Molecular characterization of Trypanosoma cruzi Mexican strains and their behavior in the mouse experimental model

INTRODUCTION: For a long time, the importance of Chagas disease in Mexico, where many regarded it as an exotic malady, was questioned. Considering the great genetic diversity among isolates of Trypanosoma cruzi, the importance of this biological characterization, and the paucity of information on the clinical and biological aspects of Chagas disease in Mexico, this study aimed to identify the molecular and biological characterization of Trypanosoma cruzi isolates from different endemic areas of this country, especially of the State of Jalisco. METHODS: Eight Mexican Trypanosoma cruzi strains were biologically and genetically characterized (PCR specific for Trypanosoma cruzi, multiplex-PCR, amplification of space no transcript of the genes of the mini-exon, amplification of polymorphic regions of the mini-exon, classification by amplification of intergenic regions of the spliced leader genes, RAPD - (random amplified polymorphic DNA). RESULTS: Two profiles of parasitaemia were observed, patent (peak parasitaemia of 4.6×10(6) to 10(7) parasites/mL) and subpatent. In addition, all isolates were able to infect 100% of the animals. The isolates mainly displayed tropism for striated (cardiac and skeletal) muscle. PCR amplification of the mini-exon gene classified the eight strains as TcI. The RAPD technique revealed intraspecies variation among isolates, distinguishing strains isolated from humans and triatomines and according to geographic origin. CONCLUSIONS: The Mexican T. cruzi strains are myotrophic and belong to group TcI.

Biological behavior of Trypanosoma cruzi stocks obtained from the state of Amazonas, Western Brazilian Amazon, in mice

INTRODUCTION: The biological diversity of circulating Trypanosoma cruzi stocks in the Amazon region most likely plays an important role in the peculiar clinic-epidemiological features of Chagas disease in this area. METHODS: Seven stocks of T. cruzi were recently isolated in the State of Amazonas, Brazil, from humans, wild mammals, and triatomines. They belonged to the TcI and Z3 genotypes and were biologically characterized in Swiss mice. Parasitological and histopathological parameters were determined. RESULTS: Four stocks did not promote patent parasitemia in mice. Three stocks produced low parasitemia, long pre-patent periods, and a patent period of 1 day or oscillating parasitemia. Maximum parasitemia ranged from 1,400 to 2,800 trypomastigotes/0.1mL blood. Mice inoculated with the T. cruzi stocks studied showed low positivity during fresh blood examinations, ranging from 0% to 28.6%. In hemoculture, positivity ranged from 0% to 100%. Heart tissue parasitism was observed in mice inoculated with stocks AM49 and AM61. Stock AM49 triggered a moderate inflammatory process in heart tissue. A mild inflammatory process was observed in heart tissue for stocks AM28, AM38, AM61, and AM69. An inflammatory process was frequently observed in skeletal muscle. Examinations of brain tissue revealed inflammatory foci and gliosis in mice inoculated with stock AM49. CONCLUSIONS: Biological and histopathological characterization allowed us to demonstrate the low infectivity and virulence of T. cruzi stocks isolated from the State of Amazonas.

Eclectic feeding behavior of Lutzomyia (Nyssomyia) intermedia (Diptera, Psychodidae, Phlebotominae) in the transmission area of American cutaneous leishmaniasis, state of Paraná, Brazil

Introduction The blood meal source of sandflies provides valuable information about the vector/host interaction and allows for an understanding of American cutaneous leishmaniasis (ACL) transmission mechanisms. The aim of this study was to identify the blood meal sources of Lutzomyia (Nyssomyia) intermedia in an endemic area of leishmaniasis in Brazil's State of Paraná using a precipitin test. Methods Sandflies were collected in the rural locality of Epitácio Pessoa within the City of Adrianópolis, State of Paraná, in southern Brazil. A total of 864 female sandflies were captured, and 862 (99.8%) were identified as L. intermedia species. However, two unidentified specimens were considered to be part of the genus Lutzomyia. Results Among the females examined, 396 specimens presented reactions to a certain type of tested antiserum, and most (67.9%) reacted to the simple type. These sandflies fed mainly on the blood of birds, opossums, and rodents, but specimens that fed on the blood of humans, dogs, horses, cattle, and cats were also found. Among the cross-reactions found (32.1%), bird/rodent, bird/opossum, bird/dog, bird/human, and horse/dog cross-reactions were the most common. Conclusions These results demonstrate a tendency in the eclectic feeding behavior of L. intermedia and support its potential role as a vector for ACL in the study area.

Description of microsporidia in simulids: molecular and morphological characterization of microsporidia in the larvae of Simulium pertinax Kollar (Diptera: Simuliidae)

IntroductionMicrosporidia constitute the most common black fly pathogens, although the species' diversity, seasonal occurrence and transmission mechanisms remain poorly understood. Infections by this agent are often chronic and non-lethal, but they can cause reduced fecundity and decreased longevity. The objective of this study was to identify microsporidia infecting Simulium (Chirostilbia) pertinax (Kollar, 1832) larvae from Caraguatatuba, State of São Paulo, Brazil, by molecular and morphological characterization.MethodsLarvae were collected at a single point in a stream in a rural area of the city and were kept under artificial aeration until analysis. Polydispyrenia spp. infection was characterized by the presence of at least 32 mononuclear spores measuring 6.9 ± 1.0 × 5.0 ± 0.7µm in persistent sporophorous vesicles. Similarly, Amblyospora spp. were characterized by the presence of eight uninucleate spores measuring 4.5 × 3.5µm in sporophorous vesicles.ResultsThe molecular analysis confirmed the presence of microsporidian DNA in the 8 samples (prevalence of 0.51%). Six samples (Brazilian larvae) were related to Polydispyrenia simulii and Caudospora palustris reference sequences but in separate clusters. One sample was clustered with Amblyospora spp. Edhazardia aedis was the positive control taxon.ConclusionsSamples identified as Polydispyrenia spp. and Amblyospora spp. were grouped with P. simulii and Amblyospora spp., respectively, corroborating previous results. However, the 16S gene tree showed a considerable distance between the black fly-infecting Amblyospora spp. and the mosquito-infecting spp. This distance suggests that these two groups are not congeneric. Additional genomic region evaluation is necessary to obtain a coherent phylogeny for this group.

Size of gastroesophageal varices: its behavior after the surgical treatment of portal hypertension

The size of gastroesophageal varices is one of the most important factors leading to hemorrhage related to portal hypertension. An endoscopic evaluation of the size of gastroesophageal varices before and after different operations for portal hypertension was performed in 73 patients with schistosomiasis, as part of a randomized trial: proximal splenorenal shunt (PSS n=24), distal splenorenal shunt (DSS n=24), and esophagogastric devascularization with splenectomy (EGDS n=25). The endoscopic evaluation was performed before and up to 10 years after the operations. Variceal size was graded according to Palmer's classification: grade 1 -- up to 3 mm, grade 2 -- from 3 to 6 mm, grade 3 -- greater than 6 mm, and were analyzed in four anatomical locations: inferior, middle or superior third of the esophagus, and proximal stomach. The total number of points in the pre-operative grading minus the number of points in the post-operative grading gave a differential grading, allowing statistical comparison among the surgical groups. Good results, in terms of disappearance or decrease of variceal size, were observed more frequently after PSS than after DSS or EGDS - 95.8%, 83.3%, and 72%, respectively. When differential grading was analyzed, a statistically significant difference was observed between PSS and EGDS, but not between proximal and distal splenorenal shunts. In conclusion, shunt surgeries were more efficient than devascularization in diminishing variceal size.

Bionomia e chave para imaturos e adultos de Simulium (Diptera: Simuliidae) na Amazônia Central, Brasil

O presente estudo teve como objetivos apresentar informações bionômicas e de distribuição sobre larvas e pupas de simulídeos na Amazônia Central, delimitada aqui como Manaus (AM) e municípios nos arredores (Presidente Figueiredo, Itacoatiara, Careiro da Várzea, Manacapum, Novo Airão), assim como apresentar chaves para todos os estágios das espécies presentes na área. Amostrou-se 79 locais, que foram caracterizados com os seguintes parâmetros físico-químicos: largura, profundidade, correnteza, vazão, pH, temperatura, vegetação predominante e presença de represamento d'água. Onze espécies foram coletadas nos igarapés dessa área (Simulium cauchense Floch & Abonnenc, Simulium daltanhani Hamada & Adler, Simulium goeldii Cerqueira & Nunes de Mello, Simulium iracouboense Floch & Abonnenc, Simulium maroniense Floch & Abonnenc, Simulium perflavum Roubaud, Simulium quadrifidum Lutz, Simulium rorotaense Floch & Abonnenc, Simulium trombetense Hamada, Py-Daniel & Adler, Simulium "6-B1" e Simulium "A"). Simulium argentiscutum Shelley & Luna Dias foi coletada apenas como fêmeas adultas, picando seres humanos no município de Itacoatiara. Chaves para larvas, pupas e adultos fêmeas e machos, com ilustrações de caracteres relevantes, das 12 espécies presentes na Amazônia Central, são fornecidas. Os imaturos de Simuliidae se distribuíram de maneira distinta, formando grupos de espécies diferenciados pelas características medidas no criadouro.

Simuliidae (Diptera, Culicomorpha) no Brasil - XII sobre o gênero Thyrsopelma Enderlein, 1934: descrição de T. jeteri Sp.N., considerações sobre o cibário e chave para as pupas

É acrescentado um novo caráter pupal para a espécie T. hirtipupa (Lutz,1910). É evidenciada a existência de dentes no cibário da fêmea de T. guianense (Wise,1911), principal espécie envolvida na transmissão da filaria Onchocerca volvulus (Leuckart,1893) no foco da região amazônica. É proposta uma nova relação para os artículos antenais da larva de T. guianense. É descrita uma nova espécie para o gênero Thyrsopelma Enderlein,1934, T. jeteri sp.n., proveniente da bacia hidrográfica do rio Uruguai, no Estado do Rio Grande do Sul. São apresentados os parâmetros físico-químicos do criadouro de onde provem o material tipo. É apresentada uma chave para diferenciação das pupas das espécies constantes neste gênero.

Simuliidae (Diptera:Culicomorpha) no Brasil XI - Shelleyellum gênero novo

É caracterizado um novo gênero neotropical de simulídeo, com base nas espécies previamente incluídas no "Grupo Siolii", do gênero Psaroniocompsa. Chaves de identificação para imaturos são apresentadas. O gênero Shelleyellum fica constituído pelas espécies: S. damascenoi (Py-Daniel,1988), S. guaporense (Py-Daniel, 1989), S. lourencoi (Py-Daniel, 1988), S. siolli (Py-Daniel, 1988), S. tergospinosum (Hamada, 2000). O gênero é caracterizado, principalmente, pela de carena longitudinal nos distímeros dos machos e a presença de tubérculos tegumentares nas larvas.

The influence of climatic parameters in the haematophagic daily activity of Cerqueirellum argentiscutum (Shelley & Luna Dias) (Diptera: Simuliidae) in Amazonas, Brazil

We studied the influence of climatic parameters in the daily haematophagic activity of Cerqueirellum argentiscutum from September/1999 to August/2000. The bite activity observed was different according to the annual rain precipitation (dry and rainy seasons). Humidity and temperature were the factors that most influenced it in both periods. During the dry season, it was greater in the beginning of the morning, showing a positive association with the humidity. However, during the rainy season, it was negatively related to that same factor. When wind speed was higher than 10 Km.h-1, it was reduced abruptly. Light intensity, atmospheric pressure and cloudiness seemed to act as secondary factors in the daily abundance of C. argentiscutum.

Haematophagic behavior in laboratory of Lutzomyia cruzi (Mangabeira) (Diptera: Psychodidae) in relation to three mammalian blood sources in Manaus, Brazil

The sand fly Lutzomyia cruzi is considered as one of vectors of visceral leishmaniasis in Brazil. This work examined optimum feeding age, feeding time, host preference, fecundity rates, and female blood meal volume taken by single females from a closed colony of L. cruzi. Mean feeding time was longer on hamsters, 6.6 minutes, than on humans, 5.7 minutes. 49.1% of the 48h-old flies fed on humans and 43.3% of 72h-old flies fed on hamsters. Of a total of 120 females, 61% fed on humans and 25% fed on hamsters. Total fecundity was significantly higher in females fed on hamster than on human or opossum. Laboratory-reared L. cruzi females fed earlier, more promptly, and preferably on humans than on hamsters when offered these blood-meal sources simultaneously. The blood-meal volume is higher in females fed on hamsters than other hosts (human and opossum).

Simuliidae (Diptera, Culicomorpha) no Brasil: XIII sobre Inaequaliumlundi sp.n.

É descrita uma nova espécie para o gênero Inaequalium (Coscarón & Wygodzinsky, 1984), Inaequaliumlundi sp.n. É proposto um novo grupo de espécies dentro de Inaequalium. Os imaturos de Inaequaliumlundi sp.n. foram coletados em cursos d'água em uma altitude de 2.800 m no complexo geológico denominado Pico da Neblina, Amazonas, Brasil.

Cladistic analysis of the Neotropical genera Cerqueirellum Py-Daniel, 1983, Coscaroniellum Py-Daniel, 1983 and Shelleyellum Py-Daniel & Pessoa, 2005 (Diptera: Simuliidae)

Females of simuliid black flies are haematophagous insects and vectors of several pathogenic agents of human diseases such as the filarial worms Mansonella ozzardi and Onchocerca volvulus. The genus Cerqueirellum is one of the most important groups of vectors of mansonellosis and onchocerciasis diseases in South America, and the genera Coscaroniellum and Shelleyellum are phylogenetically close to Cerqueirellum. There is not yet an agreement among authors about the generic classification of the species which compose these three genera, being all lumped by some taxonomists within Psaroniocompsa. A cladistic analysis of all species of Coscaroniellum, Cerqueirellum, and Shelleyellum, based on 41 morphological characters were done. Species closely related to Cerqueirellum were included in the analysis. The genera Cerqueirellum, Coscaroniellum and Shelleyellum were demonstrated as consistent basal entities and well-defined monophyletic clades.

A new species of Cerqueirellum Py-Daniel, 1983 (Diptera: Simuliidae) and proven new vector of mansonelliasis from the Ituxi River, Amazon basin, Brazil

A new species of the genus Cerqueirellum Py-Daniel, 1983 (Diptera: Simuliidae) is described. The adults are similar to the species C. oyapockense (Floch & Abonnenc, 1946) and C. roraimense (Nunes de Mello, 1974), of which the females are similar, and the males present discrete differences. The main differences of this new species to others of the genus Cerqueirellum are the integument of the larva recovered from stout spines and long cephalic trichomes in the pupa. Some females were infected with Mansonella ozzardii (Manson, 1897) (Nematoda, Onchocercidae) and probably transmit mansonelliasis in the Ituxi river, state of Amazonas, Brazil.