Gabriel Carvalho de Macedo

Tick-borne agents in domesticated and stray cats from the city of Campo Grande, state of Mato Grosso do Sul, midwestern Brazil

Anaplasmataceae agents, piroplasmids and Hepatozoon spp. have emerged as important pathogens among domestic and wild felines. The present work aimed to detect the presence of species belonging to the Anaplasmataceae family, piroplasmas and Hepatozoon spp. DNA in blood samples of domesticated and stray cats in the city of Campo Grande, state of Mato Grosso do Sul, midwestern Brazil. Between January and April 2013, whole blood samples were collected from 151 cats (54 males, 95 females and two without gender registration) in the city of Campo Grande, state of Mato Grosso do Sul, Brazil. DNA extracted from cat blood samples was submitted to conventional PCR assays for Theileria/Babesia/Cytauxzoon spp. (18S rRNA, ITS-1), Ehrlichia spp. (16S rRNA, dsb, groESL), Anaplasma spp. (16S rRNA, groESL) and Hepatozoon spp. (18S rRNA) followed by phylogenetic reconstructions. Out of 151 sampled cats, 13 (8.5%) were positive for Ehrlichia spp. closely related to Ehrlichia canis, 1 (0.66%) for Hepatozoon spp. closely related to Hepatozoon americanum and Hepatozoon spp. isolate from a wild felid, 1 (0.66%) for Cytauxzoon sp. closely related do Cytauxzoon felis, and 18 (11.9%) for Babesia/Theileria (one sequence was closely related to Babesia bigemina, eight for Babesia vogeli, five to Theileria spp. from ruminants [Theileria ovis, Theileria lestoquardi] and four to Theileria sp. recently detected in a cat). The present study showed that Ehrlichia spp., piroplasmids (B. vogeli, Theileria spp. and Cytauxzoon spp.) and, more rarely, Hepatozoon spp. circulate among stray and domesticated cats in the city of Campo Grande, state of Mato Grosso do Sul, midwestern Brazil.

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.

Pathology of dogs in Campo Grande, MS, Brazil naturally co-infected with Leishmania infantum and Ehrlichia canis

Different parasites that commonly occur concomitantly can influence one another, sometimes with unpredictable effects. We evaluated pathological aspects of dogs naturally co-infected with Leishmania infantum and Ehrlichia canis. The health status of the dogs was investigated based on histopathological, hematological and biochemical analyses of 21 animals infected solely with L. infantum and 22 dogs co- infected with L. infantum and E. canis. The skin of both groups showed chronic, predominantly lymphohistioplasmacytic inflammatory reaction. The plasmacytosis in the lymphoid tissues was likely related with the hypergammaglobulinemia detected in all the dogs. The disorganization of extracellular matrix found in the reticular dermis of the inguinal region and ear, characterized by the substitution of thick collagen fibers for thin fibers, was attributed to the degree of inflammatory reaction, irrespective of the presence of parasites. In addition, the histopathological analysis revealed that twice as many dogs in the co-infected group presented Leishmania amastigotes in the ear skin than those infected solely with Leishmania, increasing the possibility of becoming infected through sand fly vectors. Our findings highlight the fact that the health of dogs infected concomitantly with L. infantum and E. canis is severely compromised due to their high levels of total plasma protein, globulins, alkaline phosphatase and creatine kinase, and severe anemia.

Health and epidemiological approaches of Trypanosoma evansi and equine infectious anemia virus in naturally infected horses at southern Pantanal.

Equine infectious anemia virus (EIAV) and Trypanossoma evansi are endemic in Brazilian Pantanal Biome, an important area for livestock production. In this sense, we evaluated the epidemiological single and co-infection effects of T. evansi and EIAV in naturally infected horses in the southern Pantanal wetland by serological tests and hematological assays. Both higher seroprevalence and heath poor condition of the sampled animals were associated with differences in horse management between farms. We found that the negative animals for both infectious agents (NN) represented the major group in F1 (37%), and the smallest group in F2 (19%). Furthermore, we recorded higher EIAV seroprevalence (56%) in F2, compared to F1 (38%). We observed that T. evansi infection was mostly related to young horses, as seen by their higher seroprevalence, ranging from 70.7% in the beginning of the rainy season to 81% in the end of flood period, in comparison with the values of 42% and 68%, respectively, in working animals. on the other hand, working animals showed a higher seroprevalence for EIAV (48%) in both seasons than young horses. We observed that the management of working horses could be a risk factor of EIAV infection. On the other hand, as T. evansi is maintained in the study region by many species of wild mammals, the mechanical transmission through blood-sucking vectors ensures the infection to horses since early. Our results showed that single or co-infection by EIAV and T. evansi caused different degree of anemia in the infected animals. Moreover, the health of horses in Brazilian Pantanal is also influenced by differences in horse management and environmental circumstances.

Genetic Diversity of Bartonella spp. in Wild Mammals and Ectoparasites in Brazilian Pantanal

The present work aimed to investigate the genetic diversity of Bartonella in mammals and ectoparasites in Pantanal wetland, Brazil. For this purpose, 31 Nasua nasua, 78 Cerdocyon thous, 7 Leopardus pardalis, 110 wild rodents, 30 marsupials, and 42 dogs were sampled. DNA samples were submitted to a quantitative real-time PCR assay (qPCR). Positive samples in qPCR were submitted to conventional PCR assays targeting other five protein-coding genes. Thirty-five wild rodents and three Polygenis (P.) bohlsi bohlsi flea pools showed positive results in qPCR for Bartonella spp. Thirty-seven out of 38 positive samples in qPCR were also positive in cPCR assays based on ftsZ gene, nine in nuoG-cPCR, and six in gltA-cPCR. Concatenated phylogenetic analyses showed that two main genotypes circulate in rodents and ectoparasites in the studied region. While one of them was closely related to Bartonella spp. previously detected in Cricetidae rodents from North America and Brazil, the other one was related to Bartonella alsatica, Bartonella pachyuromydis, Bartonella birtlesii, Bartonella acomydis, Bartonella silvatica, and Bartonella callosciuri. These results showed that at least two Bartonella genotypes circulate among wild rodents. Additionally, the present study suggests that Polygenis (P.) bohlsi bohlsi fleas could act as possible Bartonella vectors among rodents in Pantanal wetland, Brazil.

Correction: Outcomes of Trypanosoma cruzi and Trypanosoma evansi infections on health of Southern coati (Nasua nasua), crab-eating fox (Cerdocyon thous), and ocelot (Leopardus pardalis) in the Brazilian Pantanal

The occurrence of Trypanosoma spp. in wild carnivore populations has been intensively investigated during the last decades. However, the impact of these parasites on the health of free-living infected animals has been largely neglected. The Pantanal biome is the world’s largest seasonal wetland, harboring a great diversity of species and habitats. This includes 174 species of mammals, of which 20 belong to the order Carnivora. The present study aimed to investigate the effect of Trypanosoma evansi and Trypanosoma cruzi infections and coinfections on the health of the most abundant carnivores in the Pantanal: coati (Nasua nasua), crab-eating fox (Cerdocyon thous), and ocelot (Leopardus pardalis). We captured 39 coatis, 48 crab-eating foxes, and 19 ocelots. Diagnostic tests showed T. cruzi infection in 7 crab-eating foxes and 5 coatis. Additionally, 7 crab-eating foxes, 10 coatis, and 12 ocelots were positive for T. evansi. We observed coinfections in 9 crab-eating foxes, 8 coatis, and 2 ocelots. This is the first report of T. evansi and T. cruzi infection on the health of free-living ocelots and crab-eating foxes. We showed that single T. evansi or T. cruzi infection, as well as coinfection, caused some degree of anemia in all animals, as well as an indirect negative effect on body condition in coatis and crab-eating foxes via anemia indicators and immune investment, respectively. Furthermore, the vigorous immune investment observed in sampled coatis, crab-eating foxes and ocelots infected by T. evansi, T. cruzi and coinfected can be highly harmful to their health. Overall, our results indicate that single and combined infection with T. evansi and T. cruzi represent a severe risk to the health of wild carnivores in the Pantanal region.

Maintenance of Trypanosoma cruzi, T. evansi and Leishmania spp. by domestic dogs and wild mammals in a rural settlement in Brazil-Bolivian border

Domestic dogs are considered reservoirs hosts for several vector-borne parasites. This study aimed to evaluate the role of domestic dogs as hosts for Trypanosoma cruzi, Trypanosoma evansi and Leishmania spp. in single and co-infections in the Urucum settlement, near the Brazil-Bolivian border. Additionally, we evaluated the involvement of wild mammals’ in the maintenance of these parasites in the study area. Blood samples of dogs (n = 62) and six species of wild mammals (n = 36) were collected in July and August of 2015. The infections were assessed using parasitological, serological and molecular tests. Clinical examination of dogs was performed and their feeding habits were noted. Overall, 87% (54/62) of sampled dogs were positive for at least one trypanosomatid species, in single (n = 9) and co-infections (n = 45). We found that 76% of dogs were positive for T. cruzi, four of them displayed high parasitemias demonstrated by hemoculture, including one strain types TcI, two TcIII and one TcIII/TcV. Around 73% (45/62) of dogs were positive to T. evansi, three with high parasitemias as seen by positive microhematocrit centrifuge technique. Of dogs sampled, 50% (31/62) were positive for Leishmania spp. by PCR or serology. We found a positive influence of (i) T. evansi on mucous pallor, (ii) co-infection by T. cruzi and Leishmania with onychogryphosis, and (iii) all parasites to skin lesions of sampled dogs. Finally, feeding on wild mammals had a positive influence in the Leishmania spp. infection in dogs. We found that 28% (5/18) coati Nasua nasua was co-infected for all three trypanosamatids, demonstrating that it might play a key role in maintenance of these parasites. Our results showed the importance of Urucum region as a hotspot for T. cruzi, T. evansi and Leishmania spp. and demonstrated that dogs can be considered as incidental hosts.

Terrestriality of Wild Sapajus cay (Illiger, 1815) as Revealed by Camera Traps

Although primarily arboreal, the capuchin monkey (Sapajus cay) descends to the ground for several reasons. We used terrestrial records obtained by camera-trapping surveys to investigate seasonality in the terrestriality of capuchin monkeys, reasons to descend to the ground, and periods of the day when terrestriality was more pronounced. We carried out the study in the Urucum Massif, Brazilian Pantanal. We obtained data from 2 camera-trapping surveys carried out in the dry and rainy seasons. Terrestrial behaviours were categorized, and terrestrial activity patterns were described using a kernel density approach. We observed a seasonal effect on the frequency of terrestrial behaviours of capuchin monkeys, who used the ground more during the dry season. We identified 6 different types of terrestrial behaviour, but travelling (33.3%), foraging (23.3%), and drinking water (23.3%) were the most frequently observed. All records occurred during the day, with 2 peaks in terrestrial activity. Seasonal terrestriality was mainly linked to ecological needs during periods of scarce food and water.