Molecular differentiation of Anopheles (Nyssorhynchus) benarrochi and An. (N.) oswaldoi from Southern Colombia

Anopheles (Nyssorhynchus) benarrochi, An. (N.) oswaldoi, and An. (N.) rangeli are the most common anthropophilic mosquitoes in the southern Colombian state of Putumayo. Adult females are most commonly collected in epidemiological studies, and this stage poses significant problems for correct identification, due to overlapping inter-specific morphological characters. Although An. rangeli is easy to identify, the morphological variant of An. benarrochi found in the region and An. oswaldoi are not always easy to separate. Herein we provide a rapid molecular method to distinguish these two species in Southern Colombia. Sequence data for the second internal transcribed spacer (ITS2) region of rDNA was generated for link-reared progeny of An. benarrochi and An. oswaldoi, that had been identified using all life stages. ITS2 sequences were 540 bp in length in An. benarrochi (n = 9) and 531 bp in An. oswaldoi (n = 7). Sequences showed no intra-specific variation and ungapped inter-specific sequence divergence was 6.4%. Species diagnostic banding patterns were recovered following digestion of the ITS2 amplicons with the enzyme Hae III as follows: An. benarrochi (365, 137, and 38 bp) and An. oswaldoi (493 and 38 bp). This polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay provides rapid, accurate, and inexpensive species diagnosis of adult females. This will benefit future epidemiological studies and, as PCR amplification can be achieved using a single mosquito leg, the remaining specimen can be either retained as a morphological voucher or further used in vector incrimination studies. That An. benarrochi comprises a complex of at least two species across Latin America is discussed.

Incrimination of Anopheles (Nyssorhynchus) rangeli and An. (Nys.) oswaldoi as natural vectors of Plasmodium vivax in Southern Colombia

Malaria transmission in the Southern Colombian state of Putumayo continues despite the absence of traditional vector species, except for the presence of Anopheles darlingi near the southeastern border with the state of Amazonas. In order to facilitate malaria vector incrimination in Putumayo, 2445 morphologically identified Anopheles females were tested for natural infection of Plasmodium vivax by ELISA. Specimens tested included An. apicimacula (n = 2), An. benarrochi B (n = 1617), An. darlingi (n = 29), An. mattogrossensis (n = 7), An. neomaculipalpus (n = 7), An. oswaldoi (n = 362), An. peryassui (n = 1), An. punctimacula (n = 1), An. rangeli (n = 413), and An. triannulatus (n = 6). Despite being overwhelmingly the most anthropophilic species in the region and comprising 66.1% of the mosquitoes tested, An. benarrochi B was not shown to be a vector. Thirty-five An. rangeli and one An. oswaldoi were naturally infected with P. vivax VK210. Sequence data were generated for the nuclear second internal transcriber space region of 31 of these 36 vivax positive mosquitoes (86.1%) to confirm their morphological identification. An. oswaldoi is known to be a species complex in Latin America, but its internal taxonomy remains unresolved. Herein we show that the An. oswaldoi found in the state of Putumayo is genetically similar to specimens from the state of Amapá in Brazil and from the Ocama region in the state of Amazonas in Venezuela, and that this form harbors natural infections of P. vivax. That An. rangeli and this member of the An. oswaldoi complex are incriminated as malaria vectors in Putumayo, is a novel finding of significance for malaria control in Southern Colombia, and possibly in other areas of Latin America.

Relationships between microbial activity and soil physical and chemical properties in native and reforested Araucaria angustifolia forests in the state of São Paulo, Brazil

Araucaria angustifolia (Bert.) O. Kuntze is the main component of the Mixed Ombrophilous forest and, in the State of São Paulo, it is associated with a high diversity of soil organisms, essential for the maintenance of soil quality, making the conservation of this ecosystem a major and pressing challenge. The objective of this study was to identify the physical and chemical properties that are most closely correlated with dehydrogenase enzyme activity, basal respiration and microbial biomass under native (NF) and replanted (RF) Araucaria angustifolia forests in three regions of the state of São Paulo, in winter and summer. The main differentiating factors between the areas were also determined. Each forest was represented by three true replications; at each site, from around the araucaria trees, 15 soil samples (0-20 cm) were collected to evaluate the soil physical, chemical and microbiological properties. At the same points, forest litter was sampled to assess mass and chemical properties. The following microbiological properties were evaluated: microbial biomass carbon (MBC), basal respiration (CO2-C), metabolic quotient (Q: CO2), dehydrogenase enzyme activity (DHA) as well as the physical properties (moisture, bulk density, macroporosity and total porosity), soil chemical properties [pH, organic carbon (org-C), P, Ca, K, Mg, Al, H+Al], litter dry mass, and C, N and S contents. The data were subjected to analysis of variance (TWO-WAY: ANOVA). A Canonical Discriminant Analysis (CDA) and a Canonical Correlation Analysis (CCA) were also performed. In the soil under NF, the values of K, P, soil macroporosity, and litter dry mass were higher and Q: CO2 and DHA lower, regardless of the sampling period, and DHA was lower in winter. In the RF areas, the levels of moisture, porosity and Q: CO2 were higher in both sampling periods, and DHA was higher in winter. The MBC was only higher under NF in the summer, while the litter contents of C, N and S were greater in winter. In winter, CCA showed a high correlation of DHA with CO2-C, pH and H+Al, while in the summer org-C, moisture, Mg, pH and litter C were more associated with DHA and CO2-C. The CDA indicated H+Al, available P, total porosity, litter S content, and soil moisture as the most discriminating variables between NF and RF, but moisture was the most relevant, in both seasons and CO2-C only in winter. The combined analysis of CCA and CDA showed that the contribution of the microbiological variables to a differentiation of the areas was small at both samplings, which may indicate that the period after reforestation was long enough to allow an almost complete recovery of the microbial activity.

Climate, soil and tree flora relationships in forests in the state of São Paulo, southeastern Brasil

ABSTRACT - (Climate, soil and tree flora relationships in forests in the state of São Paulo, southteastern Brasil). With the aim of verifying possible influences of abiotic features on the spatial distribution of forest tree species and families, thirteen surveys in the state of São Paulo were selected, representing different conditions (localization at the extreme coordenates and altitudes, succesional stages, surveying methods). By applying Jaccard's Index to the binary matrices of 806 synonymized specific binomina and 79 families (Cronquist's system) phenograms were constructed using the method of the unweighted pair grouping by mathematical average (UPGMA). The species formed two floristic blocks: hygrophyllous (yearly rainfall greater than 2000 mm without dry season) and mesophyllous (yearly rainfall about 1400 mm with variable dry season). The latter was divided in two other groups: the high-altitudinal (median altitudes higher than 750 m, frost average frequency greater than 3 days/year) and low-altitudinal. Both mesophyllous floristic blocks were subdivided according to soil conditions (texture, eutrophism, acid or allic dystrophism, iron content). At the family level the relations were weak, but also showed the soil nutritional status as a possible constraint to the spatial partition of families.

The relationships between selected anthropometric and socio-economic data in schoolchildren from different social strata in Rio de Janeiro, Brazil

The nutritional status according to anthropometric data was assessed in 756 schoolchildren from 5 low-income state schools and in one private school in the same part of Rio de Janeiro, Brazil. The prevalence of stunting and wasting (cut-off point: <90% ht/age and <80% wt/ht) ranged in the public schools from 6.2 to 15.2% and 3.3 to 24.0%, respectively, whereas the figures for the private school were 2.3 and 3.5%, respectively. Much more obesity was found in the private school (18.0%) than in the state schools (0.8 - 6.2%). Nutritional problems seem to develop more severely in accordance with the increasing age of the children. Therefore it appears advisable to assess schoolchildren within the context of a nutritional surveillance system.

Determinants of vaccination after the Colombian health system reform

OBJECTIVE: To assess the effects of individual, household and healthcare system factors on poor children's use of vaccination after the reform of the Colombian health system. METHODS: A household survey was carried out in a random sample of insured poor population in Bogota, in 1999. The conceptual and analytical framework was based on the Andersen's Behavioral Model of Health Services Utilization. It considers two units of analysis for studying vaccination use and its determinants: the insured poor population, including the children and their families characteristics; and the health care system. Statistical analysis were carried out by chi-square test with 95% confidence intervals, multivariate regression models and Cronbach's alpha coefficient. RESULTS: The logistic regression analysis showed that vaccination use was related not only to population characteristics such as family size (OR=4.3), living area (OR=1.7), child's age (OR=0.7) and head-of-household's years of schooling (OR=0.5), but also strongly related to health care system features, such as having a regular health provider (OR=6.0) and information on providers' schedules and requirements for obtaining care services (OR=2.1). CONCLUSIONS: The low vaccination use and the relevant relationships to health care delivery systems characteristics show that there are barriers in the healthcare system, which should be assessed and eliminated. Non-availability of regular healthcare and deficient information to the population are factors that can limit service utilization.

Analysis of genetic relatedness between populations of Aedes aegypti from different geographic regions of São Paulo state, Brazil

RAPD markers have been used for the analysis of genetic differentiation of Aedes aegypti, because they allow the study of genetic relationships among populations. The aim of this study was to identify populations in different geographic regions of the São Paulo State in order to understand the infestation pattern of A. aegypti. The dendrogram constructed with the combined data set of the RAPD patterns showed that the mosquitoes were segregated into two major clusters. Mosquitoes from the Western region of the São Paulo State constituted one cluster and the other was composed of mosquitoes from a laboratory strain and from a coastal city, where the largest Latin American port is located. These data are in agreement with the report on the infestation in the São Paulo State. The genetic proximity was greater between mosquitoes whose geographic origin was closer. However, mosquitoes from the coastal city were genetically closer to laboratory-reared mosquitoes than to field-collected mosquitoes from the São Paulo State. The origin of the infestation in this place remains unclear, but certainly it is related to mosquitoes of origins different from those that infested the West and North region of the State in the 80's.

Spatio-temporal patterns of tuberculosis incidence in Ribeirão Preto, State of São Paulo, southeast Brazil, and their relationship with social vulnerability: a Bayesian analysis

INTRODUCTION: The purpose of this ecological study was to evaluate the urban spatial and temporal distribution of tuberculosis (TB) in Ribeirão Preto, State of São Paulo, southeast Brazil, between 2006 and 2009 and to evaluate its relationship with factors of social vulnerability such as income and education level. METHODS: We evaluated data from TBWeb, an electronic notification system for TB cases. Measures of social vulnerability were obtained from the SEADE Foundation, and information about the number of inhabitants, education and income of the households were obtained from Brazilian Institute of Geography and Statistics. Statistical analyses were conducted by a Bayesian regression model assuming a Poisson distribution for the observed new cases of TB in each area. A conditional autoregressive structure was used for the spatial covariance structure. RESULTS: The Bayesian model confirmed the spatial heterogeneity of TB distribution in Ribeirão Preto, identifying areas with elevated risk and the effects of social vulnerability on the disease. We demonstrated that the rate of TB was correlated with the measures of income, education and social vulnerability. However, we observed areas with low vulnerability and high education and income, but with high estimated TB rates. CONCLUSIONS: The study identified areas with different risks for TB, given that the public health system deals with the characteristics of each region individually and prioritizes those that present a higher propensity to risk of TB. Complex relationships may exist between TB incidence and a wide range of environmental and intrinsic factors, which need to be studied in future research.

Parasite-host relationship between the tambaqui (Colossoma macropomum Cuvier 1818) and ectoparasites, collected from fish farms in the city of Rolim de Moura, State of Rondônia, Western Amazon, Brazil

In this study we investigated the relationship between tambaqui fish (Colossoma macropomumi Cuvier 1818) and parasites in two fish farms (L204S and L180N) in the state of Rondônia, Brazil, during a 1-year period. The objective of the study was to describe the relationships between parasites, hosts and the environment. From the 80 fish specimens collected, 100% were parasitized by at least one parasite species. Seven ectoparasites species were recorded, six of the class Monogenea: Anacanthorus spathulatusi, Mymarothecium spp. (Mymarothecium sp. 1, Mymarothecium sp. 2 and M. viatorum), Notozothecium sp. and Linguadactyloides brinkimanni, classified as dominants, and the copepod Perulernaea gamitanae, classified as subordinate. Despite their high prevalence, the parasites were not abundant. A. spathulatus presented positive and significant correlations between the abundance of parasitism and the standard length of the hosts in the two fish farms; Mymarothecium spp. showed significant correlations, negative in L180N, and positive in L204S; significant positive correlations were observed for Notozothecium sp. in L204S, and for L.brinkimanni in L180N. Young monogeneans were found; these parasites presented a negative correlation in L180N and a significant negative correlation in L204S. The results of the correlation between the relative condition factor (Kn) and the abundance of parasites were not significant for the recorded parasite species. Regarding the hepatosomatic relation (HSR) of fish and the abundance of parasites, Anacanthorus spathulatusi showed a significant negative correlation with the HSR in L180N, and a positive correlation in L204S. Mymarothecium spp. and Notozothecium sp. presented significant positive correlations in L204S. Considering the correlation of the fish splenosomatic relation (SSR) and the abundance of parasites, L. brinkimanni presented significant correlations, positive in L180N and negative in L204S. Despite 100% prevalence, the high water quality contributes to infracommunities with low parasite abundance and good levels of Kn, HSR and SSR, allowing good tambaqui development.

Reproductive aspects of Macrobrachium amazonicum (Decapoda: Palaemonidae) in the State of Amapá, Amazon River mouth

Macrobrachium amazonicum is an indigenous prawn vastly distributed in basins of South America, widely exploited by artisanal fisheries in northern and northeastern Brazil and, with great potential for aquaculture. This study aimed to investigate general aspects of population structure and reproductive characteristics (size at first maturity, fecundity and reproductive output) of M. amazonicum from two important areas to artisanal prawn fishing located at the mouth of the Amazon River, State of Amapá. The specimens were captured using 20 handcrafted traps called "matapi". A number of 5,179 prawns were captured, 2,975 females and 2,195 males resulting in 1.35:1 female to male ratio. Santana Island and Mazagão Velho showed females predominated in the population. A reproductive peak period was observed from January to April/2009 and in December/2010, coinciding with the period of higher rainfall. The recruitment peak occurred in June and July/2009. Egg-bearing females ranged in size (carapace length) from 11.10 to 29.6 mm. Fecundity increased with female size and reached up to 7,417 eggs. This amount of eggs is considered low if compared with other Macrobrachium estuarine species. Mean egg volume increased gradually from 0.121 to 0.24 mm³ during embryogenesis, representing 68.5% of overall increase from Stage I to Stage III. Eggs of M. amazonicum are small; this is typical for Macrobrachium species, which depends on brackish water to complete the larval development. Irrespective of female size, reproductive output of M. amazonicum varied between 4.8 and 21.85% of their body weight into eggs production.

Fecundity of the spider crab Mithraculus forceps (Decapoda, Mithracidae) from the northeastern coast of the state of São Paulo, Brazil

Fecundity is one of the most important parameters in studying the reproductive output of Pleocyemata decapods, especially because of its relationship to the efficiency of population replacement. Knowledge of fecundity provides basic elements for understanding the reproductive strategies, dynamics, and evolution of a given population. The present investigation provides informations on fecundity, egg size, egg loss, and the relationship between fecundity and selected environmental features, for the spider crab Mithraculus forceps (A. Milne Edwards, 1875). Ovigerous crabs were collected each month during 2000, by SCUBA diving, at Couves Island (23º25'25"S, 44º52'03"W) on the northeastern coast of the state of São Paulo, Brazil. A total of 40 ovigerous females with egg in early development (initial stage) and 28 final stage eggs were obtained and analyzed. Mean fecundity, from eggs of the initial stage, was 402.8 ± 240.1 eggs, ranging from 60 to 1,123 eggs. Sizes of females ranged from 9.4 to 14.0 mm carapace width. Mean egg size was 0.56 ± 0.06 mm diameter. A 20.33% rate of egg loss was estimated by comparing the fecundities of batches of eggs in early and late development (40 initial batches and 28 final stages batches). There were no significant relationships between the water temperature or salinity and variations in fecundity. As in most brachyuran species, M. forceps showed a strong conservative relationship between fecundity and body size.

The relative growth and sexual maturity of the freshwater crab Dilocarcinus pagei (Brachyura, Trichodactylidae) in the northwestern region of the state of São Paulo

The aim of the present study was to determine the size at sexual maturity in the freshwater crab Dilocarcinus pagei Stimpson, 1861, from a population located in Mendonça, state of São Paulo, Brazil. The crabs were sampled monthly (July 2005 to June 2007), at Barra Mansa reservoir. The specimens were captured manually or in sieves passed through the aquatic vegetation. The crabs were captured and separated by sex based on morphology of the pleon and on the number of pleopods. The following dimensions were measured: carapace width (CW); carapace length (CL); propodus length (PL); and abdomen width (AW). The morphological analysis of the gonads was used to identify and categorize individuals according to their stage of development. The morphological maturity was estimated based on the analysis of relative growth based on the allometric equation y = ax b. The gonadal maturity was based on the morphology of the gonads by the method CW50 which indicates the size at which 50% of the individuals in the population showed gonads morphologically mature to reproduction. The biometric relationships that best demonstrated the different patterns of growth for the juvenile and adult stages were CW vs. PL for males and CW vs. AW for females (p<0.001). Based on these relationships, the estimated value to morphological sexual maturity was 21.5 mm (CW) in males and 19.7 mm (CW) in females. The determination of the size at sexual maturity and the adjustment of the data based on the logistic curve (CW50) resulted in a size of 38.2 mm for males and 39.4 mm for females (CW). Based on the data obtained for sexual maturity for D. pagei, we can estimate a minimum size for capture of 40 mm (CW). This minimum size allows at least half of the population to reproduce and retains the juveniles and a portion of the adults in the population.

Batflies Parasitic on Some Phyllostomid Bats in Southeastern Brazil: Parasitism Rates and Host-parasite Relationships

Ectoparasitic batflies were studied on 12 species of phyllostomid bats, by making 35 nightly collections of bats using mist nets at the "Panga" Ecological Reservation near Uberlândia, State of Minas Gerais, southeastern Brazil, from August 1989 to July 1990. Eleven species of Streblidae and one of Nycteribiidae were collected on 12 species of bats. Prevalence of ectoparasitic flies was lower than those reported by other authors for the New World and may be the result of the lack of caves in the study area, causing bats to roost in less favorable locations, forming smaller colonies. The fly, Trichobius joblingi Wenzel, was found on Carollia perspicillata (Linnaeus), showing preference for adult male bats. This could be explained by the predominance of males in the bat colonies, and by the fact that females rest in isolation during the reproductive period making them less exposed to the parasites. The streblid flies, Aspidoptera falcata Wenzel and Megistopoda proxima (Séguy), were found on Sturnira lilium (Geoffroy). A. falcata occurred mainly on young and adult females, whereas M. proxima did not show any preferences relative to the reproductive condition of the host. Ecological factors are important in determining differential numbers of parasites occurring on the different sexes, ages and reproductive state of the hosts.

A Mitochondrial DNA Phylogeny Indicates Close Relationships between Populations of Lutzomyia whitmani (Diptera: Psychodidae, Phlebotominae) from the Rain-forest Regions of Amazônia and Northeast Brazil

Phylogenetic analysis of all 31 described mitochondrial (cytochrome b) haplotypes of Lutzomyia whitmani demonstrated that new material from the State of Rondônia, in southwest Amazônia, forms a clade within a lineage found only in the rain-forest regions of Brazil. This rain-forest lineage also contains two other clades of haplotypes, one from eastern Amazônia and one from the Atlantic forest zone of northeast Brazil (including the type locality of the species in Ilhéus, State of Bahia). These findings do not favour recognizing two allopatric cryptic species of L. whitmani, one associated with the silvatic transmission of Leishmania shawi in southeast Amazônia and the other with the peridomestic transmission of Le. braziliensis in northeast Brazil. Instead, they suggest that there is (or has been in the recent past) a continuum of inter-breeding populations of L. whitmani in the rain-forest regions of Brazil.

Population genetic analysis of Colombian Trypanosoma cruzi isolates revealed by enzyme electrophoretic profiles

Although Colombia presents an enormous biological diversity, few studies have been conducted on the population genetics of Trypanosoma cruzi. This study was carried out with 23 Colombian stocks of this protozoa analyzed for 13 isoenzymatic loci. The Hardy-Weinberg equilibrium, the genetic diversity and heterogeneity, the genetic relationships and the possible spatial structure of these 23 Colombian stocks of T. cruzi were estimated. The majority of results obtained are in agreement with a clonal population structure. Nevertheless, two aspects expected in a clonal structure were not discovered in the Colombian T. cruzi stocks. There was an absence of given zymodemes over-represented from a geographical point of view and the presumed temporal stabilizing selective phenomena was not observed either in the Colombian stocks sampled several times through the years of the study. Some hypotheses are discussed in order to explain the results found.