Abstract: Many high-risk pathogens that cause disease in
humans are transmitted through various food items. Food-borne
disease constitutes a major public health problem. Assessment of the
quality and safety of foods is important in human health. Rapid and
easy detection of pathogenic organisms will facilitate precautionary
measures to maintain healthy food. The Polymerase Chain Reaction
(PCR) is a handy tool for rapid detection of low numbers of bacteria.
We have designed gene specific primers for most common food
borne pathogens such as Staphylococci, Salmonella and E.coli.
Bacteria were isolated from food samples of various food outlets and
identified using gene specific PCRs. We identified Staphylococci,
Salmonella and E.coli O157 using gene specific primers by rapid and
direct PCR technique in various food samples. This study helps us in
getting a complete picture of the various pathogens that threaten to
cause and spread food borne diseases and it would also enable
establishment of a routine procedure and methodology for rapid
identification of food borne bacteria using the rapid technique of
direct PCR. This study will also enable us to judge the efficiency of
present food safety steps taken by food manufacturers and exporters.
Abstract: In the present work we report a gram negative
bacterial isolate, from soil of a dye industry, with promising
biorefining and bioremediation potential. This isolate (GBS.5) could
utilize carbazole (nitrogen containing polycyclic aromatic
hydrocarbon) as the sole source of nitrogen and carbon and utilize
almost 98% of 3mM carbazole in 100 hours. The specific activity of
our GBS.5 isolate for carbazole degradation at 30°C and pH 7.0 was
found to be 11.36 μmol/min/g dry cell weight as compared to 10.4
μmol/min/g dry cell weight, the highest reported specific activity till
date. The presence of car genes (the genes involved in
denitrogenation of carbazole) was confirmed through PCR
amplification.
Abstract: Fourty one strains of ESBL producing P.aeruginosa
which were previously isolated from burn patients in Kerman
University general hospital, Iran were subjected to PCR, RFLP and
sequencing in order to determine the type of extended spectrum β-
lactamases (ESBL), the restriction digestion pattern and possibility of
mutation among detected genes. DNA extraction was carried out by
phenol chloroform method. PCR for detection of bla genes was
performed using specific primer for each gene. Restriction Fragment
Length Polymorphism (RFLP) for ESBL genes was carried out using
EcoRI, NheI, PVUII, EcoRV, DdeI, and PstI restriction enzymes. The
PCR products were subjected to direct sequencing of both the strands
for identification of the ESBL genes.The blaCTX-M, blaVEB-1, blaPER-1,
blaGES-1, blaOXA-1, blaOXA-4 and blaOXA-10 genes were detected in the
(n=1) 2.43%, (n=41)100%, (n=28) 68.3%, (n=10) 24.4%, (n=29)
70.7%, (n=7)17.1% and (n=38) 92.7% of the ESBL producing isolates
respectively. The RFLP analysis showed that each ESBL gene has
identical pattern of digestion among the isolated strains. Sequencing
of the ESBL genes confirmed the genuinety of PCR products and
revealed no mutation in the restriction sites of the above genes. From
results of the present investigation it can be concluded that blaVEB-1
and blaCTX-M were the most and the least frequently isolated ESBL
genes among the P.aeruginosa strains isolated from burn patients. The
RFLP and sequencing analysis revealed that same clone of the bla
genes were indeed existed among the antibiotic resistant strains.