Chicken as potential contamination source of Campylocacter lari in Iquitos, Peru

In order to know the importance of chicken as natural reservoir of Campylobacter lari in Iquitos, Peru; samples were obtained by cloacal swabs from 200 chickens and immediately placed into a semisolid enrichment medium; these were streaked on modified Skirrow Agar. The organism was isolated from 21 (10.5%) samples, corresponding 58.8% to biovar I and 41.2% to biovar II (Lior scheme). The results provide evidence that chicken appear to be prominent reservoirs of Campylobacter lari in Iquitos.

Detection and identification of dengue virus isolates from Brazil by a simplified reverse transcription - polymerase chain reaction (RT-PCR) method

We show here a simplified RT-PCR for identification of dengue virus types 1 and 2. Five dengue virus strains, isolated from Brazilian patients, and yellow fever vaccine 17DD as a negative control, were used in this study. C6/36 cells were infected and supernatants were collected after 7 days. The RT-PCR, done in a single reaction vessel, was carried out following a 1/10 dilution of virus in distilled water or in a detergent mixture containing Nonidet P40. The 50 µl assay reaction mixture included 50 pmol of specific primers amplifying a 482 base pair sequence for dengue type 1 and 210 base pair sequence for dengue type 2. In other assays, we used dengue virus consensus primers having maximum sequence similarity to the four serotypes, amplifying a 511 base pair sequence. The reaction mixture also contained 0.1 mM of the four deoxynucleoside triphosphates, 7.5 U of reverse transcriptase, 1U of thermostable Taq DNA polymerase. The mixture was incubated for 5 minutes at 37ºC for reverse transcription followed by 30 cycles of two-step PCR amplification (92ºC for 60 seconds, 53ºC for 60 seconds) with slow temperature increment. The PCR products were subjected to 1.7% agarose gel electrophoresis and visualized by UV light after staining with ethidium bromide solution. Low virus titer around 10 3, 6 TCID50/ml was detected by RT-PCR for dengue type 1. Specific DNA amplification was observed with all the Brazilian dengue strains by using dengue virus consensus primers. As compared to other RT-PCRs, this assay is less laborious, done in a shorter time, and has reduced risk of contamination

Water-contact patterns and risk factors for Schistosoma mansoni infection in a rural village of Northeast Brazil

Schistosomiasis mansoni in the Serrano village, municipality of Cururupu, state of Maranhão, Brazil, is a widely spread disease. The PECE (Program for the Control of Schistosomiasis), undertaken since 1979 has reduced the prevalence of S. mansoni infection and the hepatosplenic form of the disease. Nevertheless piped water is available in 84% of the households, prevalence remains above 20%. In order to identify other risk factors responsible for the persistence of high prevalence levels, a cross-sectional survey was carried out in a systematic sample of 294 people of varying ages. Socioeconomic, environmental and demographic variables, and water contact patterns were investigated. Fecal samples were collected and analyzed by the Kato-Katz technique. Prevalence of S. mansoni infection was 24.1%, higher among males (35.5%) and between 10-19 years of age (36.6%). The risk factors identified in the univariable analysis were water contacts for vegetable extraction (Risk Ratio - RR = 2.92), crossing streams (RR = 2.55), bathing (RR = 2.35), fishing (RR = 2.19), hunting (RR = 2.17), cattle breeding (RR = 2.04), manioc culture (RR = 1.90) and leisure (RR = 1.56). After controlling for confounding variables by proportional hazards model the risks remained higher for males, vegetable extraction, bathing in rivers and water contact in rivers or in periodically inundated parts of riverine woodland (swamplands)

QUESTIONNAIRES IN THE SCREENING FOR Schistosoma mansoni INFECTION: A STUDY OF SOCIO DEMOGRAPHIC AND WATER CONTACT VARIABLES IN FOUR COMMUNITIES IN BRAZIL

The use of questionnaires has been recommended for identifying, at a lower cost, individuals at risk for schistosomiasis. In this study, validity of information obtained by questionnaire in the screening for Schistosoma mansoni infection was assessed in four communities in the State of Minas Gerais, Brazil. Explanatory variables were water contact activities, sociodemographic characteristics and previous treatment for schistosomiasis. From 677, 1474, 766 and 3290 individuals eligible for stool examination in the communities, 89 to 97% participated in the study. The estimated probability of individuals to be infected, if they have all characteristics identified as independently associated with S.mansoni infection, varied from 15% in Canabrava, to 42% in Belo Horizonte, 48% in Comercinho and 80% in São José do Acácio. Our results do not support the hypothesis that a same questionnaire on risk factors could be used in screening for S.mansoni infection in different communities.

Occurrence of Cryptosporidium oocysts and Giardia cysts in raw water from the Atibaia river, Campinas, Brazil

Cryptosporidium parvum and Giardia duodenalis are waterborne parasites that have caused several outbreaks of gastrointestinal disease associated with drinking water. Due to the lack of studies about the occurrence of these protozoa in water in the Southeast of Brazil, an investigation was conducted to verify the presence of cysts and oocysts in superficial raw water of the Atibaia River. The water samples were submitted to membrane filtration (3.0 mum) and elution was processed by (1) scraping and rinsing of membrane (RM method) and (2) acetone-dissolution (ADM method). Microbiologic and chemical parameters were analyzed. Aliquots of the pellets were examined by immunofluorescence (Merifluor, Meridian Diagnostics, Cincinnati, Ohio). All water samples were positive for Cryptosporidium and Giardia, in spite of the high turbidity. Higher recovery rates occurred in samples treated by the RM method than by the ADM technique. The goal for future work is the assessment of viability of cysts and oocysts to determine the public health significance of this finding.

Importance of hand germ contamination in health-care workers as possible carriers of nosocomial infections

The importance of hands in the transmission of nosocomial infection has been world wide admitted. However, it is difficult to induce this behavior in health-care workers. The aim of the present work was to point out the importance of hand bacteria colonization, the influence of hand washing and of patient physical examination. One hundred health-care workers were randomly divided in two groups: Group A without hand washing previous to patient physical examination or handling (PPE); group B with hand washing previous to PPE. Direct fingerprint samples in Columbia agar before and after PPE were obtained. The colonies were counted and identified by conventional techniques, and antibiograms according to NCCLS were performed. Before PPE group A participants showed a high number of bacteria regarding group B participants (73.9 Vs 20.7; p < 0.001); 44 out of 50 participants were carriers of potentially pathogen bacteria. No group B participants were carriers of potential pathogen bacteria before PPE. The latter group showed an increase in number of bacteria after PPE (20.7 CFU (before) Vs 115.9 CFU (after); p < 0.001). Sixteen group B participants were contaminated after PPE with potential pathogens such as S. aureus (50% of them meticillin resistant); Escherichia coli, Pseudomonas aeruginosa and Enterococcus faecalis, half of them multiresistant. We can conclude on the importance of these results to implement educational programs and to provide the health-care workers with the proper commodities to fulfill this practice.

Burkholderia pseudomallei: a case report of a human infection in Ceará, Brazil

Burkholderia pseudomallei has rarely been isolated from environmental and clinical specimens in South America, particularly, in Brazil. This report describes a case of melioidosis with fulminant sepsis in a 10 year old boy, from rural area, in Tejuçuoca, State of Ceará, Brazil. Blood samples were positive and, through the analysis of results from biochemical tests and of drugs susceptibility profile, identified this gram-negative bacillus as B. pseudomallei. The contamination source remains obscure in this case, as soil and water tanks samples submitted to microbiological analyses did not indicate the presence of B. pseudomallei.

The frequency rate of Toxocara species contamination in soil samples from public yards in a urban area "Payathai", Bangkok, Thailand

Toxocara species are most common roundworms of Canidae and Felidae. Human toxocariasis develops by ingesting of embryonated eggs in contaminated soil. There is no previous report of Toxocara contamination in the soil samples from the public areas in Bangkok. For this reason our study have been carried out to examine the frequency of Toxocara eggs in public yards in Bangkok, Thailand. A total of 175 sand and clay samples were collected and examined for parasite eggs. According to this study, Toxocara eggs were detected from 10 (5.71%) of 175 soil samples. The high rate of contamination in this study implies the importance of the control of this possible zoonotic disease: control of abandon of dogs and cats, is still necessary.

Environmental contamination by Toxocara sp. eggs in Ribeirão Preto, São Paulo State, Brazil

Toxocariasis is a zoonosis mainly caused by Toxocara canis, an intestinal nematode of dogs. Man acquires the infection through accidental ingestion of viable eggs, and the toxocariasis clinical manifestations may vary from an asymptomatic infection up to the Visceral Larva Migrans syndrome. Seventy eight public squares of Ribeirão Preto, São Paulo, Brazil, including Bonfim Paulista district were visited aiming to evaluate the soil contamination by Toxocara eggs. The squares were divided in five different areas corresponding to the Sanitary Districts of the city. From May to December 2003, soil samples weighting about 250 g each were collected from five distinct sites of each public square. The laboratorial analysis was done by centrifugal-flotation techniques in magnesium sulphate solutions with 5% of potassium iodide (d = 1.33) and zinc sulphate (d = 1.20), and by the sedimentation- flotation in conic chalices with zinc sulphate (d = 1.20). Toxocara sp. eggs were found on 16 (20.5%) squares, with the lowest prevalence (12%) at the central area. From these results, it is expected that the legal authority will adopt protection measures for the city public areas, reducing thus the contamination risk by Toxocara sp. eggs.

Frequency of soil contamination by Toxocara canis eggs in the south region of São Paulo municipality (SP, Brazil) in a 18 month period

Soil contamination by embryonic eggs of Toxocara canis is the main source of human infection by this ascarid larvae resulting, sometimes, in the occurrence of visceral larva migrans syndrome. The objective of the present research is to determine the frequency of T. canis eggs in soil samples monthly collected in nine public places, located at the South Region of São Paulo municipality in a 18-month period, from February 2004 to July 2005. The soil samples collected were treated with a 30% antiformine solution and with a sodium dichromate solution (d = 1.40) and microscopic slides were prepared and examined under light microscopy for searching T. canis eggs. Two peaks of higher frequency had been found, one in February - May 2004 and the other in April - July 2005.

Occurrence of Proteus mirabilis associated with two species of venezuelan oysters

The fecal contamination of raw seafood by indicators and opportunistic pathogenic microorganisms represents a public health concern. The objective of this study was to investigate the presence of enteric bacteria colonizing oysters collected from a Venezuelan touristic area. Oyster samples were collected at the northwestern coast of Venezuela and local salinity, pH, temperature, and dissolved oxygen of seawater were recorded. Total and fecal coliforms were measured for the assessment of the microbiological quality of water and oysters, using the Multiple Tube Fermentation technique. Analyses were made using cultures and 16S rRNA gene sequencing. Diverse enrichment and selective culture methods were used to isolate enteric bacteria. We obtained pure cultures of Gram-negative straight rods with fimbriae from Isognomon alatus and Crassostrea rhizophorae. Our results show that P. mirabilis was predominant under our culture conditions. We confirmed the identity of the cultures by biochemical tests, 16S rRNA gene sequencing, and data analysis. Other enterobacteria such as Escherichia coli, Morganella morganii and Klebsiella pneumoniae were also isolated from seawater and oysters. The presence of pathogenic bacteria in oysters could have serious epidemiological implications and a potential human health risk associated with consumption of raw seafood.

Environmental contamination by Toxocara spp. Eggs in a rural settlement in Brazil

In order to study the environmental contamination by Toxocara spp. eggs in a rural community from the Pontal do Paranapanema region, São Paulo State, Brazil, soil samples from 31 out of 121 plots were collected in eight different places on each house. The samples were submitted to flotation technique in sodium nitrate (d = 1.20g/cm³). Eggs of Toxocara spp. were recovered in nine (29.03%) out of the 31 plots. At least one dog was registered in 27 of the 31 plots examined (87.1%) and at least one cat in 17 (54.84%). The number of pets per plot ranged from one to six (mean of 2.3) for dogs and one to 14 (mean of 1.29) for cats. In 16 plots (51.61%), the presence of both dogs and cats was observed. There was no relation between the presence of pets in the plots and soil contamination (p > 0.05). However, the environmental contamination by Toxocara spp. eggs associated to the poor conditions of the inhabitants may be an important risk factor for the human population to ocular or visceral larva migrans.

Influence of variables on centrifuge-flotation technique for recovery of Toxocara canis eggs from soil

The purpose of this study was to evaluate the influence of variables in a flotation technique for the recovery of Toxocara canis eggs from soil. The trials were done under standardized conditions on one gram of previously sterilized soil samples contaminated with 200 eggs of T. canis. The following variables were evaluated in serial steps: sieving; type of wash; time of stirring; resuspension of sediment; solution flotation. Centrifuge-flotation in sodium nitrate (d = 1.20 g/cm³) was adopted as an initial technique, using Tween 80 (0.2%) and decinormal sodium hydroxide as solutions for washing the samples. Ten tests were done to compare the variables, using counting in triplicate. The sieving of the material reduced significantly the recovery of eggs (p < 0.001) and the number of eggs recovered was higher when the sediment was resuspended (p < 0.05). After standardization, flotation solutions sodium chloride, zinc sulfate, sodium dichromate, magnesium sulfate, and sodium nitrate (d = 1.20g/cm³) were compared. The best results were obtained by using zinc sulfate solution. In conclusion, the chances of recovering T. canis eggs from samples using flotation solutions can be increased by washing of soil twice using distilled water, and resuspension of sediment. On the other hand, the sieving procedure can drastically reduce the number of eggs.