Diferentes métodos para aislamiento y detección de Salmonella spp. en canales porcinas / Different methods for isolation and detection of Salmonella spp. in pig carcasses

RESUMEN La Organización Mundial de la Salud (OMS) define salmonelosis como una de las enfermedades transmitida por alimentos (ETA) de mayor casuística, ampliamente extendida en todo el mundo. La enfermedad es producida por Salmonella spp. y causa una de las zoonosis más frecuentes y de mayor impacto económico. El hombre adquiere la infección después de la ingestión de alimentos contaminados, aunque también puede transmitirse de persona a persona o por vía fecal-oral. Actualmente, las técnicas micro-biológicas de aislamiento convencional para detección de Salmonella spp. son establecidas por el Código Alimentario Argentino para verificar la aptitud de un producto para consumo, pero éstas requieren de 4 a 5 días para la obtención de un resultado, tiempo que juega en contra para el productor y la conservación de dichos alimentos. Por este motivo en este trabajo se analizan los métodos de diagnóstico tradicional según Normas ISO 6579:2002 con algunas modificaciones, los métodos de inmunoensayo comerciales y la Reacción en Cadena de la Polimerasa técnica (PCR) de detección del gen invA implicado en el proceso de invasión de cepas patógenas. Se analizaron 60 muestras procedentes de canales porcinas destinadas a comercialización. Se detectó un 10% de Salmonella spp. Se pudo determinar que el diagnóstico molecular por PCR posee alta sensibilidad, pero no es alentador el resultado que reflejan los test comerciales inmunocromatográficos ya que queda en evidencia la necesidad de alta carga microbiana para un diagnóstico certero.

ABSTRACT The World Health Organization (WHO) defines salmonellosis as one of the most important foodborne diseases, widely spread worldwide. The disease is produced by Salmonella spp. and causes one of the most frequent zoonoses and of greater economic impact. The infection is acquired after ingestion of contaminated food, although it can also be transmitted from person to person or by fecal-oral route. Currently, conventional isolation microbiological techniques for detection of Salmonella spp. are established by the Argentine Food Code to verify the suitability of a product for consumption. But microbiological techniques require 4 to 5 days to obtain a result, time that plays against the producer and the conservation of such foods. For this reason in this work we analyze the traditional diagnostic methods according to ISO standards 6579: 2002 with some modifications, the commercial immunoassay methods and the Polymerase Chain Reaction technique (PCR) detection of the invA gene involved in the process of invasion of pathogenic strains. Sixty samples from pigs destined for commercialization were analyzed. 10% of Salmonella spp. was detected. It was possible to determine that the molecular diagnosis by PCR has high sensitivity, but it is not encouraging the result that reflect the commercial immunochromatographic tests since it is evident the need of high microbial load for a correct diagnosis.

Transferência de imunidade passiva em bezerras alimentadas com colostro de vacas com mastite subclínica / Passive immunity transfer in calves fed with colostrum from dams with subclinical mastitis

Este trabalho avaliou a influência da infecção bacteriana da glândula mamária (GM) sobre a transferência de imunidade passiva (TIP) em bezerros recém-nascidos. Vacas holandesas (n=13) foram observadas no momento da parição e ordenhadas para a obtenção de forma asséptica das amostras de colostro (n=52) para os testes microbiológicos. Os recém-nascidos receberam 6 litros de colostro de uma ordenha nas primeiras 12 horas de vida, proveniente de suas respectivas mães. Amostras de sangue foram colhidas antes (D0) e após (D2) o manejo do colostro. A TIP foi avaliada por meio de testes bioquímicos, eletroforese e leucograma. Os bezerros foram distribuídos conforme a ausência (IB-) ou presença (IB+) de infecção mamária em pelo menos uma GM de suas respectivas mães. Todas as amostras de colostro (n=52) foram negativas ao cultivo fúngico. Das 13 fêmeas, 8 (61%) apresentaram crescimento bacteriano em ≥1 quartos mamários. Considerando-se os quartos mamários, foi obtido isolamento bacteriano em 21,15% (11/52), observando-se predomínio de espécies bacterianas do grupo Staphylococcus coagulase negativa. Não foram encontradas diferenças entre os parâmetros de acordo com os grupos experimentais. Em relação aos momentos, foi possível verificar aumento nos valores de proteína total, globulinas, atividade sérica da gama glutamiltransferase e frações eletroforéticas beta e gamaglobulina após a ingestão do colostro materno. A mastite subclínica não influencia a transferência de imunidade passiva em bezerros recém-nascidos da raça holandesa, avaliados por teste bioquímicos, eletroforese e leucograma.(AU)

This study evaluated the influence of intramammary infection under the passive immunity transfer in newborn calves. Holstein cows (n=13) were monitored at delivery, and milked to obtain sterile colostrum samples (n=52) for microbiological examination. The newborn calves received 6 liters of colostrum from their dams up to 12 hours after birth. Blood samples were harvested before (D0) and after (D2) colostrum feeding. The passive immune transfer was evaluated by biochemical tests, electrophoresis and leukogram. Calves were distributed according to absence ­(IB-) or presence (IB+) of mammary gland infection in one or more quarters. All colostrum samples (n=52) were negative for fungal culture. Eight dams (8; 61%) had bacterial growth in one or more mammary quarter. In relation to mammary gland unit, bacterial growth was observed in 21.15% (11/52) of the colostrum samples, with predominance of coagulase negative staphylococcal infections. Statistical differences were not found among variables according to the groups. According to the experimental moments, the evaluated parameters (total protein, albumin, globulins, GGT and beta and alpha electrophoretic) increased after colostrum intake. Subclinical mastitis did not influence the passive immune transfer in Holstein calves evaluated by biochemical tests, electrophoresis or leukogram.(AU)

Transferência de imunidade passiva em bezerras alimentadas com colostro de vacas com mastite subclínica / Passive immunity transfer in calves fed with colostrum from dams with subclinical mastitis

This study evaluated the influence of intramammary infection under the passive immunity transfer in newborn calves. Holstein cows (n=13) were monitored at delivery, and milked to obtain sterile colostrum samples (n=52) for microbiological examination. The newborn calves received 6 liters of colostrum from their dams up to 12 hours after birth. Blood samples were harvested before (D0) and after (D2) colostrum feeding. The passive immune transfer was evaluated by biochemical tests, electrophoresis and leukogram. Calves were distributed according to absence ­(IB-) or presence (IB+) of mammary gland infection in one or more quarters. All colostrum samples (n=52) were negative for fungal culture. Eight dams (8; 61%) had bacterial growth in one or more mammary quarter. In relation to mammary gland unit, bacterial growth was observed in 21.15% (11/52) of the colostrum samples, with predominance of coagulase negative staphylococcal infections. Statistical differences were not found among variables according to the groups. According to the experimental moments, the evaluated parameters (total protein, albumin, globulins, GGT and beta and alpha electrophoretic) increased after colostrum intake. Subclinical mastitis did not influence the passive immune transfer in Holstein calves evaluated by biochemical tests, electrophoresis or leukogram.

Este trabalho avaliou a influência da infecção bacteriana da glândula mamária (GM) sobre a transferência de imunidade passiva (TIP) em bezerros recém-nascidos. Vacas holandesas (n=13) foram observadas no momento da parição e ordenhadas para a obtenção de forma asséptica das amostras de colostro (n=52) para os testes microbiológicos. Os recém-nascidos receberam 6 litros de colostro de uma ordenha nas primeiras 12 horas de vida, proveniente de suas respectivas mães. Amostras de sangue foram colhidas antes (D0) e após (D2) o manejo do colostro. A TIP foi avaliada por meio de testes bioquímicos, eletroforese e leucograma. Os bezerros foram distribuídos conforme a ausência (IB-) ou presença (IB+) de infecção mamária em pelo menos uma GM de suas respectivas mães. Todas as amostras de colostro (n=52) foram negativas ao cultivo fúngico. Das 13 fêmeas, 8 (61%) apresentaram crescimento bacteriano em 1 quartos mamários. Considerando-se os quartos mamários, foi obtido isolamento bacteriano em 21,15% (11/52), observando-se predomínio de espécies bacterianas do grupo Staphylococcus coagulase negativa. Não foram encontradas diferenças entre os parâmetros de acordo com os grupos experimentais. Em relação aos momentos, foi possível verificar aumento nos valores de proteína total, globulinas, atividade sérica da gama glutamiltransferase e frações eletroforéticas beta e gamaglobulina após a ingestão do colostro materno. A mastite subclínica não influencia a transferência de imunidade passiva em bezerros recém-nascidos da raça holandesa, avaliados por teste bioquímicos, eletroforese e leucograma.

Pacientes em diálise peritoneal e em hemodiálise: Existe diferença em relação ao estado nutricional? / Patients in peritoneal dialysis and hemodialysis: Is there any difference in the nutritional status?

A hemodiálise (HD) e a diálise peritoneal (DP) são as terapias renais de substituição mais comumente utilizadas, sendo que o estado nutricional dos pacientes com Doença Renal Crônica (DRC) pode ser influenciado pela modalidade de diálise utilizada. Objetivo: Comparar o estado nutricional de pacientes em HD e em DP, por meio da avaliação antropométrica e bioquímica. Trata-se de um estudo transversal, realizado no setor de nefrologia do Hospital Universitário da Universidade Federal de Juiz de Fora. O estudo foi constituído por 29 pacientes em DP e 36 em HD, totalizando 65 pacientes. Os dados coletados foram: peso corporal seco, estatura, Índice de Massa Corporal (IMC), área muscular do braço (AMB), circunferência muscular do braço (CMB), prega cutânea tricipital (PCT) e circunferência do braço (CB); além dos exames bioquímicos: potássio, fósforo, uréia e albumina. Pode-se observar maior prevalência do sexo feminino na modalidade DP em relação a HD (75% X 44,4%, p = 0,01). Houve diferença nos tempos em diálise entre os grupos (DP: 41,86 ±33,2 meses X HD: 22,36 ±5,4 meses, p = 0,001). Em relação à classificação da CMB, 48,3% dos pacientes em DP e 22,2% em HD foram classificados como eutróficos (p = 0.001). Quanto à AMB, 72,5% dos pacientes em DP e 36,1% dos pacientes em HD foram classificados como eutróficos (p = 0.001). Em relação aos exames bioquímicos, somente o potássio apresentou diferença significante entre os grupos (DP: 4,9 ±0,73 mEq/L e HD: 5,64 ±0,78 mEq/L, p = 0,01). No presente estudo, os pacientes em DP apresentam melhor classificação em relação a CMB e AMB, além de melhor controle do potássio sérico, em relação aos pacientes em HD.

Hemodialysis (HD) and peritoneal dialysis (PD) are the most commonly used renal replacement therapy. The nutritional status of chronic kidney disease (CKD) patients can be influenced by the dialysis modality used. Objective: To compare the nutritional status of patients in HD and PD, through anthropometric and biochemical evaluation. This is a cross-sectional study carried out in the nephrology sector of the University Hospital of the Federal University of Juiz de Fora. The study consisted of 29 patients in PD and 36 in HD, totaling 65 patients. The data collected were: dry body weight, stature, body mass index (BMI), arm muscle area (BMA), arm muscle circumference (BMC), triceps skin fold and arm circumference. In addition, the biochemical tests: potassium, phosphorus, urea and albumin. It is possible to observe a higher prevalence of female sex in the PD modality in relation to HD (75% X 44.4%, p = 0.01). There was a difference in dialysis times between groups (SD: 41.86 ± 33.2 months X HD: 22.36 ± 5.4 months, p = 0.001). Regarding BMC classification, 48.3% of patients in PD and 22.2% in HD were classified as eutrophic (p = 0.001). As for BMA, 72.5% of patients in PD and 36.1% of patients in HD were classified as eutrophic (p = 0.001). In relation to the biochemical tests, only potassium had a significant difference between the groups (SD: 4.9 ± 0.73 mEq / L and HD: 5.64 ± 0.78 mEq / L, p = 0.01). In the present study, patients in PD present better classification in relation to BMC and BMA, in addition to better control of serum potassium, compared to patients in HD.

Comparison phenotypic and genotypic identification of Staphylococcus species isolated from bovine mastitis / Comparação da identificação fenotípica e genotípica de espécies do gênero Staphylococcus isoladas de mastite bovina

In addition to Staphylococcus aureus nowadays other coagulase-positive staphylococci (CoPS) and coagulase-negative staphylococci (CoNS), earlier considered of minor importance, are now accepted as relevant pathogens for humans and animals. The involvement of these microorganisms in bovine mastitis etiology and the possibility their transmission through milk to humans justify the requirement of developing reliable methods for identification of the most frequent species among them. The purpose of this study was to compare the phenotypic techniques with the genotypic method carried out by sequencing of the rpoB gene in identification of several species of the genus Staphylococcus isolated from bovine mastitis. A total of 300 staphylococci isolates of bovine mastitis cases from several Brazilian dairy herds were studied by phenotypic and genotypic techniques, respectively: 150 CoPS and 150 CoNS strains. A total of 18 CoNS different species and 4 CoPS species were identified. Among the CoNS the following species were recognized: 48 (32%) Staphylococcus warneri, 22(15%) S. epidermidis, 20(13%) S. hyicus, 10(7%) S. xylosus, 7(5%) S. haemolyticus, 6(4%) S. simulans, 6(4%) S. schleiferi subsp schleiferi, 6(4%) S. hominis, 5(3%) S. pasteuri, 4(2.7%) S. cohnii, 3(2%) S. saprophyticus subsp. saprophyticus 3(2%) S. chromogenes 3(2%) S. sciuri, 2(1%) S. saccharolyticus, 2(1%) S. lugdunensi, 1(0,7%) S. auricularis, 1(70%) S. saprophyticus subsp. bovis, 1(0.7%) S. capitis. And among the 150 CoPS were identified respectively: 105 (70%) S. aureus, 21(14%), S. hyicus, 19(13%) S. intermedius e 5(3%) S. schleiferi subsp coagulans. Considering the 150 CoNS isolates, the identifications performed by phenotypic and genotypic tests presented 96.7% of concordance, kappa coefficient of agreement = 0.933, SE (standard error) of kappa=0.021 (95% confidence interval: 0.893 to 0.974), Pearson's correlation coefficient (r) = 0.9977, (confidence interval 95%: 0.9938 a 0.9992) and in relation to 150 CPS isolates it was detected an agreement of 98.7%, kappa = 0.960, SE of kappa = 0.016, (95% confidence interval: 0.929 to 0.992) Pearson's correlation coefficient (r) = 0.9994 (95% confidence interval: 0.9681 to 1.0000). The verified agreement strength between the identification methods can be considered as excellent. These results assure that according to laboratory resources any of them will be suitable to perform the staphylococci identification.(AU)

Além de Staphylococcus aureus atualmente outros estafilococos coagulase positiva (SCP) e estafilococos coagulase-negativos (SCN), anteriormente considerados de menor relevância, são reconhecidos como importantes patógenos para humanos e animais. O envolvimento desses micro-organismos na etiologia da mastite bovina e a possibilidade da sua transmissão através do leite aos humanos justifica a utilização de métodos confiáveis para a identificação das espécies mais frequentes. O objetivo deste estudo foi comparar as técnicas fenotípicas com o método genotípico realizada por sequenciamento do gene rpoB na identificação de espécies do gênero Staphylococcus spp. isolados de mastite bovina. Um total de 300 estafilococos isolados de casos de mastite bovina em diferentes rebanhos leiteiros brasileiros foram estudados por técnicas fenotípicas e genotípicas, respectivamente: 150 linhagens de SCP e 150 linhagens de SCN. Foram identificados um total de 18 espécies de SCN e 4 espécies SCP. Entre os SCN as seguintes espécies identificadas: 48 (32%) Staphylococcus warneri, 22 (15%) S. epidermidis, 20 (13%) S. hyicus, 10 (7%) S. xylosus, 7 (5%) S. haemolyticus, 6 (4%) S. simulans, 6 (4%) S. schleiferi subsp schleiferi, 6 (4%) S. hominis, 5 (3%) S. pasteuri, 4 (2,7%) S. cohnii, 3 (2%) S. saprophyticus subsp. saprophyticus, 3 (2%) S. chromogenes, 3 (2%) S. sciuri, 2 (1%) S. saccharolyticus, 2 (1%) S. lugdunensi, 1 (0,7%) S. auricularis, 1 (70 %) S. saprophyticus subsp. bovis, 1 (0,7%) S. capitis. E entre as 150 SCP foram identificados, 105 (70%) S. aureus, 21 (14%), S. hyicus, 19 (13%) S. intermedius e 5 (3%) S. schleiferi subsp coagulans. Considerando-se os 150 SCN isolados, as identificações realizadas por testes fenotípicos e genotípicos apresentaram 96,7% de concordância, coeficiente de concordância kappa = 0,933, SE (erro padrão) de kappa = 0,021 (95% intervalo de confiança: 0,893-0,974), coeficiente de correlação de Pearson (r) = 0,9977, (intervalo de confiança de 95%: 0,9938 a 0,9992) e em relação a 150 SCP isolados foi observado uma concordância de 98,7%, kappa = 0,960, sE de kappa = 0,016, (95% de intervalo de confiança: 0,929 a 0,992) coeficiente de correlação de Pearson (r) = 0,9994 (95% intervalo de confiança: 0,9681-1,0000). A correlação entre os métodos de identificação pode ser considerada como excelente. Esses resultados demonstraram que de acordo com os recursos disponíveis no laboratório, poderia ser utilizada qualquer uma das metodologias.(AU)

Comparative research of blood-gas biochemical analyzer in emergency biochemistry test / 中国医学装备

Objective:To analyze statistically the results of potassium, sodium, chlorine tests by blood-gas biochemical analyzer and automatic biochemical analyzer, in order to have a reference data for blood-gas biochemical analyzer in emergency biochemistry examination.Methods: A total of 101 cases were selected in emergency department. Arterial and venous blood samples of these cases were drawn and tested in electrolyte examination by blood-gas biochemical analyzer and automatic biochemical analyzer. The results of both instruments measured were analyzed statistically.Results: The results of potassium, sodium, chloride ion concentrations tested by blood-gas analyzer ABL80 were lower than the results tested by Beckman AU5800 automatic biochemical analyzer. The correlation analysis were performed in each tested item between the two instruments, and the differences were statistically significant(r=0.997,r=0.992,r=0.989;P<0.05).Conclusion: Although there were differences between two instruments in testing potassium, sodium and chloride ion concentration, the correlation was significant in each test item between the two instruments and the testing speed of blood-gas analyzer ABL80 is faster than the other. Therefore, in order to make quick and accurate clinical judgments about patient's condition, it is worth using blood-gas biochemical analyzer for a preliminary reference in clinical application.

A cost-effective carbohydrate fermentation test for yeast using microtitre plate.

Carbohydrate fermentation test is a well-established technique for speciation of bacteria and fungi. However, long incubation period, cost and procedural complexity may limit its use. Here, we describe a simple modification of conventional carbohydrate fermentation technique using microtitre plate. Thirty-one yeast isolates were identified based on their fermentation property by microtitre plate method and results were compared with conventional method. The modified method had 85.7% sensitivity and 100% specificity. The average time taken to produce positive reaction was 24 hours. When compared with conventional method, modified method reduced turn-around time and cost of processing without significant increase in discordant results.

Early Liver and Kidney Dysfunction Associated with Occupational Exposure to Sub-Threshold Limit Value Levels of Benzene, Toluene, and Xylenes in Unleaded Petrol

BACKGROUND: Unleaded petrol contains significant amounts of monocyclic aromatic hydrocarbons such as benzene, toluene, and xylenes (BTX). Toxic responses following occupational exposure to unleaded petrol have been evaluated only in limited studies. The main purpose of this study was to ascertain whether (or not) exposure to unleaded petrol, under normal working conditions, is associated with any hepatotoxic or nephrotoxic response. METHODS: This was a cross-sectional study in which 200 employees of Shiraz petrol stations with current exposure to unleaded petrol, as well as 200 unexposed employees, were investigated. Atmospheric concentrations of BTX were measured using standard methods. Additionally, urine and fasting blood samples were taken from individuals for urinalysis and routine biochemical tests of kidney and liver function. RESULTS: The geometric means of airborne concentrations of BTX were found to be 0.8 mg m-3, 1.4 mg m-3, and 2.8 mg m-3, respectively. Additionally, means of direct bilirubin, alanine aminotransferase, aspartate aminotransferase, blood urea and plasma creatinine were significantly higher in exposed individuals than in unexposed employees. Conversely, serum albumin, total protein, and serum concentrations of calcium and sodium were significantly lower in petrol station workers than in their unexposed counterparts. CONCLUSION: The average exposure of petrol station workers to BTX did not exceed the current threshold limit values (TLVs) for these chemicals. However, evidence of subtle, subclinical and prepathologic early liver and kidney dysfunction was evident in exposed individuals.

Pruebas bioquímicas tradicionales y de alta resolución para identificación manual de enterobacterias / Traditional and high-resolution biochemical tests for manual identification of enterobacteria / Provas bioquimicas tradicionais e de alta resolução para identificação manual de enterobacterias

Se comparó una serie tradicional de pruebas bioquímicas con una de alta resolución para identificar 500 cepas de enterobacterias utilizando un método probabilístico para interpretar los resultados. La serie tradicional estuvo formada por 10 pruebas (producción de ornitina descarboxilasa, lisina descarboxilasa, lisina desaminasa, ácido sulfhídrico, indol y gas, hidrólisis de urea, utilización de citrato y de malonato, y movilidad). La serie de alta resolución, también con 10 pruebas, se integró con las primeras 4 mencionadas en la serie tradicional y 6 de fermentación de hidratos de carbono (adonitol, L-arabinosa, celobiosa, L-ramnosa, rafinosa y sorbitol). Con la serie de alta resolución se asignaron identidades únicas a 445 cepas (351 con probabilidad de 1,0 y 94 con probabilidades entre 0,010 y 0,999), y de las restantes 55 cepas, a 53 y 2 se asignaron dos y tres identidades probables respectivamente. Con la serie tradicional se asignaron identidades únicas a 306 cepas (110 con probabilidad de 1,0 y 196 con probabilidades entre 0,001 y 0,999) y a 179 y 5 se asignaron dos y tres identidades probables respectivamente. Diez cepas no se pudieron identificar. Todos los indicadores analizados revelaron la superioridad de la serie de alta resolución. El método probabilístico permitió la comparación objetiva de ambas series.

A traditional series of biochemical tests-was compared to a high-resolution one in order to identify 500 strains of enterobacteria, using a probabilistic method for the interpretation of experimental results. The traditional series was formed by 10 tests (ornithine decarboxylase, lysine decarboxylase, lysine deaminase, sulfhydric acid, indol and gas production, urea hydrolysis, citrate and malonate utilization, and motility). The high-resolution one was also formed by 10 tests, including the first 4 tests mentioned above and 6 carbohydrate fermentation tests (adonitol, L-arabinose, cellobiose, L-rhamnose, raffinose, and sorbitol). With the high-resolution series, single identities were assigned to 445 strains (351 with a probability of 1.0 and 94 with probabilities in the range 0.010-0.999), and for the remaining strains two and three probable identities were assigned to 53 and 2 strains, respectively. With the traditional series, single identities were assigned to 306 strains (110 with a probability of 1.0 and 196 with probabilities in the 0.001-0.999 range), two and three probable identities were assigned to 179 and 5 strains respectively; 10 strains turned out to be non-identifiable. Every parameter of comparison used revealed the superiority of the high-resolution series. The probabilistic method for interpretation of experimental results allowed an objective comparison of both series.

Foi comparada uma série tradicional de testes bioquímicos com uma outra de alta resolução para identificar 500 cepas de enterobactérias usando uma abordagem probabilística para a interpretação dos resultados. A série tradicional consistiu em 10 testes (produção de ornitina descarboxilase, lisina descarboxilase, lisina desaminase, ácido sulfídrico, indol e gás, hidrólise da ureia, utilização de citrato e de malonato, e mobilidade). A série de alta resolução, também com 10 ensaios, integrou-se com as primeiras 4 mencionadas na série tradicional e 6 de fermentação de hidratos de carbono (adonitol, L-arabinose, celobiose, L-ramnose, rafinose e sorbitol). Com a série de alta resolução foram atribuídas identidades únicas a 445 cepas (351, com probabilidade 1,0, e 94, com probabilidade entre 0,010 e 0,999), e das restantes 55 cepas, a 53 e 2 foram atribuídas duas e três identidades possíveis, respectivamente. Com a série tradicional foram atribuídas identidades únicas a 306 cepas (110 com probabilidade de 1,0 e 196 com probabilidades entre 0,001 e 0,999) e a 179 e 5 foram atribuídas duas e três identidades prováveis, respectivamente. Dez cepas não puderam ser identificadas. Todos os indicadores analisados demonstraram a superioridade da série de alta resolução. O método probabilístico permitiu a comparação objetiva de ambas as séries.

Application of evidence-based laboratory medicine in quality control of biochemical tests in Chinese peacekeeping secondary hospital of United Nations Interim Force in Lebanon / 国际检验医学杂志

Objective To seek a operation method of quality control of biochemical tests in peacekeeping mission via perspective of evidence-based laboratory medicine .Methods Literatures related to quality control of biochemical tests in international peace-keeping were referred ,compared and discriminated .Combined with 5-year peacekeeping laboratory experience ,a set of feasible ap-proaches were worked out and a practice evaluation was performed in a task period .Results Under the supervision of the quality control system ,providing accurate and reliable test results to the clinical ,developing and implementing quality control of biochemical tests were economical ,practical and reliable ,and could guarantee the effectiveness of reports of biochemical test in peacekeeping medical institutions .Conclusion Evidence-based laboratory medicine is important for quality control of biochemical tests in peace-keeping medical institutions .

Suplementação com creatina associada ao treinamento resistido não altera as funções renal e hepática / Creatine supplementation associated with resistance training does not alter renal and hepatic functions

A creatina é o suplemento nutricional mais popular utilizado para melhorar o desempenho em atividades que envolvem exercícios de curta duração e alta intensidade. Porém, as possíveis intercorrências advindas do seu uso não estão totalmente elucidadas. O objetivo do estudo foi avaliar os efeitos de duas dosagens de suplementação com creatina nas funções renal e hepática de adultos saudáveis durante oito semanas de treinamento de musculação. Exames bioquímicos foram realizados em 35 praticantes de musculação distribuídos aleatoriamente em três grupos, placebo (PLA, n = 12), creatina (CRE1, n = 12) e creatina 2 (CRE2, n = 11), antes e após oito semanas de treinamento com exercícios resistidos. Em desenho duplo-cego, os voluntários foram suplementados (20g/dia) com creatina (CRE1 e CRE2) ou placebo (PLA) por sete dias e nos 53 dias subsequentes com 0,03g/kg de massa corporal de creatina (CRE1) e placebo (PLA) e com 5g/dia o grupo CRE2. Não houve intervenção na composição de suas dietas, que foram registradas e analisadas. Os resultados dos exames bioquímicos realizados permaneceram dentro das faixas de normalidade. Os valores de creatinina aumentaram 12,2 por cento no grupo CRE1 e 9,0 por cento, no CRE2, enquanto que no grupo PLA diminuiu 4,7 por cento, entretanto, esses valores não ultrapassaram os índices de normalidade. Os valores dos exames da função hepática diminuíram em quase todas as frações, em todos os tratamentos, contudo, sem significância estatística. Conclui-se que a suplementação com creatina nas dosagens utilizadas (0,03g/kg e 5g/dia) para indivíduos saudáveis por oito semanas não altera a função hepática ou renal, sendo assim, nas condições deste estudo, foi considerada segura.

Creatine is the most popular nutritional supplement widely used to improve performance in activities that involve exercise of short duration and high intensity. However, the complications arising from its use are not fully elucidated. The aim of this study was to evaluate the effects of two doses of creatine supplementation on renal and hepatic function in healthy adults during eight weeks of resistance exercise training. Biochemical tests were performed on 35 athletes randomly distributed into three groups, placebo (PLA, n = 12), creatine (CRE1, n = 12) and creatine2 (CRE2, n = 11) before and after eight weeks of resistance training. In a double-blind design, the volunteers were supplemented (20 g/day) with creatine (CRE1, CRE2) or placebo (PLA) for seven days and at the 53 subsequent days with 0.03g/kg of body weight of each supplement (CRE1, PLA) and 5g/day for CRE2. There was no intervention in the composition of their diets, which were recorded and analyzed. The results of biochemical tests conducted remained within normal ranges. Creatinine values increased by 12.2 percent for CRE1 and 9.0 percent for CRE2, whereas decreased by 4.7 percent in PLA; however, these values did not exceed normal rates. The values of liver function tests declined in nearly all fractions in all treatments, not being statistically significant, though. It is concluded that creatine supplementation at the dosages used (0.03g/kg and 5g/day) for healthy subjects during eight weeks does not alter hepatic or renal function, hence under the conditions of this study, creatine was considered safe.

Isolation, characterization and identification of bacteria from coolant oils.

Coolants are hydrocarbons, used to lubricate parts of machines for smooth performance. While in use, a coolant quickly gets contaminated with foreign materials, making it less effective and unpleasant odors are developed due to microbial action. Hence coolants need to be replaced frequently. The expense of disposing used coolants and replacing it with fresh coolants adds significantly to the manufacturing cost. The present study is focused on isolation, identification and characterization of coolant oil contaminating bacteria as an initial step to solve these contamination problems. Used and unused samples of coolant were collected from oil stations, auto mechanic workshops and steel industry for the isolation of the contaminants. Ten dominant bacterial isolates of the genus Pseudomonas, Staphylococcus, Micrococcus, Salmonella, Cellobiococcus and Pneumonia were identified by morphology, biochemical tests and PIB tool. Isolates were subjected to four different media, various pH and temperatures for characterization of optimal conditions of growth. Pseudomonas pseudomallei, Micrococcus luteus 3, Micrococcus varians and Salmonella ferlac were observed in mineral, synthetic and aerobic media, Staphylococcus hyicus, Cellobiococcus species and Staphylococcus intermedius in synthetic and aerobic fermentation media and Pseudomonas cepacia, Pseudomonas piketti in mineral salt and aerobic fermentation media. The ten isolates showed optimal growth at different temperatures between 20°C and 90°C and different pH, ranging from acidic to alkaline. In conclusion, the used coolants harbor hazardous pathogens such as Pseudomonas species which multiply rapidly and survive high temperatures. These isolates could be targeted for further studies on development of antidotes as a solution to the coolant contamination problems.

Perfil de adolescentes usuários de crack e suas consequências metabólicas / Profile of teenage crack users and metabolic consequences

Introdução: O crack, droga derivada da cocaína, está se disseminando entre os jovens de diferentes classes sociais do Brasil. O objetivo do estudo foi caracterizar o perfil dos usuários de crack e verificar suas consequências metabólicas. Métodos: Estudo descritivo exploratório, do tipo quantitativo. Aplicaram-se questionários e analisaram-se os exames bioquímicos dos participantes através de prontuários. Foram entrevistados 22 adolescentes internados para tratamento de desintoxicação. Resultados: O perfil do usuário de crack foi jovem, nível de escolaridade baixo e sem emprego formal. O crack foi eleito a droga de preferência pela maioria dos pacientes (86,3%), seguida do pitico e da maconha. A maioria (95,4%) relatou mudanças quanto aos hábitos alimentares. A forma em que o crack era mais utilizado foi em cachimbos (50%). Em relação às alterações metabólicas, foi observado apenas o aumento na fosfatase alcalina, que pode indicar dano hepático. Os outros exames não apresentaram alterações significativas. Conclusão: O presente estudo demonstra que o crack causa prejuízos sociais, econômicos, além de danos à saúde do dependente, como alterações no estado nutricional pela inapetência e possíveis danos hepatotóxicos. Portanto, observa-se que esses sujeitos estão expostos a diversas situações de risco e vulnerabilidade, o que indica grave problema de saúde pública.

Introduction: Crack, a drug derived from cocaine, is spreading among young people from different social classes in Brazil. The aim of this study was to characterize the profile of crack users and assess its metabolic consequences. Methods: A descriptive exploratory study of the quantitative type. Questionnaires were administered and the participants’ biochemical tests in medical records were analyzed. We interviewed 22 adolescents admitted for detoxification treatment. Results: The profile of the crack user was young, low education level and without formal occupation. Crack was the most used drug by most patients (86.3%), followed by pitico and marijuana. The majority (95.4%) reported changes regarding eating habits. The most common way of consuming crack was through pipes (50%). Concerning metabolic changes, only an increase in alkaline phosphatase was detected, which may indicate liver damage. The other tests did not show significant changes. Conclusion: This study demonstrates that crack has adverse social and economic consequences and causes damage to the health of the user, such as changes in nutritional status and appetite for possible hepatotoxic damage. Therefore, it is observed that these subjects are exposed to various situations of risk and vulnerability.

Clostridium perfringens isolate typing by multiplex PCR

Clostridium perfringens is an important pathogen that provokes numerous different diseases. This bacterium is classified into five different types, each of which capable of causing a different disease. There are various methods for the bacterial identification, many are labor-intensive, time-consuming, expensive and also present low sensitivity and specificity. The aim of this research was to identify the different types of C. perfringens using PCR molecular method. In this study, 130 sheep-dung samples were randomly collected from areas around the city of Kerman, southeastern Iran. After processing and culturing of samples, the produced colonies were morphologically studied, gram stain test was also carried out and the genera of these bacteria were identified through biochemical tests. DNA extracted from isolated bacteria for genotyping was tested by multiplex PCR with specific primers. Based on length of synthesized fragments by PCR, toxin types and bacterial strains were detected. C. perfringens isolated types were divided as follows: 17.39 percent type A, 21.74 percent type B, 34.78 percent type C and 26.09 percent type D. It should be emphasized that, up to the present moment, C. perfringens type A has not been reported in Iran.(AU)

Evaluation of a simplified key for the identification of coagulase-positive Staphylococcus isolated from bovine mastitis / Avaliação de uma chave de identificação simplificada para Staphylococcus coagulase-positivos isolados de mastite bovina

Evaluation of a simplified key for the identification of coagulase-positive Staphylococcus isolated from bovine mastitis. Three hundred fourty four strains of coagulase-positive Staphylococcus (CPS), isolated from mastitis cases, underwent phenotypic and genotypic tests to evaluate the efficiency of a simplified key, based on phenotypic tests for the discrimination of these microorganisms. The tests consisted of amplification of the femA gene and hemolysis in blood agar, production of acetoin and fermentation of maltose, mannitol and trehalose. Strains that showed negative results in the amplification test of the femA gene or that were not identified as Staphylococcus aureus (S. aureus) by phenotypic tests were tested with the APISTAPH kit (Biomériux-France), for precise identification of species. Phenotypic tests revealed 338 strains (98.25%) as S. aureus, three strains (0.86%) as Staphylococcus hyicus, and three microorganisms (0.86%) as Staphylococcus intermedius. PCR demonstrated that 338 (98.25%) strains belonged to the S. aureus species, confirming the results for 336 strains from 338 identified, through a simplified phenotypic key. A high rate of correlation (98.83%) was verifeid between the results of genotypic and phenotypic tests for the identification of S. aureus, demonstrating the applicability of the proposed key, for the discrimination of this microorganism in CPS isolated from bovine mastitis.

Visando testar a eficiência de uma chave simplificada baseada em testes fenotípicos para a discriminação de Staphylocococcus coagulase positivos (SCP) isolados de infecções intramamárias de bovinos, 344 amostras destes microrganismos foram submetidas a testes fenotípicos e genotípicos. Estes consistiram na amplificação do gene femA, na observação de hemólise em ágar sangue, produção de acetoína e fermentação de maltose, manitol e trealose. Amostras que apresentaram resultado negativo na amplificação do gene femA ou que foram identificadas com não Staphylococcus aureus (S. aureus) por meio dos testes fenotípicos foram submetidas ao kit APISTAPH (Biomériux-França) para identificação mais precisa. Os testes fenotípicos utilizados na chave simplificada permitiram identificar 338 amostras (98,25%) como S. aureus, três amostras (0,86%) como Staphylococcus hyicus e três (0,86%) como Staphylococcus intermedius. Por meio da reação em cadeia da polimerase (PCR) 338 (98,25%) amostras foram identificadas como S. aureus, ratificando os resultados para 336 das 338 amostras identificadas por meio da chave fenotípica simplificada. Observou-se elevada concordância (98,83%) entre os resultados dos testes genotípicos e fenotípicos para a identificação de S. aureus, demonstrando a aplicabilidade da chave de identificação proposta para a discriminação deste microrganismo entre SCP isolados de casos de mastite bovina.

Identificación de micobacterias no tuberculosas: comparación de métodos bioquímicos y moleculares / Identification of non-tuberculosis mycobacteria: comparison between biochemical and molecular methods

Las infecciones causadas por micobacterias no tuberculosas (MNT) o atípicas constituyen en la actualidad un grave problema de salud, especialmente en pacientes inmunocomprometidos. Estas micobacterias presentan patrones de susceptibilidad a antibióticos particulares y distintos a M. tuberculosis, por lo que la administración del tratamiento adecuado requiere de un método rápido, sencillo y sensible de identificación. La técnica de PRA (Análisis de Restricción de Productos de PCR), basada en la digestión enzimática del producto de amplificación del gen hsp65, ha mostrado ser un método adecuado de identificación de micobacterias. En el presente trabajo se comparó la técnica de PRA con el estándar de identificación de micobacterias representado por las pruebas bioquímicas en 30 aislados provenientes del Laboratorio de Tuberculosis del Instituto de Biomedicina. La técnica de PRA permitió identificar 96% de las cepas analizadas, en comparación con 92.% de cepas identificadas por las técnicas bioquímicas. Los resultados obtenidos fueron idénticos en 18 de 22 cepas, correspondiendo al 82% de los resultados. Se concluye que el PRA es un método rápido, sencillo y económico que produce resultados concordantes con las técnicas tradicionales, con un menor grado de error. Basados en estos resultados se recomienda el uso del PRA en los laboratorios clínicos como método de identificación de rutina para micobacterias.

Infections caused by atypical mycobacteria at present constitute a serious health problem, especially in immunocompromised patients. These mycobacteria present particular susceptibility patterns, different from M. tuberculosis, due to which the administration of an adequate treatment requires a fast, simple and sensitive identification method. The PRA technique (PCR Restriction), based on the enzymatic digestion of the amplification product of the hsp65 gene has shown to be an adequate method for the identification of mycobacteria. In this study we compared the PRA technique with the standard mycobacterial identification method, represented by biochemical tests, in 30 isolates from the Tuberculosis Laboratory of the Instituto de Biomedicina. The PRA technique allowed the identification of 96% of the strains analyzed, as compared with 92% of strains identified through biochemical methods. The results obtained were identical in 18 of 22 strains, corresponding to 82% of the results. It is concluded that the PRA technique is a fast, simple and economical method that produces results in concord with traditional techniques, with a lesser degree of error. Based in these results, the use of PRA as routine identification technique for mycobacteria is recommended for clinical laboratories.

Perfil hematológico, bioquímico sérico e sorológico de Felis domesticus com lagochilascariose experimental / Hematological, serum biochemical and serological profile of Felis domesticus with experimental lagochilascariosis

No presente trabalho, avaliou-se o hemograma, diversas proteínas e enzimas séricas ou plasmáticas e a produção de anticorpos específicos em Felis domesticus, experimentalmente infectados por Lagochilascaris minor. Verificou-se nos animais infectados aumento de leucócitos totais, principalmente eosinófilos; queda do número de plaquetas; aumento de aspartato-aminotransferase e alanina-aminotransferase; e principalmente a presença de anticorpos IgG específicos para antígenos do parasita. A reação com extrato bruto de parasitas adultos mostrou-se mais específica, permitindo a discriminação de soros de animais: não infectados, com infecção por outros parasitas, e com lagochilascariose. Esta é a primeira descrição da padronização de uma reação sorológica para diagnóstico da lagochilascariose em Felis domesticus.

The present study evaluated the hemogram, different proteins, plasma enzymes, serum enzymes and specific antibody production of Felis domesticus experimentally infected by Lagochilascaris minor. The infected animals were seen to present increased total leukocytes (particularly eosinophils), decreased platelet counts, increased aspartate-aminotransferase and alanine-aminotransferase and, especially, the presence of specific IgG antibodies against antigens of the parasite. The reaction with crude extract of adult parasites was shown to be more specific, thereby enabling serum discrimination between the animals: non-infected, infected with other parasites and infected with lagochilascariosis. This is the first description of the standardization of a serological reaction for diagnosing lagochilascariosis in Felis domesticus.

A METHOD FOR BLOOD EXAMINATIONS IN OVERSEAS MOBILE CLINICS: CLINICAL APPLICATION IN THE RURAL AREAS OF LAO PDR

We conducted an overseas mobile clinic in Lao PDR and examined the results of biochemical tests and blood examinations. For the biochemical tests, we used a method involving plasma skimming film and filter paper, called “plasma separation plate.” In this study, we report the usefulness of the plasma separation plate, which is a newly developed method for the screening of biochemical tests in Japan. We were able to apply this method in a mobile clinic in Lao PDR. The quantitative concentration of plasma from blood collected from pricked fingers was measured by this method. Using the plasma skimming film and filter paper, the non-cell components in the blood were separated, and a quantity of the plasma was adsorbed by filter paper. Neither elaborate equipment nor electricity was required. The results of our test data indicated that few inhabitants suffered from adult diseases for which we were screening. We concluded, therefore, that this method is suitable for use in mobile clinics in the rural areas of developing countries.

Antropometric and Health Status of the Elderly Women Attending a Health Promotion Program in an Urban Community / 대한지역사회영양학회지

This study was conducted to evaluate the health status of elderly women who attended in a Health Promotion Program of the Seo-gu Health Center in Daegu. The study subjects were 158 elderly women (over 65 years) in an urban community. The subjects were investigated by means of individual interviews using a questionnaire, Blood tests for analyzing their biochemical status were carried out. The average age of the study subjects was 70.9 +/- 2.3 years. Of the subject group 79.1% ranged in age from 65 to 74 year and 20.9% were over 75 years. With respect to health related factors, 23.4% of subjects drank alcoholic beverages and 15.2% of sujects currently smoker. The prevalence with chronic diseases was 51.9%, and 26.6% of the subjects were healthy. The diseases most frequently reported as having been or being treated were arthritis (38.1%), hypertension (21.4%), and diabetes (17.9%). The average height of subjects was below the standard established in the Korean Recommended Dietary Allowances, while the average weight was close to the standard. The means of systolic and diastolic blood pressure were 130.2 mmHg and 71.9 mmHg, respectively. The mean serum albumin level was 4.5 mg/dl, and the value of hemoglobin and hematocrit was 12.5 mg/dl and 0.7%, respectively. The mean serum cholesterol level was 207.1 mg/dl and the mean triglyceride level was 187.7 mg/dl. The serum lipid levels were higher then in those reported in research. The serum cholesterol levels of 71.5%of subjects were within the normal range. The GOT and GTP levels were within the moderate range. In conclusion, the health status of the elderly who attended the Health Promotion Program in the Seo-gu Health Center were very average. However, it was necessary to prepare a health management program to deal with the serum lipids so as to establish and maintain good health. When we carry out the health promotion program in a community, individual program of adequate to health status should be developed more.

Streptococcus mutans isolates identified by colony morphology,biochemical tests and DNA G+C mol% / 实用口腔医学杂志

砄bjective:To investigate the consistency of Streptococcus mutans and Streptococcus sobrinus isolates identified by colony morphology,biochemical tests and DNA base content analysis.Methods:46 clinical isolates (30 of S.mutans and 16 of S.sobrinus ) were identified by colony morphology,biochemical tests and determination of DNA base contents with reversed phase high performance liquid chromatography. S.mutans Ingbritt and S.sobrinus 6715 were used as the controls. Results:The DNA contents (G+C mol%) of S.mutans and S.sobrinus clinical isolates, identified by colony morphdogy and biochemical test, were 37.52 and 45.19 respectively,consistent with the expected 36～38 and 44～46 mol percent of G+C.Conclusion: S.mutans and S.sobrinus isolates,identified by colony morphology and biochemical tests ,were consistent with the DNA G+C content analysis. S.mutans and S.sobrinus can be easily and reliably identified by colony morphology and biochemical tests.