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      OCORRÊNCIA DE GENES CODIFICADORES DE ENTEROTOXINAS ESTAFILOCÓCICAS EM AMOSTRAS DE LEITE DE VACAS Translated title: OCCURRENCE OF GENES ENCODING STAPHYLOCOCCAL ENTEROTOXINS ISOLATED FROM COW´S MILK SAMPLES

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          Abstract

          Resumo Objetivou-se com este trabalho identificar a ocorrência de genes codificadores de enterotoxinas estafilocócicas (sea, seb, sec e seg) e do gene da toxina 1 responsável pela síndrome do choque tóxico (tst) em isolados de Staphylococcus aureus procedentes de casos de mastite bovina, no estado de Pernambuco, Brasil. Foram analisados 93 isolados e observou-se a presença de genes toxigênicos em 20 (21,6%) deles, dos quais 11 (55,0%) foram positivos para o gene tst, sete (35,0%) para o gene sec e dois (10,0%) para o gene seg. Dentre os 20 isolados que amplificaram na PCR para presença dos genes sec, seg e tst, 16 (80,0%) foram positivos apenas para um gene e quatro (20,0%) foram positivos para dois genes (sec e tst). Das 17 propriedades de onde as amostras tiveram origem, sete (41,2%) apresentaram amostras positivas para pelo menos um dos genes sec, seg e tst. Este é primeiro registro de ocorrência dos genes codificadores das enterotoxinas SEC e TST-1 em amostras de leite de vacas com mastite no estado de Pernambuco, Brasil.

          Translated abstract

          Abstract The objective of this study was to identify the occurrence of genes expressing staphylococcal enterotoxin (sea, seb, sec, and seg) and the toxin gene 1 of toxic shock syndrome (tst) in Staphylococcus aureus coming from cases of bovine mastitis in the state of Pernambuco, Brazil. We analyzed 93 isolates and we observed the presence of toxigenic gene in 20 (21.6%) samples, of which 11 (55.0%) were positive for tst gene, seven (35.0%) for the sec gene, and two (10.0%) for the seg gene. Of the 20 isolates amplified in PCR for the presence of the sec gene, seg and tst, 16 (80.0%) were positive for only one gene and four (20.0%) were positive for two genes (sec and tst). Of the 17 properties from which the samples originated, seven (41.2%) had positive samples for at least one of the genes sec, seg, and tst. This is the first record of the occurrence of the genes encoding the enterotoxin SEC and TST-1 in milk samples from cows with mastitis in the state of Pernambuco, Brazil.

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          Identification of Staphylococcus aureus: DNase and Mannitol salt agar improve the efficiency of the tube coagulase test

          Background The ideal identification of Staphylococcus aureus clinical isolates requires a battery of tests and this is costly in resource limited settings. In many developing countries, the tube coagulase test is usually confirmatory for S. aureus and is routinely done using either human or sheep plasma. This study evaluated Mannitol salt agar and the deoxyribonuclease (DNase) test for improving the efficiency of the tube coagulase test in resource limited settings. The efficiency of human and sheep plasma with tube coagulase tests was also evaluated. Methods One hundred and eighty Gram positive, Catalase positive cocci occurring in pairs, short chains or clusters were subjected to growth on Mannitol salt agar, deoxyribonuclease and tube coagulase tests. Of these, isolates that were positive for at least two of the three tests (n = 60) were used to evaluate the performance of the tube coagulase test for identification of S. aureus, using PCR-amplification of the nuc gene as a gold standard. Results Human plasma was more sensitive than sheep plasma for the tube coagulase test (sensitivity of 91% vs. 81% respectively), but both plasmas had very low specificity (11% and 7% respectively). The sensitivity and specificity of the tube coagulase test (human plasma) was markedly improved when Mannitol salt agar and DNase were introduced as a tri-combination test for routine identification of Staphylococcus aureus (100% specificity and 75% sensitivity). The specificity and sensitivity of Mannitol salt agar/DNase/tube coagulase (sheep plasma) combination was 100% and 67%, respectively. Conclusion The efficiency of the tube coagulase test can be markedly improved by sequel testing of the isolates with Mannitol salt agar, DNase and Tube coagulase. There is no single phenotypic test (including tube coagulase) that can guarantee reliable results in the identification of Staphylococcus aureus.
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            Staphylococcal superantigens in colonization and disease

            Superantigens (SAgs) are a family of potent immunostimulatory exotoxins known to be produced by only a few bacterial pathogens, including Staphylococcus aureus. More than 20 distinct SAgs have been characterized from different S. aureus strains and at least 80% of clinical strains harbor at least one SAg gene, although most strains encode many. SAgs have been classically associated with food poisoning and toxic shock syndrome (TSS), for which these toxins are the causative agent. TSS is a potentially fatal disease whereby SAg-mediated activation of T cells results in overproduction of cytokines and results in systemic inflammation and shock. Numerous studies have also shown a possible role for SAgs in other diseases such as Kawasaki disease (KD), atopic dermatitis (AD), and chronic rhinosinusitis (CRS). There is also now a rich understanding of the mechanisms of action of SAgs, as well as their structures and function. However, we have yet to discover what purpose SAgs play in the life cycle of S. aureus, and why such a wide array of these toxins exists. This review will focus on recent developments within the SAg field in terms of the molecular biology of these toxins and their role in both colonization and disease.
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              Use of multiplex PCR to detect classical and newly described pyrogenic toxin genes in staphylococcal isolates.

              Staphylococcus aureus may contain one or more genes that encode a variety of immunomodulatory pyrogenic toxins (PTs), including the staphylococcal enterotoxins and toxic shock syndrome toxin (TSST). The PTs interact with several cellular targets to produce disease, such as food poisoning and toxic shock syndrome. At present, nine serologically distinct enterotoxins and one immunoreactive form of TSST have been identified and characterized. As isolates of S. aureus are further assessed, it is anticipated that this number will increase. To facilitate screening, a multiplex PCR was designed to simultaneously determine which of these 10 currently known PT genes an individual S. aureus isolate possesses. We show here, using S. aureus isolates with characterized PT phenotypes, that this novel PCR technique reliably detects each of the known PTs in a single reaction.
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                Author and article information

                Contributors
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Journal
                cab
                Ciência Animal Brasileira
                Ciênc. anim. bras.
                Universidade Federal de Goiás (Goiânia, GO, Brazil )
                1518-2797
                1809-6891
                July 2018
                : 19
                : 0
                : e43108
                Affiliations
                [1] Recife Pernambuco orgnameUniversidade Federal Rural de Pernambuco Brazil
                Article
                S1809-68912018000100314
                10.1590/1809-6891v19e-43108
                27c9115e-2572-49f8-9c53-590b692e8dc5

                This work is licensed under a Creative Commons Attribution 4.0 International License.

                History
                : 02 September 2016
                : 08 March 2018
                Page count
                Figures: 0, Tables: 0, Equations: 0, References: 25, Pages: 0
                Product

                SciELO Brazil

                Categories
                Medicina Veterinária

                Staphylococcus aureus,PCR,patogênese,mastite,pathogenesis,mastitis

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