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The role of the genus Acanthamoeba in the process of maintaining, multiplying and transmitting human pathogens has not been well recognized. The aim of our study was to evaluate the association of potentially pathogenic Acanthamoeba spp. with bacteria in strains recently isolated from Lake Malta near the city of Poznań, Poland. Most of the isolated amoebae belonged to the species A. castellanii and A. rhysodes. The majority of the isolates were pathogenic for mice and amoebae could be recovered from their brains and lungs. In those cases when amoebae could not be recovered from the autopsy material histopathological analysis showed changes in tissues indicating bacterial infection. We found that approximately 50% of the isolates were associated with endosymbiotic or endocytobiotic bacteria, e.g., Proteus sp., Micrococcus sp., Chryseobacterium meningosepticum, Clostridium perfringens, Escherichia coli, Salmonella sp., Legionella pneumophila and other species. The presence of bacteria residing inside free-living amoebae possess a great challenge in terms of disease control and sanitation of contaminated water sources.
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Heat shock response in gastrointestinal tract

72%
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Frequency of Helicobacter pylori infection in children under 4 years of age

72%
The work aimed at establishment of frequency of Helicobacter pylori (Hp) infection in children under 4 years of age. One hundred ninety-eight children (6 month to 4 year) were tested using urea breath test (UBT) with the non-radioactive isotope 13C (50 mg of urea). The air was collected before and in the 20th and 30th minute after standard meal. The results of measurements (mass spectrometry IRMS) were given as a quotient 13CO2 /CO2 (), and a positive value was set at >3,5‰. Parents of tested children were asked to fill in a questionnaire on a somatic development, the digestive tract symptoms of a child and family members as well as socioeconomic conditions. The data were analyzed to establish the risk factors in Hp infection in children. Hp infection was found in 18,38% of children. It was not related to child’s sex nor age. The statistical significance was found in the occurrence of Hp infection among children whose family members had infection and among those attending créches or kindergartens. Non-radioactive 13C UBT is very useful and easy method to use in epidemiological studies even in youngest children. The course of infection was asymptomatic and had no impact on their somatic development. Factors increasing the risk of Hp infection were occurrence of Hp among other family members and contact with other children in educational facilities.
The aim of the present research work to investigate antimicrobial activity of some honey samples six winter honeys six summer honeys collected from different regions of Western Ghats. The microbes used in this study are Staphylococcus aureus, Streptococcus pyogenes, Escherichia coli, Pseudomonas aeruginosa, and Proteus mirabilis. Antibacterial activity of the honeys was assayed using the Disc diffusion method. Noticeable variations in the antibacterial activity of the different honey samples were observed. Among the microbes Staphylococcus aureus is the most sensitive against all honey samples shows the maximum inhibitor zone compare to summer honeys.
Leptospirosis, caused by pathogenic Leptospira, is one of the most important zoonoses in the world. Several molecular techniques have been developed for detection and differentiation between pathogenic and saprophytic Leptospira spp. The aim of this study was to develop a rapid and simple assay for specific detection and differentiation of pathogenic Leptospira spp. by multiplex real-time PCR (TaqMan) assay using primers and probes targeting Leptospira genus specific 16S ribosomal RNA gene, the pathogen specific ligA/B genes and non-pathogen Leptospira biflexa specific 23S ribosomal RNA gene. Sixteen reference strains of Leptospira spp. including pathogenic and nonpathogenic and ten other negative control bacterial strains were used in the study. While the 16S primers amplified target from both pathogenic and non-pathogenic leptospires, the ligA/B and the 23S primers amplified target DNA from pathogenic and non-pathogenic leptospires, respectively. The multiplex real-time PCR (TaqMan) assay detection limit, that is, the sensitivity was found approximately 1 x 10² cells/ml for ligA/B gene and 23S ribosomal RNA gene, and 10 cells/ml 16S ribosomal RNA. The reaction efficiencies were 83-105% with decision coefficients of more than 0.99 in all multiplex assays. The multiplex real-time PCR (TaqMan) assay yielded negative results with the ten other control bacteria. In conclusion, the developed multiplex real-time PCR (TaqMan) assay is highly useful for early diagnosis and differentiation between pathogenic and non-pathogenic leptospires in a reaction tube as having high sensitivity and specificity.
The application of random amplified polymorphic DNA- polymerase chain reaction (RAPDPCR) was found to be a simple, cheap and rapid tool to discriminate human pathogenic bacterial isolates especially at intraspecific level. This molecular biological technique relies on the use of random oligonucleotide primers that arbitrarily amplifies specific regions of the genome which gives rise to a unique genomic fingerprint of the strains under investigations. With continued development of novel molecular-based technologies for rapid, high-throughput detection of food borne pathogenic bacteria, the future of conventional microbiological methods such as viable cell enumeration, selective isolation of bacteria on commercial media, and immunoassays seems tenuous. Approaches that enhance recovery of sub lethally injured bacteria, differentiation among species, differentiation among bacteria of interest using biochemical profiling, enumeration using impedance technology, techniques to confirm the presence of target pathogens using immunological methods, and bioluminescence applications for hygiene monitoring are of utmost need in identifying and combating the human pathogenic isolates. The aim of this study is to estimate the efficiency of RAPD-PCR technique in assessing the genetic diversity of diseases causing bacterial isolates. The use of RAPD-PCR in evaluating the genomic variability among the pathogenic strains belonging to different genus are also been discussed in the present report.
Microbial contamination of a water distribution system was examined. The number and the taxonomy of non-pigmented and pigmented heterotrophic bacteria (HB), number of bacteria (Pseudomonas sp., Enterococcus sp., Campylobacter sp., Yersinia sp., representatives of the Enterobacteriaceae, coagulase-positive staphylococci, and C. perfringens) in the bulk water phase, biomass of zoogloeal aggregates of bacteria, fungi, algae, protozoa and rotifers (ZABFAPR) (separated from the above on 5 pm pore size fdters) and in pipe sediments was determined. An increased number of HB occurred at the sampling sites situated as close as 4.2 km to the Water Treatment Plant (WTP), and was especially significant at 10.3 km. It was shown that the main reservoir of hygienically relevant bacteria did not occur in the water phase which is monitored in routine control analyses carried out by the WTP laboratories, but in the ZABFAPR biomass not monitored so far.
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