Dental caries are more likely to impact on children's quality of life than malocclusion or traumatic dental injuries.

AIM: To measure the impact of dental caries, malocclusion, and traumatic dental injuries (TDI) on the oral health-related quality of life (OHRQoL) of Brazilian children. STUDY DESIGN: population-based cross-sectional study. MATERIALS AND METHODS A representative sample of 1,204 8-to-10-year-old children randomly selected from 19 public and private schools in Belo Horizonte (Brazil) was evaluated. The Decayed, Missing and Filled Teeth Index (DMFT), the Dental Aesthetic Index (DAI), and Andreasen's classification were used by two calibrated examiners to diagnose dental caries, malocclusion, and TDI, respectively. Children were clinically examined at school. The Brazilian version of the Child Perceptions Questionnaire for ages 8 to 10 years (CPQ8-10) was used to assess the impact on OHRQoL. RESULTS: There were statistically significant differences (p<0.001) between groups (dental caries, malocclusion, and TDI) in all subscales and the CPQ8-10 total score. The presence of dental caries alone and its association with TDI and malocclusion were associated with all CPQ8-10 subscales (p<0.05). STATISTICS: the Poisson regression model with a robust variance estimator was utilised for the multivariate analysis. Adjusted prevalence ratios were obtained for the association between oral conditions and the total score on the CPQ8-10 and its subscales. CONCLUSION: Dental caries seems to be the oral condition most commonly associated with a higher impact on the OHRQoL of Brazilian 8-to-10-year-olds.

Toxicological in vitro and subchronic evaluation of LASSBio-596.

LASSBio-596, 2-[4-(1,4-tiazinan-4-ylsulfonyl) phenylcarbamoyl] benzoic acid, is an achiral compound containing a subunit carboxylic amide, was capable of preventing induced mechanical and morphological changes in the lungs that commonly caused the onset of asthma. Previous studies to determine the acute toxicity of oral LASSBio-596 at dose of 2000mg/kg caused no deaths in any of the tested animals. To further evaluate the safety of LASSBio-596, in vitro and in vivo tests were carried out. Regarding to in vitro test were used renal, hepatic, pulmonary, cardiac, neurologic and intestinal cell lines. They were evaluated using neutral red (NR) and [3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] (MTT) assays. Micronuclei also was performed. Concerning to in vivo was performed subchronic on Wistar rats at doses of 10, 50, and 250mg/kg and zebrafish test. The in vitro tests results showed the safety of LASSBio-596. However, subchronic toxicity study results revealed changes in the blood parameters of amylase, alanine aminotransferase (ALT), aspartate aminotransferase (AST), glucose and creatine kinase (CK) which is used for cardiotoxicity evaluation, although, did not identify any histopathological alterations. However, zebrafish test demonstrated cardiac damage. It was impossible to estimate the no-observed-adverse-effect-levels and lowest observed-adverse-effect level due to the presence of cardiotoxicity in all tested doses.

Factors associated with dental caries in Brazilian children: a multilevel approach.

OBJECTIVES: The aims of the present study were to assess caries experience among Brazilian children aged eight to 10 years and determine social factors of this disease, through a multilevel approach. METHODS: This is a cross-sectional study in which a representative sample of 1204 eight-to-ten-year-old children were randomly selected from 19 public and private schools in the city of Belo Horizonte, Brazil. After formal authorization from the parents, the children were clinically examined at school by two calibrated examiners (Kappa=0.78). Individual socioeconomic status was determined using household income, number of residents in the home and parents'/caregivers' schooling. The social vulnerability index and type of school were used as contextual variables. The outcome variables were severity score of dental caries in permanent teeth (DMFT index), dental caries experience in permanent teeth (DMFT = 0 or ≠ 0), severity score of dental caries in primary teeth (dmft index) and dental caries experience in primary teeth (dmft = 0 or ≠ 0). RESULTS: The minority of children (37.2%) had dental caries experience. Chi-squared and Mann-Whitney tests showed that age, parents'/caregivers' schooling, household income and type of school were associated with dental caries experience in permanent and primary teeth (P < 0.05). Mann-Whitney, Kruskal-Wallis tests and Spearman correlation coefficient showed that age, parents'/caregivers' schooling, household income, and type of school were associated with severity score of dental caries in permanent and primary teeth (P < 0.05). In multilevel approach, the individual variables, age and household income demonstrated association with dental caries experience and with severity score of dental caries in primary teeth (P < 0.05). Type of school was also associated with dental caries experience and with severity score of dental caries in these teeth (P < 0.05). In the Poisson multilevel approach, the analysis of the final estimation of variance components of the null model revealed a significant difference among the 19 schools with regard to severity score of dental caries in permanent teeth too (P < 0.001). Younger children and attending private schools showed lower severity score of dental caries in permanent teeth. CONCLUSION: Dental caries experience and severity score of dental caries in primary teeth was influenced by age, household income, and the type of school The dental caries experience in permanent teeth was influenced by age, parents'/caregivers' schooling, household income, and type of school, whereas the severity score of dental caries in permanent teeth was influenced by age and type of school.

High-throughput multiclass screening method for antibiotic residue analysis in meat using liquid chromatography-tandem mass spectrometry: a novel minimum sample preparation procedure.

A multiresidue and multiclass method based on liquid chromatography-tandem mass spectrometry for the determination of antibacterials was developed and validated for screening purposes. This method can be applied to commonly used drugs in veterinary medicine such as tetracyclines, quinolones and sulfonamides. Sample preparation consists in cell disruption with sand (previously purified and washed with EDTA 100 mM) followed by protein precipitation with acidified acetonitrile. Validation was conducted in accordance to European Union requirements (2002/657/EC) for qualitative methods covering detection capability (CCß), selectivity, specificity and stability. The method enabled the detection of 21 different drugs and had a false-compliant rate of <5% (ß error) at between 25% and 50% of the maximum residue levels established by legal authorities. The methodology was successfully applied to incurred poultry samples.

Campomanesia lineatifolia Ruiz & Pav. as a gastroprotective agent.

ETHNOPHARMACOLOGICAL RELEVANCE: Campomanesia lineatifolia Ruiz and Pav. (Myrtaceae) is a native edible species found in the Amazon Rainforest, commonly known as gabiroba. In Brazil, Campomanesia species are frequently used in traditional medicine for gastrointestinal disorders. MATERIALS AND METHODS: The present study performed phytochemical analyses and determined both the in vitro antioxidant activity of the ethanolic extract of Campomanesia lineatifolia leaves (EEC) as well as its ethyl acetate fraction (EAFC). In this analysis, quercetin was used as a positive control. Gastroprotective activity was also investigated at different oral doses in two experimental models in rats - gastric lesion induced by ethanol and gastric lesion induced by indomethacin. In this analysis, cimetidine and sucralfate were used as positive controls. The area of gastric lesion underwent macroscopic and histomorphometric evaluations, while the mucus content was estimated by applying the periodic acid-Schiff stain. Oral acute toxicity was also assessed. RESULTS: Phytochemical studies revealed the presence of flavonoids and tannins. Catechin and quercitrin were isolated by bioguided chromatographic fractionation of EAFC. EEC and EAFC presented in vitro antioxidant activity. The oral administration of EEC and EAFC at doses 100-400 mg/kg (ethanol model) and at doses of 400-1200 mg/kg (indomethacin model) proved to be effective in preventing gastric ulcerations in rats. Pretreatment with EAFC (400mg/kg) significantly increased the gastric mucus content in the ethanol model. No animals died during the acute oral toxicology test. CONCLUSIONS: Results confirm the Brazilian ethnopharmacological use of Campomanesia lineatifolia as a gastroprotective agent and indicate that the anti-ulcer effect is most likely mediated by scavenging free radicals due to the polyphenol content and, at least in part, by increasing the mucus secretion and the mucosal defense. In addition, EEC and EAFC were found to be safe when applied to a 2000 mg/kg single oral dose.

Lavado traqueobrônquico por via nasotraqueal como metodologia de colheita de células do trato respiratório de ovinos sadios e portadores de afecções pulmonares / Tracheobronchial lavage by nasotracheal via to collect cells of respiratory tract of health and broncopneumonic ovines

Os estudos das secreções traqueobrônquicas são amplamente utilizados nas pesquisas de doenças pulmonares nas diversas espécies animais, inclusive no homem. Os objetivos desta pesquisa foram a viabilização da técnica de colheita de lavado traqueobrônquico na espécie ovina e o estudo da relação clínico-citológica do lavado de ovinos portadores de broncopneumonia e sadios. Foram utilizados 33 ovinos, 18 sadios e 15 portadores de enfermidade respiratória com sinais clínicos de envolvimento das vias aéreas, divididos nos respectivos grupos, GS e GD. Após o exame físico foi realizado o lavado traqueobrônquico por via nasotraqueal. A colheita do lavado foi feita com a inoculação e aspiração de solução fisiológica estéril. As amostras foram processadas citologicamente através de citocentrifugação e coradas pelos métodos Wright-Giemsa e Shorr. Tanto a contagem total de células epiteliais quanto o número de hemácias por mililitro foi maior no grupo de animais com broncopneumonia. Nos animais sadios notou-se predomínio de macrófagos, seguido por células epiteliais cilíndricas, neutrófilos e linfócitos. No grupo de animais doentes havia menor número de macrófagos, e predomínio da população de neutrófilos. Por ser de fácil realização, pouco dispendiosa e pela obtenção representativa de material, a técnica estudada mostrou-se eficaz na obtenção de fluidos traqueobrônquicos e, portanto um bom método de colheita de células para uso nas pesquisas de vias aéreas.

Exams of tracheobronchial secretions are widely used in research of lung diseases in several animal species, including men. The objective of this paper was to investigate feasibility of the technique for harvesting trachea-bronchial lavage in sheep and to verify its clinic-cytological relationship in healthy sheep and others suffering from pneumonia. In this study 33 sheep were used, 18 healthy and 15 sheep with clinical signs of respiratory disease, divided into groups GS and GD. Clinic examination was performed by obtaining tracheobronchial lavage through the nasal route. The crop was washed with the addition and aspiration of sterile saline. The samples were processed by cytospin and staining with Wright-Giemsa and Shorr. The total cell count and the number of red blood cells per milliliter was higher in the group of sheep with bronchopneumonia. In healthy animals predominance of macrophages was noted, followed by the number of epithelial cells, neutrophils and lymphocytes. In the group of sick animals fewer macrophages and predominance of the neutrophil population were noted. Because of its easy performance to obtain representative material, the technique studied was considered effective to obtain tracheobronchial fluid and therefore is a good method to harvest cells for research of the airways.

Qualidade espermática de sêmen de cães naturalmente infectados por leishmania sp / Semen quality of dogs naturally infected by leishmania sp

Avaliaram-se alterações espermáticas associadas à infecção por leishmaniose no sêmen de cães naturalmente infectados, utilizando-se, durante oito semanas consecutivas, ejaculados de seis cães soronegativos e seis cães soropositivos. As amostras foram colhidas uma vez por semana e avaliadas quanto ao volume, concentração, motilidade, vigor, morfologia espermática, integridade da cromatina, avaliação simultânea da integridade da membrana plasmática, acrossoma e potencial mitocondrial. Concomitantemente foram dosadas a proteína total do plasma seminal e sanguíneo. A leishmaniose visceral causou aumento dos defeitos maiores e menores nos espermatozoides dos animais acometidos pelo estágio moderado a severo da doença. Em estágios mais avançados da enfermidade, a integridade das membranas acrossomal e plasmática foi afetada negativamente. Não foi possível estabelecer um critério quanto à avaliação do potencial mitocondrial. A incidência de alterações morfológicas nos animais acometidos não promoveu aumento de injurias à cromatina. Todos os animais com leishmaniose apresentaram hiperproteinemia do sêmen.

The spermatic changes associated with the natural infection in dogs by Leishmania sp was evaluated during eight consecutive weeks, using ejaculates of six seronegative and six seropositive dogs. The samples were collected once a week and evaluated for volume, concentration, motility, vigor, sperm morphology, chromatin integrity, simultaneous evaluation of the plasmatic membrane integrity, acrosome, and mitochondrial potential. The total proteins of the seminal plasma and blood were measured. The visceral leishmaniasis caused increase of major and minor defects in spermatozoa of animals attacked by moderate to severe stages of the disease. In more advanced stages of the illness, the acrosomal and plasmatic membranes integrity was adversely affected. It was not possible to establish a pattern refering the evaluation of the mitochondrial potential. The incidence of morphological changes in the seropositive animals did not promote an increase of injuries to the chromatin. All animals with leishmaniasis presented hyperproteinemia of the semen.

Preliminary validation of the Brazilian version of the Child Perceptions Questionnaire 8-10.

AIM: The aim of the study was to perform a preliminary validation of the Brazilian version of the Child Perceptions Questionnaire 8-10 (CPQ 8-10). METHODS: The sample used to test the measurement equivalence was made up of 59 children from 8 to 10 years of age at the Federal University of Minas Gerais, Brazil. The CPQ 8-10 was administered, and clinical exams were performed by a single examiner, calibrated for the diagnosis of dental caries (Kappa= 0.98) and malocclusion (weighted Kappa and intraclass correlation coefficient = 0.66-1.00). The children were divided into three groups according to their clinical condition (22 with cavitated carious lesions; 20 with malocclusion; 17 with both oral conditions). STATISTICS: The internal consistency of the instrument was assessed by Cronbach's alpha Coefficient. The Intraclass correlation coefficient (ICC) was used for the test-retest reliability (40 children). Discriminant validity was determined using the Kruskal-Wallis test. RESULTS: An assessment of the psychometric properties revealed the instrument to be reliable (Cronbach's alpha=0.92 for the total scale and 0.63-0.85 for the subscales), exhibiting excellent stability (ICC=0.96 for the total scale and 0.79-0.95 for the subscales). Construct validity was demonstrated through the significant correlations between the global indicators and subscales. There were statistically significant differences between the clinical groups regarding the total scale, thereby demonstrating discriminant validity (p=0.03). CONCLUSIONS: The preliminary validation of the Brazilian version of the CPQ 8-10 showed valid and reliable for use on Brazilian children.

Maspin expression in carcinoma ex pleomorphic adenoma.

AIMS: To investigate the presence and distribution of the protein maspin in carcinoma ex pleomorphic adenoma (CXPA). METHODS: Maspin expression was studied by means of immunohistochemistry in 16 cases of CXPA, using the labelled polymer method. RESULTS: According to the extent of invasion, the tumours were subdivided into: intracapsular (five cases), minimally invasive (four cases), and invasive (seven cases). Twelve patients had carcinoma with only epithelial differentiation, whereas four had a malignant myoepithelial component. Non-luminal cells in the duct-like structures of the remnant pleomorphic adenoma were strongly positive for maspin, whereas only a few luminal cells were immunopositive. A few positive cells were seen in the frequent hypocellular and hyalinised areas. Maspin was abundantly expressed, mainly in non-luminal cells, in transitional areas of CXPA with only epithelial differentiation. In frankly carcinomatous areas there was a gradual decrease in maspin expression. Almost all cells were maspin positive in CXPA with a myoepithelial component. When present, luminal cells were in general negative for maspin. CONCLUSIONS: When only epithelial cells undergo malignant transformation, maspin expression is gradually lost. In cases with a myoepithelial component, maspin expression is high, and this might be related to the tumour suppressor activity attributed to this cell.

Carcinoma ex pleomorphic adenoma (CXPA): immunoprofile of the cells involved in carcinomatous progression.

AIMS: To characterize the cellular component in pleomorphic adenoma (PA) that undergoes malignant transformation in carcinoma ex pleomorphic adenoma (CXPA). METHODS AND RESULTS: A panel of antibodies against cytoskeletal proteins was applied in 16 cases of CXPA: intracapsular carcinoma (five cases), minimally invasive (four cases) and frankly invasive (seven cases). The CXPAs were classified into two main groups according to their predominant cellular component as detected by the panel of antibodies: (i) carcinomas with only epithelial differentiation (75% of the cases), and (ii) carcinomas with a myoepithelial component (25%). CXPA with only epithelial differentiation showed two types of malignant areas in the part of the tumour that was confined by the PA capsule: (i) intraductal carcinoma areas characterized by ductal structures containing both benign myoepithelial cells positive for alpha-smooth muscle actin (alpha-SMA), vimentin and cytokeratin (CK)14 and proliferating atypical luminal cells reactive for CK7, CK8 and CK19, and (ii) carcinoma areas composed only of epithelial cells reactive for CK7, CK8 and CK19. In the latter, the cells presented morphological and immunohistochemical characteristics similar to those found in areas of invasive carcinoma outside the PA capsule. CXPAs with a myoepithelial component were composed mainly or exclusively of cells that expressed vimentin and alpha-SMA. In this group, ductal structures reminiscent of PA filled by malignant cells were not identified. CONCLUSION: Most CXPAs consist only of epithelial cells that have an immunoprofile comparable to ductal luminal cells of PA. These malignant luminal cells arise in the duct-like structures as intraductal carcinoma and probably only at this early stage of development should the lesion be considered as a non-invasive carcinoma.

Expression of integrin subunits alpha2, alpha3, alpha5, alphav, beta1, beta3 and beta4 in different histological types of ameloblastoma compared with dental germ, dental lamina and adult lining epithelium.

OBJECTIVE: To analyze integrin expression and distribution in different histological types of ameloblastoma, compared with dental germ, dental lamina and adult lining epithelium. MATERIALS AND METHODS: Three-micrometer sections from paraffin-embedded specimens were evaluated employing a streptavidin-biotin immunohistochemical method and anti-integrin alpha2, alpha3, alpha5, alphav, beta1, beta3 and beta4 antibodies. RESULTS: All integrins were present in all specimens, exhibiting different patterns. In follicular ameloblastoma, the integrin staining was stronger in the periphery while integrin alpha2 was not present in the central cells. Acanthomatous ameloblastoma showed a similar pattern, with positive staining for integrins alpha3, alpha5, alphav, beta1 and beta4 in the metaplastic cells. In the unicystic, integrin staining was uniform except for integrins alpha5 and beta3 which showed weaker staining in the upper layers. In the plexiform ameloblastoma, dental germ and lamina integrin staining was uniform. In the adult lining epithelium, staining for integrins alpha2, alpha5 and beta4 was confined to the basal layer, while integrins alphav and beta3 were present in the basal and parabasal, with integrins alpha3 and beta1 in the upper layers. CONCLUSION: Acanthomatous, follicular and unicystic ameloblastomas showed integrin staining patterns similar to the adult lining epithelium while the plexiform ameloblastoma was similar to the dental germ and lamina.

Leiomyomatous hamartoma of the incisive papilla.

A case of unusual hamartoma in a six-year-old otherwise healthy Brazilian girl is reported, with emphasis on histological and immunohistochemical features. A mass observed in the incisive papilla was detected whose appearance was similar to congenital epulis or fibroma. Histological findings showed interlacing fascicles of large spindle cells resembling smooth muscle cells. Immunohistochemical staining for desmin and for smooth-muscle actin was positive. The histological diagnosis was leiomyomatous hamartoma, based on clinical and microscopic observations.

Immunohistochemical Mdm2 expression in minor salivary gland tumours and its relationship to p53 gene status.

Mdm2 protein is a cellular regulator of p53 protein activity. Minor salivary gland tumours were investigated for immunohistochemical expression of Mdm2 protein and for p53 gene status. Formalin-fixed sections were submitted to monoclonal antibody anti-Mdm2 through use of the streptavidin-biotin method. Nuclear immunoreactivity was scored 1 (0-25% nuclei positive), 2 (26-50%), 3 (51-75%) and 4 (> 75%). The scores found were: PLGA = 1-4; ACC = 3 and 4; ACA = 2 and 4; PA = 3. Genomic DNA of p53 gene exons 5-8 was examined by polymerase chain reaction and no alterations were detected. The strong immunohistochemical Mdm2 expression may represent an alternative mechanism to the development of salivary gland tumours.

Extranodal follicular dendritic cell sarcoma of the palate.

Follicular dendritic cell tumors are uncommon and usually occur in lymph nodes. We report the case of a follicular dendritic cell tumor that occurred in the palate of a 14-year-old boy and manifested itself as a nodular mass. Histologically, the neoplasm consisted of spindle-shaped or oval-shaped cells with eosinophilic cytoplasms and nuclei with delicate, dispersed chromatin. The lesional cells were principally arranged in diffuse, fascicular patterns with vaguely whorled or storiform areas. Focal multinucleate tumor giant cells and lymphocytes were observed throughout the neoplasm. Immunohistochemically, tumor cells were positive for the follicular dendritic cell markers CD21, CD35, and CD23 and for S-100 protein, CD68, and muscle-specific actin. Tumor cells were negative for LCA, CD20, EMA, CK (AE1/AE3), HMB45, and CD34. Lymphocytes were positive for LCA and CD45RO. Although follicular dendritic cell sarcoma is a very uncommon tumor, it should be included in the differential diagnosis of tumors in this location.

Plasma kinetics of a chylomicron-like emulsion in patients with coronary artery disease.

Chylomicron catabolism in the bloodstream consists of lipolysis by lipoprotein lipase and uptake of remnants by the liver. In rats, triglyceride-rich emulsions can mimic chylomicron metabolism. To further validate this model in man, the emulsion was injected intravenously into fasting and into subjects previously fed a test fatty meal. The plasma kinetic curves of the emulsion 3H-triglyceride and 14C-cholesteryl ester were determined. The fractional clearance rate (FCR) of both labels was markedly reduced in the fed subjects (triglycerides: fed = 0.018 +/- 0.007; fasting = 0.105 +/- 0.013 min-1, P < 0.001; cholesteryl ester: fed = 0.016 +/- 0.001; fasting = 0.040 +/- 0.006 min-1; P < 0.05) indicating that the emulsion and chylomicrons generated from the testinal lipid absorption compete for the same catabolic processes, confirming the validity of the method. The emulsion was injected into 11 patients with CAD and into 11 controls. All had plasma cholesterol < 240 and triglycerides < 250 mg/dl. FCR of triglycerides was 5-fold smaller in CAD compared to controls (0.028 +/- 0.004 and 0.141 +/- 0.069 min-1, respectively, P < 0.01). FCR of cholesteryl ester was 4-fold smaller in CAD than in controls (0.015 +/- 0.004 and 0.056 +/- 0.067 min-1 respectively, P < 0.05). These results indicate that both chylomicron lipolysis and remnant removal are diminished in CAD.

Mutagenicity of airborne particulate organic material from urban and industrial areas of São Paulo, Brazil.

The mutagenicity of airborne particulate matter at three different sites within the São Paulo urban area and the Cubatão industrial area, São Paulo State, Brazil, was evaluated using the Salmonella mutagenicity assay over a year's period (June 1990-May 1991). Total suspended particles (TSP) were collected using a Hi-vol sampler and extracted with methylene chloride by ultrasonication. Pooled seasonal extracts were tested using S. typhimurium TA98 and TA100 with and without metabolic activation (S9) and TA98NR and TA98/1,8-DNP6 without metabolic activation. Organic extracts of the samples with the highest monthly TSP concentration were also assayed for mutagenicity. All samples collected at São Paulo and Cubatão showed mutagenicity with strain TA98, and in general the addition of S9 did not modify the mutagenic response, suggesting the prevalence of direct-acting frameshift mutagens in the atmosphere of these sites. The mutagenicity detected in the urban areas of São Paulo (mainly downtown) was much higher than in the Cubatão industrial area and similar to the more urbanized cities in the world, where vehicle emissions are the major pollution source. Results obtained with the strains TA98NR and TA98/1,8-DNP6 suggested the presence of mononitro- and dinitroarenes contributed to the mutagenicity of these atmospheric samples. A seasonal variation was observed with higher levels of frameshift mutagens during Spring in São Paulo and Summer in Cubatão. During the Winter, more significant responses were found with strain TA100 in the presence of S9 at all sites. Monthly samples presented high levels of mutagenicity during the period of June to November. Results from this research provide support for Air Pollution Control Programs in the detection of the more potent organic mutagenic compounds in the atmosphere and may help in the establishment of priorities for control and regulatory actions.

Enterobacterial repetitive intergenic consensus sequences and the PCR to generate fingerprints of genomic DNAs from Vibrio cholerae O1, O139, and non-O1 strains.

Enterobacterial repetitive intergenic consensus (ERIC) sequence polymorphism was studied in Vibrio Cholerae strains isolated before and after the cholera epidemic in Brazil (in 1991), along with epidemic strains from Peru, Mexico, and India, by PCR. A total of 17 fingerprint patterns (FPs) were detected in the V. cholerae strains examined; 96.7% of the toxigenic V. cholerae O1 strains and 100% of the O139 serogroup strains were found to belong to the same FP group comprising four fragments (FP1). The nontoxigenic V. cholerae O1 also yielded four fragments but constituted a different FP group (FP2). A total of 15 different patterns were observed among the V. cholerae non-O1 strains. Two patterns were observed most frequently for V. cholerae non-01 strains, 25% of which have FP3, with five fragments, and 16.7% of which have FP4, with two fragments. Three fragments, 1.75, 0.79, and 0.5 kb, were found to be common to both toxigenic and nontoxigenic V. cholerae O1 strains as well as to group FP3, containing V. cholerae non-O1 strains. Two fragments of group FP3, 1.3 and 1.0 kb, were present in FP1 and FP2 respectively. The 0.5-kb fragment was common to all strains and serogroups of V. cholerae analyzed. It is concluded from the results of this study, based on DNA FPs of environmental isolates, that it is possible to detect an emerging virulent strain in a cholera-endemic region. ERIC-PCR constitutes a powerful tool for determination of the virulence potential of V. cholerae O1 strains isolated in surveillance programs and for molecular epidemiological investigations.

Detection of cholera (ctx) and zonula occludens (zot) toxin genes in Vibrio cholerae O1, O139 and non-O1 strains.

Vibrio cholerae O1 and V. cholerae non-O1 strains isolated from environmental samples collected in São Paulo, Brazil, during cholera epidemics and pre-epidemic periods were examined for the presence of toxin genes. V. cholerae O1 strains isolated from clinical samples in Peru and Mexico, and V. cholerae O139 strains from India were also examined for the presence of ctx (cholera toxin gene) and zot (zonula occludens toxin gene) by polymerase chain reaction (PCR). A modified DNA-extraction method applied in this study yielded satisfactory recovery of genomic DNA from vibrios. Results showed that strains of V. cholerae O1 isolated during the preepidemic period were ctx (-)/zot (-) whereas strains isolated during the epidemic were ctx (+)/zot (+). All V. cholerae non-O1 strains tested in the study were ctx (-)/zot (-), whereas all V. cholerae O139 strains were ctx (+)/zot (+). Rapid detection of the virulence genes (ctx and zot) can be achieved by PCR and this can serve as an important tool in the epidemiology and surveillance of V. cholerae.

Potentially pathogenic vibrios associated with mussels from a tropical region on the Atlantic coast of Brazil.

Mussels (Perna perna) harvested on the coast of Ubatuba, in three different stations in the State of São Paulo, Brazil, were examined for Vibrio spp. over a 1 year period. The ranges of most probable number (MPN 100 g-1) were: Vibrio alginolyticus (< 3-24,000), V. parahaemolyticus (< 3-24,000), V. fluvialis (< 3-1100), V. cholerae non-O1 (< 3-23), V. furnissii (< 3-30), V. mimicus (< 3-9) and V. vulnificus (< 3-3). The highest incidence was observed for V. alginolyticus (92-100%), followed by V. parahaemolyticus (67-92%), V. fluvialis (34-67%), V. vulnificus (8-17%), V. furnissii (0-17%), V. mimicus (0-17%) and V. cholerae non-O1 (0-8%). Tests for virulence factors were positive in 34.1% of the vibrios in the rabbit ileal loop and 31.7% in the Dean test. Positive results in the Kanagawa test were obtained with 0.51% of V. parahaemolyticus strains. The mean values (MPN 100 g-1) of faecal coliforms in mussels from the three regions varied from 1100 to 44,000, and seawater collected at the same stations gave average values for faecal coliforms in the range 18-3300 MPN 100 ml-1. These results highlight the potential risks of food poisoning associated with raw or undercooked seafood.

Assessment of media using B-D-Glucoronidase activity for the detection of Escherichia coli in water

Quatro meios de cultura contendo B-D-glucoronide MUG (Lauril Sulfato Fluorocult, ECD Fluorocult Agar, Caldo Lauril Triptose e Agar nutriente suplementados com MUG) foram comparados com meios convencionais (caldo Lauril Triptose, Caldo E.C. e Agar M-Endo LES) para verificar se havia diferenças entre eles na detecçäo de E. coli. As amostras de água foram contaminadas artificialmente com E.coli isoladas tanto do ambiente quanto de espécimes clínicos e analisados pelas técnicas dos tubos múltiplos e de membrana filtrante usando os meios acima descritos. Entre as cepas previamente analisadas para o preparo das amostras artificialmente contaminadas, 6,4 por cento näo apresentaram atividade B-D-Glucoronidase quando testadas com meio contendo MUG. Os resultados obtidos foram submetidos ao Teste de Wilcoxon que mostrou que Lauril Sulfato Fluorocult, caldo Lauril Triptose com MUG e Agar Nutriente com MUG foram täo sensíveis quanto o Caldo Lauril Triptose e meio E.C. e Agar M-Endo LES, e que o ECD Fluorocult Agar apresentou uma sensibilidade muito baixa. Esta técnica é de grande importância pois pode ser aplicada em métodos mais rápidos para detecçäo de E.coli e em automaçäo