Spatiotemporal changes in exposition risk to leishmaniases vector in residences within a fishing tourism area of Pantanal wetland.

Miranda Municipality of Mato Grosso do Sul, borders the Pantanal wetland, a famous fishing destination visited by tourists from all over the world, and is a location where visceral leishmaniasis has been reported. To assess the risk of Leishmania infantum transmission, we studied the sandfly community, focusing on known vector and parasite presence. We conducted light trap collections twice per month at nine sites within the city (including two forested areas) for one year. We collected a total of 12,727 sand flies, 10,891 males and 1,836 females belonging to 11 species: Brumptomyia avellari, Evandromyia aldafalcaoae, Ev. evandroi, Ev. lenti, Ev. sallesi, Ev. walkeri, Lu. longipalpis, Nyssomyia whitmani, Psathyromyia bigeniculata, Pa. hermanlenti and Pa. punctigeniculata. Lutzomyia longipalpis, the proven vector of Leishmania infantum, was captured each month, and was the most abundant species observed, accounting for more than 99% of sand flies captured in most sites, especially where chicken coops were present. Evidence of Leishmania infantum infection was detected in 0.40% of Lu. longipalpis tested. We developed a generalized mixed multilevel model for Lu. longipalpis, that includes within-year seasonality, location of capture (indoors vs. outdoors), vector abundance, and sex ratio. The VL vector was abundant both inside and outside houses. Large numbers of Lu. longipalpis were observed in outdoor sites where domestic animals were present but were absent from forest sites. Our findings suggest high vector populations and Le. infantum presence in a city where tourists could be exposed to visceral leishmaniasis, with significant implications for more surveillance and control activities.

Antileishmanial Activity, Toxicity and Mechanism of Action of Complexes of Sodium Usnate with Lanthanide Ions: Eu(III), Sm(III), Gd(III), Nd(III), La(III) and Tb(III).

Leishmaniases are neglected diseases with limited therapeutic options. Diffuse cutaneous leishmaniasis can occur in Brazil due to Leishmania amazonensis. This study details the antileishmanial activity and cytotoxicity of complexes of sodium usnate (SAU) with lanthanide ions ([LnL3 (H2O)x] (Ln = La(III), Nd(III), Gd(III), Tb(III), Eu(III) and Sm(III); L = SAU). All lanthanide complexes were highly active and more potent than SAU against L. amazonensis promastigotes and intracellular amastigotes (Pro: IC50 < 1.50 µM; Ama: IC50 < 7.52 µM). EuL3·3H2O and NdL3·3H2O were the most selective and effective on intracellular amastigotes, with a selectivity index of approximately 7.0. In silico predictions showed no evidence of mutagenicity, tumorigenicity or irritation for all complexes. Treatment with EuL3·3H2O triggered NO release even at the lowest concentration, indicating NO production as a mechanism of action against the parasite. Incubating promastigotes with the lanthanide complexes, particularly with SmL3·4H2O and GdL3·3H2O, led to a change in the mitochondrial membrane potential, indicating the ability of these complexes to target this essential organelle. The same complexes caused cell death through cell membrane disruption, but their relationship with early or late apoptotic processes remains unclear. Thus, the inclusion of lanthanide ions in SAU improves selectivity with a promising mechanism of action targeting the mitochondria.

Raillietia auris (Mesostigmata: Raillietiidae) in cattle in the state of Mato Grosso do Sul, Brazil.

Parasitic otitis in cattle, caused by mites, has been reported from several continents. The present study aimed to determine the distribution, prevalence, intensity, mean intensity, and range of the agent in cattle in the state of Mato Grosso do Sul, Brazil. The samples were designed at random, with an acceptable margin of error of 3% and a confidence interval of 99%. A total of 449 animals were sampled immediately after slaughter from 34 different municipalities in the state using the technique of flushing both ear canals. Only Raillietia auris (Leidy, 1872) were found, with a prevalence of 98.6%, mean intensity of 53.78 mites/animal, and a range of 1-323. Impressively, the prevalence found was identical to another survey carried out 39 years ago in the same region. Details about the parasite intensity in different age categories of the animals are presented. The study demonstrates that the prevalence and intensity of infestation by Raillietia auris are high, and in older cattle are higher than young ones.

Canine Visceral Leishmaniasis in an Area of Sporadic Transmission in Brazil.

Canine visceral leishmaniasis is a zoonotic disease caused by the protozoan Leishmania infantum in Latin America. Visceral leishmaniasis (VL) diagnosis in Brazil includes two serological tests according to the Ministry of Health (MH) protocol. Sensitivity and specificity of diagnostic tests, as well as clinical signs of VL, are usually reported in disease-endemic areas; however, it is known that local epidemiological factors can influence these results. This study aimed to evaluate the clinical features, sensitivity, and specificity of TR-DPP® and EIE-LVC in naturally infected dogs in a region of sporadic VL transmission to humans in Brazil. A total of 288 dogs were clinically evaluated and serological and parasitological (lymph node aspirates) samples were collected for VL diagnosis. TR-DPP and EIE-LVC showed poor sensitivity (0.62 and 0.44, respectively) to detect infected animals, compared with the direct parasitological examination, which is considered a gold standard method. Thus, the protocol of MH presented low sensitivity (0.42) to estimate prevalence and control measures in this region. TR-DPP presented a high negative predictive value (0.89), resulting in its indication as a confirmatory test in sporadic transmission areas. Classical clinical signs of VL were not frequently observed; therefore, clinical scoring systems might not be useful in this region. Veterinarians of nonendemic areas should be alert for asymptomatic dogs, especially those presenting lymph adenomegaly.

Improving the serodiagnosis of canine Leishmania infantum infection in geographical areas of Brazil with different disease prevalence.

Serodiagnosis of Leishmania infantum infection in dogs relies on the detection of antibodies against leishmanial crude extracts or parasitic defined antigens. The expansion of canine leishmaniasis from geographical areas of Brazil in which the infection is endemic to regions in which the disease is emerging is occurring. This fact makes necessary the analysis of the serodiagnostic capabilities of different leishmanial preparations in distinct geographical locations. In this article sera from dogs infected with Leishmania and showing the clinical form of the disease, were collected in three distinct Brazilian States and were tested against soluble leishmanial antigens or seven parasite individual antigens produced as recombinant proteins. We show that the recognition of soluble leishmanial antigens by sera from these animals was influenced by the geographical location of the infected dogs. Efficacy of the diagnosis based on this crude parasite preparation was higher in newly endemic regions when compared with areas of high disease endemicity. We also show that the use of three of the recombinant proteins, namely parasite surface kinetoplastid membrane protein of 11 kDa (KMP-11), and two members of the P protein family (P2a and P0), can improve the degree of sensitivity without adversely affecting the specificity of the diagnostic assays for canine leishmaniasis, independently of the geographical area of residence. In addition, sera from dogs clinically healthy but infected were also assayed with some of the antigen preparations. We demonstrate that the use of these proteins can help to the serodiagnosis of Leishmania infected animals with subclinical infections. Finally, we propose a diagnostic protocol using a combination of KMP-11, P2a y P0, together with total leishmanial extracts.

Abiotic factors and population dynamic of Aedes aegypti and Aedes albopictus in an endemic area of dengue in Brazil.

Aedes aegypti and Aedes albopictus are exotic species in the Americas with high epidemiological relevance as they are vectors of many pathogens. This study aimed at understanding the population dynamics of A. aegypti and A. albopictus and the influence of abiotic factors in an endemic area of dengue. The study was conducted in the urban area of Campo Grande, Mato Grosso do Sul, Brazil, over one year. In seven regions of the city, 50 ovitraps were installed in each neighborhood. The development of the larvae was monitored under controlled laboratory conditions until they reached the adult phase. A total of 50,900 eggs of Aedes sp. were collected, 26,073 of which reached adulthood: 25,496 (97.8 %) A. aegypti and 540 (2.1%) A. albopictus. A. aegypti was observed in all months during the study. The highest number of A. albopictus eggs were collected in June, while in August and September, an absence of this species was noted. Abiotic factors such as temperature, humidity and rainfall were responsible for the observed fluctuations in the mosquito population. The presence of A. albopictus in the urban area of the city is concerning because it could become a potential vector for other arboviruses that afflict human populations. The occurrence of these species in Campo Grande reinforces the need for constant entomological and epidemiological surveillance so that informed actions could be taken to decrease potential breeding sites.

Sandfly fauna (Diptera: Psychodidae) in an urban area, Central-West of Brazil.

Biological and ecological relations among vectors and their pathogens are important to understand the epidemiology of vector-borne diseases. Camapuã is an endemic area for visceral and tegumentary leishmaniasis. The aim of this study was to characterize the sandfly fauna present in Camapuã , MS, Brazil. Sand flies were collected every fortnight from May 2014 to April 2015 using automatic light traps in the domicile and peridomicile of twelve neighborhoods and forest. The collected specimens were identified based on morphology according to the valid identification keys. In total, 2005 sandflies of five genera and nine species were collected. Nyssomyia whitmani and Lutzomyia cruzi were the most abundant species. Males were more abundant, with a male-to-female ratio of 2.14. The highest diversity was observed in peripheral neighborhood, with abundant plant cover. The peridomicile presented greater abundance of sandflies, with the predominance of Ny. whitmani . No significant correlation between the absolute frequencies of the most abundant species and the precipitation variable was observed; however, there was a predominance of Lu. cruzi in the rainy season. We observed a high frequency of sandflies in urban area, especially vector species. The presence of Nyssomyia whitmani and Lutzomyia cruzi indicate the necessity for health surveillance in the municipality. Additional method of collection such as sticky trap is also recommended for appropriate faunestic study.

Molecular detection of Hepatozoon spp. in domestic dogs and wild mammals in southern Pantanal, Brazil with implications in the transmission route.

Hepatozoon parasites comprise intracellular apicomplexan parasites transmitted to vertebrate animals by ingestion of arthropods definitive hosts. The present work aimed to investigate the occurrence of Hepatozoon spp. in wild animals, domestic dogs and their respective ectoparasites, in southern Pantanal region, central-western Brazil, by molecular techniques. Between August 2013 and March 2015, 31 coatis (Nasua nasua), 78 crab-eating foxes (Cerdocyon thous), seven ocelots (Leopardus pardalis), 42 dogs (Canis lupus familiaris), 110 wild rodents (77 Thichomys fosteri, 25 Oecomys mamorae, and 8 Clyomys laticeps), 30 marsupials (14 Thylamys macrurus, 11 Gracilinanus agilis, 4 Monodelphis domestica and 1 Didelphis albiventris), and 1582 ticks and 80 fleas collected from the sampled animals were investigated. DNA samples were submitted to PCR assays for Hepatozoon spp. targeting 18S rRNA gene. Purified amplicons were directly sequenced and submitted to phylogenetic analysis. A high prevalence of Hepatozoon among carnivores (C. thous [91.02%], dogs [45.23%], N. nasua [41.9%] and L. pardalis [71.4%]) was found. However, ticks and fleas were negative to Hepatozoon PCR assays. By phylogenetic analysis based on 18S rRNA sequences, Hepatozoon sequences amplified from crab-eating foxes, dogs, coatis and ocelots clustered with sequences of H. canis, H. americanum and H. felis. The closely related positioning of Hepatozoon sequences amplified from wild rodents and T. macrurus marsupial to Hepatozoon from reptiles and amphibians suggest a possible transmission of those Hepatozoon species between hosts by ectoparasites or by predation. Hepatozoon haplotypes found circulating in wild rodents seem to present a higher degree of polymorphism when compared to those found in other groups of animals. Although rodents seem not to participate as source of Hepatozoon infection to wild carnivores and domestic dogs, they may play an important role in the transmission of Hepatozoon to reptiles and amphibians in Pantanal biome.

Life cycle of Ornithodoros rostratus (Acari: Argasidae) under experimental conditions and comments on the host-parasite relationship in the Pantanal wetland region, Brazil.

The genus Ornithodoros is represented by 15 species in Brazil, on which no detailed life cycle studies have been published, except for O. talaje and O. mimon. The aim of the present study was to evaluate life cycle parameters of O. rostratus based on ticks collected in the Pantanal wetland region of the state of Mato Grosso do Sul, Brazil, using domestic rabbits as experimental hosts. The periods of pre-attachment and feeding of the larvae lasted an average of 39 min (range 15-76 min). Five or six nymphal instars were found. The emergence of adults started with N3 in the following sequence: N3-two males; N4-13 males; N5-three males and 16 females; and N6-two females. Mean weight of N4 that molted to males was 31.7 ± 13.6 mg, whereas mean weight of N5 that molted to females was 100.1 ± 36.2 mg. The overall sex ratio was 1:1. Oviposition lasted 14 days, with a sharp decline beginning with the 7th day. The overall duration of the life cycle of O. rostratus ranged from approximately 66 to 136 days. Comments on the tick-host relationship in the Pantanal region are offered.

Assessment of weight gain and biological parameters of Rhipicephalus sanguineus females fed artificially via capillary tubes / Avaliação do ganho de peso e dos parâmetros biológicos de fêmeas Rhipicephalus sanguineus alimentadas artificialmente por meio de tubos capilares

This study assesses the weight gain of partially engorged Rhipicephalus sanguineus females that were artificially fed via capillary tubes and the influence of capillary tube feeding on the biological parameters of the non-parasitic stage of the species. The ticks were sorted into four groups, each containing ten females of a homogeneous weight. The groups were each treated for different feeding times, 2, 6, 12 and 24 hours. The weight gain of the artificially fed females was measured, and the biological parameters of the non-parasitic stage of the tick were observed for each treatment group. The statistical non-parametrical Dunn and Kruskal-Wallis tests were used to compare the results. The mean weights (mg) were 0.2±2.4; 4.3±5.8; 7.4±5.8 and 12.0±11.2 for the 2, 6, 12 and 24 hours feeding groups, respectively. The weight of the fed groups increased as the capillary feeding time increased, and this relationship was highly significant (P<0.05) between the groups fed for 2 and 24 hours. No statistically significant differences (P>0.05) were observed in the parameters of the non-parasitic stage for the artificially fed groups. It can be concluded that artificial feeding via capillary tubes provides an efficient and easy method for the artificial intake of blood by R. sanguineus. Furthermore, it was noted that the ticks fed in vitro were able to establish a new generation. The experimental method shows great promise in studies that aim to investigate biological disease agents.

O objetivo deste trabalho foi avaliar o ganho de peso de fêmeas Rhipicephalus sanguineus parcialmente ingurgitadas alimentadas artificialmente por meio de tubos capilares e verificar a sua influência nos parâmetros biológicos da fase não parasitária. Os carrapatos foram separados em quatro grupos de peso homogêneo compostos por 10 fêmeas cada. Os grupos foram submetidos a diferentes tempos de alimentação: 2, 6, 12 e 24 horas. Para comparação dos resultados, foram utilizados os testes estatísticos não paramétricos Dunn e Kruskal-Wallis. Os pesos médios (mg) adquiridos foram de 0,2±2,4; 4,3±5,8; 7,4±5,8 e 12,0±11,2 nos grupos de 2, 6, 12 e 24 horas, respectivamente. O peso dos grupos alimentados foi maior à medida que o tempo de exposição ao tubo capilar aumentou, observando-se diferença significativa (P<0,05) entre os grupos alimentados por 2 e 24 horas. Não houve diferenças estatísticas significativas (P>0,05) nos parâmetros relativos à fase não parasitária entre os grupos submetidos à alimentação artificial. Concluiu-se que a alimentação artificial por meio de tubos capilares é um método de fácil execução e eficiente para ingestão de sangue por carrapatos R. sanguineus. Além disso, verificou-se que os carrapatos alimentados in vitro foram capazes de estabelecer uma nova geração. Esse método tem potencial para o desenvolvimento de estudos que visem a transmissão de bioagentes.

Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) as a parasite of pampas deer (Ozoctoceros bezoarticus) and cattle in Brazil's Central Pantanal.

In the Pantanal, domestic and wild animals share the same habitats; their parasites and environment interact in a complex and dynamic way. The Rhipicephalus (Boophilus) microplus is the most important cattle tick in Brazil. In the past two decades, some traditional management practices are being replaced with the aim of intensifying cattle production. Forested areas are being replaced by exotic pasture and ranch owners are replacing Nelore for European breeds. The pampas deer (Ozoctoceros bezoarticus) is a medium-sized cervid that occurs in grasslands of South America. Between December 2005 and January 2007, we captured and collected ticks from 15 pampas deer and 172 bovines. The abundance, intensity, and prevalence of ticks found in cattle were lower than those in pampas deer (p < 0.05), indicating that pampas deer is more susceptible than cattle. The high number of engorged females collected from pampas deer, their capacity to lay eggs, and the percentage of eclosion indicates that the pampas deer population in the Pantanal wetlands is sufficient to maintain the R. (B.) microplus, thus playing a important role in the transmission of ticks and tick-borne diseases. The recent deforestation to introduce Brachiaria sp. and European cattle needs to be carefully evaluated to prevent economic losses to the cattle industry and damage to the wildlife in Pantanal.

Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) as a parasite of pampas deer (Ozoctoceros bezoarticus) and cattle in Brazil's Central Pantanal / Rhipicephalus (Boophilus) microplus como parasita de veado-campeiro (Ozoctoceros bezoarticus) e do gado bovino no Pantanal Central do Brasil

In the Pantanal, domestic and wild animals share the same habitats; their parasites and environment interact in a complex and dynamic way. The Rhipicephalus (Boophilus) microplus is the most important cattle tick in Brazil. In the past two decades, some traditional management practices are being replaced with the aim of intensifying cattle production. Forested areas are being replaced by exotic pasture and ranch owners are replacing Nelore for European breeds. The pampas deer (Ozoctoceros bezoarticus) is a medium-sized cervid that occurs in grasslands of South America. Between December 2005 and January 2007, we captured and collected ticks from 15 pampas deer and 172 bovines. The abundance, intensity, and prevalence of ticks found in cattle were lower than those in pampas deer (p < 0.05), indicating that pampas deer is more susceptible than cattle. The high number of engorged females collected from pampas deer, their capacity to lay eggs, and the percentage of eclosion indicates that the pampas deer population in the Pantanal wetlands is sufficient to maintain the R. (B.) microplus, thus playing a important role in the transmission of ticks and tick-borne diseases. The recent deforestation to introduce Brachiaria sp. and European cattle needs to be carefully evaluated to prevent economic losses to the cattle industry and damage to the wildlife in Pantanal.

No Pantanal Brasileiro, onde animais domésticos e selvagens compartilham os mesmos habitats; hospedeiros, parasitas e ambientes encontram-se em interação complexa e dinâmica. Rhipicephalus (Boophilus) microplus é o carrapato bovino mais importante no Brasil. Nas últimas duas décadas práticas de manejo vem sendo substituídas com objetivo de aumentar a produtividade. Áreas de florestas estão sendo substituídas por pastagens exóticas e os produtores começam a utilizar raças européias no lugar do Nelore. O veado-campeiro é um cervídeo que ocorre em áreas de campo na América do sul. Entre dezembro de 2005 e janeiro de 2007, foram coletados carrapatos de 20 veados-campeiros e 172 bovinos. Os valores de abundância, intensidade parasitária e prevalência de carrapatos no gado foram menores que os valores encontrados no veado-campeiro (p < 0,05), indicando que o veado-campeiro é mais susceptível que o bovino. O elevado número de fêmeas ingurgitadas coletadas, sua capacidade de ovipor e o percentual de eclosão indicam que a população veado-campeiro na região de estudo é suficiente para manter o R. (B.) microplus, desta maneira participando na epidemiologia dos carrapatos e das doenças por eles transmitidas. O recente desmatamento, a introdução de Brachiaria sp. e do gado europeu precisam ser cuidadosamente avaliados, para prevenir perdas na industria do gado e vida selvagem.

Desempenho reprodutivo de Nasonia vitripennis Walker (Hymenoptera: Pteromalidae) em pupas crioconservadas de Chrysomya megacephala Fabricius (Diptera: Calliphoridae): avaliaçäo preliminar / Reproductive performance of Nasonia vitripennis Walker (Hymenoptera: Pteromalidae) on Chrysomya megacephala Fabricius (Diptera: Calliphoridae) cryopreserved pupae: preliminary evaluation

Avaliou-se o desempenho reprodutivo de Nasonia vitripennis Walker (Hymenoptera: Pteromalidae) em pupas de Chrysomya megacephala Fabricius (Diptera: Calliphoridae) previamente armazenadas a - 20ºC de temperatura, durante 77 dias, com e sem passagem prévia em nitrogênio líquido (NL) por um, três e 15 minutos. O grupo controle foi caracterizado por pupas hospedeiras frescas. Os muscóides foram expostos aos parasitóides durante 72 horas. Utilizou-se a relação de uma pupa muscóide por fêmea parasitóide. A amostra de pupas frescas permitiu a emergência de 15 parasitóides/ pupa, em média, enquanto 10 parasitóides / pupa emergiram dos espécimens prévia e diretamente armazenados em freezer (-20ºC). Observou-se um acentuado decréscimo do desempenho reprodutivo dos microhimenópteros que exploraram os substratos previamente submetidos ao NL durante um, três e 15 minutos (: 6,1; 5,5 e 5,7, respectivamente). A dissecação das pupas hospedeiras revelou um expressivo número de pteromalídeos imaturos, nas amostras que foram expostas ao NL, e de adultos faratos, em todos os tratamentos.