Subject(s)
Bass , Fish Diseases , Histocompatibility Antigens Class II/genetics , Signal Transduction/genetics , Spleen/immunology , Vibrio Infections/veterinary , Animals , Bass/genetics , Bass/immunology , Fish Diseases/immunology , Fish Diseases/physiopathology , Gene Expression Profiling , Gene Expression Regulation , Vibrio Infections/immunology , Vibrio Infections/physiopathology , Vibrio parahaemolyticus/physiologySubject(s)
Fish Diseases/physiopathology , Signal Transduction , Spleen/physiopathology , Vibrio Infections/veterinary , Animals , Bass/immunology , Bass/microbiology , Fish Diseases/immunology , Signal Transduction/immunology , Spleen/immunology , Transcriptome , Vibrio Infections/immunology , Vibrio Infections/physiopathology , Vibrio parahaemolyticus/immunologyABSTRACT
We investigated the potential of USA300 MRSA emergence in Malaysia by examining 268 MSSA isolates from both community (110) and healthcare (158) settings. Nine isolates from both the environments were similar to the USA300 MRSA background based on MLST, spa and PFGE type. These results underscore the importance of continued surveillance to monitor the emergence of USA300 MRSA in Malaysia.
Subject(s)
Methicillin Resistance/genetics , Methicillin-Resistant Staphylococcus aureus/isolation & purification , Methicillin/pharmacology , Staphylococcal Infections/microbiology , Staphylococcus aureus/drug effects , Bacterial Typing Techniques , DNA, Bacterial/genetics , Electrophoresis, Gel, Pulsed-Field , Humans , Malaysia/epidemiology , Methicillin-Resistant Staphylococcus aureus/genetics , Multilocus Sequence Typing , Staphylococcal Infections/epidemiology , Staphylococcus aureus/genetics , Staphylococcus aureus/isolation & purificationABSTRACT
Vancomycin-resistant Enterococcus faecium (VREF) in human infections mostly belong to the high-risk, epidemic, clonal complex-17 (CC17) group. Treatment limitation and high conjugation frequency makes it dominant in hospitals worldwide. We investigated positive cultures by Pulse-field gel electrophoresis (PFGE), multi locus sequence typing (MLST). DNA of two strains (A2 and C) appeared to be clonally related by PFGE. Three strains were of ST 18 type (A1, B and C) and strain A2 is of a new ST 596. This ST 18 type strain found in our study is crucial and is believed to be the first in Malaysia.
Subject(s)
Enterococcus faecium , Vancomycin Resistance , Bacterial Proteins , DNA, Bacterial/genetics , Electrophoresis, Gel, Pulsed-Field , Gram-Positive Bacterial Infections , Hospitals , Humans , Malaysia , Multilocus Sequence Typing , VancomycinSubject(s)
Anti-Bacterial Agents/therapeutic use , Drug Resistance, Bacterial , Fusidic Acid/therapeutic use , Methicillin-Resistant Staphylococcus aureus , Staphylococcal Infections/microbiology , Humans , Malaysia , Methicillin-Resistant Staphylococcus aureus/drug effects , Methicillin-Resistant Staphylococcus aureus/isolation & purification , Staphylococcal Infections/drug therapyABSTRACT
AIMS: To evaluate a live recombinant Lactococcus lactis vaccine expressing aerolysin genes D1 (Lac-D1ae) and/or D4 (Lac-D4ae) in protection against Aeromonas hydrophila in tilapia (Oreochromis niloticus). METHODS AND RESULTS: The polymerase chain reaction (PCR)-amplified 250- and 750-bp sequences coding for domains D1 and D4 of aerolysin were individually cloned into pNZ8048 and electrotransformed into L. lactis. The recombinant vaccine candidates were then either orally fed or injected intraperitoneally into tilapia. The development of antibodies in sampled fish compared to control groups implied that the recombinant epitopes expressed in L. lactis were able to elicit an immunogenic response in tilapia. Interestingly, the lower doses of both Lac-D1ae and Lac-D4ae gave higher antibody levels over the study period. Fish immunized with Lac-D1ae and Lac-D4ae together showed the highest level of protection, and the mortality was reduced significantly compared to control strains in both modes of vaccination. CONCLUSIONS: The recombinant L. lactis strain expressing D1 and D4 produced aerolysin-specific serum IgM in tilapia. Both D1 and D4 promoted 55-82% relative per cent survival (RPS) against Aeromonas infection through intraperitoneal injection, whereas the RPS following oral feeding of the vaccine was 70-100%. SIGNIFICANCE AND IMPACT OF THE STUDY: The D1 and D4 regions of the aerolysin protein have been successfully identified as immunogenic regions that can elicit antibody production in tilapia and protect against challenge with Aer. hydrophila. A promising oral vaccine using L. lactis harbouring the D1 and D4 regions has been developed to control Aer. hydrophila.
Subject(s)
Bacterial Toxins/genetics , Fish Diseases/prevention & control , Gram-Negative Bacterial Infections/veterinary , Lactococcus lactis/genetics , Lactococcus lactis/immunology , Pore Forming Cytotoxic Proteins/genetics , Vaccines, Synthetic/genetics , Vaccines, Synthetic/immunology , Aeromonas hydrophila/physiology , Animals , Cichlids/genetics , Cichlids/immunology , Genes, Bacterial/genetics , Gram-Negative Bacterial Infections/prevention & controlABSTRACT
Methicillin-resistant Staphylococcus aureus (MRSA) from Malaysia were shown to possess staphylococcal cassette chromosome mec (SCCmec)-III and IIIA. Spa sequencing and multi-locus sequence typing (MLST) documented t037 and ST 239 (CC8) for 83.3% of the isolates. This confirms observations in several other Far Eastern countries and corroborates the epidemicity of this clone.
Subject(s)
Bacterial Typing Techniques , DNA Fingerprinting , Methicillin-Resistant Staphylococcus aureus/classification , Methicillin-Resistant Staphylococcus aureus/genetics , Staphylococcal Infections/epidemiology , Staphylococcal Infections/microbiology , Cluster Analysis , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , Genotype , Humans , Malaysia/epidemiology , Methicillin-Resistant Staphylococcus aureus/isolation & purification , Molecular EpidemiologyABSTRACT
The ctxB gene, the causative agent of cholera epidemic was successfully cloned from V. cholerae in E. coli. The insertion of the gene was confirmed by PCR as well as restriction digestion analyses. The sequencing results for the gene confirmed that the insert was in the correct orientation and in-frame with the P(BAD) promoter and it showed that the gene was 99% homologous to the published ctxB sequence. The CTB protein was successfully expressed in E. coli using the pBAD/His vector system. The expected protein of approximately 14 kDa was detected by SDS-PAGE and Western blot. The use of pBAD/His vector to express the cholera toxin gene in E. coli would facilitate future study of toxin gene products.
Subject(s)
Cholera Toxin/metabolism , Escherichia coli/metabolism , Gene Expression , Vibrio cholerae/metabolism , Cholera Toxin/genetics , Escherichia coli/genetics , Genetic Vectors/genetics , Vibrio cholerae/geneticsABSTRACT
A small plasmid designated pAR141 was isolated from Lactococcus lactis subsp. lactis M14 and its complete 1,594 base pair nucleotide sequence was determined. Analysis of the sequence indicated that this plasmid does not carry any industrially important determinants besides the elements involved in plasmid replication and control. The transcriptional repressor CopG and replication initiation protein RepB appeared as a single operon. A small countertranscribed RNA (ctRNA) coding region was found between the copG and repB genes. The double strand origin (dso) and single strand origin (sso) of rolling circle replicating (RCR) plasmids were also identified in pAR141, suggesting that this plasmid replicates by rolling circle (RC) mode. This observation was supported by S1 nuclease and Southern hybridization analyses.
Subject(s)
Lactococcus lactis/genetics , Plasmids/metabolism , Sequence Analysis, DNA , Amino Acid Sequence , Base Sequence , Blotting, Southern , Cloning, Molecular , DNA Restriction Enzymes/pharmacology , DNA, Single-Stranded/chemistry , Lactococcus/metabolism , Molecular Sequence Data , Open Reading Frames , Sequence Homology, Nucleic AcidABSTRACT
The cholera enterotoxin (CT) has been considered a major virulence factor of Vibrio cholerae. The accessory cholera enterotoxin (ace) gene is the third gene of V. cholerae virulence cassette. The gene coding for the Ace toxin was amplified from V. cholerae isolates producing a single band of 314 bp. The presence of ace gene was confirmed by hybridization as well as by sequencing. The gene was successfully expressed in Escherichia coli (LMG194) using expression, pBAD/Thio-TOPO vector. Optimal conditions for expression included choice of host strain, temperature used for culturing, and concentration of antibiotic and arabinose inducer. The Ace protein was obtained from the cell supernatant as a fusion protein with a molecular mass 34 kDa which was detected using an anti V5-HRP epitope tagged antibody.
