TSST-1, enterotoxin and bacteriocin-like substance production by Staphylococcus aureus isolated from foods / TSST-1, enterotoxinas e substâncias tipo bacteriocina produzidas por Staphylococcus aureus isolados de alimentos

The production of Toxic Shock Syndrome Toxin-1 (TSST-1), enterotoxins and bacteriocin-like substances was evaluated in 95 strains of Staphylococcus aureus recovered from raw bovine milk (n=31) and from food samples involved in staphylococcal food poisoning (n=64). Enterotoxigenicity tests with the membrane over agar associated to optimal sensibility plate assays were performed and showed that 96.77% of strains recovered from milk and 95.31% from food samples produced enterotoxins A, B, C, D or TSST-1. Reference strains S. epidermidis, Bacillus cereus, Listeria monocytogenes, Lactobacillus casei, Pseudomonas aeruginosa, S. aureus, Salmonella Typhimurium, Escherichia coli, Enterococcus faecalis and Bacteroides fragilis were used as indicator bacteria in the antagonistic assays, the first five being sensitive to antagonistic substances. Brain heart infusion agar, in pH values ranging from 5.0 to 7.0 in aerobic atmosphere showed to be the optimum condition for antagonistic activity as evaluated with the best producer strains against the most sensitive indicator bacterium, L. monocytogenes. Sensitivity to enzymes confirmed the proteinaceous nature of these substances. Neither bacteriophage activity nor fatty acids were detected and the antagonistic activity was not due to residual chloroform. Results did not establish a positive correlation between the bacteriocinogenic profile and toxigenicity in the tested S. aureus strains.

Avaliou-se a produção de toxina-1 da síndrome do choque tóxico (TSST-1), enterotoxinas e substâncias antagonistas tipo bacteriocina em 95 amostras de Staphylococcus aureus recuperadas de leite bovino in natura (n=31) e de alimentos envolvidos em surto de intoxicação (n=64). Testes de enterotoxigenicidade pelo método da membrana sobre ágar, associado à técnica da sensibilidade ótima em placa, revelaram que 96,77% das amostras do leite e 95,31% daquelas dos alimentos produziram enterotoxinas estafilocócicas tipos A, B, C, D ou TSST-1. Nos ensaios de antagonismo, foram utilizadas como reveladoras amostras de referência de S. epidermidis, Bacillus cereus, Listeria monocytogenes, Lactobacillus casei, Pseudomonas aeruginosa, S. aureus, Salmonella typhimurium, Escherichia coli, Enterococcus faecalis e Bacteroides fragilis, sendo as cinco primeiras sensíveis às substâncias produzidas. As condições ótimas para a atividade antagonista, avaliadas com as melhores produtoras contra a indicadora mais sensível, L. monocytogenes, foram observadas em aerobiose, em ágar infuso de cérebro-coração, nos valores de pH entre 5,0 e 7,0. A sensibilidade a enzimas confirmou a natureza proteica destas substâncias. Não foram detectadas atividades de bacteriófagos nem de ácidos graxos, e a atividade antagonista não foi devido ao clorofórmio residual. Os resultados não mostraram correlação entre o perfil bacteriocinogênico e a toxigenicidade nas amostras de Staphylococcus testadas.

The use of mice as animal model for testing acute toxicity (LD-50) of toxic shock syndrome toxin / Utilização de camundongos como modelo animal para a verificação da toxicidade aguda da toxina-1 da síndrome do choque tóxico

Acute toxicity test (LD-50) using toxic shock syndrome toxin (TSST-1) was tested in BALB/c, C57BL/6 and Swiss mice. Animals (n = 10) were intraperitoneally injected with TSST-1 (0.01-10.0µg/mouse) followed 4h later by potentiating dose of lipopolysaccharide (75.0µg of LPS - E. coli O111:B4) and cumulative mortality was recorded over 72h. Control animals received either TSST-1 or LPS alone. The data were submitted to qui-Square test and acute toxicity test was calculated by probit analysis (confidence limits expressed as µg toxin/kg). BALB/c mice was the most sensitive (20.0µg/kg, 95 percent confidence limits: 9.0-92.0) followed by C57BL/6 (38.5µg/kg, 95 percent confidence limits: 9.11- 401.6). Data from Swiss mice was not conclusive, indicating only low sensitivity. Selection of the animal model and standardization of the experiment are fundamental for the development of serum neutralization tests used for final quality control of vaccine production.

A toxicidade aguda (DL-50) da toxina da síndrome do choque tóxico (TSST-1) foi testada em linhagens de camundongos BALB/c, C57BL/6 e Suíça. Os animais (n=10) inoculados intraperitoneal com doses crescentes de toxina (0,01 - 10,0µg/animal) receberam 4h após 75µg de LPS (E. coli O111: B4). A toxicidade aguda (DL50) foi observada por um período de 72h e os dados submetidos ao teste de qui- quadrado. Os resultados e os limites de confiança foram expressos em µg de toxina/kg. A linhagem BALB/c apresentou maior sensibilidade (20µg/kg - limite de confiança a 95 por cento entre 9,0- 92,0), seguida da C57BL/6 (38,5µg/kg - limite de confiança a 95 por cento entre 9,11 - 401,6). A amplitude dos limites de confiança deve-se à natureza da toxina, ao mecanismo de ação, a via de inoculação e ao animal utilizado. A seleção do modelo animal e a padronização do experimento são fundamentais para o desenvolvimento de testes de soro neutralização para fins de controle de qualidade do processo de produção de vacinas.

Cinética do fósforo em tecidos de ovinos em crescimento alimentados com diferentes fontes de cálcio / Kinetics of phosphorus in tissues of lambs fed different sources of calcium

Foram utilizados 12 cordeiros machos, inteiros, com peso médio de 31,6kg. Os cordeiros foram alimentados com dietas à base de milho e farelo de soja, suplementadas com diferentes fontes de cálcio: calcário calcítico (CC), feno de alfafa (FA), polpa cítrica (PC) e farinha de conchas de ostras (FCO). Os animais receberam injeção de 7,4 MBq do radiofósforo (32P) e após sete dias foram abatidos para posteriores análises. Utilizou-se o delineamento em blocos ao acaso, com quatro tratamentos e três repetições. A taxa de retenção de 32P, as atividades específicas relativas e padronizadas, e a quantidade de P inorgânico nos tecidos foram iguais para todos os tratamentos (P>0,05). A comparação entre os tecidos mostrou que a concentração de P inorgânico e a taxa de retenção de 32P (P<0,01) foram mais altas nos ossos em relação aos tecidos moles (P<0,01). A retenção de 32P apresentou a seguinte ordem decrescente: rins, fígado, coração, músculos e ossos (P<0,01). A reabsorção óssea de P foi maior que a deposição de P, levando ao balanço negativo do elemento para a maioria dos animais. Concluiu-se que as diferentes fontes de cálcio não interferiram na cinética do P e que a quantidade de P ingerida não atendeu às exigências de P dos animais.

Sixteen growing rams, averaging 31.6kg were studied. They were fed diets based on corn and soya meal, supplemented with different sources of calcium: limestone (CC), alfalfa hay (FA), citrus pulp (PC) and oyster shell meal (OSM). The animals received an injection of 7.4 MBq of radioactive P (32P) and after 7 days they were slaughtered for further analyses. Experimental measurements were analyzed as a completely randomized design with four treatments and three repetitions. The rate of 32P retention, the relative specific activity, the standardized specific activity and the inorganic P in tissue were similar for all treatments (P>0.05). The comparison between tissues showed that the concentration of inorganic P and rate of 32P retention was greater in bone (P<0.01) than in soft tissue. The 32P retention presented the following decreasing order: kidney, liver, heart, muscle and bone (P<0.01). P resorption in bone was higher than P accreted in bone, leading to a negative balance of P in bone for the majority of animals. The means for P accreted in bone were 1.56, 3.29, 2.74 and 2.06g/animal and the means for P resorption in bone were 2.82, 4.51, 4.09 and 3.05g/animal for limestone (CC), alfalfa hay (FA), citrus pulp (PC) and oyster shell meal (OSM), respectively. P resorption values were higher than P accretion in bone leading to a negative balance of P for the majority of animals. It was concluded that the different sources of calcium did not interfere in the kinetics of phosphorus and the amount of P absorbed did not attend P demands of the animals.

Social disability of Brazilian mood disorder patients

Mood disorders cause many social problems, often involving family relationships. Few studies are available in the literature comparing patients with bipolar, unipolar, dysthymic, and double depressive disorders concerning these aspects. In the present study, demographic and disease data were collected using a specifically prepared questionnaire. Social adjustment was assessed using the Disability Adjustment Scale and family relationships were evaluated using the Global Assessment of Relational Functioning Scale. One hundred patients under treatment for at least 6 months were evaluated at the Psychiatric Outpatient Clinic of the Botucatu School of Medicine, UNESP. Most patients were women (82 percent) more than 50 (49 percent) years old with at least two years of follow-up, with little schooling (62 percent had less than 4 years), and of low socioeconomic level. Logistic regression analysis showed that a diagnosis of unipolar disorder (P = 0.003, OR = 0.075, CI = 0.014-0.403) and dysthymia (P = 0.001, OR = 0.040, CI = 0.006-0.275) as well as family relationships (P = 0.002, OR = 0.953, CI = 0914-0.992) played a significant role in social adjustment. Unipolar and dysthymic patients presented better social adjustment than bipolar and double depressive patients (P < 0.001), results that were not due to social class. These patients, treated at a teaching hospital, may represent the severest mood disorder cases. Evaluations were made knowing the diagnosis of the patients, which might also have influenced some of the results. Social disabilities among mood disorder patients are very frequent and intensive.

Relation of cervical length at 22-24 weeks of gestation to demographic characteristics and obstetric history

Preterm delivery is the main cause of neonatal death and ultrasonographic cervical assessment has been shown to be more accurate than digital examination in recognizing a short cervix. This is a cross-sectional study, involving 1131 women at 22-24 weeks of pregnancy, designed to determine the distribution of cervical length and to examine which variables of demographic characteristics and obstetric history increase the risk of a short cervix (15 mm or less). The distribution of maternal demographic and obstetric history characteristics among patients with cervical length ú15 mm was analyzed and compared to the findings for the general population. Risk ratios (RR) between subgroups were generated from this comparison. Median cervical length was 37 mm and in 1.5 percent of cases it was 15 mm or less. The proportion of women with a short cervix (<=15 mm) was significantly higher among patients with a low body mass index (RR = 3.5) and in those with previous fetal losses between 16-23 weeks (RR = 33.1) or spontaneous preterm deliveries between 24-32 weeks (RR = 14.1). We suggest that transvaginal sonographic measurement of cervical length be performed as part of a routine midtrimester ultrasound evaluation. There are specific variables of demographic characteristics and obstetric history which increase the risk of detecting a short cervix at 22-24 weeks.

Staphylococcal food poisoning in Minas Gerais State, Brazil

Foram examinados oito surtos de toxinfecção alimentar em Belo Horizonte, entre novembro de 1991 e outubro de 1992. Em um dos surtos, várias pessoas foram acometidas e dentre estas, 38 foram hospitalizadas. Os alimentos envolvidos foram queijo tipo Minas Frescal, bolo recheado, um prato a base de arroz e outro de carne. A contagem de Staphylococcus variou entre 10 elevado à sexta e 10 elevado à oitava ufc/g de alimento. Estafilococos isolados dos alimentos envolvidos nesses surtos produziram enterotoxina A em um dos surtos, enterotoxinas A e B em quatro surtos, enterotoxinas A, B e C em um surto e enterotoxinas A e C em um surto. Linhagens näo enterotoxigênicas foram isoladas de um dos surtos. A enterotoxina A foi detectada pelo método de ELISA em três dos quatro alimentos envolvidos nos surtos

Protective effect of Saccharomyces boulardii against the cholera toxin in rats

The effect of orogastric administration of Saccharomyces boulardii on the anatomopathological aspect of the jejunal villi was studied in male Fischer rats (weighing about 40 g) orogastrically infected with a culture of Vibrio cholerae. Experimental and control groups received lyophilized S. boulardii (25 mg suspended in 0.5 ml saline) or 0.5 ml saline, respectively, three times a day for 10 days by gastric intubation. On day 5 of treatment, 0.5 ml of a culture of V. cholerae containing 10(8) viable cells was inoculated by gastric intubation into both groups. Histopathological examination of the jejunal mucosa showed extensive lesions of the superficial epithelium of the villi from the control group whereas few lesions of this superficial epithelium were observed in the experimental group. These data show that the inhibition of the action of the cholera toxin on enterocytes by S. boulardii suggested by recent results in vitro can be demonstrated in vivo