Molecular Detection of Rickettsia parkeri Strain Atlantic Rainforest in Ticks Parasitizing Small Mammals in Northeastern Brazil.

PURPOSE: Small mammals are important reservoirs of ticks and their pathogens in nature. However, studies reporting these associations are still rare in Brazil. In the present study, we investigated the presence of Rickettsia DNA in ticks parasitizing rodents and marsupials captured in different areas throughout the Atlantic rainforest biome, Bahia, Northeastern (NE), Brazil. METHODS: The study was conducted in five municipalities within of the Atlantic Forest biome, Bahia state, in NE Brazil. Two campaigns were done in each municipality. For host captures Sherman and Tomahawk traps were used, and pitfall traps. After being captured, the hosts were anesthetized and their entire body examined for ticks. When ticks were detected, they were manually removed and stored in eppendorf tubes (1.5 ml) containing absolute PA ethanol for future laboratory analysis (identification of ticks and detection of Rickettsia spp.). RESULTS: A total of 609 mammals were captured. Overall, 208 ticks of the genus Amblyomma and Ixodes were collected: A. ovale, I. loricatus and A. varium. Rickettsia DNA was detected in A. ovale and it was 99-100% of identity to the sequence deposited in GenBank as Rickettsia parkeri strain Atlantica rainforest. CONCLUSION: These results suggest that R. parkeri strain Atlantic rainforest occurs in the region, and A. ovale is likely the vector.

Records and altitudinal assessment of Amblyomma aureolatum and Amblyomma ovale (Acari: Ixodidae) in the State of Rio de Janeiro, southeast Brazil.

Information on the altitudinal distribution of the hard ticks Amblyomma aureolatum and Amblyomma ovale in Brazil is scarce and mainly limited to occasional records. In this study we report our evaluation of records on the altitudinal distribution of A. aureolatum and A. ovale collected from dogs and humans and directly from the environment (host-questing ticks), based on active or passive procedures. The collections were conducted in rural areas of municipalities in the state of Rio de Janeiro, Brazil between 2013 and 2017. Active procedures consisted of dragging or flagging, visual examination of vegetation and removal of ticks present on the authors' clothing or on infested dogs. Overall, 222 ticks were collected. The altitudes at the collection sites ranged from 98 to 1220 m a.s.l. We noted a significant difference in the altitudinal distribution of A. aureolatum and A. ovale (Mann-Whitney U-test, U = 518.5, P < 0.001). The overlap of these two species occurred at altitudes of between 650 and 900 m a.s.l. The results indicated that the higher the altitude, the greater the probability for the occurrence of A. aureolatum and, conversely, the lower the likelihood for the occurrence of A. ovale. The findings of this study improve currrent knowledge on the bioecology of these tick species and have implications for studies on the epidemiology of spotted fever in Brazil.

A longitudinal survey of gastrointestinal parasites of the black-eared opossum Didelphis aurita at an urban-sylvatic interface area in Southeast Brazil: a morphological and ecological characterization based on helminth eggs.

Parasites are important organisms in ecosystem dynamics and take part in the regulation and structure of host populations. The prevalence, abundance, and distribution of parasites in their host species may be affected by biotic and abiotic factors. Most studies of helminths of Brazilian marsupials are taxonomic descriptions or re-descriptions and records of occurrence. The use of noninvasive techniques for studying parasitic worms of vertebrate hosts is more common in large or threatened species. The aims of this study were to describe and identify the helminth morphotypes and to analyse the parasitological parameters of gastrointestinal helminth eggs obtained in faecal samples of marsupial Didelphis aurita at the Fiocruz Atlantic Forest Biological Station and surroundings, municipality of Rio de Janeiro, southeast Brazil. The common opossums were sampled during a capture-mark-recapture study. Faecal samples collected from each animal were analysed for helminth egg diagnosis using the methods of flotation in sugar solution and sedimentation. Eggs were compared with samples obtained from the uterus of adult worms obtained from a previous study carried out in the same area. Eleven helminth egg morphotypes were found. Among them, seven were identified at the species level. The highest values of parasitic load and prevalence were observed for the families Viannaiidae and Trichuridae. A significant relationship was found between the number of morphotypes and host body size in females. The influence of parasite load on host condition factor or body size was not observed.

Histological assessment of granulomas in natural and experimental Schistosoma mansoni infections using whole slide imaging.

The pathology of schistosomiasis mansoni, a neglected tropical disease of great clinical and socioeconomic importance, results from the parasite eggs that become trapped in host tissues, particularly in the liver and intestines. Continuous antigenic stimulation from these eggs leads to recruitment of inflammatory cells to the sites of infection with formation of periovular granulomas. These complex structures have variable size and composition and are the most striking histopathological feature of schistosomiasis mansoni. However, evaluation of granulomas by conventional microscopy methods is time-consuming and limited, especially in large-scale studies. Here, we used high resolution Whole Slide Imaging (WSI), which allows fast scanning of entire histological slides, and multiple morphometric evaluations, to assess the granulomatous response elicited in target organs (liver, small and large intestines) of two models of schistosomiasis mansoni. One of the advantages of WSI, also termed virtual microscopy, is that it generates images that simultaneously offer high resolution and a wide field of observation. By using a model of natural (Nectomys squamipes, a wild reservoir captured from endemic areas in Brazil) and experimental (Swiss mouse) infection with Schistosoma mansoni, we provided the first detailed WSI characterization of granulomas and other pathological aspects. WSI and quantitative analyses enabled a fast and reliable assessment of the number, evolutional types, frequency and areas of granulomas and inflammatory infiltrates and revealed that target organs are differentially impacted by inflammatory responses in the natural and experimental infections. Remarkably, high-resolution analysis of individual eosinophils, key cells elicited by this helminthic infection, showed a great difference in eosinophil numbers between the two infections. Moreover, features such as the intestinal egg path and confluent granulomas were uncovered. Thus, WSI may be a suitable tool for detailed and precise histological analysis of granulomas and other pathological aspects for clinical and research studies of schistosomiasis.

Natural Schistosoma mansoni Infection in the Wild Reservoir Nectomys squamipes Leads to Excessive Lipid Droplet Accumulation in Hepatocytes in the Absence of Liver Functional Impairment.

Schistosomiasis is a neglected tropical disease of a significant public health impact. The water rat Nectomys squamipes is one of the most important non-human hosts in the schistosomiasis mansoni transmission in Brazil, being considered a wild reservoir. Cellular mechanisms that contribute to the physiological adaptation of this rodent to the Schistosoma mansoni parasite are poorly understood. Here we identified, for the first time, that a hepatic steatosis, a condition characterized by excessive lipid accumulation with formation of lipid droplets (LDs) within hepatocytes, occurs in response to the natural S. mansoni infection of N. squamipes, captured in an endemic region. Significant increases of LD area in the hepatic tissue and LD numbers/hepatocyte, detected by quantitative histopathological and ultrastructural analyses, were paralleled by increased serum profile (total cholesterol and triglycerides) in infected compared to uninfected animals. Raman spectroscopy showed high content of polyunsaturated fatty acids (PUFAs) in the liver of both groups. MALDI-TOFF mass spectroscopy revealed an amplified pool of omega-6 PUFA arachidonic acid in the liver of infected animals. Assessment of liver functional activity by the levels of hepatic transaminases (ALT and AST) did not detect any alteration during the natural infection. In summary, this work demonstrates that the natural infection of the wild reservoir N. squamipes with S. mansoni elicits hepatic steatosis in the absence of liver functional harm and that accumulation of lipids, markedly PUFAs, coexists with low occurrence of inflammatory granulomatous processes, suggesting that lipid stores may be acting as a protective mechanism for dealing with the infection.

An ecological field study of the water-rat Nectomys squamipes as a wild reservoir indicator of Schistosoma mansoni transmission in an endemic area

Small mammals are found naturally infected by Schistosoma mansoni, becoming a confounding factor for control programs of schistosomiasis in endemic areas. The aims of this study were: to investigate the infection rates by S. mansoni on the water-rat Nectomys squamipes during four years in endemic areas of Sumidouro, state of Rio de Janeiro, using mark-recapture technique; to compare two diagnostic methods for schistosomiasis; and to evaluate the effects of the chemotherapy in the human infected population on the rodent infection rates. The rodent infection rates of S. mansoni increased when rodent population sizes were lower. Coprology and serology results presented the same trends along time and were correlated. Serology could detect recent infection, including the false negatives in the coprology. The chemotherapy in the humans could not interrupt the rodent infection. Rodents can increase the schistosomiaisis transmission where it already exists, they probably maintain the transmission cycle in the nature and can be considered as biological indicators of the transmission sites of this parasite since they are highly susceptible to infection. The water-rats may present different levels of importance in the transmission dynamics of S. mansoni infection cycle for each area, and can be considered important wild-reservoirs of this human disease.

An ecological field study of the water-rat Nectomys squamipes as a wild reservoir indicator of Schistosoma mansoni transmission in an endemic area.

Small mammals are found naturally infected by Schistosoma mansoni, becoming a confounding factor for control programs of schistosomiasis in endemic areas. The aims of this study were: to investigate the infection rates by S. mansoni on the water-rat Nectomys squamipes during four years in endemic areas of Sumidouro, state of Rio de Janeiro, using mark-recapture technique; to compare two diagnostic methods for schistosomiasis; and to evaluate the effects of the chemotherapy in the human infected population on the rodent infection rates. The rodent infection rates of S. mansoni increased when rodent population sizes were lower. Coprology and serology results presented the same trends along time and were correlated. Serology could detect recent infection, including the false negatives in the coprology. The chemotherapy in the humans could not interrupt the rodent infection. Rodents can increase the schistosomiaisis transmission where it already exists, they probably maintain the transmission cycle in the nature and can be considered as biological indicators of the transmission sites of this parasite since they are highly susceptible to infection. The water-rats may present different levels of importance in the transmission dynamics of S. mansoni infection cycle for each area, and can be considered important wild-reservoirs of this human disease.