Técnicas moleculares aplicadas à microbiologia de alimentos

Gandra, Eliézer Ávila; Gandra, Tatiane Kuka Valente; Mello, Willians Sebastião de; Godoi, Huana da Silva

doi:10.4025/actascitechnol.v30i1.3245

Cited by 17 publications

(19 citation statements)

References 9 publications

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“…Cohen et al (1996) observed that positive samples in PCR were negative in culture; however the majority of the positive cultures were also positive in PCR. Although traditional techniques of culture and bacterial identification are used as official for analysis, it is fundamental to complement molecular techniques, since they present a higher sensitivity in a shorter time (Gandra et al 2008). Samples from five animals that were PCR positive (samples from the TT broth) were not confirmed by bacteriological culture which may indicate different differentiated bacterial profiles.…”

Section: Discussionmentioning

confidence: 99%

The urban and rural capybaras (Hydrochoerus hydrochaeris) as reservoir of Salmonella in the western Amazon, Brazil

et al. 2019

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The capybara (Hydrochoerus hydrochaeris) is the largest rodent in the world. In the state of Acre, Brazil, populations of capybaras have been increasing significantly. The role of capybaras in the transmission of certain bacterial zoonotic infections is not well understood, including bacteria of the genus Salmonella. Salmonella spp. generally cause enteritis or septicemia in mammals, however many mammalian species can carry the bacteria asymptomatically and shed it in their feces. To better understand the possible role of capybaras as reservoirs of Salmonella spp., we conducted a study of Salmonella within fecal samples from capybara in Acre. In a convenience sample, 54 capybaras from two urban and two rural areas of Acre were captured and kept for three to four days for sampling. None of the animals were symptomatic of any intestinal illness. Three separate fecal samples were collected from each animal, during their stays in captivity. Each sample was cultured for the presence of Salmonella spp. at the bacteriology laboratory of the Veterinary College of the Federal University of Acre. Samples were seeded in tetrationate pre-enrichment broth and in pre-enrichment broth peptone. After a 24 hour of incubation all samples were streaked on MacConkey Agar (MC) and Salmonella-Shigella Agar (SS). Suggestive colonies were submitted to biochemical analysis. Salmonella compatible colonies according to biochemical profile were submitted to serotyping (Sorokit for Salmonella - Probac do Brasil). In addition, the first sample from each of the 54 capybara was tested for Salmonella spp. using PCR targeting gene hilA. Eight (5%) of the 162 samples examined by bacterial culture were positive for Salmonella spp., while four (7%) of the 54 examined by PCR were positive. From the eight positive animals on culture, five were from urban area and three from rural area. On PCR, only one positive animal was from urban area and four were from rural area. Overall, by either test, one of the 54 animals was positive. All samples were collected in free - living animals with no apparent clinical signs of salmonellosis, indicating the potential of capybara as reservoir on this ecosystem.

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Section: Discussionmentioning

confidence: 99%

The urban and rural capybaras (Hydrochoerus hydrochaeris) as reservoir of Salmonella in the western Amazon, Brazil

et al. 2019

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“…The cost of most PCR techniques is relatively high; however, there are low cost systems that involve a single PCR reaction, followed by electrophoresis in agarose or polyacrylamide gel, such as ARMS-PCR (Amplification Refraction Mutation System), which makes the technique more accessible (Gandra et al, 2008;Guimarães et al, 2009). ARMS-PCR differs from an RFLP-PCR for allele determination since it does not require the use of a restriction enzyme in order to cleave the amplified DNA.…”

Section: Introductionmentioning

confidence: 99%

Research Article RFLP-PCR is more efficient than ARMS-PCR for identifying CYP2C19*2 polymorphism in atherosclerotic patients

Costa¹,

Santos²,

Bento³

et al. 2019

Genet. Mol. Res.

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The CYP2C19*2 polymorphism is the result of a point mutation leading to an alternative splicing defect, generating an enzyme with a reduced function. The enzyme CYP2C19 is responsible for the activation of the prodrug Clopidogrel and the loss of its function leads to an increase in platelet aggregation. We compared the ARMS-PCR (Amplification Refraction Mutation System) and RFLP-PCR (Restriction Fragment Length Polymorphism) to determine which of the techniques was more efficient for identifying the CYP2C19*2 polymorphism. We analyzed 200 samples from patients with atherosclerosis and 100 samples from a control group, free from the disease. The polymorphic allele was not observed in any of the samples analyzed by the ARMS-PCR technique, but was present in more than 70% of the samples when analyzed by RFLP-PCR. Thus, RFLP-PCR generated more satisfactory results for the analysis of CYP2C19*2 polymorphism, and it should be used, despite the lower cost of ARMS-PCR, as the latter did not detect this polymorphism.

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“…Various PCR modifications have been described, among which Rep-PCR, which involves the amplification of areas between repetitive elements, generating specific profiles (Gandra et al, 2008). This technique allows the determination of the origins of pathogenic microorganisms' contamination in food catering services such as those of hotels, thus filling the gap caused by the lack of information in this field of activity.…”

Section: Introductionmentioning

confidence: 99%

Good hygienic practices and identification of contamination sources in Hotel Food and Beverage sector

et al. 2018

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This study aimed to evaluate the application of good practices in hotel restaurants in Southern Brazil, relate these practices to the microbiological quality of the food offered, in addition to identifying contamination sources. The food and beverage sector of four hotels was evaluated by means of a checklist based on the Resolution 216/2014 of Agência Nacional de Vigilância Sanitária and microbiological analyzes. Surface, utensil, equipment and handlers' hand samples were collected for coagulase positive Staphylococcus and thermotolerant coliform count. Osolates obtained from these culture counts were compared by band profiles obtained by rep-PCR. According to non-conformities observed, only one hotel was rated as excellent in good practices avaliation. Some microbiological analysis exceeded the allowed limit for the analyzed microorganism count, revealing failure in the hygiene-sanitary process. Genetic similarities between some food samples and handlers' hands and utensils were identified. These results showed inadequacies in the process of food care and handling which may compromise the quality of food offered to custumers.

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Técnicas moleculares aplicadas à microbiologia de alimentos

Cited by 17 publications

References 9 publications

The urban and rural capybaras (Hydrochoerus hydrochaeris) as reservoir of Salmonella in the western Amazon, Brazil

The urban and rural capybaras (Hydrochoerus hydrochaeris) as reservoir of Salmonella in the western Amazon, Brazil

Research Article RFLP-PCR is more efficient than ARMS-PCR for identifying CYP2C19*2 polymorphism in atherosclerotic patients

Good hygienic practices and identification of contamination sources in Hotel Food and Beverage sector

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