Further evidences on a new diagnostic approach for monitoring human Leishmania (L.) infantum chagasi infection in Amazonian Brazil.

This was a prospective study carried out during a period over 2 years (May/2006-September/2008) with a cohort of 1,099 individuals of both genders, aged 1 year old and older, from an endemic area of American visceral leishmaniasis (AVL) in Pará state, Brazil. The object was to analyze the prevalence and incidence of human Leishmania (L.) infantum chagasi infection as well as the dynamics evolution of its clinical-immunological profiles prior identified: (1) asymptomatic infection (AI); (2) symptomatic infection (SI = AVL); (3) sub-clinical oligosymptomatic infection (SOI); (4) sub-clinical resistant infection (SRI) and; (5) indeterminate initial infection (III). The infection diagnosis was performed by using both the indirect fluorescent antibody test and leishmanin skin test with amastigotes and promastigotes antigens of L. (L.) i. chagasi, respectively. A total of 187 cases of infection were recorded in the prevalence (17%), 117 in the final incidence (6.9%), and 304 in the accumulated prevalence (26.7%), which provided the following distribution into the clinical-immunological profiles: AI, 51.6%; III, 22.4%; SRI, 20.1%; SOI, 4.3%; and SI (=AVL), 1.6%. The major finding regarding the dynamics evolution of infection was concerned to III profile, from which the cases of infection evolved to either the resistant profiles, SRI (21 cases, 30.8%) and AI (30 cases, 44.1%), or the susceptible SI (=AVL; 1 case, 1.5%); the latter 16 cases remained as III till the end of the study. These results provided the conclusion that this diagnostic approach may be useful for monitoring human L. (L.) i. chagasi infection in endemic area and preventing the high morbidity of severe AVL cases.

Proven and putative vectors of American cutaneous leishmaniasis in Brazil: aspects of their biology and vectorial competence.

The aim of the present review is to give relevant information on aspects of the biology and ecology, including the vectorial competence of Lutzomyia sand fly species suggested as vectors of American cutaneous leishmaniasis in Brazil. The disease, due to Leishmania (Viannia) braziliensis, has been registered in most municipalities in all the Brazilian states and its transmission is associated with more than one sand fly species in each geographical region. A variety of Leishmania species can be found in the Amazon basin, where different epidemiological chains have been detected with the participation of different phlebotomine vectors. Finally, a discussion is presented on some sand fly species found naturally infected by Leishmania, but for which there is as yet no evidence regarding their epidemiological importance.

Ultrastructural study of the gametocytes and merogonic stages of Fallisia audaciosa (Haemosporina: Garniidae) that infect neutrophils of the lizard Plica umbra (Reptilia: Iguanidae).

Little is known regarding the ultrastructure of the genus Fallisia (Apicomplexa: Haemosporina: Garniidae). This report describes the fine structure of some developmental stages of Fallisia audaciosa that infect neutrophils in the peripheral blood of the Amazonian lizard Plica umbra (Reptilia: Iguanidae). The parasites lie within a parasitophorous vacuole and exhibit the basic structures of members of the Apicomplexa, such as the pellicle and the cytostome. Invaginations of the inner membrane complex were seen in the gametocytes and may be concerned with nutrition. The meronts were irregularly shaped before division, a feature unusual among members of the Apicomplexa. The unusual presence of a parasitic protozoan within neutrophils, in some way interfering with or modulating the microbicidal activity of such cells, is discussed.

Natural Leishmania (Viannia) spp. infections in phlebotomine sand flies (Diptera: Psychodidae) from the Brazilian Amazon region reveal new putative transmission cycles of American cutaneous leishmaniasis.

In Amazonian Brazil the etiological agents of American cutaneous leishmaniasis (ACL) belong to at least seven Leishmania species but little is known about the putative phlebotomine sand fly vectors in different biomes. In 2002-2003 a survey of the phlebotomine fauna was undertaken in the "Floresta Nacional do Tapajós", Belterra municipality, in the lower Amazon region, western Pará State, Brazil, where we recently confirmed the presence of a putative hybrid parasite, L. (V.) guyanensis × L. (V.) shawi shawi. Sand flies were collected from Centers for Disease Control (CDC) light traps, Shannon traps and by aspiration on tree bases. Females were dissected and attempts to isolate any flagellate infections were made by inoculating homogenized midguts into Difco B(45) medium. Isolates were characterized by monoclonal antibodies and isoenzyme electrophoresis. A total of 9,704 sand flies, belonging to 68 species or subspecies, were collected. Infections were found in the following sand flies: L. (V.) naiffi with Psychodopygus hirsutus hirsutus (1) and Ps. davisi (2); and L. (V.) shawi shawi with Nyssomyia whitmani (3) and Lutzomyia gomezi (1). These results provide strong evidence of new putative transmission cycles for L. (V.) naiffi and L. (V.) s. shawi.

Ultrastructural aspects of Fallisia effusa (Haemosporina: Garniidae) in thrombocytes of the lizard Neusticurus bicarinatus (Reptilia: Teiidae).

The fine structure of the different stages of the Fallisia effusa (Haemosporina: Garniidae), infecting the thrombocytes of the semi-aquatic Amazonian lizard Neusticurus bicarinatus (Reptilia: Teiidae) is described. Gametocytes, meronts, and merozoites of Fallisia effusa were found within a parasitophorous vacuole (PV). Multiple infections of micro- and macrogametocytes were observed. A circumferential coil of microtubules was seen in the cytoplasm of the infected host cell and this microtubule array was pronounced in cells harboring gametocytes. A deep invagination of the inner membrane complex of gametocytes may be involved in nutrition. The non-pigmented parasites underwent both merogony and gametogony in thrombocytes of the peripheral blood. No infection of the erythrocytes was observed. These observations confirm that Fallisia effusa displays characteristic features distinguishing it from other members of the Haemosporidian families, and that it has the ability to modulate microtubule assembly.

Comparative studies of populations of Lutzomyia umbratilis (Diptera: Psychodidae) in Brazil and Venezuela.

We analyzed 64 quantitative and qualitative morphological characters in different populations of Lutzomyia (Nyssomyia) umbratilis Ward & Fraiha from Venezuela (Gran Sabana, Bolívar/BO) and Brazil (Serra do Navio, Amapá/AP Manaus, Amazonas/AM, Monte Dourado, Pará/ PA, and Peixoto de Azevedo, Mato Grosso/MT). Based on an analysis of qualitative characters, no differences were observed that could distinguish between the populations. Parametric tests were used to verify differences between sample means and variance homogeneity. The analysis-of-variance (ANOVA) test showed the existence of differences between character means in the five populations. In the graphic representation of the Student-Newman-Keuls test (Student-Newman-Keuls), we observed that 77% of the analyzed characters displayed disjunctions between the Venezuelan and Brazilian populations. Considering only the Brazilian population averages, we concluded that they are statistically different. When the Student-Newman-Keuls test was applied, no pattern was found that could separate them. However, in part, the phenetic analysis showed some evidences of heterogeneity in MT population.

Fine structure of erythrocytic stages of a Plasmodium tropiduri-like malaria parasite found in the lizard Kentropyx calcarata (Teiidae) from north Brazil.

The fine structure is described of the merogonic stages and gametocytes of a Plasmodium tropiduri Aragão et Neiva, 1909-like parasite infecting the teiid lizard Kentropyx calcarata Spix from North Brazil. The trophozoites are bordered by two membranes, and with growth a pellicle is formed by the addition of an inner, thick double layer and fragmented membrane. The same type of inner membrane occurs in the pellicle of the merozoites differentiating from the meronts. Merozoites contained a large electron-dense body, sometimes seen to be embraced by a tubular mitochondrion with a dense matrix. Micro- and macrogametocytes are bounded by a double membrane, closely apposed by the detached wall of the parasitophorous vacuole. Both contain osmiophilic bodies. The microgametocyte contains an electron-dense aggregate, and the macrogametocyte has a large mitochondrion and a complex of tubuli and cisternae. These features are compared with those described in other malarial parasites.

Phenotypic characterization of Leishmania spp. causing cutaneous leishmaniasis in the lower Amazon region, western Pará state, Brazil, reveals a putative hybrid parasite, Leishmania (Viannia) guyanensis × Leishmania (Viannia) shawi shawi.

We phenotypically characterized 43 leishmanial parasites from cutaneous leishmaniasis by isoenzyme electrophoresis and the indirect immunofluorescence antibody test (23 McAbs). Identifications revealed 11 (25.6%) strains of Leishmania (V.) braziliensis, 4 (9.3%) of L. (V.) shawi shawi, 7 (16.3%) of L. (V.) shawi santarensis, 6 (13.9%) of L. (V.) guyanensis and L. (V.) lainsoni, 2 (4.7%) of L. (L.) amazonensis, and 7 (16.3%) of a putative hybrid parasite, L. (V.) guyanensis/L. (V.) shawi shawi. McAbs detected three different serodemes of L. (V.) braziliensis: I-7, II-1, and III-3 strains. Among the strains of L. (V.) shawi we identified two populations: one (7 strains) expressing the B19 epitope that was previously considered to be species-specific for L. (V.) guyanensis. We have given this population sub-specific rank, naming it L. (V.) s. santarensis. The other one (4 strains) did not express the B19 epitope like the L. (V.) shawi reference strain, which we now designate as L. (V.) s. shawi. For the first time in the eastern Brazilian Amazon we register a putative hybrid parasite (7 strains), L. (V.) guyanensis/L. (V.) s. shawi, characterized by a new 6PGDH three-band profile at the level of L. (V.) guyanensis. Its PGM profile, however, was very similar to that of L. (V.) s. shawi. These results suggest that the lower Amazon region - western Pará state, Brazil, represents a biome where L. (V.) guyanensis and L. (V.) s. shawi exchange genetic information.

Hematological parameters of Ameiva ameiva (Reptilia: Teiidae) naturally infected with hemogregarine: confirmation of monocytosis.

Little is known on how hematozoan infection changes reptile hematology. The lizard Ameiva ameiva is widely distributed in the Americas and is infected by hematozoan parasites. Previous studies on this lizard have shown that the parasite of monocytes causes a variety of ultrastructural changes in infected host cells. The present study reports that this infection does not cause any change to the erythrocytic values. However, a marked increase in the number of leukocytes (especially monocytes) was detected. This indicates that the hemogregarine not only modulates the infected monocyte, but also increases the blood pool of this leukocyte. A Plasmodium sp was also found infecting erythrocytes of one lizard.

Canine visceral leishmaniasis due to Leishmania (L.) infantum chagasi in Amazonian Brazil: comparison of the parasite density from the skin, lymph node and visceral tissues between symptomatic and asymptomatic, seropositive dogs.

Canine visceral leishmaniasis (CVL) is recognizable by characteristic signs of disease and is highly lethal. The infection, however, may be quite inapparent in some seropositive dogs, and this has raised the polemic question as to whether or not such animals can be a source of infection for Lutzomyia longipalpis, the vector of American visceral leishmaniasis (AVL). In this study we have examined 51 dogs with acute CVL from an AVL area in Pará State, northern Brazil, and compared the parasite density, amastigotes of Leishmania (L.) infantum chagasi, in the skin, lymph node and viscera of symptomatic with that of nine asymptomatic but seropositive dogs (IFAT-IgG). Post-mortem biopsy fragments of these tissues were processed by immunohistochemistry, using a polyclonal antibody against Leishmania sp. The X² and Mann Whitney tests were used to evaluate the means of infected macrophage density (p < 0.05). There was no difference (p > 0.05) in the skin (10.7/mm² x 15.5/mm²) and lymph node (6.3/mm² x 8.3/mm²), between asymptomatic and symptomatic dogs, respectively. It was higher (p < 0.05), however, in the viscera of symptomatic (5.3/mm²) than it was in asymptomatic (1.4/mm²) dogs. These results strongly suggest that asymptomatic or symptomatic L. (L.) i. chagasi-infected dogs can serve as a source of infection, principally considering the highest (p < 0.05) parasite density from skin (10.7/mm² x 15.5/mm²), the place where the vetor L. longipalpis takes its blood meal, compared with those from lymph node (6.3/mm² x 8.3/mm²) and viscera (1.4/mm²x 5.3/mm²).

A prospective study on the dynamics of the clinical and immunological evolution of human Leishmania (L.) infantum chagasi infection in the Brazilian Amazon region.

This prospective study was carried out from October 2003 to December 2005 and involved a cohort of 946 individuals of both genders, aged 1-89 years, from an endemic area for American visceral leishmaniasis (AVL), in Pará State, Brazil. The aim of the study was to analyze the dynamics of the clinical and immunological evolution of human Leishmania (L.) infantum chagasi infection represented by the following clinical-immunological profiles: asymptomatic infection (AI); symptomatic infection (SI=AVL); subclinical oligosymptomatic infection (SOI); subclinical resistant infection (SRI); and indeterminate initial infection (III). Infection diagnosis was determined by the indirect fluorescent antibody test and leishmanin skin test. In total, 231 cases of infection were diagnosed: the AI profile was the most frequent (73.2%), followed by SRI (12.1%), III (9.9%), SI (2.6%) and SOI (2.2%). The major conclusion regarding evolution dynamics was that the III profile plays a pivotal role from which the cases evolve to either the resistant, SRI and AI, or susceptible, SOI and SI, profiles; only one of the 23 III cases evolved to SI, while most evolved to either SRI (nine cases) or SOI (five cases) and eight cases remained as III.

A cross-sectional study on the clinical and immunological spectrum of human Leishmania (L.) infantum chagasi infection in the Brazilian Amazon region.

The objectives of this study were to identify individuals with symptomatic and/or asymptomatic infection due to Leishmania (L.) infantum chagasi; to study the two types of infection, both clinically and immunologically, and to determine the prevalence rate of infection at the beginning of the study. This was a cross-sectional study with a cohort of 946 individuals, of both genders, from the age of 1 year, living in the municipality of Barcarena, PA, Brazil, an area endemic for American visceral leishmaniasis (AVL). The leishmanin skin test (LST) and the indirect fluorescent test (IFAT), were used for the diagnosis of infection. One hundred and twenty cases of infection were diagnosed, with a prevalence rate of 12.6%; eight cases showed high seroreactivity (1280-10240, IgG) in IFAT and no LST reaction; four of these cases were typical AVL and four had subclinical oligosymptomatic infection. Using two immunological methods with a clinical examination of the infected individuals enabled the identification of five clinical-immunological profiles which may promote a better understanding of the interaction between L. (L.) i. chagasi and the human immune response: asymptomatic infection (AI) 73.4%; subclinical resistant infection (SRI) 15%; subclinical oligosymptomatic infection (SOI) 3%; symptomatic infection (AVL) 3% and indeterminate initial infection (III) 5%.

A longitudinal study on the transmission dynamics of human Leishmania (Leishmania) infantum chagasi infection in Amazonian Brazil, with special reference to its prevalence and incidence.

This was a longitudinal study carried out during a period over 2 years with a cohort of 946 individuals of both sexes, aged 1 year and older, from an endemic area of American visceral leishmaniasis (AVL) in Pará State, Brazil. The object was to analyze the transmission dynamics of human Leishmania (Leishmania) infantum chagasi infection based principally on the prevalence and incidence. For diagnosis of the infection, the indirect fluorescent antibody test (IFAT) and leishmanin skin test (LST) were performed with amastigote and promastigote antigens of the parasite, respectively. The prevalence by LST (11.2%) was higher (p < 0.0001) than that (3.4%) by IFAT, and the combined prevalence by both tests was 12.6%. The incidences by LST were also higher (p < 0.05) than those by IFAT at 6 (4.7% x 0.6%), 12 (4.7% x 2.7%), and 24 months (2.9% x 0.3%). Moreover, there were no differences (p > 0.05) between the combined incidences by both tests on the same point surveys, 5.2%, 6.3%, and 3.6%. During the study, 12 infected persons showed high IFAT IgG titers with no LST reactions: five children and two adults developed AVL (2,560-10,120), and two children and three adults developed subclinical oligosymptomatic infection (1280-2560). The combined tests diagnosed a total of 231 cases of infection leading to an accumulated prevalence of 24.4%.

Reviewing the role of the dendritic Langerhans cells in the immunopathogenesis of American cutaneous leishmaniasis.

The role of dendritic Langerhans cells (LCs) in the immunopathogenesis of American cutaneous leishmaniasis (ACL) was reviewed in the light of more recent clinical and immunological features of ACL, caused by the principal human pathogenic leishmanial parasites found in Brazil: Leishmania (Viannia) braziliensis and L. (L.) amazonensis. The report shows a species-specific correlation between the LC density and the CD4+ and CD8+ T-cell profiles in the cellular infiltrate of skin lesions of ACL patients, providing the conclusion that LCs might be influencing the dichotomy of interaction between L. (V.) braziliensis and L. (L.) amazonensis with the human T-cell immune response. While L. (V.) braziliensis shows a clear tendency to direct infection to the hypersensitivity pole of the ACL clinical-immunological spectrum marked by a strong Th1-type immune response, L. (L.) amazonensis shows the opposite, directing infection to the hyposensitivity pole associated with a marked Th2-type immune response. These are probably the main immunological mechanisms of LCs regarding the immune response dichotomy that modulates infection outcome by these Leishmania parasites.

In vitro infectivity of species of Leishmania (Viannia) responsible for American cutaneous leishmaniasis.

There is little available information regarding the infectivity of New World Leishmania species, particularly those from the Amazonian Brazil, where there are six species of the subgenus Viannia causing American cutaneous leishmaniasis (ACL). The aim of this study was to compare, in vitro, the potential infectivity of the following Leishmania (Viannia) spp.: L. (V.) braziliensis from localized cutaneous leishmaniasis (LCL) and mucocutaneous leishmaniasis (MCL) patients, L. (V.) guyanensis, L. (V.) shawi, L. (V.) lainsoni and L. (V.) naiffi from LCL patients only, in cultured BALB/c mice peritoneal macrophage, as well as the production of NO by the infected cells. The infectivity of parasites was expressed by the infection index and, the nitric oxide (NO) production in the macrophage culture supernatant was measured by the Griess method. It was found that L. (V.) braziliensis from MCL, the more severe form of disease, showed the highest (p

Theileria electrophorin.sp., a parasite of the electric eel Electrophorus electricus (Osteichthyes: Cypriniformes: Gymnotidae) from Amazonian Brazil.

The name Theileria electrophori n.sp. is proposed for a small parasite described in the erythrocytes of the electric eel, Electrophorus electricus, from Amazonian Brazil. Division of the organism in the erythrocyte produces only four bacilliform daughter cells which become scattered in the host cell, without a cruciform or rosette-shaped disposition. Exoerythrocytic meronts producing a large number of merozoites were encountered in Giemsa-stained impression smears of the internal organs, principally in the liver, and are presumably the source of the intraerythrocytic forms of the parasite. This developmental pattern is characteristic of piroplasms within the family Theileriidae, where the author considers the parasite of E. electricus to most appropriately belong. It effectively distinguishes the organism from the dactylosomatid parasites Babesiosoma Jakowska and Nigrelli, 1956 and Dactylosoma Labbé, 1894 also found in fishes. This appears to be the second report of Theileria Bettencourt, Franca and Borges, 1907 in a fish.

Observations on Eimeria species of Dasyprocta leporina (Linnaeus, 1758)(Rodentia: Dasyproctidae) from the state of Pará, North Brazil.

Redescriptions are given of the mature oocysts of Eimeria aguti Carini 1935, E. cotiae Carini, 1935 and E. paraensis Carini, 1935, in the faeces of five specimens of the rodent Dasyprocta leporina (Rodentia: Dasyproctidae) from the state of Pará, North Brazil. New information is provided on the sporulation time of these parasites and the prepatent period in experimentally infected D. leporina. Some endogenous stages of E. cotiae are described in the epithelial cells of the ileum, and the absence of any oocysts in the gall-bladder contents of the infected animals indicates that the intestine is also the site of development of E. aguti and E. paraensis. Difficulties in separating E. cotiae and E. paraensis on morphology of the oocysts are discussed. The oocysts of both parasites share many structural features and have a wide size range. It is concluded that although it is at present best to maintain these names, the possibility exists that they were separately given to oocysts of smaller dimensions (E. cotiae) and larger dimensions (E. paraensis) of a single parasite. Location of an endogenous site of development for E. paraensis that is distinctly separate from that of E. cotiae might establish more definitely the separate specific status of the two parasites.

Lutzomyia (Nyssomyia) whitmani s.l . (Antunes & Coutinho, 1939)(Diptera: Psychodidae: Phlebotominae): geographical distribution and the epidemiology of American cutaneous leishmaniasis in Brazil--mini-review.

The aim of the present study, in view of the widespread geographical distribution of Lutzomyia (Nyssomyia) whitmani s.l. in Brazil, in close association with the regions of transmission of Leishmania (Viannia) braziliensis and L. (V.) shawi, is to investigate the distribution of this sand fly species and American cutaneous leishmaniasis (ACL) in relationship to vegetation and landscape ecology throughout Brazil. Thematic maps were elaborated by the MapInfo programme, giving information on the spatial distribution of L. whitmani s.l., in accordance with types of vegetation and foci of ACL. With regards to the known areas of transmission of ACL in Brazil, it is notable that L. (N.) whitmani s.l. occurs in most of them, where it has been implicated as a possible vector of L. (V.) braziliensis. The presence of L. whitmani s.l. has been registered in 26 states, the one exception being Santa Catarina; in some states such as Roraima, Acre, Tocantins, and Mato Grosso do Sul this sand fly species has been recorded in a large number of municipalities. L. whitmani s.l. has been found in association with a variety of vegetation types, including the Amazonian forest, savanna ("campos cerrados"), and northeastern savanna ("caatingas nordestinas" or "savana estépica").

The genus Cyclospora (Apicomplexa: Eimeriidae), with a description of Cyclospora schneideri n.sp. in the snake Anilius scytale scytale (Aniliidae) from Amazonian Brazil--a review.

A review is made of the recorded species of the coccidian genus Cyclospora and major events leading up to the discovery of C. cayetanensis, which is responsible for serious outbreaks of diarrhoea in man and is one of the aetiological agents of "traveller's diarrhoea". Humans appear to be the specific hosts, with the entire life-cycle in the intestine: to date there is no convincing evidence that the disease is a zoonosis. A description is given of oocysts and endogenous stages of C. schneideri n.sp., in the snake Anilius scytale scytale. Sporulation is exogenous and completed after about one week at 24-26 degrees. Mature oocysts 19.8 x 16.6 (15.1 x 13.8-25.7 x 20.1), shape-index 1.2 (1.0-1.3): no oocyst residuum or polar bodies. Oocyst wall a single colourless, smooth layer with no micropyle: it is rapidly deformed or broken. Sporocysts 13.6 x 9.4 (11.3 x 8.3-15.1 x 9.9), shape-index 1.4 (1.2-1.5) with an inconspicuous Stieda body. Sporozoites 11-13 x 2.5-3. Endogenous stages are intracytoplasmic in the epithelial cells of the small intestine and with the characters of the Eimeriorina.

The Neotropical Leishmania species: a brief historical review of their discovery, ecology and taxonomy / 117

This paper is a review of the major historical events leading to our present classification of the Neotropical Leishmania species, and apart from indicating the basic type of disease these different parasites may cause in humans, it does not discuss the clinical or epidemiological features of the leishmaniases. For each of these species, information is given on the known geographical distribution, recorded phlebotomine sand fly host(s) and the secondary, wild or domestic mammalian hosts. Reasons are given for regarding the parasite referred to as Leishmania (L.) infantum chagasi, the causative agent of American visceral leishmaniasis, as most probably indigenous to the Neotropics rather than imported during the Iberian colonisation...

Este artigo apresenta uma revisão dos mais importantes eventos históricos que levaram à atual classificação das espécies neotropicais de Leishmania e indica as doenças básicas causadas em seres humanos por estes diferentes parasitos, sem discutir os aspectos clínicos e epidemiológicos das leishmanioses. Para cada uma das espécies descritas, são fornecidas informações a respeito de sua conhecida distribuição geográfica, dos flebotomíneos hospedeiros registrados e de seus reservatórios mamíferos secundários, selvagens ou domésticos. Os dados apresentados levam à conclusão de que o parasito Leishmania (L.) infantum chagasi, agente causador da leishmaniose visceral americana, é provavelmente autóctone da região neotropical, e não importada durante a colonização ibérica...