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Item Emprego das técnicas de pcr e vidas na determinação da escherichia coli o157:H7 em queijos minas frescal em feiras livres e estabelecimentos sob inspeção federal(Universidade Federal de Goiás, 2009-08-31) CARVALHO, Rosangela Nunes; NUNES, Iolanda Aparecida; http://lattes.cnpq.br/2555738917312914; OLIVEIRA, Jaison Pereira de; http://lattes.cnpq.br/7342298421214478; OLIVEIRA, Antonio Nonato de; http://lattes.cnpq.br/8261502405023882Escherichia coli O157:H7 is a serotype that belongs to Shiga toxin-producing Escherichia coli group and it has been incriminated in outbreaks of food poisoning and also as cause of sporadic cases of hemorrhagic colitis and hemolytic uremic syndrome in many countries. In Brazil, there is no data available about outbreaks caused by E. coli O157:H7, but this pathogen has been found often in cattle feces. Taking into consideration the high susceptibility of Minas Frescal cheese to the contamination by E. coli O157:H7 and also considering that milk is frequently associated with outbreaks, the aim of this research was to determinate the occurrence of the pathogen in Minas Frescal cheese, and also to compare techniques for this bacterium detection, such as PCR (Polymerase Chain Reaction) and VIDAS ECO O157® (bioMérieux, Lyon, France) test. Thirty cheese samples were collected from open-air markets in Goiânia and thirty samples were collected from establishments under Federal Inspection located in Goiás that trade their products in supermarkets in Goiânia. The MPN (More probable number) of Thermo-tolerant Coliforms and MPN of E. coli were also determined from the samples. From those samples collected in open air markets, E. coli O157:H7 was detected in 6,67% and 23,33% when the technique used was VIDAS ECO O157® and PCR, respectively. From those samples collected in establishments under Federal Inspection, the bacterium was not detected. There was no significant difference (p>0,05) between VIDAS ECO O157® and PCR for pathogen detection in samples from open air market and establishments under Federal Inspection. No significant difference (p> 0,05) was observed in samples collected from open air market and industries under Federal Inspection when VIDAS ECO O157® was used to E.coli O157:H7 detection. When PCR technique was applied, there was significant difference (p< 0,05) between samples origin. In 86,67% of samples collected from open air markets, Thermo-tolerant Coliforms were detected over the level established in Brazilian legislation, but in samples collected from industries under Federal Inspection, this value was 10%. The Minas Frescal cheese samples collected from open air market were more contaminated by E.coli than those collected from establishments under Federal Inspection and commercialized in supermarkets.Item Determinação da imunogenicidade de proteínas de Mycobacterium bovis isolados de bovinos naturalmente infectados no Estado de Goiás(Universidade Federal de Goiás, 2010-02-08) FLEURY, Cinthya Marques Fortes; KIPNIS, André; http://lattes.cnpq.br/4434965360286741; FIORAVANTI, Maria Clorinda Soares; http://lattes.cnpq.br/8772502020076257; KIPNIS, Ana Paula Junqueira; http://lattes.cnpq.br/1252262903952987The purpose of this work is to contribute to the research in new diagnostic tests for bovine tuberculosis. To this end a study was performed to identify Mycobacterium bovis immunogenic proteins isolated from bovines in the field. Total extracted protein, were initially fractioned by electrophoresis in a poliacrylamide gel and then identified by Western Blotting, using serum from reagent and non-reagent animals to the purified protein derivate (PPD) intradermal diagnostic test. Protein immunogenicity was assessed according to the molecular weight profile of the positive bands. 93 serum samples were tested from PPD positive and PPD negative animals from Goiás, Distrito Federal and Minas Gerais, as well as tuberculosis free regions of the USA. By means of the Western Blotting technique it was possible to identify the 26kDa protein as the most immunogenic among the positive PPD tested animals.Item Listeria spp. e L. monocytogenes em carne bovina refrigerada e embalada a vácuo, equipamentos e ambiente de matadouros frigoríficos(Universidade Federal de Goiás, 2008-08-28) FRANÇA, Leonardo; MESQUITA, Albenones José de; http://lattes.cnpq.br/3180029815183858; MEIRELES, Marcelo Vasconcelos; http://lattes.cnpq.br/0903513897615274; NUNES, Iolanda Aparecida; http://lattes.cnpq.br/2555738917312914Intending to stipulate the parcel of Listeria spp. and L. monocytogenes in several sources, using the PCR techicic, 360 samples were analyzed in the period between november 2006 and february 2008, proceeding from 25 commercial abattoirs of five companies all licensed to export from some Brazilian States. Primers U1/LI1 for Listeria spp. and LM1/LM2 and LL5/LL6 for L. monocytogenes. Listeria spp. was detected in 19,72% of the samples, with percentage of 23% in the bone rooms, cold chamber and half carcass, 15% in beef cuts and 10% in the slaughter rooms. In the slaughterhouses it was detected in 19,42% in the A, 17,72% in the B, 22,95% in the C and 20% in the D and E. Analyzing the states, 16,34% in São Paulo, 23,94% in Mato Grosso do Sul, 14,06% in Goiás, 32% in Mato Grosso, 27,27% in Bahia and 14,28% in Pará were positive for its genus. No matter the pair of ‟primers‟‟ used for amplication, L. monocytogenes were detected in 14,72% of the samples, 20% in cold chambers, 15,83% in bone rooms, 13.33% in halfcarcass and in beef cuts and 10% in slaughter rooms. In the slaughterhouse A, 12,57% of the samples were detected positive‟, 11,39% in B, 21,31% in C and 13,33% in D e E. The bacterium was detected in São Paulo in 12,42% of positive samples, 14,08% in Mato Grosso do Sul, 12,30% in Goiás, 24%, in Mato Grosso, 27,27% in Bahia and 14,28% in Pará. The pair of primer LM1/LM2 revealed better efficience than the LL5/LL6 and although this last one has presented low efficiency to detection, the total of positive samples of L. monocytogenes of this research only was possible by the association of both. Listeria spp. and L. monocytogenes are largely propagated in the sources and studied places, and, therefore, disseminated in all slaughterhouses and analyzed States, excepting Minas Gerais. The results of this research revealed the necessariness of adoption of urgent providences about the control of the pathogen in the bovine slaughterhouses in Brazil and must alert the possibility of occurrence of outbreak of listeriosis of nutritious transmission, considering the occurrence and the frequency repetition here verified in half-carcass and beef cutsItem Impacto da assistência técnica sobre a qualidade do leite(Universidade Federal de Goiás, 2008-08-21) GUIMARÃES, Caroline Paixão do Amaral; NICOLAU, Edmar Soares; http://lattes.cnpq.br/9601723963736071; LAGE, Moacir Evandro; http://lattes.cnpq.br/2053334263459111; MESQUITA, Albenones José de; http://lattes.cnpq.br/3180029815183858In a globalization context, producing food only focused on quantity makes no more sense. At the same time, the quality becomes the most important step on food processing chain, since it s essential to guarantee life quality. Then, we purposed this study to verify the effect of private technical assistance on milk quality. The study was carried out during the dry and rain periods of the year, on 40 dairy farms located at the counties of Bela Vista de Goiás, São Miguel do Passa Quatro, Piracanjuba and Cristianópolis, in Goiás State, Brazil. The farms were randomly selected from herds with high results of somatic cell count (SCC) and total bacterial count (TBC). The effect of technical assistance on milk quality was evaluated using the results of milk analyses before and after visits of technicians on the farms. Milk samples were taken from bulk milk tanks and analyzed to somatic cell count (SCC) and total bacterial count (TBC) in electronic devices. We observed a significant decrease on the SCC and TBC after farmers had received technical assistance on milking procedures and mastitis control. The rain period didn t affect the observed results. As the SCC decreased, we observed an increase on SNF values of milk. So, we confirmed that technical assistance leaves to better milk quality, in any time of the yearItem Dinâmica da carga microbiana da sala de desossa em um matadouro frigorífico de Goiânia-GO, durante a jornada de trabalho(Universidade Federal de Goiás, 2009-07-30) MARRA, Kelly Nobre; PRADO, Cristiano Sales; http://lattes.cnpq.br/1661902818272650; LAGE, Moacir Evandro; http://lattes.cnpq.br/2053334263459111; MESQUITA, Albenones José de; http://lattes.cnpq.br/3180029815183858The meat is a food of high nutrition value and easy to damage. The deboning process, if it´s not realized in appropriated hygienic-sanitary can compromise the microbiologic quality of this noble product. The present study was realized with the major objective of monitor the dynamics of microbial load in the deboning room of a slaughterhouse - Goiânia´s - GO cold store, during a work journey. The samples were collected in different times being the first, before the beginning of the activities, the second two hours after the first, the third two hours after the second, the fourth two hours after the third and the fifth two hours after the fourth. The sampling sites were: tables surface, knives and knifegrinders, meat pieces, water from sterilizers and air environment. At the end of each day work there was collected a sample of each of the 6 points sampled at 5 different times, totaling 30 samples per day and 180 at the end of the experiment. It was analyzed the counting of mesophylic, psychrotrophic, Enterobacteriaceae, anaerobic bacteria and lactic acid, in addition to the determination of the most probable number of fecal coliforms and Escherichia coli, as 62 of IN MAP (BRAZIL, 2003). The tables, knives, knifegrinders surfaces, as well as, meat pieces, water sterilizers and the air from the deboning room, in general, presented low levels of bacterial contamination. Except the counting of mesophylic in tables, knives and knifegrinders which showed 80%, 40% and 100%, respectively, the results above the limits set by Decision 471 of MCE 2001. The levels of contamination in the deboning room by mesophylic microorganisms, psychrotrophic, fecal coliforms, Escherichia coli, Enterobacteriaceae, lactic acid and anaerobic bacteria, do not differ, within each species or group of indicators, regardless of the time of the day work. Relatively, on the counting of mesophylic in tables, 74% of bacterial growth is related to the time of the day work. In knives, 85% of Enterobacteriaceae in knifegrinders, 61% of mesophylic, 74% of psychrotrophic and 98% of the anaerobes also have this contact. In meat pieces 97% of the MPN of Escherichia coli and the water of sterilizers, 82% of mesophiles can be explained by the time of the day workItem Associação entre polimorfismos de nucleotídeo único (SNPs) no gene codificador do Toll-like receptor 4 (TLR4) e contagem celular somática(Universidade Federal de Goiás, 2010-07-30) MESQUITA, Adriano Queiroz de; ANDRADE, Maria Auxiliadora; http://lattes.cnpq.br/9441751521255467; DIAS FILHO, Francisco de Carvalho; http://lattes.cnpq.br/3160458392345919; REZENDE, Cíntia Silva Minafra e; http://lattes.cnpq.br/5841210447886226Item Aplicação da espectroscopia de infravermelho próximo para a determinação do caseinomacropeptídeo em leite UAT(Universidade Federal de Goiás, 2010-02-26) OLIVEIRA, Raphael Rocha de; PRADO, Cristiano Sales; http://lattes.cnpq.br/1661902818272650; OLIVEIRA, Antonio Nonato de; http://lattes.cnpq.br/8261502405023882; LAGE, Moacir Evandro; http://lattes.cnpq.br/2053334263459111This work was carried out to develop a method for the determination of CMP in UHT milk using near-infrared spectroscopy. UHT milk from eight different brands were used for the construction of calibration models. In each of these brands were obtained eight treatments based on the addition of reconstituted whey powder to milk, to 0 (sample with no added whey), 1%, 2%, 3%, 4%, 6% 8% and 10%, giving a total of 64 treatments. The spectra were obtained by NIRS in three replicates, giving a total of 192 spectra. The HPLC results showed a rate of CMP in the samples obtained in the market, ranging between 33.64 and 398.78 mg/L. These results indicate fraud by the addition of whey or low quality of raw milk used. Altogether, 48 models were obtained, showing that the development of a calibration model suitable for the detection of CMP in UHT milk should be used with the PLS method, the method of selecting maximum distance and the pretreatment 2nd derivative and standard normal variate. The near-infrared regions most closely correlated with the vibrational motion of the amino acids present in the CMP were: 1100-1310, 1400-1430, 1490-1550, 1640-1680, 1780-1970, 2020- 2100 and 2310-2350 nm. When these regions were grouped with the selected chemometric tools, have been found the best results from R2 (0,8946) and PRESS (1323569,25), using 6 factors, and eliminating only 1.56% of the spectra (outliers). The results of the samples used for validation, indicates that the addition of whey, or the use of materials of poor quality for the production of UHT milk can be a routine. The results also showed that the calibration model was fine adjusted, as the ratio SEC/SEV was within the range of similarity (0.74). Moreover, the calibration model showed the ability to distinguish, because it recognized the 48 spectra of validation, but did not recognized spectra of four brands of skim milk UHT. However, there is a need for a calibration set wider so that be representative of the entire populationItem Efeitos toxicológicos agudos de extratos de Melia azedarach (MELIACEAE) em ratos (Rattus novergicus), camundongos (Mus musculus) e Artemia salina(Universidade Federal de Goiás, 2010-02-24) PIRES JÚNIOR, Hélio Bernardes; CUNHA, Luiz Carlos da; http://lattes.cnpq.br/6349547031976679; SILVA, Andrea Caetano da; http://lattes.cnpq.br/1432918585717355; BORGES, Lígia Miranda Ferreira; http://lattes.cnpq.br/2591106734434348The revival of research involving botanical insecticides owes itself to the need to make available new products devoid of environmental degradation, with lesser residual affects and development of resistant insects to synthetic products. Phytochemicals from the family Meliaceae have been considered preeminent pesticides. Among the Maliaceae, one of the plants that stands out is Melia azedarach. Diverse studies report the effects of this plant on insects of importance in public health, agriculture and livestock sectors. In spite of M. azedarach being considered a medicinal plant, its use as a natural insecticide needs to be cautious because the plant contains various toxic components to mammals. The objective of this work was to verify the toxicity of extract from unripe fruits of M. azedarach against Wistar rats, Swiss mice and Artemia salina, as well as dermal and ocular irritation in rabbits. Unripe fruits were harvested and processed at the Chemistry Department of UFG. For the acute toxicity test, fruit extracts were administered by gavage at doses of 300 and 2000 mg/kg, in rats and mice of both sexes. For evaluation, animals were observed both for the presence of clinical signs or their death for 14 days after the administration, afterwards all the animals underwent euthanasia and their organs collected for use in histopathology analysis. Data on ration and water consumption, weight gain and weight of the organs were also analysed. Dermal and ocular irritation was evaluated on rabbits using a crude extract on the skin and on the conjunctive sac. The bioassay with A. salina involved eight repeats in the following dilutions, 125, 250, 500 and 1000 μg/ml and an untreated control. The dishes containing the groups were incubated in the dark at 25 ºC and readings taken after 24 hours, by counting the number of surviving individuals. The acute toxicity results obtained demonstrated that the M. azedarach extract did not induce mortality or clinical alteration, just as there was no change in weight gain, ration consumption and organs weight when the treatments and controls were compared. From histopathology analyses, some alterations were detected especially in the liver and lungs, however these lesions were common to the treatments and controls. The assay done in rabbits demonstrated that the extract was not irritative to the skin and eye. The LC50 of the M. azedarach extract was 669 ug/ml, as obtained from the A. salina assay, the extract being considered moderately toxic to this microcrustacean. In spite of these results to have produced evidence that this plant s extract is safe to mammals, further studies such as modes of action of this plant, sub-acute and chronic toxicity assays, effects on specific organisms and on the environment, are necessary towards the definition of this plant as being of low risk to the environment, domestic animals and to man.Item Qualidade do leite produzido no estado de Goiás - ocorrência de resíduos de antimicrobianos e acidez titulável(Universidade Federal de Goiás, 2011-02-21) SILVA, Thaysa dos Santos; OLIVEIRA, Antonio Nonato de; http://lattes.cnpq.br/8261502405023882; LAGE, Moacir Evandro; http://lattes.cnpq.br/2053334263459111; NICOLAU, Edmar Soares; http://lattes.cnpq.br/9601723963736071Milk is considered one of the nutritionally complete foods, consumers and inspection organizations require that this product has innocuousness and good quality. The veterinary medicinal products, in particular antimicrobials have been widely used in stockfarming to treat and preventi diseases (mastitis). However, when they are used in an inappropriate manner, the presence of residues of antimicrobials such as sanitizers, antibiotics or chemotherapic drugs, endanger animals´ and humans´ health, predisposing the population to allergic reactions, carcinogenic activity, microbial resistance, in addition to causing economic losses to industries, producers and governments. Furthermore, the analysis of titratable acidity in milk is one of the parameters used by dairy industry for the indirect determination of its quality, being compulsory in the milk reception platforms the industries. In this study 992 samples (441 of raw, 327 of pasteurized and 224 of Ultra High Temperature (UHT) milk) were collected from April to November 2010 in the properties and places of commercialization, and analyzed in the State of Goias. The commercial kit Delvotest® SP-NP was used to carry out the antimicrobials research, a methodology according the federal law for the titratable acidity, and the software SPSS version 15.0 for data analysis. Out of the analyzed samples 8.47% (5.34% of raw milk, 1.92% of pasteurized milk and 1.21% UHT milk) were positive for the presence of antimicrobials residues. As for titratable acidity, 25.30% of the samples (12.10 % raw milk, 9.57% of pasteurized milk and 3.63% UHT milk) were not within standard values. Among these samples, 10.48% were below the recommendation and 14.82% were above. The analyzes results confirm that the quality of raw, pasteurized and UHT milk produced and consumed in Goias is not within the standard values recommended by federal law, as regards the presence of antimicrobial residue and titratable acidity, which represents risk to public health, besides causing economic losses in milk processing.Item Otimização de ensaios de PCR para a detecção específica de Leishmania chagasi(Universidade Federal de Goiás, 2010-02-24) SILVEIRA NETO, Osvaldo José da; BORGES, Lígia Miranda Ferreira; http://lattes.cnpq.br/2591106734434348; JAYME, Valéria de Sá; http://lattes.cnpq.br/0603234425928309; LINHARES, Guido Fontgalland Coelho; http://lattes.cnpq.br/6261928164195145The visceral leishmaniasis (LV) is a very important disease in public health, including zoonosis caused by members of Leishmania gender, occurring in many regions of the world. Several methods are being used to diagnose these diseases; among these methods are the Enzyme Linked Immuno Sorbent Assay (ELISA), indirect immunofluorescence reaction (RIFI) and direct examination by microscope. As none of these techniques allows a fast and sensitive diagnostic, methods of molecular diagnosis based on polymerase chain reaction (PCR) were developed. Recently, many groups have shown that PCR is a sensitive and specific method to Leshmania DNA detection in a variety of samples of humans, dogs and other animals. The protocols developed until these days to the parasite diagnose by PCR, although they be effective in the identification of the genomic DNA target, promote some inespecific amplifications or are inconclusive in the distinction of L.chagasi from other species of the gender. This work`s objective was the construction of speciesspecific iniciadores to detection of L.chagasi, and evaluation of protocols described by other authors, aiming LV diagnosis. The primers were selected from the alignment of 18S rRNA gene sequences and also of the LSU rRNA. Were selected six new pairs of species-specific primers for L. chagasi. Besides these new pairs were assessed seven other pairs of primers described in the literature. The results showed that the new primer pairs LCS2/LCS3 LCS1/LCS3 were efficient for the species-specific amplification of DNA fragments of L. chagasi of 259 bp and 820 bp, respectively. In the other hand the two new PCR assays optimized in this study using the primer pairs and LCS1/LCS3 LCS2/LCS3 were effective for species-specific detection of up to 1 pg / mL of DNA from L. chagasi. The pairs of primers MC1/MC2 as well as the pair of primers PIA-94 were effective for the discrimination of species-specific L. chagasi.Other pairs of primers evaluated didn´t showed a satisfactory results regarding the specificity for L. chagasi.Item Ocorrência de Prototheca zopfii em propriedades leiteiras do estado de Goiás e avaliação do limite de sensibilidade para o seu isolamento em leite(Universidade Federal de Goiás, 2008-08-05) TUNDELA, Eveline Silva Xavier; MESQUITA, Albenones José de; http://lattes.cnpq.br/3180029815183858; NICOLAU, Edmar Soares; http://lattes.cnpq.br/9601723963736071; OLIVEIRA, Antonio Nonato de; http://lattes.cnpq.br/8261502405023882Mastitis is the most frequent disease in dairy cows, it causes great economic losses in herds and dairy industries and represents a hazard for public health. Among the etiologic agents identified, there is a predominance of bacteria species. However, several other groups of microorganisms may be present, one of those is a unicellular alga of the Prototheca genus, the Prototheca zopfii, an environmental agent of bovine mastitis. The infected cows usually have granulomatous changes in mammary tissue. Moreover microorganisms do not respond to routine therapy, resulting in the elimination of the infected animals. The main purposes of this study was to determine the occurrence of Prototheca zopfii on bulk tank milk in the state of Goiás and identify the limit of detection for Prototheca zopfii by two different approaches. We also wanted to know the samples pre-incubated with small concentration of Protoheca zopfii permits us to obtain positive growth in microbiological culture. We analyzed 473 samples from farms with high somatic cell count patterns but we did not detect Protoheca zopfii in any of those samples. The limit of sensitivity of the Prototheca zopfii was analyzed, in the approach of isolation the limit of detection was between 130 to 3,400 UFC/mL. In the approach of tally the limit was between 25 to 300 UFC/mL. It wasn t observed significant difference (p>0,05) between the approaches analyzed. However, significant statistical difference (p<0,05) occurred between the five types analyzed. After the pre-incubation at 37ºC/24h, the limit of sensibility was analyzed by the approach isolation, the limit was 30 UFC/mL, what means that the pre-incubation is so important to decrease the possibility of occurrence of false negativeItem Inquéritos soroepidemiológicos em eqüinos da região Sul do Brasil para detecção de anticorpos anti-flavivírus de interesse em saúde pública(Universidade Federal de Goiás, 2010-07-02) VIANNA, Ricardo da Silva Teixeira; JAYME, Valéria de Sá; http://lattes.cnpq.br/0603234425928309; BRITO, Wilia Marta Elsner Diederichsen de; http://lattes.cnpq.br/7605775995731168; ANDRADE, Maria Auxiliadora; http://lattes.cnpq.br/9441751521255467The arboviruses are diseases that affect humans, horses and other animal species causing in the majority of cases an asymptomatic infection to neurological disorders. The Flaviviruses are important arbovirus found in Brazil. A descriptive study was conducted from serological surveys in 1775 horses for detection of anti-Flavivirus antibodies in the State of Paraná (Foz do Iguaçú, Maringá and Barracão), Santa Catarina (São Miguel do Oeste, Guaraciaba, Iraceminha, Dionisio Cerqueira, Guarujá do Sul and São José do Cedro) and Rio Grande do Sul (Uruguaiana, São Borja, Itaqui Alegrete and Porto Alegre) in the years of 2007 and 2008. By testing hemagglutination inhibition (HI) were detected HI antibodies of Saint Louis and Ilhéus and other Flaviviruses included in the tests, as well as cross-reactivity for Flaviviruses. By HI test, 14.3% (254/1775) of animals were positive for Flavivirus, monospecific reactions were observed in 42.9% (109/254) serum samples, being that 78.9% (86/109) for St. Louis, 17.4% (19/109) for Ilhéus and 3.7% (4 / 109) for Rocio and cross-reactions were detected in 57.1% (145/254). Among the positives, there was no difference between the sexes. The age group ≥ 10 years old was the most affected with 35.4% (73/206). The animals used for the practice of sport were positive in 34.3% (87/254). The state of Paraná showed 16.3% (107/657) of reacting animals, followed by Rio Grande do Sul with 15.1% (142/939) and Santa Catarina with 2.8% (5/179). This study brings new data regarding the immunity of horses against Flaviviruses in Brazil, and confirms the wide distribution of St. Louis and Ilhéus and the diversity of Flavivirus in the country, as well as the apparent absence of clinical disease in horses infected with the virus studied.