Typing of Clinical and EnvironmentalAeromonassp. Strains by Random Amplified Polymorphic DNA PCR, Repetitive Extragenic Palindromic PCR, and Enterobacterial Repetitive Intergenic Consensus Sequence PCR

Abstract

A collection of 120 strains isolated from stool specimens collected from humans suffering from gastroenteritis and from environmental samples were analyzed by random amplified polymorphic DNA PCR (RAPD), repetitive extragenic palindromic PCR (REP-PCR), and enterobacterial repetitive intergenic consensus sequence PCR (ERIC-PCR). Species of Aeromonoas hydrophila, A. bestiarum, A. salmonicida, A. caviae, A. media, and A. veronii revealed clonal structure. There was no dominant clone causing gastroenteritis in humans. Moreover, there was no genetic similarity between clinical and environmental strains of Aeromonas sp. isolated from different geographical areas as well as from the same geographical area. Some clones colonized specific ecosystems, e.g., drinking water distribution systems. RAPD and ERIC-PCR methods had the same discriminatory power and proved to be useful for epidemiological investigation and population genetic analysis of Aeromonas spp., whereas REP-PCR was less effective for differentiating the isolates of Aeromonas spp.Bacteria of Aeromonas sp. are gram-negative, straight cells (rod-shaped to coccoid) with rounded ends. They are oxidase and catalase positive, reduce nitrate to nitrite, and ferment D-glucose. These bacteria are widely spread in the environment, especially in surface water and sewage; they also occur in untreated and treated drinking water (1, 2, 4, 18). In humans, Aeromonas spp. are responsible for gastroenteritis, chronic diarrhea, wound infections, respiratory tract infections, peritonitis, urinary tract infections, and septicemia (2, 17). Among Aeromonas-associated infections of humans A. hydrophila, A. caviae, and A. veronii are the predominating species, whereas A. eucrenophila, A. popoffii, (2), and A. culicicola (28) have never been found in clinical samples. Some Aeromonas species are associated with a wide variety of diseases in cold-and warm-blooded animals, including fish, frogs, water buffaloes, reptiles, birds, and cattle (16, 28). A. veronii, A. hydrophila, and A. salmonicida are capable of causing septicemia in freshwater and marine fish (3, 9). Clinical and environmental Aeromonas sp. isolates secrete many extracellular products, such as hemolysins, enterotoxins, and proteases. Studies conducted by Kühn et al. (22) showed that some isolates of a given species produce virulence factors more frequently than others. These findings indicate that the virulence within the genus Aeromonas might be a clonal property and only some clones may be responsible for progressive disease. However, there have been no studies that would have determined clonal structure within Aeromonas spp. and the spread of specific clones in human population and in the environment. Our study was undertaken to recognize the clonal relatedness of strains derived from diarrheal stool specimens collected from humans living in different geographical areas. We also compared genetic similarities of clones recovered from stool and the environment. In addition, we attempted to determine the genetic relati...

Multilocus sequence typing revealed a clonal lineage of Aeromonas hydrophila caused motile Aeromonas septicemia outbreaks in pond-cultured cyprinid fish in an epidemic area in central China

“…Many studies have investigated the genetic diversity of human diarrhea and environmental isolates of A. hydrophila (Aguilera-Arreola et al, 2007;Sechi et al, 2002;Szczuka and Kaznowski, 2004). All these studies revealed a high intra-specific genetic diversity within A. hydrophila, and no dominant clones were found.…”

“…We found that A. hydrophila was responsible for these MAS Aquaculture 432 (2014) 1-6 outbreaks. A. hydrophila isolates were previously studied to be phenotypically, serologically, and genetically diverse (Abbott et al, 2003;Aguilera-Arreola et al, 2005;Szczuka and Kaznowski, 2004;Thomas et al, 1990). Thus, we used a recently developed precise method of multilocus sequence typing (MLST) to evaluate the population structure of MAS outbreak isolates of A. hydrophila.…”

β-lactamase-producing Gram-negative bacteria in an intensive care unit in southern Brazil

“…Bacterial genomic DNA was extracted by the heat boil method from the overnight bacterial growth (Swanenburg et al, 1998). The PCRs were performed using primers ERIC1R (5'-ATGTAAGCTCCTGGGGATTCAC-3') and ERIC2 (5'-AAGTA AGTGACTGGGGTGAGCG-3'), as described by Szczuka and Kaznowski (2004). The ERIC-PCR fragments obtained were examined by electrophoresis in 2% agarose gels and stained with ethidium bromide.…”

Analysis of major band of Enterobacter sakazakii by ERIC-PCR and development of a species-specific PCR for detection of Ent. sakazakii in dry food samples

“…sakazakii ATCC51329 (Fig. 1) using the similarity index criteria (Szczuka and Kaznowski, 2004) of ≥90%.…”

“…Recently, ERIC-PCR was used for the evaluation of genetic analysis and epidemiology investigation of some pathogens such as Vibrio parahaemolyticus, Campylobacter, Escherichia coli, Salmonella enterica and Aeromonas sp. strains (Versalovic et al, 1991;Wong and Lin, 2001;Da Silverra et al, 2002;Mclellan et al, 2003;Szczuka and Kaznowski, 2004;Houf et al, 2005;Mouwen et al, 2005). However, little attempt has been focused on the typing of Ent.…”