one sample from Dodoma had Mycobacterium tuberculosis. Knowledge on health risks associated with milk consumption was low (20.8%). It is concluded that lack of awareness on health risks associated with milk consumption amongst rural communities needs to be addressed in order to safeguard their health.
Animals ; Cattle ; Female ; Humans ; Mastitis, Bovine/*epidemiology ; Milk/*microbiology/*standards ; Prevalence ; Public Health ; Tanzania/epidemiology ; Tuberculosis, Bovine/*epidemiology

In this study, we investigated the impact of mastitisinfection on the quality of milk composition in small-scaledairy bovine herds. The purpose of this study was to finda milk quality somatic cell count (SCC) standard thatcould be used as an integral component of a controlprogram. In all, 396 quarter milk samples from lactatingcross-bred cows (Holstein & Zebu) were analyzed; 56% ofthese quarters were experiencing intramammary infection,with an overall mean SCC of 5.46x10(5)+/-2.30x10(4)cells/ml. Infected quarters had significantly (p<0.05) highermean SCC levels (6.19x10(5)+/-4.40x10(4)cells/ml) comparedto healthy quarters (2.65x10(5)+/- 2.40x10(4)cells/ml). Inhigh SCC milk and infected quarters, the concentrationsof non-casein fractions, sodium, chloride, and free fattyacid were higher (p<0.05), while the casein content,lactose, casein-to-total protein, potassium, and calciumwere lower (p<0.05) compared to normal quarters. Thesefindings suggest a mean SCC threshold limit of 5.46x10(5)cells/ml for the region. It was concluded that the resultscould be used to propose a milk quality SCC standard thatcan be used as an integral component of a control program.
Animals ; Cattle ; Cell Count ; Cross-Sectional Studies ; Dairying ; Female ; Mastitis, Bovine/*metabolism/microbiology ; Milk/chemistry/*metabolism/microbiology ; Rural Population ; Statistics, Nonparametric

Our recent study has provided that the in vitro SEC-induced proliferation of bovine T cells is preceded by a period of a non-proliferative immunoregulation of T cells that may be associated with cytokine production regulated by type 1 or type 2 T cells. Inversion of CD4+:CD8+ T cell ratio and induction of CD8+T cells with immunoregulatory activity could increase the probability of intracellular survival of Staphylococcus aureus (S. aureus). The increase of activated CD8+(ACT2+ BoCD8+) T cells in cows with mastitis caused by S. aureus may be associated with immune-regulatory function in the bovine mammary gland. The difference and similarity between bovine activated CD8+ T cells (CD8+ CD26+)and well-established human CD4+ CD25+ T regulatory (Tr)cells may help to reveal their unique immune regulatory system in the host infected with S. aureus.
Animals ; Cattle ; Cell Proliferation ; Female ; Lymphocyte Activation/immunology ; Mastitis, Bovine/*immunology/microbiology ; Staphylococcal Infections/immunology/*veterinary ; Staphylococcus/*immunology ; *Superantigens ; T-Lymphocytes/*immunology

A simplified polymerase chain reaction (PCR) assay was developed for fast and easy screening of mycoplasma mastitis in dairy cattle. Species of major mycoplasma strains [Mycoplasma (M.) bovis, M. arginini, M. bovigenitalium, M. californicum, M. bovirhinis, M. alkalescens and M. canadense] in cultured milk samples were detected by this simplified PCR-based method as well as a standard PCR technique. The minimum concentration limit for detecting mycoplasma by the simplified PCR was estimated to be about 2.5 x 10(3) cfu/mL and was similar to that of the standard PCR. We compared the specificity and sensitivity of the simplified PCR to those of a culture method. Out of 1,685 milk samples cultured in mycoplasma broth, the simplified PCR detected Mycoplasma DNA in 152 that were also positive according to the culture assay. The sensitivity and specificity of the simplified PCR were 98.7% and 99.7%, respectively, for detecting mycoplasma in those cultures. The results obtained by the simplified PCR were consistent with ones from standard PCR. This newly developed simplified PCR, which does not require DNA purification, can analyze about 300 cultured samples within 3 h. The results from our study suggest that the simplified PCR can be used for mycoplasma mastitis screening in large-scale dairy farms.
Animals ; Cattle ; Colony Count, Microbial/veterinary ; DNA, Bacterial/chemistry/genetics ; Disease Outbreaks/prevention & control/veterinary ; Female ; Mastitis, Bovine/diagnosis/*microbiology ; Milk/cytology/*microbiology ; Mycoplasma/genetics/*isolation & purification ; Mycoplasma Infections/diagnosis/microbiology/*veterinary ; Polymerase Chain Reaction/veterinary

The present study determined the genetic relationships between 41 Staphyloccocus (S.) aureus isolates from bovines, humans, and food using a single enzyme amplified fragment length polymorphism (AFLP) technique. We evaluated the prevalence of staphylococcal enterotoxin (SE) genes and other virulence gene determinants by PCR. The identification of S. aureus was based on culturing and biochemical tests, and by amplifying a specific section of the 23S rRNA gene. PCR amplification of the SE genes (sea, seb, sec, see, seg, seh, and sei) singly or in combination was observed. Most isolates of bovine origin harbored hla (84%) and cap5 (74%), while most isolates from humans harbored hla (73%), cap8 (91%), and fnbA (100%). Strains from food sources were positive for hla (100%), cap5 (100%), and cap8 (64%) unlike isolates from humans or bovines. A single enzyme AFLP analysis revealed a correlation between AFLP clusters of some strains and the source of the isolates The genotypic results of the present study might help to better understand the distribution of prevalent S. aureus clones among humans, bovines, and food and will help control S. aureus infections in Indonesia.
Amplified Fragment Length Polymorphism Analysis ; Animals ; Bacterial Proteins/genetics/metabolism ; Cattle ; Female ; *Food Microbiology ; Gene Expression Regulation, Bacterial ; Humans ; Indonesia/epidemiology ; Mastitis, Bovine/epidemiology/microbiology ; Phylogeny ; Staphylococcal Infections/epidemiology/microbiology/*veterinary ; Staphylococcus aureus/*genetics

OBJECTIVEIn this study, milk from a cow with mastitis was analyzed to determine the presence of mycobacterial infection. Milk quality and security problems pertaining to the safe consumption of dairy products were also discussed in this study.

METHODSMilk was preprocessed with 4% NaOH. Then, mycobacteria were isolated from the milk sample on L-J medium. The isolate was identified using multiple loci Polymerase Chain Reaction (PCR) and multi-locus sequence analysis with 16S rRNA, sodA, hsp65, and ITS genes. The drug sensitivity of the isolate to 27 antibiotics was tested through alamar blue assay.

RESULTSSmooth, moist, pale yellow colonies appeared on the L-J medium within a week after inoculation. Based on the results of multiple loci PCR analysis, the isolate was preliminarily identified as non-tuberculous mycobacteria. The 16S rRNA, SodA, hsp65, and ITS gene sequences of the isolate exhibited 99%, 99%, 99%, and 100% similarities, respectively, with those of the published reference strains of Mycobacterium elephantis (M. elephantis). The drug sensitivity results showed that the strain is resistant to isoniazid, p-aminosalicylic acid, and trimesulf but is sensitive to ofloxacin, rifampicin, amikacin, capreomycin, moxifloxacin, kanamycin, levofloxacin, cycloserine, ethambutol, streptomycin, tobramycin, rifabutin, ciprofloxacin, linezolid, cefoxitin, clarithromycin, and minocycline.

CONCLUSIONTo the best of our knowledge, this study is initially to report the isolation of M. elephantis from the milk of a cow with mastitis in China.

Animals ; Anti-Bacterial Agents ; pharmacology ; Cattle ; China ; Drug Resistance, Bacterial ; Female ; Mastitis, Bovine ; epidemiology ; microbiology ; Milk ; microbiology ; Mycobacterium ; drug effects ; genetics ; isolation & purification ; Mycobacterium Infections ; epidemiology ; microbiology ; veterinary ; Phylogeny ; Polymerase Chain Reaction

Staphylococcus (S.) aureus is a common infectious agent of bovine chronic mastitis, a disease that is difficult to eradicate. The abilities of Staphylococci to be internalized and form a biofilm can contribute to host immunological defence evasion that subsequently impairs antimicrobial therapy. The invasive capability of six S. aureus field isolates with different biofilm-forming profiles was compared in vitro using a bovine mammary epithelial cell line. This was further confirmed in primary cell cultures using fluorescent rRNA probes against S. aureus. The results suggest that S. aureus invasion levels are not related to biofilm formation.
Animals ; *Biofilms ; Cattle ; Cell Line ; Colony Count, Microbial/veterinary ; Epithelial Cells/microbiology ; Female ; In Situ Hybridization, Fluorescence ; Mastitis, Bovine/*microbiology ; Portugal ; Staphylococcal Infections/*veterinary ; Staphylococcus aureus/classification/genetics/immunology/*physiology ; Virulence Factors/i

Cows infected with Escherichia (E.) coli usually experience severe clinical symptoms, including damage to mammary tissues, reduced milk yield, and altered milk composition. In order to investigate the host response to E. coli infection and discover novel markers for mastitis treatment, mammary tissue samples were collected from healthy cows and bovines with naturally occurring severe E. coli mastitis. Changes of mammary tissue proteins were examined using two-dimensional gel electrophoresis and label-free proteomic approaches. A total of 95 differentially expressed proteins were identified. Of these, 56 proteins were categorized according to molecular function, cellular component, and biological processes. The most frequent biological processes influenced by the proteins were response to stress, transport, and establishment of localization. Furthermore, a network analysis of the proteins with altered expression in mammary tissues demonstrated that these factors are predominantly involved with binding and structural molecule activities. Vimentin and alpha-enolase were central "functional hubs" in the network. Based on results from the present study, disease-induced alterations of protein expression in mammary glands and potential markers for the effective treatment of E. coli mastitis were identified. These data have also helped elucidate defense mechanisms that protect the mammary glands and promote the pathogenesis of E. coli mastitis.
Animals ; Cattle ; Electrophoresis, Gel, Two-Dimensional/veterinary ; Escherichia coli/*physiology ; Escherichia coli Infections/genetics/immunology/microbiology/*veterinary ; Female ; Mammary Glands, Animal/*immunology/pathology ; Mastitis, Bovine/*genetics/immunology/microbiology ; Proteome/*genetics/metabolism ; *Proteomics

Sensitivity to commercial teat dips (nonoxinol-9 iodine complex and chlorhexidine digluconate) of 56 Staphylococcus (S.) aureus strains isolated from quarter milk samples of various German dairy herds treated with different teat dipping schemes was investigated in this study. The minimum inhibitory concentration was determined using a broth macrodilution method according to the German Veterinary Association guidelines. The main objective of the current study was to induce in vitro resistance induction of S. aureus to chemical disinfectants. Ten different strains were repeatedly passed ten times in growth media with sub-lethal concentrations of disinfectants. Nine strains showed a significant reduction in susceptibility to the nonoxinol-9 iodine complex but only one strain developed resistance to chlorhexidine digluconate. Stability of the acquired resistance was observed in all S. aureus strains adapted to the nonoxinol-9 iodine complex and chlorhexidine digluconate. In contrast, simultaneous resistance to different antibiotics was not observed in any of the ten investigated S. aureus strains. However, the isolates exhibited a high degree of resistance to penicillin G. Based on these results, resistance of S. aureus to chemical disinfectants may be more likely to develop if the chemicals are used at concentrations lower than that required for an optimal biocidal effect.
Animals ; Anti-Bacterial Agents/pharmacology ; Cattle ; Chlorhexidine/*pharmacology ; Disinfectants/pharmacology ; Dose-Response Relationship, Drug ; Drug Resistance, Bacterial ; Female ; Germany/epidemiology ; Iodine/chemistry/*pharmacology ; Mastitis, Bovine/epidemiology/*microbiology ; Nonoxynol/*pharmacology ; Staphylococcal Infections/epidemiology/microbiology/*veterinary ; Staphylococcus aureus/classification/*drug effects

In the present study, 35 Staphylococcal strain isolated from milk samples of 16 cows from eight farms of three different geographic locations in Central Java, Indonesia, and from milk samples of 19 cows from 19 farms of different geographic locations in Hesse, Germany, were compared pheno- and genotypically. On the basis of cultural and biochemical properties as well as by amplification of the 23S rRNA specific to Staphylococcus aureus, all isolates could be identified as S. aureus. In addition, all S. aureus isolates harboured the genes clfA and coa encoding staphylococcal clumping factor and coagulase, and the gene segments encoding the immunoglobulin G binding region and the X-region of protein A gene spa. By PCR amplification, the genes seb, seg, seh, and sei was observed for the S. aureus cultures isolated in Central Java, Indonesia and the genes sec, sed, seg, seh, sei, sej and tst for the S. aureus cultures isolated in Hesse, Germany. None of the S. aureus of both origins harboured the genes sea, see, eta and etb. All isolates were additionally positive for the genes nuc, fnbA, hla, and set1. The gene hlb was found for 6 cultures from Central Java, Indonesia and 16 cultures from Hesse, Germany. However, the gene fnbB and the gene segments cnaA and cnaB were not present among the strains isolated in Central Java, Indonesia and rare among the strains isolated in Hesse, Germany. It was of interest that most of the S. aureus isolated in Central Java, Indonesia harboured the gene cap5 and most of the strains isolated in Hesse, Germany the gene cap8. The phenotypic and genotypic results of the present study might help to understand the distribution of prevalent S. aureus clones among bovine mastitis isolates of both countries and might help to control S. aureus infections in dairy herds.
Animals ; Bacterial Typing Techniques ; Cattle ; Female ; Genes, Bacterial ; Genotype ; Germany ; Indonesia ; Mastitis, Bovine/*microbiology ; Phenotype ; Polymerase Chain Reaction ; RNA, Bacterial/genetics ; RNA, Ribosomal, 23S/genetics ; Staphylococcal Infections/microbiology/*veterinary ; Staphylococcus aureus/classification/*genetics/isolation&purification