Genetic diversity of the entomopathogenic fungus Cordyceps tenuipes in forests and butterfly gardens in Quindío,Colombia

Cordyceps tenuipes is an entomopathogenic fungus that infects mostly pupae of several lepidopteran families. In Colombia the species has been reported in non-disturbed tropical rain forests and more recently in butterfly gardens. The aim of this study was to assess the genetic diversity in populations of C. tenuipes present in natural (forests) and artificial (e.g. butterfly gardens) environments in the department of Quindío, Colombia, using three molecular nuclear markers ITS, TEF-1α and RPB1. All the samples evaluated corresponded morphologically and phylogenetically to C. tenuipes. The butterfly garden of Quindio Botanical Garden (QBG) showed the highest genetic diversity among all sampling localities and was very similar to that of its adjacent forest. The Amaranta Butterfly Garden (ABG), located north of QBG, showed lower genetic diversity as well as little genetic differentiation with QBG, consistent with the hypothesis of a pathogen transfer from QBG to ABG. Higher F_ST values were observed for TEF-1α and ITS, revealing genetic differentiation between all demes and the southern forest population. Our research constitutes the first study of the intraspecific diversity of C. tenuipes in Colombia and can serve as the first step in identifying diversity reservoirs and management of epizootic episodes caused by this fungal species.     

Environmental variables and intertidal beach annelids of São Sebastião Channel (State of São Paulo,Brazil)

Benthic annelid communities were studied during a one-year period (August/95 to July/96) in two sectors of the beaches Engenho d'Agua and S?o Francisco, S?o Sebasti?o Channel (S?o Paulo, Brazil), where the substrate is composed by a mixture of sand and rock fragments. Abiotic parameters such as salinity of interstitial water and sediment properties were used to characterize the environment. The polychaetes were well represented in the two sectors and their distribution was related with sediment type. The density of individuals and the number of taxa was higher at S?o Francisco, while the diversity and the evenness were higher at Engenho d'Agua. This difference can be a consequence of organic enrichment caused by domestic input, and of the lower and more variable salinity at S?o Francisco. Due to these factors, the high density of opportunistic species, like Capitella capitata ssp., Scolelepis squamata, Laeonereis acuta and several oligochaetes, represented 75.5% of total abundance at this sector.     

Elmidae (Coleoptera, Byrrhoidea) larvae in the state of São Paulo, Brazil: Identification key, new records and distribution

The family Elmidae Curtis, 1830 has cosmopolitan distribution and most species inhabit riffles on streams and rivers, hence the name “riffle beetle”. In recent years, this family has been featured in papers addressing the assessment and environmental monitoring of water quality. In Brazil, studies on the family remain scarce and the present investigation is a pioneering study in the state of São Paulo. This study aims to propose a taxonomic key for the identification of larvae of Elmidae genera known to occur in the State, as well as to report new records and the distribution of these genera. The material analyzed was collected from various locations in each of 15 drainage basins from 2005 to 2010. The identification key includes 12 genera (Austrolimnius Carter & Zeck, 1929, Heterelmis Sharp, 1882, Hexacylloepus Hinton, 1940, Hexanchorus Sharp, 1882, Huleechius Brown, 1981, Macrelmis Motschulsky, 1859, Microcylloepus Hinton, 1935, Neoelmis Musgrave, 1935, Phanocerus Sharp, 1882, Potamophilops Grouvelle, 1896, Stegoelmis Hinton, 1939 and Xenelmis Hinton, 1936) known in Brazil as well as three morphotypes designated herein as Genus A, Genus M and Genus X. The genus Hexanchorus is recorded for the first time in the state of São Paulo.     

An aryl β-d-glucosidase of the aquatic fungus Lagenidium giganteum,a parasite of mosquito larvae

Lagenidium giganteum Couch, a watermold parasitic on mosquito larvae possesses an intracellular -d-glucosidase (-d-glucoside glucohydrolase, E. C. 3.2.1.21). The enzyme was purified 132 fold and, with the substrate p-nitrophenyl -d-glucoside, was shown to have the following properties: The pH of optimum enzyme stability and activity lay between 5.5 and 6.0, and the enzyme was inactivated at temperatures above 50°C. The K _m was 4.6×10^-5 M and the Arrhenius activation energy was 8.35 Kcal·mole^-1. Elution from Sephadex G-200 gave an approximate molecular weight of 120000. The enzyme was inhibited by Pb²⁺, Ag²⁺ and Hg²⁺, by glucose and by p-chloromercuribenzoate. The latter inhibition was overcome cy cysteine. Chromatographic studies demonstrated transferase as well as hydrolase properties.This paper represents a portion of a Ph.D. dissertation presented by T. M. McInnis, Jr., to the University of North Carolina at Chapel Hill. The work was supported in part by grants from the American Cancer Society, the University of North Carolina Research Council, the Society of Sigma Xi, and a United States Army Medical Research DevelopmentGrant DADA 17-72-C-2168.     

The isolation and characterization of virulence factors of Cryptococcus spp. from saprophytic sources in the city of Ribeirão Preto, São Paulo, Brazil

Yeasts of the Cryptococcus genus are distributed in nature associated to animal and vegetal organic residues. Occasionally, species other than C. neoformans may be responsible for infectious diseases in human and animals. This study aims to determine the occurrence of Cryptococcus species in the atmosphere and bird droppings in the city of Ribeir?o Preto, S?o Paulo, Brazil, and to evaluate three virulence factors: capsule formation, growth at 37 degrees C and melanin production. We analyzed 86 environmental samples (54 droppings and 32 air). Of the 41 strains isolated, 15 were C. neoformans var. neoformans (12 droppings and 3 air), 15 C. albidus (12 droppings and 3 air), 9 C. laurentii (7 droppings and 2 air) and 2 C. uniguttulatus (from droppings). Capsules were produced by 93.3% of C. neoformans var. neoformans, 66.7% of C. albidus, 88.9% of C. laurentii and 50% (1/2) of C. uniguttulatus. All strains of C. neoformans, 20% of C. albidus and 44.4% of C. laurentii were able to grow at 37 degrees C. The melanin production on DOPA agar was verified in C. neoformans (93.3%), C. albidus (26.7%) and C. laurentii (66.7%). We concluded that different Cryptococcus species coexist in the same ecological niche and they are able to produce virulence factors.     

Cassava shoot infestation by larvae of Neosilba perezi (Romero & Ruppell) (Diptera: Lonchaeidae) in São Paulo State, Brazil

Among the pests of cassava, the shoot fly, Neosilba perezi (Romero & Ruppell), is one of the most prevalent. It attacks mainly the terminal shoots and causes infested plants to produce lateral shoots. Reports on this species are rare or inexistent; thus, the purpose of this study was to assess three different areas for N. perezi infestation. The survey began in March 2008 and finished in February 2009. Fortnightly analyses were performed starting 45 days after planting, calculating the rate of infestation by N. perezi larvae in each study area. The areas were correlated separately for each parameter: fortnightly mean temperature, fortnightly mean rainfall, and plant age. The N. perezi larvae occurrence rate was higher in area 1 - which presented the highest population peaks in autumn and winter. There was only a single population peak in area 2, in winter; and area 3 presented the weakest population peak among the three, in November. The shoot fly population dynamics in the studied region is separately correlated to temperature, rainfall and plant age: temperatures above 23oC, relatively high rainfall and older plants seem to have a negative effect on populations of this insect.     

Description of the male of Tityus kuryi Louren?o, 1997 and notes about males of Tityus stigmurus (Thorell, 1877) and Tityus serrulatus Lutz & Mello, 1922 (Scorpiones,Buthidae)

The male of Tityus kuryi Lourenço, 1997 is described for the first time. Despite being very similar to the female, the male presents more robust metasomal segments. Additionally, the distribution of the sexual populations of another two species of the T. stigmurus complex is reported herein: T. serrulatus Lutz & Mello, 1922 and T. stigmurus (Thorell, 1877). Males of T. serrulatus were, until now, restricted to the Minas Gerais State (Southwestern region of Brazil), and with new records reported here, its known distribution now encompasses the Northeastern region of Brazil. Males of T. stigmurus were previously recorded only for two municipalities in the State of Bahia, and here we present eight new records for Bahia State and one for Pernambuco State. We present a key to related species of the T. stigmurus complex based on morphology and coloration pattern.     

Investigation of biota associated and natural infection by sporulated bacteria in Culicidae larvae from São Paulo state, Brazil

The aim of this study was to investigate the naturally occurring bacteria from Culicidae larvae found in São Paulo state between 2006 and 2008 from the Tiete Ecological Park in the municipality of Caraguatatuba. Bacterial strains were obtained after surface sterilization of larvae followed by thermal treatment and incubation in bacteriological media under laboratory conditions. Identification was determined using cytomorphology, biochemical and physiological tests. Strains were characterized by qualitative evaluation of biological activity against Culicidae larvae, comparing protein profiles obtained by electrophoresis of crystal protoxins, electrophoresis of Lysinibacillus sphaericus isoenzymes, detection of the L₂ fraction of Hemolytic BL enterotoxin and amplification of DNA using Multiplex-PCR to detect HBL, NHE, CytK, BceT and EntFM enterotoxins. The results obtained in this study demonstrated that the endosymbiotic bacterial diversity belonging to Bacillus and related genera associated with Culicidae larvae is limited to a number of species that does not vary throughout the seasons.     

Prevalence of Toxocara infection in schoolchildren from the Butantã region,São Paulo,Brazil

Visceral larva migrans syndrome by Toxocara affects mainly children between 2 and 5 years of age, it is generally asymptomatic, and the seroprevalence varies from 3 to 86% in different countries. A total of 399 schoolchildren from 14 public schools of the Butant? region, S?o Paulo city, Brazil, were evaluated by Toxocara serology (enzyme-linked immunosorbent assay). Epidemiological data to the Toxocara infection obtained from a protocol were submitted to multiple logistic regression analysis for a risk profile definition. Blood was collected on filter paper by finger puncture, with all samples tested in duplicate. Considering titers > or = 1/160 as positive, the seroprevalence obtained was 38.8%. Among infected children, the mean age was 9.4 years, with a similar distribution between genders. A significant association was observed with the presence of onychophagia, residence with a dirty backyard, living in a slum, previous wheezing episodes, school attended, and family income (p < 0.05). All data, except "living in a slum", were considered to be determinant of a risk profile for the acquisition of Toxocara infection. A monthly income > or = 5 minimum salaries represented a protective factor, although of low relevance. Toxocara eggs were found in at least one of the soil samples obtained from five schools, with high prevalence of Toxocara infections, indicating the frequent soil contamination by this agent.     

Influence of mating and feeding on reproduction pattern of haematophagous bug Triatoma williami Galvão,Souza & Lima, 1965 (Hemiptera,Triatominae)

The bloodsucking bug Triatoma williami is the main vector of Trypanosoma cruzi, the etiological agent of Chagas disease in State of Mato Grosso, Brazil. It occurs in sylvatic areas of Brazilian savanna with often records of adult specimens in domestic and peridomestic habitats. We describe, for the first time, the reproductive parameters of T. williami females such as fecundity, fertility and lifespan. We also evaluate whether mating and nourishment status of females of T. williami may modulate the reproductive parameters. As a result, the presence of male corroborated to increase female lifespan from 5.14±1,77 to 34.57±10,86 weeks. The oviposition rate in mated female increased 275% when we compared to the fecundity values in unmated females. Finally, our results indicate that mating is crucial to improve lifespan and fecundity of T. williami females. This characteristic may be related to the environmental conditions under T. williami develops, because the sylvatic ecotopes form small scattered colonies with infrequent access to blood sources.     

The biodiversity and conservation of the birds of São Tomé and Príncipe

At present the majority of the endemic bird species occurring on São Tomé and Príncipe remain common, the rarer species being those largely confined to primary rainforest. In 1990 the dwarf olive ibis (Bostrychia bocagei), São Tomé fiscal shrike (Lanius newtoni), São Tomé short-tail (Amaurocichla bocagii) were rediscovered and in 1991 the São Tomé grosbeak (Neospiza concolor) was seen for the first time since 1888. Lowland primary forest is the only habitat on São Tomé in which all the endemic species are found. Primary forest on Príncipe remains largely unsurveyed since the beginning of the century. Due to the decline in the cocoa industry and poor infra-structure post-independence the extent of secondary forest is probably at its greatest since the late 1800s. This habitat is an important buffer zone against development for the remaining primary forest and also contains important populations of many of the endemic species, in particular the São Tomé scops owl (Otus hartlaubi), São Tomé oriole (Oriolus crassirostris) and giant weaver (Ploceus grandis). On both islands the remaining areas of primary forest need immediate protection and suitable boundaries have been designated under the Zona Ecológica plan. Fortunately, except for an important area around Lagôa Amélia, primary forest is not under immediate threat, although a variety of pressures are likely to increase. In conjunction with the protection of primary forest, a plant for managing the remaining timber resource on the islands will be a vital requirement, particularly if areas of shade forest, an important habitat for endemic bird species and a potentially valuable economic resource, are to be conserved and sustainably managed.     

Temperature and salinity effects on the respiratory metabolism of the first zoeal stage of Macrobrachium holthuisi Genofre & Lobão (decapoda: Palaemonidae)

Oxygen consumption rates of stage I Macrobrachium holthuisi Genofre & Lobão zoeae were measured in 24 different temperature and salinity combinations using Cartesian diver microrespirometers. Metabolic rates varied little with salinity at 15°C while at 20°C a marked elevation occurred in 0 and 35‰ At 25°C, a slight elevation occurred in 0‰; rates remained constant, however, in the other salinities. At 30°C, respiratory rates were similar to those recorded at 25°C except for decreases at 0 and 28‰ salinity. Q₁₀ values in the different salinities were usually highest between 15 and 20°C. Statistical analyses showed that while both temperature, salinity and their interaction significantly influenced larval respiratory rates, temperature had the more pronouced effect. Larval metabolism is salinity independent over the salinity range encountered in the larval biotope (7–21‰) at temperatures of 15–30°C.     

Rickettsia infection in five areas of the state of São Paulo, Brazil

This study investigated rickettsial infection in animals, humans, ticks, and fleas collected in five areas of the state of S?o Paulo. Eight flea species (Adoratopsylla antiquorum antiquorum, Ctenocephalides felis felis, Polygenis atopus, Polygenis rimatus, Polygenis roberti roberti, Polygenis tripus, Rhopalopsyllus lugubris, and Rhopalopsyllus lutzi lutzi), and five tick species (Amblyomma aureolatum, Amblyomma cajennense, Amblyomma dubitatum, Ixodes loricatus, and Rhipicephalus sanguineus) were collected from dogs, cats, and opossums. Rickettsia felis was the only rickettsia found infecting fleas, whereas Rickettsia bellii was the only agent infecting ticks, but no animal or human blood was shown to contain rickettsial DNA. Testing animal and human sera by indirect immunofluorescence assay against four rickettsia antigens (R. rickettsii, R. parkeri, R. felis, and R. bellii), some opossum, dog, horse, and human sera reacted to R. rickettsii with titers at least four-fold higher than to the other three rickettsial antigens. These sera were considered to have a predominant antibody response to R. rickettsii. Using the same criteria, opossum, dog, and horse sera showed predominant antibody response to R. parkeri or a very closely related genotype. Our serological results suggest that both R. rickettsii and R. parkeri infected animals and/or humans in the studied areas.     

Comparative morphology of eggs and nymphs of Triatoma vandae Carcavallo, Jurberg, Rocha, Galvão, Noireau & Lent, 2002 and Triatoma williami Galvão, Souza & Lima, 1965

Eggs and all nymphs of these species were studied employing light microscopy (LM) and scanning electron microscopy (SEM). The major differences observed by LM in the eggs were related to the presence and the distribution of pores on the surface of their chorion. Morphological differences among three nymphal stages (1st, 3rd, and 5th) development of each species were observed. The differential characteristics are chromatic and in the shape of connexival spots. The ultrastructure of the ventral region of the head and the IX, X, and XI abdominal segments (anal tube) of the both species were described demonstrating morphological differences that can be used for diagnosis of the species.     

Ticks collected on birds in the state of São Paulo,Brazil

The present study reports a collection of Amblyomma spp. ticks in birds from several areas of the state of São Paulo, Brazil. A total of 568 tick specimens (404 larvae, 164 nymphs) were collected from 261 bird specimens. From these ticks, 204 (36%) specimens (94 larvae, 110 nymphs) were reared to the adult stage, being identified as Amblyomma longirostre (94 larvae, 90 nymphs), Amblyomma calcaratum (13 nymphs), Amblyomma nodosum (2 nymphs), and Amblyomma cajennense (5 nymphs). Additionally, 39 larvae reared to the nymphal stage and 8 nymphs that died before reaching the adult stage were identified as A. longirostre according to peculiar characters inherent to the nymphal stage of this species: scutum elongate, and hypostome pointed. The remaining 271 larvae and 46 nymphs were identified as Amblyomma sp. Ticks were collected from 51 species of birds distributed in 22 bird families and 6 orders. The order Passeriformes constituted the vast majority of the records, comprising 253 (97%) out of the 261 infested birds. Subadults of A. longirostre were identified from 35 species of Passeriformes, comprising 11 families (Cardinalidae, Dendrocolaptidae, Fringillidae, Furnariidae, Parulidae, Pipridae, Thamnophilidae, Thraupidae, Turdidae, Tyrannidae, and Vireonidae), and from 1 species of a non-passerine bird, a puffbird (Bucconidae). Subadults of A. calcaratum were identified from 5 species of Passeriformes, comprising 5 families (Cardinalinae, Conopophagidae, Pipridae, Thamnophilidae and Turdidae). Subadults of A. nodosum were identified from 2 species of Passeriformes, comprising two bird families (Thamnophilidae and Pipridae). Subadults of A. cajennense were identified from 2 species of non-passerine birds, belonging to 2 different orders (Ciconiiformes: Threskiornithidae, and Gruiformes: Cariamidae). Birds were usually infested by few ticks (mean infestation of 2.2 ticks per bird; range: 1–16). Currently, 82 bird species are known to be infested by immature stages of A. longirostre, with the vast majority [74 (90%)] being Passeriformes. Our results showed that Passeriformes seems to be primary hosts for subadult stages of A. longirostre, A. calcaratum, and A. nodosum. However, arboreal passerine birds seem to be the most important hosts for A. longirostre whereas ground-feeding passerine birds seem to be the most important for both A. calcaratum and A. nodosum. In contrast, the parasitism of birds by subadults of A. cajennense has been restricted to non-passerine birds.     

Spatial Distribution of the Risk of Dengue and the Entomological Indicators in Sumaré, State of S?o Paulo,Brazil

Dengue fever is a major public health problem worldwide, caused by any of four virus (DENV-1, DENV-2, DENV-3 and DENV-4; Flaviviridae: Flavivirus), transmitted by Aedes aegypti mosquito. Reducing the levels of infestation by A. aegypti is one of the few current strategies to control dengue fever. Entomological indicators are used by dengue national control program to measure the infestation of A. aegypti, but little is known about predictive power of these indicators to measure dengue risk. In this spatial case-control study, we analyzed the spatial distribution of the risk of dengue and the influence of entomological indicators of A. aegypti in its egg, larva-pupa and adult stages occurring in a mid-size city in the state of São Paulo. The dengue cases were those confirmed by the city''s epidemiological surveillance system and the controls were obtained through random selection of points within the perimeter of the inhabited area. The values of the entomological indicators were extrapolated for the entire study area through the geostatistical ordinary kriging technique. For each case and control, the respective indicator values were obtained, according with its geographical coordinates and analyzed by using a generalized additive model. Dengue incidence demonstrated a seasonal behavior, as well as the entomological indicators of all mosquito''s evolutionary stages. The infestation did not present a significant variation in intensity and was not a limiting or determining factor of the occurrence of cases in the municipality. The risk maps of the disease from crude and adjusted generalized additive models did not present differences, suggesting that areas with the highest values of entomological indicators were not associated with the incidence of dengue. The inclusion of other variables in the generalized additive models may reveal the modulatory effect for the risk of the disease, which is not found in this study.