Spatial and Temporal Distribution of Aedes aegypti and Aedes albopictus Oviposition on the Coast of Paraná, Brazil, a Recent Area of Dengue Virus Transmission.

Aedes aegypti and Aedes albopictus are considered the most important vectors of arboviruses in the world. Aedes aegypti is the primary vector of dengue, urban yellow fever, chikungunya and zika in Brazil, and Ae. albopictus is considered a potential vector. Distribution patterns and the influence of climatic variables on the oviposition of Ae. aegypti and Ae. albopictus were evaluated in Morretes, a tourist city in the coastal area of Paraná State, Brazil, which has recently been experiencing cases of dengue fever. Eggs were collected using ovitraps over a period of one year (September 2017 to September 2018) and reared from hatching until the emergence of the adults. Both Aedes species were found in anthropized areas with a high human density index. Findings suggest that the monthly average temperature (LRT = 16.65, p = 0.001) had significant positive influences on the oviposition of the Aedes species. Considering the wide distribution of DENV around the Paraná coast and the presence of Ae. albopictus alongside Ae. aegypti, studies on natural arbovirus infection patterns and seasonality are recommended in the region.

Predictive modeling of sand fly distribution incriminated in the transmission of Leishmania (Viannia) braziliensis and the incidence of Cutaneous Leishmaniasis in the state of Paraná, Brazil.

Southern Brazil concentrates a considerable number of cases of cutaneous leishmaniasis reported since 1980, and Paraná is the state that most records CL cases in the region. The main sand fly species incriminated as vectors of Leishmania (Viannia) braziliensis (Vianna,1911) are Migonemyia (Migonemyia) migonei (França, 1920), Nyssomyia (Nyssomyia) neivai (Pinto, 1926) and Nyssomyia (Nyssomyia) whitmani (Antunes & Coutinho, 1936). In this study, we evaluated areas with climatic suitability for the distribution of these vectors and correlated these data with CL incidence in the state. The occurrence points of Mg. migonei, Ny. neivai, and Ny. whitmani were extracted from a literature review and field data. For CL analysis in the state of Paraná, data were obtained from the Informatics Department of the Unified Health System of Brazil (DATASUS), covering the period from 2001 to 2019. The layers of bioclimatic variables from the WorldClim database were used in the study. Species distribution modeling was developed using the MaxEnt Software version 3.4.4. ArcGIS software version 10.5 was used to develop suitability maps and the graphical representation of disease incidence. The AUC values were acceptable for all models (> 0,8). Bioclimatic variables BIO13 and BIO14 were the most influential in the distribution of Mg. migonei, while BIO19 and BIO6 were the variables that most influenced the distribution of Ny. neivai, and Ny. whitmani was most influenced by variables BIO5 and BIO9. During 19 years, 4992 cases of CL were reported in the state by 286 municipalities (71,6%). Northern Paraná showed the highest number of areas with very high and high climatic suitability for the occurrence of these species, coinciding with the highest number of CL cases. The modeling tools allowed analyzing the association between climatic variables and the geographical distribution of CL in the state. Moreover, they provided a better understanding of the climatic conditions related to the distribution of different species, favoring the monitoring of risk areas, the implementation of preventive measures, risk awareness, early and accurate diagnosis, and consequent timely treatment.

Cutaneous leishmaniosis in naturally infected dogs in Paraná, Brazil, and the epidemiological implications of Leishmania (Viannia) braziliensis detection in internal organs and intact skin.

Environmental changes have occurred over the years, altering the eco-epidemiological pattern of leishmaniosis in the State of Paraná, Brazil, involving the pillars of the cycle (parasite, vectors, reservoir, and environment) and their interaction. Much has been discussed about the dog's role as a reservoir of the Leishmania (Viannia) braziliensis Vianna, 1911 transmission cycle. However, this question remains unanswered. The purpose of this study was to investigate, using parasitological and molecular methods, different samples in eight naturally infected dogs from an endemic rural locality where only L. (V.) braziliensis is present, and where human cases have been previously notified. Blood and biopsied organ samples from naturally infected dogs were analyzed by culture media, PCR, random amplified polymorphic DNA and sequencing methodologies. Only skin lesions from all dogs yielded positive cultures and when PCR was performed, L. (V.) braziliensis DNA was amplified from intact skin, peripheral blood, bone marrow, spleen, liver and lymph nodes. RAPD was also applied to isolates from the skin lesions, exhibiting the genetic variability of the parasite identified. To confirm which species of Leishmania was amplified in PCR, the sequencing method was performed, verifying 100% similarity with the Viannia subgenus. This study showed that L. (V.) braziliensis can spread to other sites besides the ulcerous lesions, such as intact skin, peripheral blood and internal organs, making it possibility for dogs to serve as active sources of parasite transmission. For definitive proof, xenodiagnostic test on intact skin of infected dogs, should be done.

Can equids be a reservoir of Leishmania braziliensis in endemic areas?

In this study, we detected Leishmania (Viannia) braziliensis infection in equids living in endemic regions of cutaneous leishmaniasis. To determine the role of these animals in the Leishmania cycle, we used two approaches: serological and molecular methods. Antibodies to the parasite were assayed using the Enzyme Linked Immunosorbent Assay (ELISA). Blood samples were collected and tested by polymerase chain reaction (PCR), and the positive products were sequenced. The results showed that 11.0% (25/227) of the equids were seropositive for Leishmania sp, and 16.3% (37/227) were PCR positive. Antibodies were detected in 20 horses, 3 donkeys, and 2 mules, and the parasite DNA was detected in 30 horses, 5 donkeys, and 2 mules. Sequencing the amplified DNA revealed 100% similarity with sequences for Viannia complex, corroborating the results of PCR for L. braziliensis. Our results show that equids are infected with L. braziliensis, which could be food sources for phlebotomines in the peridomiciliary environment and consequently play a role in the cutaneous leishmaniasis cycle.

Detection of Neospora caninum DNA in capybaras and phylogenetic analysis.

The role of rodents in the sylvatic cycle of Neospora sp. and in the neosporosis epidemiology is still uncertain. The aim of the present work was to detect Neospora caninum and to determine its prevalence in capybaras (Hydrochaeris hydrochaeris), to help elucidate the role of this rodent in the life cycle of the parasite. N.caninum DNA was detected by PCR using 4 different sets of primers specific to the Nc5 and ITS1 sequences. The parasite was found in the lymph nodes, heart, liver, and blood of 23% of the twenty-six capybaras studied. Sequencing the amplified DNA revealed 98% of similarity with N. caninum sequences deposited in the Genbank. Our findings provide the first molecular evidence of N. caninum infection in capybaras, supporting the hypothesis that these rodents can act as reservoirs of N. caninum and play a role in the life cycle of this parasite.

Spinocerebellar ataxias: microsatellite and allele frequency in unaffected and affected individuals.

The diagnosis and incidence of spinocerebelar ataxias (SCA) is sometimes difficult to analyze due the overlap of phenotypes subtypes and are disorders of mutations caused by CAG trinucleotide repeat expansion. To investigate the incidence of the SCA in Southern Brazil, we analyzed the trinucleotide repeats (CAG)n at the SCA1, SCA2, SCA3, SCA6 and SCA7 loci to identify allele size ranges and frequencies. We examined blood sample from 154 asymptomatic blood donors and 115 individuals with progressive ataxias. PCR products were submitted to capillary electrophoresis. In the blood donors, the ranges of the five loci were: SCA1, 19 to 36 (CAG)n; SCA2, 6 to 28 (CAG)n; SCA3, 12 to 34 (CAG)n; SCA6, 2 to 13 (CAG)n; and SCA7, 2 to 10 (CAG)n. No deviations from Hardy-Weinberg equilibrium were detected. In the ataxia group, we found (CAG)n above the range of the asymptomatic blood donors in SCA3 (21.74%) followed by SCA2 (5.22%), SCA7 (2.61%), SCA6 (0.87%), and no cases of SCA1. The remaining 80 cases (69.56%) have different diagnoses from the type here studied. These data defined the alleles and their frequencies, as well as demonstrated their stability in the population not affected. The molecular diagnosis test confirmed the clinical diagnosis in 28/45 cases and classified another 7/70 from the clinical unclassified ataxias group.

Low gene flow of Aedes aegypti between dengue-endemic and dengue-free areas in southeastern and southern Brazil.

We present a population genetic study of Aedes aegypti in Brazil using isoenzyme markers. Four polymorphic loci were used to examine 11 mosquito collections at four periods in 2003. Samples from a dengue-endemic area (southeastern region) and a dengue-free area (southern region) connected by an important network of roads and railways were analyzed. The degree of genetic differentiation observed between populations is consistent with limited gene flow between them. There was no evidence of passive dispersion of Ae. aegypti by vehicles among the different routes linking metropolitan areas.

Higher genetic variation estimated by microsatellites compared to isoenzyme markers in Aedes aegypti from Rio de Janeiro.

Aedes aegypti populations from five districts in Rio de Janeiro were analyzed using five microsatellites and six isoenzyme markers, to assess the amount of variation and patterns of gene flow at local levels. Microsatellite loci were polymorphic enough to detect genetic differentiation of populations collected at small geographic scales (e.g. within a city). Ae. aegypti populations were highly differentiated as well in the city center as in the outskirt. Thus, dengue virus propagation by mosquitoes could be as efficient in the urban area as in the outskirt of Rio de Janeiro, the main entry point of dengue in Brazil.