Presence of anti-Leishmania (Viannia) braziliensis antibodies in blood donors in the West-Central region of the State of Paraná, Brazil

ABSTRACTINTRODUCTION:Serological screening in blood banks does not include all transmittable diseases. American cutaneous leishmaniasis (ACL) has a high detection rate in the municipalities of the State of Paraná.METHODS:This study analyzed the presence of anti- Leishmania braziliensisantibodies in 176 blood donors who live in these endemic areas. The variables were analyzed with the χ2 test and Stata 9.1 software. RESULTS: Twenty (11.4%) samples were positive for the presence of anti- L. braziliensisantibodies. CONCLUSIONS: The high percentage of donors with anti- Leishmania spp. antibodies indicates the need to study the risk of ACL transmission through blood donors.

Seroprevalence for hepatitis E virus (HEV) infection among volunteer blood donors of the Regional Blood Bank of Londrina, State of Paraná , Brazil

A cross-sectional study was carried out among 996 volunteer blood donors enrolled from May 1999 to December 1999 to determine the seroprevalence of hepatitis E virus (HEV) infection among volunteer blood donors of the Regional Blood Bank of Londrina, State of Paraná, Brazil, and to evaluate whether the rate of seroprevalence of IgG anti-HEV antibodies is associated with sociodemographic variables and with seropositivity for hepatitis A virus (HAV) infection. All participants answered the questionnaire regarding the sociodemographic characterisitcs. Serum samples were tested for IgG antibodies to HEV (anti-HEV) by an enzyme linked immunoassay (ELISA). All serum samples positive for anti-HEV IgG and 237 serum samples negative for anti-HEV were also assayed for IgG anti-HAV antibodies by ELISA. Anti-HEV IgG was confirmed in 23/996 samples, resulting in a seroprevalence of 2.3% for HEV infection, similar to previous results obtained in developed countries. No significant association was found between the presence of anti-HEV IgG antibodies and the sociodemographic variables including gender, age, educational level, rural or urban areas, source of water, and sewer system (p > 0.05). Also, no association with seropositivity for anti-HAV IgG antibodies was observed (p > 0.05). Although this study revealed a low seroprevalence of HEV infection in the population evaluated, the results showed that this virus is circulating among the population from Londrina, South Brazil, and point out the need of further studies to define the clinical and epidemiological importance of HEV infection and to identify additional risk factors involved in the epidemiology and pathogenesis of this infection in this population.

Induction of phagocytic activity and nitric-oxide production in natural populations of Trypanosoma Cruzi I and II from the state of Paraná, Brazil

Twelve strains of Trypanosoma cruzi isolated from wild reservoirs, triatomines, and chronic chagasic patients in the state of Paraná, southern Brazil, and classified as T. cruzi I and II, were used to test the correlation between genetic and biological diversity. The Phagocytic Index (PI) and nitric-oxide (NO) production in vitro were used as biological parameters. The PI of the T. cruzi I and II strains did not differ significantly, nor did the PI of the T. cruzi strains isolated from humans, triatomines, or wild reservoirs. There was a statistical difference in the inhibition of NO production between T. cruzi I and II and between parasites isolated from humans and the strains isolated from triatomines and wild reservoirs, but there was no correlation between genetics and biology when the strains were analyzed independently of the lineages or hosts from which the strains were isolated. There were significant correlations for Randomly Amplified Polymorphic Deoxyribonucleic acid (RAPD) and biological parameters for T. cruzi I and II, and for humans or wild reservoirs when the lineages or hosts were considered individually.

Study of infection by Rickettsiae of the spotted fever group in humans and ticks in an urban park located in the City of Londrina, State of Paraná, Brazil

INTRODUCTION: Spotted fevers are emerging zoonoses caused by Rickettsia species in the spotted fever group (SFG). Rickettsia rickettsii is the main etiologic agent of Brazilian spotted fever (BSF) and it is transmitted by Amblyomma spp. ticks. METHODS: The study aimed to investigate SFG rickettsiae in the Arthur Thomas Municipal Park in Londrina, PR, by collecting free-living ticks and ticks from capybaras and blood samples from personnel working in these areas. Samples from A. dubitatum and A. cajennense were submitted for PCR in pools to analyze the Rickettsia spp. gltA (citrate synthase gene). RESULTS: All the pools analyzed were negative. Human sera were tested by indirect immunofluorescence assay with R. rickettsii and R. parkeri as antigens. Among the 34 sera analyzed, seven (20.6%) were reactive for R. rickettsii: four of these had endpoint titers equal to 64, 2 titers were 128 and 1 titer was 256. None of the samples were reactive for R. parkeri. An epidemiological questionnaire was applied to the park staff, but no statistically significant associations were identified. CONCLUSIONS: The serological studies suggest the presence of Rickettsiae related to SFG that could be infecting the human population studied; however, analysis of the ticks collected was unable to determine which species may be involved in transmission to humans.

Sandfly frequency in a clean and well-organized rural environment in the state of Paraná, Brazil

INTRODUCTION: Sandflies caught in Santa Juliana Farm in Sarandi, State of Paraná, Brazil, were assessed in terms of their fauna, seasonality, and frequency in the homes and in shelters of domestic animals around the homes, as well as in the nearby forest. METHODS: In Santa Juliana Farm, there are no records of cases of ACL, differing from other relatively clean and organized areas where surveys of sandflies have been conducted in Paraná. Samples were collected with Falcão light traps, fortnightly from 22:00 to 02:00 hours, from November 2007 to November 2008. RESULTS: A total of 4,506 sandflies were captured, representing 13 species, predominantly Nyssomyia whitmani (71.8%). More sandflies were collected in the forest (52.6%) than outside the forest (residences and pigsty) (47.4%). However, Ny. whitmani was collected in greater numbers outside (38.3%) than inside the forest (33.5%). Most sandflies were collected in the warmer months and during periods with regular rainfall. CONCLUSIONS: The results suggest that cleaning and organization around the houses could reduce sandfly population in peridomicile. Constructing shelters for animal at a distance of approximately 100m from domiciles is recommended to prevent the invasion of sandflies, as this farm has an area of preserved forest, with wild animals and sandflies present to maintain the enzootic cycle of Leishmania.

Geographic analysis on the occurrence of human and canine leptospirosis in the city of Maringá, state of Paraná, Brazil

INTRODUCTION: Leptospirosis is a zoonotic disease, the primary hosts of which are wild, synanthropic, and household animals. Humans behave as terminal and accidental hosts. The prevalence of leptospirosis depends on carrier animals that disseminate the agent, on the environmental survival of this agent, and on the contact of susceptible individuals. Each serovar has one or more hosts with different adaptation levels. The focuses of leptospirosis are infected, sick, and asymptomatic animals, which are considered to be sources of environmental infection. This study aimed to determine the risk areas for leptospiral infection in stray dogs and patients diagnosed with leptospirosis from 2006 to 2008 in Maringá, State of Paraná, Brazil. METHODS: Three hundred and thirty-five stray dogs and 25 patients were studied. Serum from both animals and patients was examined by the microscopic serum agglutination test to study anti-leptospiral antibodies. To determine the risk areas and the spatial distribution of the disease, thematic maps were designed. RESULTS: Forty-one (12.2%) dogs positive for one or more leptospire serovars were observed, the most frequent serovars being Pyrogenes (43.9%), Canícola (21.9%), and Copennhageni (19.5%). Among the humans, 2 (8%) were positive for serovars Pyrogenes and Hardjo Prajitno and for Pyrogenes and Cynopteri. CONCLUSIONS: Spatial analysis showed that the risk for dogs and humans in the City of Maringá to become infected with leptospires exists in both the central and the peripheral areas, a fact that reinforces the relevance of this study and of continuous epidemiological and environmental surveillance actions to control the disease in animals and in humans.

Genetic diversity of dengue virus serotypes 1 and 2 in the State of Paraná, Brazil, based on a fragment of the capsid/premembrane junction region

INTRODUCTION: The precise identification of the genetic variants of the dengue virus is important to understand its dispersion and virulence patterns and to identify the strains responsible for epidemic outbreaks. This study investigated the genetic variants of the capsid-premembrane junction region fragment in the dengue virus serotypes 1 and 2 (DENV1-2). METHODS: Samples from 11 municipalities in the State of Paraná, Brazil, were provided by the Central Laboratory of Paraná. They were isolated from the cell culture line C6/36 (Aedes albopictus) and were positive for indirect immunofluorescence. Ribonucleic acid (RNA) extracted from these samples was submitted to the reverse transcription polymerase chain reaction (RT-PCR) and nested PCR. RESULTS: RT-PCR revealed that 4 of the samples were co-infected with both serotypes. The isolated DENV-1 sequences were 95-100% similar to the sequences of other serotype 1 strains deposited in GenBank. Similarly, the isolated DENV-2 sequences were 98-100% similar to other serotype 2 sequences in GenBank. According to our neighbor-joining tree, all strains obtained in this study belonged to genotype V of DENV-1. The DENV-2 strains, by contrast, belonged to the American/Asian genotypes. CONCLUSIONS: The monitoring of circulating strains is an important tool to detect the migration of virus subtypes involved in dengue epidemics.

Ecological aspects of the sandfly fauna (Diptera, Psychodidae) in an American cutaneous leishmaniasis endemic area under the influence of hydroelectric plants in Paranapanema river, State of Paraná, Brazil

INTRODUCTION: An epidemiological study was undertaken to identify determinant factors in the occurrence of American cutaneous leishmaniasis in areas under the influence of hydroelectric plants in Paranapanema river, State of Paraná, Brazil. The ecological aspects of the phlebotomine fauna were investigated. METHODS: Sandflies were sampled with automatic light traps from February 2004 to June 2006 at 25 sites in the urban and rural areas of Itambaracá, and in Porto Almeida and São Joaquim do Pontal. RESULTS: A total of 3,187 sandflies of 15 species were captured. Nyssomyia neivai predominated (34.4%), followed by Pintomyia pessoai (32.6%), Migonemyia migonei (11.6%), Nyssomyia whitmani (8.8%), and Pintomyia fischeri (2.7%), all implicated in the transmission of Leishmania. Males predominated for Ny. neivai, and females for the other vector species, with significant statistical differences (p < 0.001). Nyssomyia neivai, Pi. pessoai, Ny. whitmani, Brumptomyia brumpti, Mg. migonei, and Pi. fischeri presented the highest values for the Standardized Species Abundance Index (SSAI). The highest frequencies and diversities were found in the preserved forest in Porto Almeida, followed by forests with degradation in São Joaquim do Pontal and Vila Rural. CONCLUSIONS: Sandflies were captured in all localities, with the five vectors predominating. Ny. neivai had its highest frequencies in nearby peridomestic environments and Pi. pessoai in areas of preserved forests. The highest SSAI values of Ny. neivai and Pi. pessoai reflect their wider dispersion and higher frequencies compared with other species, which seems to indicate that these two species may be transmitting leishmaniasis in the area.

The first record of Lutzomyia longipalpis (Lutz & Neiva, 1912) (Diptera: Psychodidae: Phlebotominae) in the State of Paraná, Brazil

INTRODUCTION: We report the first find of Lutzomyia longipalpis in the State of Paraná, Brazil. METHODS: The specimens were captured in the urban area of the municipality of Foz do Iguaçu, with Falcão light traps, in domiciliary and peridomiciliary areas of 61 properties, on two consecutive nights from 18:00 to 06:00hs in March 2012. RESULTS: We captured 40 specimens of Lu. longipalpis and 54 specimens of other sandfly species. CONCLUSIONS: This find expands knowledge of the geographical distribution of this sandfly in Brazil.

Polymerase chain reaction and nested-PCR approaches for detecting Cryptosporidium in water catchments of water treatment plants in Curitiba, State of Paraná, Brazil

Introduction Cryptosporidium is an important protozoan cause of waterborne disease worldwide of concern to public health authorities. To prevent outbreaks of cryptosporidiosis, the monitoring of this parasite in drinking water is necessary. In the present work, the polymerase chain reaction (PCR) and nested-PCR techniques were used to detect Cryptosporidium in raw water from catchment points of four water treatment plants (WTP) in Curitiba, Paraná, Brazil. Methods First, DNA extraction techniques were tested in samples containing decreasing amount of oocysts in reagent water, and PCR and nested-PCR with specific primers for 18SSU rDNA of Cryptosporidium were conducted to determine their sensitivity. In reagent water, a commercial extraction kit provided the best analytical sensitivity, and PCR and nested-PCR allowed the detection of five and two oocysts, respectively, with the primers XIAOR/XIAOF and XIAO1F/XIAO2R. Results In the spiking experiments, only the PCR with the primers AWA995F/AWA1206R was successful at detecting concentrations of 0.1 oocysts/mL. Two catchments samples of raw water and/or water sludge from four WTPs were contaminated with Cryptosporidium. Conclusions The application of the techniques to monitor Cryptosporidium in water and detect contamination in water catchments of WTPs in Curitiba are discussed in the present work.

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.

Toxoplasmosis: an examination of knowledge among health professionals and pregnant women in a municipality of the State of Paraná

Introduction The aim of this study was to investigate the knowledge of toxoplasmosis among professionals and pregnant women in the public health services in Paraná, Brazil. Methods A cross-sectional observational and transversal study of 80 health professionals (44 nurses and 36 physicians) and 330 pregnant women [111 immunoglobulin M (IgM)- and IgG-non-reactive and 219 IgG-reactive] was conducted in 2010. An epidemiological data questionnaire was administered to the professionals and to the pregnant women, and a questionnaire about the clinical aspects and laboratory diagnosis of toxoplasmosis was administered to the professionals. Results The participants frequently provided correct responses about prophylactic measures. Regarding the clinical and laboratory aspects, the physicians provided more correct responses and discussed toxoplasmosis with the pregnant women. The professionals had difficulty interpreting the avidity test results, and the physicians stated that they referred pregnant women with high-risk pregnancies to a county reference center. Of the professionals, 53 (91.4%) reported that they instructed women during prenatal care, but only 54 (48.6%) at-risk pregnant women and 99 (45.2%) women who were not at risk reported receiving information about preventive measures. The physicians provided verbal instructions to 120 (78.4%) women, although instructional materials were available in the county. The pregnant women generally lacked knowledge about preventive measures for congenital toxoplasmosis, but the at-risk pregnant women tended to respond correctly. Conclusions This study provides data to direct public health policies regarding the importance of updating the knowledge of primary care professionals. Mechanisms should be developed to increase public knowledge because prophylactic strategies are important for preventing congenital toxoplasmosis.

Ectoparasites Ixodida Leach, 1817 on wild mammals in the state of Paraná, Brazil

A taxonomical and ecological study was made on 264 samples of 12 species of ectoparasite ixodides collected on wild mammals from several natural regions of Paraná state, Brazil. These species of ticks as their hosts are listed by their identified evolutive forms and capture locations. A review of the early studies on the Ixodida from South Brazil is made considering the frequency of the parasite species on each host concerning the geographic distribution of such species. This paper is a contribution to the knowledge of the Ixodidae ectoparasites species of wild mammals in Brazil.

A Preliminary Analysis of Insects of Medico-legal Importance in Curitiba, State of Paraná

A survey of the carrion fauna was made at two sites in Curitiba, State of Paraná, with the objective of describing the insects associated with carrion and setting up a preliminary data-base for medico-legal purposes in south Brazil. Vertebrate exclusion experiments were carried out in each season between 1994 and 1995 with a 250 g laboratory-bred rat (Rattus norvegicus). Five stages of decomposition were identified: fresh, bloated, decaying, dry and adipocere-like. Some species showed seasonal and site preference and so could be used to identify the probable place and season where death took place. Sarconesia chlorogaster (Diptera, Calliphoridae) was restricted to an open field site and to cooler months. Hemilucilia semidiaphana (Diptera, Calliphoridae) and Pattonella resona (Diptera, Sarcophagidae) were restricted to the forest site and warmer months. Phaenicia eximia (Diptera, Calliphoridae) and Oxyletrum discicolle (Coleoptera, Silphidae) were present at both sites throughout the year and could be useful for population level analysis. Dissochaetus murray (Coleoptera, Cholevidae) was present throughout the year at the forest site and was associated with the adipocere-like stage. Ants played an important role producing post-mortem injuries to the carcasses. Insects of 32 species are reported as being useful in community level approaches