Occurrence of phytoplasma associated with sugarcane yellow leaf syndrome in three regions of Sao Paulo State, Brazil

Sugarcane yellow leaf syndrome caused serious damage to crops in the Sao Paulo State, Brazil, in the 1990`s. The syndrome was also reported in other countries and investigations into the etiology revealed an association between the disease and virus and/or phytoplasma. The disease is potentially important and occurs endemically in that State, and for this reason the present study was conducted in order to demonstrate the occurrence of phytoplasma in three traditional sugarcane-producing areas, in Sao Paulo State, through molecular detection From naturally infected plants. Symptomatic plants belonging to varieties SP71-6163, SP71-6180 and SP89-1115 were sampled from Piracicaba, Jau and Ribeirao Preto, and total DNA was extracted from foliar tissues. Nested PCR was conducted with primer pairs P1/ Tint-16F2n/R2, and the amplified products were analyzed by electrophoresis on agarose gels. Amplified DNA fragments of 1.2 kb evidenced the presence of phytoplasma in 36% of symptomatic plants and revealed its Occurrence in all sampled regions. The results demonstrated that phytoplasma is associated with the disease and that it is important to keep a safe inspection of nurseries and monitoring plants in the field, as well as to Select Sugarcane genotypes with a good level of resistance in breeding programs.

South American leaf blight intensity evaluated in six clones of young and adult rubber trees in the Vale do Ribeira region, Sao Paulo state, Brazil

Rubber tree clones present different intensity of symptoms, depending on their age. This is mostly clearly seen in the presence or absence of old leaves with ascopores, corresponding to flows of new and susceptible leaves. The objective of this work was to evaluate the intensity of symptoms of south American leaf blight (SALB) in six rubber tree clones, one and eight years old, in the Vale do Ribeira region, Sao Paulo state. The results Showed that clones FX 3864, RRIM 600, IAN 873 and IAN 717 Suffered more attacks when young, and RRIM 600, IAN 717 and FX 3864 when adult. The clone IAN 873 showed the smallest amounts of disease in the adult phase, due to uniform change in the leaves and a compact flow of new leaves, which happened during a season that was unfavorable to pathogen infection, presenting the phenomenon of avoidance or evasion in time.

Occurrence and distribution of Vibrio parahaemolyticus in retail oysters in Sao Paulo State, Brazil

Vibrio parahaemolyticus is a potentially pathogenic bacterium that occurs naturally in estuarine environments worldwide, and is often associated with gastroenteritis in humans following consumption of raw bivalve mollusks, especially raw oysters. The occurrence of total and pathogenic V. parahaemolyticus in 74 samples of raw oysters collected in restaurants, supermarkets, groceries and beach huts in Sao Paulo State, was monitored between February 2006 and January 2007. Enumeration of V. parahaemolyticus was performed according to the most probable number (MPN) procedure. Five to ten typical colonies were selected from thiosulfate-citrate-bile salts-sucrose (TCBS) agar plates for confirmation by the presence of the species-specific gene tlh and the virulence genes tdh and trh by multiplex PCR. V. parahaemolyticus was detected in 100% of samples. The densities of total V. parahaemolyticus varied from 1.78 to 6.04 logio (MPN/g), with higher densities being detected in fall and summer, and lower densities in winter (P < 0.05). There was no statistical difference among densities of V parahaemolyticus regarding the site of collection. None of the 1943 V parahaemolyticus isolates contained tdh and/or trh. These data provide information for the assessment of exposure to V. parahaemolyticus in oysters consumed in Sao Paulo, State, Brazil. (C) 2010 Elsevier Ltd. All rights reserved.

Correlation between Environmental Factors and Prevalence of Vibrio parahaemolyticus in Oysters Harvested in the Southern Coastal Area of Sao Paulo State, Brazil

The presence of Vibrio parahaemolyticus in 123 oyster samples collected from an estuary on the southern coast of Sao Paulo state, Brazil, was investigated. Of the 123 samples, 99.2% were positive with densities ranging from <3 to 10(5) most probable number (MPN)/g. Densities correlated significantly with water temperature (r = 0.48; P < 0.001) but not with salinity (r = -0.09; P = 0.34). The effect of harvest site on counts was not significant (P > 0.05). These data provide information for the assessment of exposure of V. parahaemolyticus in oysters at harvest.

Sero-epidemiology of toxocariasis in a rural settlement in Sao Paulo state, Brazil

The seroprevalence of Toxocara canis and risk factors for infection with this parasite were explored in a rural settlement in Sao Paulo state, Brazil. Total IgA and IgE levels in 79 subjects were determined by turbidimetry and chemiluminescence, respectively. Total counts of leucocytes and erythrocytes and differential counts of leucocytes were made by flow cytometry. ELISA for the detection of anti-Toxocara IgG, IgA and IgE were standardized using Toxocara excretory-secretory antigens (TES) obtained from the cultured second-stage larvae of T. canis. Seventeen (21.5%) of the subjects were found positive for anti-Toxocara IgG, with no significant differences in such seropositivity with age or gender. Thirty (38%) of the subjects showed eosinophilia and 70 (89%) had elevated levels of total IgE. Among the 17 subjects found seropositive for anti-Toxocara IgG, the percentage of leucocytes represented by eosinophils (P=0.0069) and total levels of IgE (P=0.0452) were positively correlated with the levels of anti-TES IgE. Although anti-TES IgA was detected in 10 (59%) of the subjects, there was no significant correlation between the levels of total IgA and those of Toxocara-specific IgA. Only one of the 17 subjects found positive for anti-Toxocara IgG had attended a secondary school and all but two belonged to households with monthly incomes of < U.S.$100. In the study community at least, seropositivity may be related to poor living standards and lack of basic sanitary conditions. The presence of anti-Toxocara IgE and IgA may facilitate the diagnosis of toxocariasis and may well be useful for monitoring the success of treatment.

Assessment of Ametryn Contamination in River Water, River Sediment, and Mollusk Bivalves in Sao Paulo State, Brazil

Sao Paulo state, Brazil, is one of the main areas of sugar cane agriculture in the world. Herbicides, in particular, ametryn, are extensively used in this extensive area, which implies that this herbicide is present in the environment and can contaminate the surface water by running off. Thereby, residues of ametryn were analyzed in samples of river water an river sediment and in freshwater bivalves obtained from the rivers Sapucai, Pardo and Mogi-Guacu in Sao Paulo State, Brazil. Samples were taken in the winter of 2003 and 2004 in two locations in each river. The specimens of freshwater bivalves collected and analyzed were Corbicula fluminea, an exotic species, and Diplodon fontaineanus, a native species. Additionally, the evaluation of the ability of bioconcentration and depuration of ametryn by the freshwater bivalve Corbicula fluminea was also performed. Ametryn concentrations in the samples were measured by liquid chromatography coupled to mass spectrometry. Residues of ametryn in water (50 ng/L) and in freshwater bivalves (2-7 ng/g) were found in the Mogi-Guacu River in 2004, and residues in river sediments were found in all rivers in 2003 and 2004 (0.5-2 ng/g). The observation of the aquatic environment through the analysis of these matrixes, water, sediment, and bivalves, revealed the importance of the river sediment in the accumulation of the herbicide ametryn, which can contaminate the biota.

Triphora uniflora AC Ferreira, Baptista & Pansarin (Orchidaceae: Triphorcae): a new species and the first record of the genus Triphora Nutt. for Sao Paulo state, Brazil

(Triphora uniflora A.C. Ferreira, Baptista & Pansarin (Orchidaccae: Triphorcae): a new species and the first record of the genus Triphora Nutt. for Sao Paulo state, Brazil). Triphora uniflora A. C. Ferreira, Baptista & Pansarin, a new species of Orchidaceae, is described and illustrated. Furthermore, this is the first report of the genus Triphora for Sao Paulo state, Brazil. The relationship of this new species to other taxa of the genus and the need to preserve the natural habitat of this Triphora species are discussed.

Anacroneuria (Plecoptera, Perlidae) from the Mantiqueira Mountains, Sao Paulo State, Brazil

Seven species of Anacroneuria Klapalek are considered; of these 4 were known, A. debilis (Pictet, 1841), A. flintorum Froehlich, 2002, A. toriba Froehlich, 2002 (the female is described herein), and A. vanini Froehlich, 2004. Three are new: A. mantiqueirae, A. simulans, and A. tabatae.

Toxoplasma gondii isolates: Multi locus RFLP-PCR genotyping from human patients in Sao Paulo State, Brazil identified distinct genotypes

This study investigated the genetic characteristics of Toxoplasma gondii samples collected from 62 patients with toxoplasmosis in Sao Paulo State, Brazil. DNA samples were isolated from blood, cerebrospinal fluid and amniotic fluids of 25 patients with cerebral toxoplasmosis and AIDS, two patients with acute toxoplasmosis, 12 patients with ocular toxoplasmosis, six newborns with congenital toxoplasmosis and 17 pregnant women with acute infection. Diagnosis of toxoplasmosis was based in clinical, radiological and laboratory features. Genotyping was performed using multilocus PCR-RFLP genetic markers including SAG1, SAG2, 5`- and 3`-SAG2, alt.SAG2, SAG3, BTUB, GRA6, C22-8, c29-2, L358, PK1 and Apico. Among the 62 clinical samples, 20 (32%) were successfully genotyped at eight or more genetic loci and were grouped to three distinct genotypes. Eighteen samples belonged to ToxoDB Genotype #65 and the other two samples were identified as ToxoDB Genotypes #6 and #71, respectively (http://toxodb.org/toxo/). Patients presenting Genotypes #6 and #71 had severe and atypical cerebral toxoplasmosis, characterized by diffuse encephalitis without extensive brain lesions. These results indicate that T. gondii Genotype #65 may have a high frequency in causing human toxoplasmosis in Sao Paulo State, Brazil. This unusual finding highlights the need to investigate the possible association of parasite genotypes with human toxoplasmosis. (C) 2011 Elsevier Inc. All rights reserved.

Analysis of marine bivalve shellfish from the fish market in Santos city, Sao Paulo state, Brazil, for Toxoplasma gondii

The aim of this study was to determine if Toxoplasma gondii are present in oysters (Crassostrea rhizophorae) and mussels (Mytella guyanensis) under natural conditions using a bioassay in mice and molecular detection methods. We first compared two standard protocols for DNA extraction, phenol-chloroform (PC) and guanidine-thiocyanate (GT), for both molluscs. A total of 300 oysters and 300 mussels were then acquired from the fish market in Santos city, Sao Paulo state, Brazil, between March and August of 2008 and divided into 60 groups of 5 oysters and 20 groups of 15 mussels. To isolate the parasite, five mice were orally inoculated with sieved tissue homogenates from each group of oysters or mussels. For molecular detection of T. gondii, DNA from mussels was extracted using the PC method and DNA from oysters was extracted using the GT method. A nested-PCR (Polymerase Chain Reaction) based on the amplification of a 155 bp fragment from the B1 gene of T. gondii was then performed. Eleven PCR-RFLP (Restriction Fragment Length Polymorphism) markers, SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, CS3 and Apico, were used to genotype positive samples. There was no isolation of the parasite by bioassay in mice. T. gondii was not detected in any of the groups of mussels by nested-PCR. DNA of T. gondii was apparently detected by nested-PCR in 2 groups of oysters (3.3%). Genotyping of these two positive samples was not successful. The results suggest that oysters of the species C. rhizophorae, the most common species from the coast of Sao Paulo, can filter and retain T. gondii oocysts from the marine environment. Ingestion of raw oysters as a potential transmission source of T. gondii to humans and marine mammals should be further investigated. (C) 2010 Elsevier B.V. All rights reserved.

Ehrlichia canis in dogs attended in a veterinary hospital from Botucatu, Sao Paulo State, Brazil

This study investigated the etiology of canine ehrlichiosis and possible clinical and epidemiological data associated with the infection in 70 dogs suspect of ehrlichiosis attended at the Veterinary Hospital of the Sao Paulo State University in Botucatu city during 2001 and 2002. Dogs were evaluated by clinical-epidemiological and hematological data and molecular analysis by partial amplification and DNA sequencing of the ehrlichial dsb gene. E. canes DNA was amplified and sequenced in 28 (40.0%) dogs. Dogs younger than 12 months old showed significantly higher infection rates (65.0%; P < 0.05). Diarrhea, apathy, and anorexia were the major clinical signs observed in 55.2% (P = 0.05), 47.0% (P > 0.05), and 42.4% (P > 0.05) of the PCR-positive dogs, respectively. Twenty-five anemic (<5.5 x 10(6) RBC.mu L(-1)), and 8 leukopenic (<5.5 x 10(3) WBC.mu L(-1)) dogs were PCR-positive (P > 0.05). All 28 PCR-positive dogs showed thrombocytopenia (<175 x 10(3) platelets.mu L(-1)) and revealed statistical significance (P < 0.05). E. canis was the only Ehrlichia species found in dogs in the studied region, with higher infection rates in younger dogs, and statistically associated with thrombocytopenia.

Aedes aegypti entomological indices in an endemic area for dengue in Sao Paulo State, Brazil

OBJECTIVE: To evaluate the most productive types of properties and containers for Aedes aegypti and the spatial distribution of entomological indices.METHODS: Between December 2006 and February 2007, the vector's immature forms were collected to obtain entomological indices in 9,875 properties in the Jaguare neighborhood of Sao Jose do Rio Preto, SP, Southeastern Brazil. In March and April 2007, a questionnaire about the conditions and characteristics of properties was administered. Logistic regression was used to identify variables associated with the presence of pupae at the properties. Indices calculated per block were combined with a geo-referenced map, and thematic maps of these indices were obtained using statistical interpolation.RESULTS: The properties inspected had the following Ae. aegypti indices: Breteau Index = 18.9, 3.7 larvae and 0.42 pupae per property, 5.2 containers harboring Ae. aegypti per hectare, 100.0 larvae and 11.6 pupae per hectare, and 1.3 larvae and 0.15 pupae per inhabitant. The presence of yards, gardens and animals was associated with the presence of pupae.CONCLUSIONS: Specific types of properties and containers that simultaneously had low frequencies among those positive for the vector and high participation in the productivity of larvae and pupae were not identified. The use of indices including larval and pupal counts does not provide further information beyond that obtained from the traditional Stegomyia indices in locations with characteristics similar to those of São José do Rio Preto. The indices calculated per area were found to be more accurate for the spatial assessment of infestation. The Ae. aegypti infestation levels exhibited extensive spatial variation, indicating that the assessment of infestation in micro areas is needed.

Detection of assemblages A and B of Giardia duodenalis in water and sewage from Sao Paulo state, Brazil

Giardia duodenalis is a protozoan that parasitizes humans and other mammals and causes giardiasis. Although its isolates have been divided into seven assemblages, named A to G, only A and B have been detected in human faeces. Assemblage A isolates are commonly divided into two genotypes, AI and AII. Even though information about the presence of this protozoan in water and sewage is available in Brazil, it is important to verify the distribution of different assemblages that might be present, which can only be done by genotyping techniques. A total of 24 raw and treated sewage, surface and spring water samples were collected, concentrated and purified. DNA was extracted, and a nested PCR was used to amplify an 890 bp fragment of the gdh gene of G. duodenalis, which codes for glutamate dehydrogenase. Positive samples were cloned and sequenced. Ten out of 24 (41.6%) samples were confirmed to be positive for G. duodenalis by sequencing. Phylogenetic analysis grouped most sequences with G. duodenalis genotype AII from GenBank. Only two raw sewage samples presented sequences assigned to assemblage B. In one of these samples genotype AII was also detected. As these assemblages/genotypes are commonly associated to human giardiasis, the contact with these matrices represents risk for public health.

Precipitation variability in Sao Paulo State, Brazil

The State of Sao Paulo is the richest in Brazil, responsible for over 30% of the Brazilian gross rate. It has a population of around 30 million and its economy is based on agriculture and industrial products. Any change in climate can have a profound influence on the socio-economics of the State. In order to determine changes in total and extreme rainfall over Sao Paulo State, climate change indices derived from daily precipitation data were calculated using specially designed software. Maps of trends for a subset of 59 rain gauge stations were analysed for the period 1950-1999 and also for a subset of this period, 1990-1999, representing more recent climate. A non-parametric Mann-Kendall test was applied to the time series. Maps of trends for six annual precipitation indices (annual total precipitation (PRCPTOT), very heavy precipitation days (R20mm), events greater than the 95th percentile (R95p), maximum five days precipitation total (RX5day), the length of the largest wet spell (CWD) and the length of the largest dry spell (CDD)) were analysed for the entire period. These exhibited statistically significant trends associated with a wetter climate. A significant increase in PRCPTOT, associated with very heavy precipitation days, were observed at more than 45% of the rain gauge stations. The Mann-Kendall test identified that the positive trend in PRCPTOT is possibly related to the increase in the R95p and R20mm indices. Therefore, the results suggest that there has been a change in precipitation intensity. In contrast, the indices for the more recent shorter time series are significantly different to the longer term indices. The results indicate that intense precipitation is becoming concentrated in a few days and spread over the period when the CDD and R20mm indices show positive trends, while negative ones are seen in the RX5day index. The trends found could be related to many anthropogenic aspects such as biomass burning aerosols and land use.

Floristic inventory of cerrado at Pedregulho, Sao Paulo State, Brazil

Pedregulho municipality in the far northeastern region of Sao Paulo state has fragments of cerrado vegetation thought to be floristically distinct from other cerrado remmants in the state. In this study, a floristic survey was carried out at Pedregulho, focusing on two geologically distinct regions: Furnas do Bom Jesus State Park and the Estreito district. In each region, 50x50 m plots were set up for a total sample area of 1 hectare, where 65 families and 379 angiosperm species were recorded. Collecting efforts in adjacent areas resulted in a total of 71 families and 443 species. The two study areas had low floristic similarity. Geographic species distribution was analyzed and eight patterns were defined for those species restricted to the Pedregulho region in Sao Paulo.