Rapd/pcr and phage typing of salmonella enteritidis isolated from poultry and food poisoning outbreaks

ABSTRACT Salmonella Enteritidis (SE) is an important pathogen, causing both food poisoning outbreaks in humans and economic losses to the poultry industry, being also widely spread in the environment. This work aimed to identify SE phage types and to standardize the Random Amplified Polymorphic DNA (RAPD) for evaluating SE isolates obtained from different origins. To do so, 238 SE strains were selected, of which 104 were isolated from broiler carcasses, 106 from food samples and human biological materials involved in food poisoning outbreaks and 28 from different poultry materials. Among these 238 SE isolates, 111 were phage typed, and 57.7% (64/ 111) corresponded to phage type (PT) 4, 32.4% (36/111) to PT 4a, 3.6% (4/111) to PT 6a and 0.9% (1/111) to PT 7, whereas 5.4% .6/111) of the strains were not typeable (RDNC, reacts but does not conform). After the standardization of amplification conditions, all 238 SE isolates were submitted to RAPD/PCR. Among these, 91.8% (217/238) were classified as pattern A. Twenty-one isolates were differentiated into four patterns and into seven subtypes with the use of primer 1254, and into four patterns and ten subtypes using primer OPB 17. The combination of phage typing and RAPD/PCR proved to be a useful tool in epidemiological investigations. RAPD/PCR can be easily used as a routine laboratory method, thus helping with the monitoring of SE isolates and contributing to the establishment of effective Salmonella Enteritidis control and preventive programs.

RESUMO Salmonella Enteritidis (SE) é um patógeno de importância destacada como causa de toxinfecções alimentares em humanos, prejuízos ao setor produtivo e ampla disseminação no ambiente. Este trabalho objetivou identificar fagotipos e padronizar a RAPD/PCR (DNA polimórfico amplificado ao acaso) para a avaliação de isolados de SE. Foram selecionadas 238 amostras, sendo 104 oriundas de carcaças de frango, 106 de alimentos e material biológico de humanos isolados em episódios de toxinfecções alimentares e 28 de materiais de origem avícola. Foram fagotipadas 111 amostras sendo 57,7% (64/111) do fagotipo 4, 32,4% (36/111) fagotipo 4a, 3,6% (4/111) fagotipo 6a e 0,9% (1/111) fagotipo 7, enquanto 5,4% (6/111) não foram fagotipáveis (RDNC, reagent do not conform). Para a RAPD/PCR foram utilizados 238 isolados de SE. Destes, 91,8% (217/238) foram enquadrados no padrão A e 21 isolados (8,8%) foram diferenciados em quatro padrões e sete subtipos com o primer 1254 e em quatro padrões e dez subtipos com o primer OPB 17. A facilidade de execução da RAPD/PCR, após padronizada, habilita a sua implantação em uma rotina laboratorial, auxiliando na monitoria dos isolados de SE e, conseqüentemente, contribuindo para a elaboração de programas efetivos de controle e prevenção de S. Enteritidis.

Litigation in Argentina: challenging the tobacco industry.

OBJECTIVE: To evaluate the processes and outcomes of tobacco litigation in Argentina and to analyse the strategies of the tobacco industry to oppose litigation using tobacco industry documents. METHODS: A systematic search of tobacco industry documents on the internet dating from 1978 to 2002. Law library searches using Argentinean official and unofficial reports systems were combined with computerised online searches. RESULTS: There have been at least 15 failed litigation cases in Argentina and the tobacco industry presented a concerted defence in every claim regardless of cost. We categorised 11 cases as product liability and nicotine addiction, two as health care reimbursement, and two as criminal law and secondhand smoke. Industry strategies included hiring legal consultants from prestigious international and Argentinean law firms and developing litigation prevention programmes. Industry monitored legal academic meetings, controlled the development of new product liability legislation, obtained favourable opinions from experts, and closely observed the development of litigation in Argentina. CONCLUSION: The strategies used by the industry have been successful in preventing recovery for tobacco injuries through litigation. Argentinean health advocates and lawyers need to be aware of the roles and strategies of the tobacco industry in order to develop effective litigation in Argentina.

Polymerase chain reaction (PCR) for the detection of Salmonella in artificially inoculated chicken meat.

The aim of this study was to develop a polymerase chain reaction (PCR) protocol for the detection of Salmonella in artificially contaminated chicken meat. Tests were performed with different dilutions of Salmonella Typhimurium or Salmonella Enteritidis cells (10(-7), 10(-8) or 10(-9) CFU/mL) inoculated in chicken meat samples, in order to establish the limits of detection, incubation times (0, 6, 8 and 24 hours of pre-enrichment in PBW 1%) and three DNA extraction protocols (phenol-chloroform, thermal treatment and thermal treatment and Sephaglass). The assay was able to detect until 10(-9) CFU/mL of initial dilution of Salmonella cells inoculated in chicken meat, which allows detection of Salmonella within 48 hours, including 24 hours of pre-enrichment and using the phenol-chloroform DNA extraction protocol. As the results are obtained in a shorter time period than that of microbiological culture, this procedure will be useful in the methodology for detection of Salmonella in chicken.

Studies on the skeletal cell wall of the cystocarpic stage of the red seaweed Iridaea undulosa B. Part II. Fractionation of the cell wall and methylation analysis of the inner core-fibrillar polysaccharides.

In order to determine the structure of the fibrillar cell wall, the material isolated from cystocarpic thalli of the red seaweed Iridaea undulosa was fractionated using different media. While classical methods produced a scarce solubilization of material, the use of lithium salts in polar aprotic solvents (dimethylsulfoxide or N, N-dimethylacetamide), had successfully extracted higher amounts of material. The final residue from the Li(+)/DMSO extraction contains cellulose and a mannan, while that from the Li(+)/DMAc extraction contains only cellulose and traces of a galactan. Methylation analyses of both residues confirm the presence of those polysaccharides, and shows that the mannan is (1-->4)-linked. Treatment with proteases suggests that the protein is efficiently shielded from digestion. All the extracts and residues contain major amounts of (glyco)proteins and/or proteins, in agreement with a previous suggestion that they are of major importance in the structure of the cell wall.

[Self-medication in a urban population of Cuernavaca, Morelos]. / Automedicación en población urbana de Cuernavaca, Morelos.

This work contains the results of a study on the prevalence of family drug consumption. In our country, self medication should be considered a public health problem due to the population's lack of medical education. Nevertheless, this problem has not been properly studied. For this reason, we completed an exploratory study in an urban population of Cuernavaca, Morelos, México. Its purpose was to gather information about the situation of drug consumption and self medication. A survey was given to 373 domestic groups from the Carolina neighborhood in Cuernavaca. These groups were selected through systematic and probabilistic sampling. A total of 1,537 individuals of all ages were surveyed with an average of four participants per domestic group. Thirty one per cent of those surveyed had consumed drugs during the two weeks prior to the survey. Of these consumers, 53.3 per cent did it through self medication, and 64.9 per cent of them were females. If classified by age group, the highest consumer rate, 7.6 per cent, belonged to the users between 25 to 44 years. If the classification is done by sex and age, women between 25 to 44 years consumed more antibiotics and analgesics which were obtained over the counter. Among the surveyed population, the consumption of medicines was done primarily through self medication, mostly administered by women, and most frequently consumed by infants under one year of age. This phenomenon occurred regardless of the availability and accessibility of public and private health services. The data collected do make us identify women as a fundamental element in the consumption of drugs and self medication.(ABSTRACT TRUNCATED AT 250 WORDS)