Standardization of in-house polymerase chain reaction for the identification of Mycobacterium tuberculosis at the reference tropical disease hospital in the State of Goiás, Brazil

This study compares smear, growth in Lowenstein-Jensen medium, and in-house polymerase chain reaction (PCR) techniques for the detection of Mycobacterium tuberculosis. A total of 72 specimens from 72 patients with clinical symptoms of tuberculosis, including 70 sputum and two bronchial aspirate samples, were tested in parallel by smear, culture, and in-house PCR techniques. From these, 48 (66.6%) were negative by the 3 methods, 2 (2.8%) were smear positive and negative by culture and in-house PCR, 11 (15.3%) were both smear and culture negative, and in-house PCR positive, 7 (9.7%) were positive by the 3 methods, 2 (2.8%) were positive by smear and culture, and negative by PCR, 2 (2.8%) were positive by culture and PCR, but smear negative. After the resolution of discrepancies in PCR results, the sensitivity and specificity for in-house PCR technique to M. tuberculosis relative to the culture, were 81.8% and 81.9%, respectively. These results confirm that this method, in-house PCR, may be a sensitive and specific technique for M. tuberculosis detection, occurring in both positive and negative smear and negative cultures.

Isolation and characterization of the full-length cDNA encoding a member of a novel cytochrome p450 family (CYP320A1) from the tropical freshwater snail, Biomphalaria glabrata, intermediate host for Schistosoma mansoni

Cytochrome p450s (cyp450s) are a family of structurally related proteins, with diverse functions, including steroid synthesis and breakdown of toxins. This paper reports the full-length sequence of a novel cyp450 gene, the first to be isolated from the tropical freshwater snail Biomphalaria glabrata, an important intermediate host of Schistosoma mansoni. The nucleotide sequence is 2291 bp with a predicted amino acid sequence of 584aa. The sequence demonstrates conserved cyp450 structural motifs, but is sufficiently different from previously reported cyp450 sequences to be given a new classification, CYP320A1. Initially identified as down-regulated in partially resistant snails in response to S. mansoni infection, amplification of this gene using RT-PCR in both totally resistant or susceptible snail lines when exposed to infection, and all tissues examined, suggests ubiquitous expression. Characterization of the first cyp450 from B. glabrata is significant in understanding the evolution of these metabolically important proteins.

The feature of papers and citation analysis of eleven journals in tropical medicine indexed by Science Citation Index Expanded

To determine the features of papers, authors, and citation of eleven journals in tropical medicine indexed by Science Citation Index Expanded, the database of the Institute for Scientific Information, we analyzed original articles, editorials, reviews, corrections, letters, biographies, and news published in these journals. The results show that these journals covered 107 countries or regions on six continents. The average number of reference was 23.05, with 87.89% of the references from periodicals. The Price Index was 31.43% and the self-citing rate was 7.02%. The references in the first 20 journals ranked by the amount of citation accounted for 36.71% of the total citations. Brazil, United States, India, and England are more advanced in tropical medicine research. The conclusion is that these journals covered most research done in these countries or regions. Most researches were done by cooperation of the researchers, but many of the publications used outdated articles and should include newer information.

Lack of tax diversity for tropical spastic paraparesis/human T-cell lymphotropic virus type 1 (HTLV-I) associated myelopathy development in HTLV-I-infected subjects in São Paulo, Brazil

The product of human T-cell lymphotropic virus type 1 (HTLV-1) tax gene has a transactivating effect of the viral and cellular gene expression. Genetic variations in this gene have been correlated with differences in clinical outcomes. Based upon its diversity, two closely related substrains, namely tax A and tax B, have been described. The tax A substrain has been found at a higher frequency among human T-cell leukemia virus type 1 (TSP/HAM) patients than among healthy HTLV-I-infected asymptomatic subjects in Japan. In this study, we determined the distribution of tax substrains in HTLV-I-infected subjects in the city of São Paulo, Brazil. Using the ACCII restriction enzyme site, we detected only tax A substrain from 48 TSP/HAM patients and 28 healthy HTLV-I carriers. The sequenced tax genes from nine TSP/HAM patients and five asymptomatic HTLV-I carriers showed a similar pattern of mutation, which characterizes tax A. Our results indicate that HTLV-I tax subtypes have no significant influences on TSP/HAM disease progression. Furthermore, monophyletic introduction of HTLV-I to Brazil probably occurred during the African slave trade many years ago.

Molecular and morphological characterization of heterorhabditid entomopathogenic nematodes from the tropical rainforest in Brazil

Despite massive losses of primary forest, the Amazonian rainforest remains an extremely rich source of biodiversity. In recent years, entomopathogenic nematodes (EPNs) have been isolated from soil in various parts of the world and used successfully as biological control agents against numerous insect pests. Therefore, a sampling in the rainforest of Monte Negro, Rondônia, Brazil was conducted with the aim of discovering new strains and/or species of EPNs for future development as biological control agents. From 156 soil samples taken at nine collecting sites, 19 isolates were obtained, all of them belonging to the genus Heterorhabditis. Four strains were subjected to detailed morphological and molecular evaluation. Based on morphometrics and internal transcribed spacer (ITS) sequence data, the strains LPP1, LPP2 and LPP4 were identified as Heterorhabditis indica, whereas LPP7 was considered Heterorhabditis baujardi. Comparative analysis of the ITS1 sequence of H. indica and H. baujardi isolates showed a polymorphic site for the restriction enzyme Tth 111 that could be used to distinguish the two species. Consequently, strains LPP1, LPP2, LPP3, LPP4, and LPP9 were identified as H. indica, whereas LPP5, LPP7, LPP8 and LPP10 were identified as H. baujardi.

Diversity and microdistribution of black fly (Diptera: Simuliidae) assemblages in the tropical savanna streams of the Brazilian cerrado

We describe the abiotic factors affecting the distribution of black flies at a microhabitat scale, rather than at the regional scale usually present in the literature on the Neotropics. Black fly larvae were sampled from the Tocantins River and three tributaries, located in the Brazilian savanna (state of Tocantins, Brazil) during six bi-monthly sampling periods from October 2004-August 2005. At each sampling site, 15 random quadrats (30 x 30 cm) were sampled each period and for each quadrat were determined mean water velocity, predominant substrate type (rocks, riffle litter or riparian vegetation) and depth detrended correspondence analysis (DCA) was used to determine associations with current velocity, whereas correspondence analysis (CA) was used to estimate site specific current velocity associations. Canonical correspondence analysis (CCA) was used to identify general microhabitat associations. The CCA showed that most species had a trend towards riffle litter, except for Simulium nigrimanum associated with rocky substrate and Simulium cuasiexiguum associated with riparian vegetation. The DCA showed a well defined pattern of water velocity associations. The CA revealed that the species showed different speed associations from one site to another, suggesting different competitive pressures resulting in the occurrence of different realized niches.

Upregulation of hsa-miR-125b in HTLV-1 asymptomatic carriers and HTLV-1-associated myelopathy/tropical spastic paraparesis patients

The retrovirus human T lymphotropic virus type 1 (HTLV-1) promotes spastic paraparesis, adult T cell leukaemia and other diseases. Recently, some human microRNAs (miRNAs) have been described as important factors in host-virus interactions. This study compared miRNA expression in control individuals, asymptomatic HTLV-1 carriers and HTLV-1 associated myelopathy (HAM)/tropical spastic paraparesis patients. The proviral load and Tax protein expression were measured in order to characterize the patients. hsa-miR-125b expression was significantly higher in patients than in controls (p = 0.0285) or in the HAM group (p = 0.0312). Therefore, our findings suggest that miR-125b expression can be used to elucidate the mechanisms of viral replication and pathogenic processes.

Leprosy, a neglected disease that causes a wide variety of clinical conditions in tropical countries

Leprosy is an ancient disease that remains endemic and continues to be a major public health problem in some tropical countries, where it has been internationally recognized as being linked to the underdevelopment conditions. The natural course of the disease covers a wide variety of clinical conditions with systemic involvement. In this paper, we review the findings obtained in studies of the pathological mechanisms of leprosy, including a survey of the literature and of our own work. The understanding and control of the wide variety of clinical conditions should help improve patient care and thus prevent the onset of physical impairment and the stigma of the disease.

Comparison of automatic traps to capture mosquitoes (Diptera: Culicidae) in rural areas in the tropical Atlantic rainforest

In several countries, surveillance of insect vectors is accomplished with automatic traps. This study addressed the performance of Mosquito Magnet® Independence (MMI) in comparison with those of CDC with CO2 and lactic acid (CDC-A) and CDC light trap (CDC-LT). The collection sites were in a rural region located in a fragment of secondary tropical Atlantic rainforest, southeastern Brazil. Limatus durhami and Limatus flavisetosus were the dominant species in the MMI, whereas Ochlerotatus scapularis was most abundant in CDC-A. Culex ribeirensis and Culex sacchettae were dominant species in the CDC-LT. Comparisons among traps were based on diversity indices. Results from the diversity analyses showed that the MMI captured a higher abundance of mosquitoes and that the species richness estimated with it was higher than with CDC-LT. Contrasting, difference between MMI and CDC-A was not statistically significant. Consequently, the latter trap seems to be both an alternative for the MMI and complementary to it for ecological studies and entomological surveillance.

Effectiveness of Mosquito Magnet® trap in rural areas in the southeastern tropical Atlantic Forest

Traps are widely employed for sampling and monitoring mosquito populations for surveillance, ecological and fauna studies. Considering the importance of assessing other technologies for sampling mosquitoes, we addressed the effectiveness of Mosquito Magnet® Independence (MMI) in comparison with those of the CDC trap with CO2 and Lurex3® (CDC-A) and the CDC light trap (CDC-LT). Field collections were performed in a rural area within the Atlantic Forest biome, southeastern state of São Paulo, Brazil. The MMI sampled 53.84% of the total number of mosquitoes, the CDC-A (26.43%) and CDC-LT (19.73%). Results of the Pearson chi-squared test (χ2) showed a positive association between CDC-LT and species of Culicini and Uranotaeniini tribes. Additionally, our results suggested a positive association between CDC-A and representatives of the Culicini and Aedini tribes, whereas the MMI was positively associated with the Mansoniini and Sabethini as well as with Anophelinae species. The MMI sampled a greater proportion (78.27%) of individuals of Anopheles than either the CDC-LT (0.82%) or the CDC-A traps (20.91%). Results of the present study showed that MMI performed better than CDC-LT or CDC-A in sampling mosquitoes in large numbers, medically important species and assessing diversity parameters in rural southeastern Atlantic Forest.

Molecular diagnosis of strongyloidiasis in tropical areas: a comparison of conventional and real-time polymerase chain reaction with parasitological methods

This study aimed to evaluate the use of conventional polymerase chain reaction (cPCR) and real-time quantitative PCR (qPCR) in the diagnosis of human strongyloidiasis from stool samples in tropical areas. Stool samples were collected from individuals and were determined to be positive for Strongyloides stercoralis (group I), negative for S. stercoralis (group II) and positive for other enteroparasite species (group III). DNA specific to S. stercoralis was found in 76.7% of group I samples by cPCR and in 90% of group I samples by qPCR. The results show that molecular methods can be used as alternative tools for detecting S. stercoralis in human stool samples in tropical areas.

Gall inducing arthropods from a seasonally dry tropical forest in Serra do Cipó, Brazil

Highly diverse forms of galling arthropods can be identified in much of southeastern Brazil's vegetation. Three fragments of a Seasonally Dry Tropical Forest (SDTF) located in the southern range of the Espinhaço Mountains were selected for study in the first survey of galling organisms in such tropical vegetation. Investigators found 92 distinct gall morphotypes on several organs of 51 host plant species of 19 families. Cecidomyiidae (Diptera) was the most prolific gall-inducing species, responsible for the largest proportion of galls (77%) observed. Leaves were the most frequently galled plant organ (63%), while the most common gall morphotype was of a spherical shape (30%). The two plant species, Baccharis dracunculifolia (Asteraceae) and Celtis brasiliensis (Cannabaceae), presented the highest number of gall morphtypes, displaying an average of 5 gall morphotypes each. This is the first study of gall-inducing arthropods and their host plant species ever undertaken in a Brazilian SDTF ecosystem. Given the intense human pressure on SDTFs, the high richness of galling arthropods, and implied floral host diversity found in this study indicates the need for an increased effort to catalogue the corresponding flora and fauna, observe their intricate associations and further understand the implications of such rich diversity in these stressed and vulnerable ecosystems.

Attractivity of omnivore, carnivore and herbivore mammalian dung to Scarabaeinae (Coleoptera, Scarabaeidae) in a tropical Atlantic rainforest remnant

In this study, performed in a remnant of Brazilian Atlantic Forest, three types of dung from animals with distinct alimentary habits were utilized, in order to verify possible differences of attractivity of these dungs to the Scarabaeinae and the influence of seasonality in the attractivity. Three habitats were sampled: edge, clearing and forest core, each with 40 pitfall traps. A total of 2,137 beetles were collected from August 2005 to July 2006. Canthidium sp. 1 (43%) and Dichotomius sericeus (41%) were the most abundant species. From the total number of beetles collected, 80.5% were attracted to human dung, 11% to jaguar dung, 7.8% to waterbuck dung and 0.7% to the control. The species Canthidium sp.1, Canthidium sp. 2, Ateuchus sp., Canthon nigripenne, Canthonella sp. and D. sericeus came to all three bait types. Eight species were found in the baits with human dung, where Canthidium sp.1 (49%) and D. sericeus (39%) were the most common. A significant difference in attractiveness of the different baits was observed; the highest abundance found in traps baited with human dung (F = 36.59; g.l. = 3; p < 0.0001). A significant difference in richness was observed between rainy and dry seasons (F = 12.29; g.l. = 1; p < 0.001), the highest richness found in the dry season.

Synanthropy and ecological aspects of Muscidae (Diptera) in a tropical dry forest ecosystem in Colombia

The synanthropic index and other ecological aspects of the Muscidae family were evaluated through simultaneous monthly sampling in three different environments (urban, rural and forest) using van Someren-Rydon traps baited with human faeces, chicken viscera and decomposing fish and onion. Four traps were set up in each environment (one per bait item) for 48 hours per month, with samples taken every 12 hours. A total of 5726 specimens were collected, belonging to 19 species and 13 genera. Brontaea normata (+99,9), Brontaea quadristigma (+96,9), Synthesiomyia nudiseta (+96,5), Ophyra aenescens (+96,2), Musca domestica (+95,7) and Atherigona orientalis (+93,8) had the highest synanthropic indices, showing a marked preference for human environments. The most abundant species were B. normata (24,31%), Biopyrellia bipuncta (20,60%) and Pseudoptilolepis nigripoda (15,82%), the latter two showed a preference for uninhabited areas. A total of 11 new records for Colombia were found: Ophyra aenescens, Cyrtoneuropsis pararescita, Morellia basalis, Neomuscina dorsipuncta, Biopyrellia bipuncta, Pseudoptilolepis nigripoda, Neomuscina instabilis, Neomuscina currani, Polietina orbitalis, Neomuscina pictipennis and Cyrtoneuropsis maculipennis. Except for the first four species, the remainder presented negative synantrophy indexes (from minor to major), which would allow to use them as ecological indicators of the disturbance degree of dry forests in Colombia.