Abstract: Microsporidia comprises various pathogenic species can infect humans by means of water. Moreover, chlorine disinfection of drinking-water has limitations against this protozoan pathogen. A total of 48 water samples were collected from two drinking water treatment plants having two different filtration systems (slow sand filter and rapid sand filter) during one year period. Samples were collected from inlet and outlet of each plant. Samples were separately filtrated through nitrocellulose membrane (142 mm, 0.45 µm), then eluted and centrifuged. The obtained pellet from each sample was subjected to DNA extraction, then, amplification using genus-specific primer for microsporidia. Each microsporidia-PCR positive sample was performed by two species specific primers for Enterocytozoon bieneusi and Encephalitozoon intestinalis. The results of the present study showed that the percentage of removal for microsporidia through different treatment processes reached its highest rate in the station using slow sand filters (100%), while the removal by rapid sand filter system was 81.8%. Statistically, the two different drinking water treatment plants (slow and rapid) had significant effect for removal of microsporidia. Molecular identification of microsporidia-PCR positive samples using two different primers for Enterocytozoon bieneusi and Encephalitozoon intestinalis showed the presence of the two pervious species in the inlet water of the two stations, while Encephalitozoon intestinalis was detected in the outlet water only. In conclusion, the appearance of virulent microsporidia in treated drinking water may cause potential health threat.
Abstract: Artemia is one of the most conspicuous invertebrates
associated with aquaculture. It can be considered as a model
organism, offering numerous advantages for comprehensive and
multidisciplinary studies using morphologic or molecular methods.
Since DNA extraction is an important step of any molecular
experiment, a new and a rapid method of DNA extraction from adult
Artemia was described in this study. Besides, the efficiency of this
technique was compared with two widely used alternative techniques,
namely Chelex® 100 resin and SDS-chloroform methods. Data
analysis revealed that the new method is the easiest and the most cost
effective method among the other methods which allows a quick and
efficient extraction of DNA from the adult animal.
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.