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      Genotyping of fimbrial adhesins in Escherichia coli strains isolated from slovak piglets suffering from diarrhea

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          Abstract

          One-hundred sixty Escherichia coli isolates obtained from piglets with diarrhea from different parts of Slovakia were examined for the presence of genes coding for F4, F5, F6 and F41 fimbrial adhesins, and hemolytic activity. According to polymerase chain reaction tests 74 (46 %) E. coli isolates were positive for primers that detected genes coding for fimbrial adhesins. Of these 74 isolates, 64 were positive for genes encoding for F4 +, four for F5 +, five for F6 +, and one for both F41 + and F5 + adhesins.

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          Most cited references35

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          Enterotoxigenic Escherichia coli (ETEC) in farm animals.

          Animal diseases due to enterotoxigenic Escherichia coli (ETEC) typically appear as severe watery diarrhoea during the first few days of life (also a few days after weaning in pigs). ETEC adhere to the small intestinal microvilli without inducing morphological lesions and produce enterotoxins acting locally on enterocytes. This action results in the hypersecretion (of water and electrolytes) and reduced absorption. Adhesins and toxins are the two prominent virulence attributes of ETEC and the level of knowledge of these factors determines the chances for successful prevention and therapy of the disease. For animal ETEC the most common adhesins are the fimbriae (pili) on the surface: F4(K88), F5(K99), F6(987P), F41, F42, F165, F17 and F18. Enterotoxins (extracellular proteins or peptides) of animal ETEC are classified as heat-labile (LT) and heat-stable (ST) enterotoxins with further subdivisions (LTh-I, LTp-I, LTIIa, LTIIb, STaH, STaP, STb) according to antigenic and biological differences. Fimbriae and LT enterotoxins are made up of large molecular weight proteins which facilitate their utilisation in vaccines and their detection using immunodiagnostic systems. The adhesive fimbriae and enterotoxins of animal ETEC are plasmid determined (except F41 and F17). Virulence gene probes (DNA hybridisation, PCR) are specific and sensitive diagnostic and epidemiologic tools for the detection of ETEC. Based on genetic typing, the ETEC, in spite of limited serogroups, seem to represent a population of E. coli with a diverse genetic background.
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            Multiplex PCR assay and simple preparation method for stool specimens detect enterotoxigenic Escherichia coli DNA during course of infection.

            Infection with enterotoxigenic Escherichia coli (ETEC) is a common cause of diarrhea among travelers and residents of developing countries. ETEC produces either a heat-stable toxin or a heat-labile toxin, or both, encoded by plasmid-borne ST and LT genes, respectively. Diagnosis of infection with this subclass of E. coli can be performed with oligonucleotide hybridization probes; however, the sensitivity and specificity of this method are insufficient. A nonradioactive multiplex PCR assay that provides a sensitive and specific method for detecting the presence of either or both toxin genes has been developed. A simple procedure that removed inhibitors of the PCR while efficiently releasing ETEC DNA from stool specimens for subsequent amplification was used. The results for samples from a human volunteer study of ETEC infection indicated that this method of sample preparation results in greater clinical sensitivity than conventional total nucleic acid extraction and ethanol precipitation. Detection of ETEC by a multiplex PCR assay in stool specimens directly processed with a glass matrix and chaotropic solution had greater sensitivity than culture.
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              Prevalence of virulence factors of Escherichia coli strains isolated from diarrheic and healthy piglets after weaning.

              J Osek (1999)
              This study determined the prevalence of F4, F5, F6, F17 and F41 fimbriae and the genes for FedA (F18 fimbriae), LT and ST enterotoxins, and Shiga toxins Stx1, Stx2 and Stx2e among E. coli isolated from 372 weaned pigs with diarrhea and 46 healthy pigs of the same age. Agglutination tests showed that most isolates were negative for all five fimbrial antigens. The F4 antigen was found in 71 (19.1%) and the F5, F6, or F41 antigen was detected in 6.4% of isolates from diseased pigs. Genes for the F18 fimbriae were detected in 10 (2.7%) strains from diarrheic pigs and in 1 of 46 isolates from healthy pigs. Most isolates (213, 57.3%) from pigs with diarrhea were positive for LTI only or for LTI and STI or Stx2e toxin genes. Fifteen strains (13.7%) possessed only the STI or STII toxin genes. All F4-positive bacteria had genes for LTI or LTI and STI, whereas F18-positive isolates had genes for LTI, LTI/STI, or LTI/Stx2e. Of the strains isolated from diseased pigs, 264 (71.0%) were negative for the fimbrial antigens (genes) examined in this study. The fimbria-negative isolates frequently possessed genetic determinants for LTI (118, 31.7%) or for STII (16, 4.3%) enterotoxins.
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                Author and article information

                Journal
                Folia Microbiol (Praha)
                Folia Microbiol. (Praha)
                Folia Microbiologica
                Springer Netherlands (Dordrecht )
                0015-5632
                1874-9356
                2004
                : 49
                : 1
                : 59-63
                Affiliations
                [1 ]GRID grid.412971.8, ISNI 0000000122346772, Institute of Microbiology and Immunology, Department of Food Hygiene and Technology, , University of Veterinary Medicine, ; 041 81 Košice, Slovakia
                [2 ]GRID grid.412971.8, ISNI 0000000122346772, Center for Analysis of DNA, , University of Veterinary, Medicine, ; 041 81 Košice, Slovakia
                [3 ]Department of Bacteriology, Center of Viet Nam Veterinary Research Institute, Nha Trang, Viet Nam
                [4 ]Department of Bacteriology, State Veterinary Institute, 949 01 Nitra, Slovakia
                [5 ]Department of Bacteriology, State Veterinary Institute, 842 52 Bratislava, Slovakia
                Article
                BF02931647
                10.1007/BF02931647
                7090526
                15114867
                d233a456-c10e-428c-acc9-6c95bbf6d800
                © Institute of Microbiology, Academy of Sciences of the Czech Republic 2004

                This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.

                History
                : 13 June 2003
                : 9 September 2003
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                © Institute of Microbiology, Academy of Sciences of the Czech Republic 2004

                Microbiology & Virology
                hemolytic activity,field isolate,veterinary research institute,multiplex polymerase chain reaction assay,polymerase chain reaction amplification product

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