A simple and specific method for direct detection of bovine mastitis pathogens (Streptococcus agalactiae (GBS), Staphylococcus aureus and Escherichia coli) in milk products, bacterial samples from milk and isolated bacterial DNA was developed. The method is based on polymerase chain reaction (PCR) using sequence-specific primers only for GBS and species-specific primers derived from 16S and 23S rRNA for all chosen species. The presence of the gene of surface immunogenic protein (Sip) in bovine GBS isolates, described previously only in human GBS isolates was confirmed. The GBS detection was performed with the sequence coding for surface immunogenic protein from GBS human isolates designated as Sip specific sequence (SSS); this sequence was selected for specific primer design. The sequence is unique for GBS and was designed from a consensus of all known sip genes. The specific identification was shown on a collection of 75 GBS bovine isolates from different localities in Slovakia. All isolates were positive to SSS, 16S and 23S rRNA sequence. The 16S and 23S rRNA PCR detection was also performed with S. aureus and E. coli isolates and specific PCR products were also detected. The detection limit of this assay for milk products was 6 CFU/microL (i.e. 6000 CFU/mL) for GBS and E. coli, and 16 CFU/microL for S. aureus. This rapid, sensitive and specific diagnostic method can be performed within hours and represents an innovative diagnostic tool for the detection of milk pathogens in dairy products.

• MeSH
• Antigens, Bacterial genetics   MeSH
• Bacteriological Techniques * MeSH
• DNA, Bacterial analysis   genetics   MeSH
• DNA Primers MeSH
• Electrophoresis, Agar Gel MeSH
• Escherichia coli genetics   isolation & purification   MeSH
• Dairy Products microbiology   MeSH
• Molecular Sequence Data MeSH
• Polymerase Chain Reaction methods   MeSH
• Food Microbiology MeSH
• DNA, Ribosomal analysis   genetics   MeSH
• RNA, Ribosomal, 16S genetics   MeSH
• RNA, Ribosomal, 23S genetics   MeSH
• Base Sequence MeSH
• Sensitivity and Specificity MeSH
• Staphylococcus aureus genetics   isolation & purification   MeSH
• Streptococcus agalactiae genetics   isolation & purification   MeSH
• Publication type
• Journal Article MeSH
• Evaluation Study MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Antigens, Bacterial MeSH
• DNA, Bacterial MeSH
• DNA Primers MeSH
• DNA, Ribosomal MeSH
• RNA, Ribosomal, 16S MeSH
• RNA, Ribosomal, 23S MeSH
• SIP protein, Streptococcus group B MeSH Browser

Possible correlation between Toll-like receptor (TLR)-gene mutations and the susceptibility of the mammary gland to bacterial infections and also the associate breed-dependent aspects of somatic cell concentration (SCC), bacterial infection and TLR-gene mutations in sheep are described. In Polish Lowland Sheep (PLS), milk samples exceeding the level of 500/microL (i.e. 5 x 10(5) per mL) of SCC were recorded almost twice more frequently than in Polish Heath Sheep (PHS) (40 and 22.3%, respectively). The frequency of bacterial infections was also found in a similar ratio (20 and 12.7%, respectively). During detection of the TLR-gene mutation we recorded 2 alleles of TLR1, 6 alleles of TLR2 and 10 alleles of TLR4 genes in PHS sheep, while PLS sheep possessed 2, 4 and 6 alleles, respectively. Statistical analyses revealed a relationship between the specified TLR alleles, SCC and the frequency of incidence of bacterial inflammations of mammary gland. The data may serve as a benchmark for further study of TLR-gene mutation-dependent predisposition of mammary gland defensive cells to recognize the pathogen properly and initiate the immunological response, and may help in identifying one of the markers of natural resistance against sheep mastitis.

• MeSH
• Alleles MeSH
• Bacterial Infections genetics   veterinary   MeSH
• Gene Frequency MeSH
• Genetic Predisposition to Disease * MeSH
• Mammary Glands, Animal microbiology   MeSH
• Milk cytology   microbiology   MeSH
• Sheep Diseases genetics   MeSH
• Sheep MeSH
• Polymorphism, Genetic * MeSH
• Toll-Like Receptors genetics   MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Toll-Like Receptors MeSH

A chromosomal DNA fragment of 8992 bp in size that has not been previously identified in Streptococcus agalactiae, was cloned and sequenced from strain 98-D60C. In particular, this 8992-bp fragment contained genes homologous to the sensor histidine kinase gene and the DNA-binding response-regulator gene of Streptococcus pneumoniae, and S. agalactiae bac gene. Structural and genetic features of the 8992-bp fragment were highly similar to those specific for bacterial pathogenicity islands. Analysis of epidemiologically unrelated S. agalactiae strains revealed that this fragment was present only in bac gene-positive strains. The possible origin of the 8992-bp fragment in S. agalactiae and its significance for molecular mechanisms of "bacteria-host" interactions are discussed.

• MeSH
• Antigens, Bacterial genetics   MeSH
• Bacterial Proteins genetics   MeSH
• DNA, Bacterial analysis   MeSH
• Virulence Factors genetics   MeSH
• Genomic Islands genetics   MeSH
• Humans MeSH
• Molecular Sequence Data MeSH
• Gene Expression Regulation, Bacterial genetics   MeSH
• Sequence Homology, Nucleic Acid MeSH
• Streptococcus agalactiae * genetics   pathogenicity   MeSH
• Streptococcus pneumoniae genetics   MeSH
• Pregnancy MeSH
• Check Tag
• Humans MeSH
• Pregnancy MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Antigens, Bacterial MeSH
• Bacterial Proteins MeSH
• DNA, Bacterial MeSH
• Virulence Factors MeSH

Temperate bacteriophages were induced in and released from isolates of Staphylococcus aureus and Streptococcus agalactiae using mitomycin C. Various specific indicator cultures were tested for providing clear plaques after phage infection. Specific lytic mixture of bacteriophages was prepared using the induced, modified and laboratory variants of phages. Under laboratory conditions, the mixture eliminated all isolates from the tested collection of microorganisms. The restriction barrier of some bacterial isolates to bacteriophage infection was overcome either by UV irradiation or in vitro modification of bacteriophage DNA with specific methyltransferases. Conjugative R plasmids, capable of replication in G+ and G- bacteria, were detected and isolated from S. aureus and S. agalactiae antibiotic-resistant strains.

• MeSH
• Virus Activation MeSH
• DNA, Viral genetics   isolation & purification   MeSH
• Humans MeSH
• Drug Resistance, Multiple, Bacterial genetics   MeSH
• R Factors genetics   MeSH
• Staphylococcus Phages physiology   MeSH
• Staphylococcus aureus drug effects   genetics   isolation & purification   virology   MeSH
• Staphylococcus drug effects   genetics   isolation & purification   virology   MeSH
• In Vitro Techniques MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• DNA, Viral MeSH

Group B streptococcal (GBS) gene encoding the putative lipoprotein and adherence factor ScaAB was cloned and expressed in E. coli. Recombinant ScaAB protein was isolated. Signal sequence of ScaAB was found to be cleaved in the E. coli host. ScaAB recombinant protein was immunogenic in mice and antibodies against this protein were discovered in mice sera after GBS infection. The perspectives of the use of ScaAB protein in GBS vaccine are discussed.

• MeSH
• Bacterial Adhesion MeSH
• Bacterial Proteins chemistry   genetics   immunology   metabolism   MeSH
• Immunization MeSH
• Membrane Proteins chemistry   genetics   immunology   metabolism   MeSH
• Molecular Sequence Data MeSH
• Mice MeSH
• Antibodies, Bacterial blood   MeSH
• Recombinant Proteins chemistry   genetics   immunology   metabolism   MeSH
• Amino Acid Sequence MeSH
• Streptococcus agalactiae immunology   isolation & purification   pathogenicity   MeSH
• Streptococcal Infections microbiology   mortality   prevention & control   MeSH
• Animals MeSH
• Check Tag
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Bacterial Proteins MeSH
• Membrane Proteins MeSH
• Antibodies, Bacterial MeSH
• Recombinant Proteins MeSH