Survey of Rickettsia spp. in ticks (Acari: Ixodidae) infesting opossums (Didelphis albiventris) and capybaras (Hydrochoerus hydrochaeris) from an urban park in southeastern Brazil.

Ticks are hematophagous arthropods and, during feeding, may transmit pathogens to vertebrate hosts, including humans. This study aimed to investigate the presence of Rickettsia spp. in ticks collected between 2010 and 2013 from free-ranging capybaras (Hydrochoerus hydrochaeris) and opossums (Didelphis albiventris) that inhabit Sabiá Park in Uberlândia, Brazil. Overall, 1,860 ticks were collected: 1,272 (68.4%) from capybaras (487 of the species Amblyomma sculptum, 475 adults and 12 nymphs; 778 Amblyomma dubitatum, 727 adults and 51 nymphs; and seven larva clusters of the genus Amblyomma); and 588 (31.6%) from opossums (21 A. sculptum, one adult and 20 nymphs; 79 A. dubitatum, all nymphs; 15 Ixodes loricatus, 12 adults and three nymphs; 457 Amblyomma sp. larva clusters; 15 Ixodes sp. larva clusters; and one Argasidae larva cluster). Out of 201 DNA samples tested for the presence of Rickettsia spp. DNA using polymerase chain reaction (PCR) 12 showed amplification of a gtlA gene segment that was specific to Rickettsia bellii, a bacterium non-pathogenic to humans. As there has been a report showing serological evidence of infections caused by Rickettsia species of the spotted fever group (SFG) in capybaras and opossums in the park, including Rickettsia rickettsii, the etiological agent of Brazilian spotted fever, and considering the presence of A. sculptum ticks, which are aggressive to humans, as well as these vertebrate hosts, which are amplifiers of R. rickettsii, it is important to monitor the presence of SFG rickettsiae in the Sabiá Park, which is visited daily by thousands of people.

Evaluation of a mimotope of the Rickettsia outer membrane protein A (OmpA) as an antigen in enzyme-linked immunosorbent assay to detect rickettsiosis in capybaras (Hydrochoerus hydrochaeris), horses (Equus caballus), and opossums (Didelphis sp.).

Rickettsia rickettsii is the etiological agent of Rocky Mountain spotted fever, which is an important tick-borne zoonosis and, in Brazil, it causes Brazilian spotted fever, which has high lethality rate. This study aimed to evaluate a synthetic peptide corresponding to a segment of the outer membrane protein A (OmpA) as an antigen in a serological test for the diagnosis of rickettsial infections. The amino acid sequence of the peptide was selected by predicting B cell epitopes using B Cell Epitope Prediction (Immune Epitope Database and Analysis Resource) and Epitopia and OmpA sequences of Rickettsia rickettsii strain 'Brazil' and Rickettsia parkeri strains 'Maculatum 20' and 'Portsmouth'. A peptide with amino acid sequence common to both Rickettsia species was synthesized and arbitrarily named OmpA-pLMC. To evaluate this peptide in enzyme-linked immunosorbent assay (ELISA), serum samples of capybara (Hydrochoerus hydrochaeris), horse (Equus caballus), and opossum (Didelphis albiventris) that had been previously tested by indirect immunofluorescence assay (IFA) for rickettsial infection were separated into IFA-positive and IFA-negative groups and used in the assay. There were no significant differences in ELISA optical density (OD) values between IFA-positive and IFA-negative groups with horse samples. The mean OD values were significantly higher in the IFA-positive capybara serum samples (IFA-pos vs. IFA-neg = 2.389 ± 0.761 vs. 1.760 ± 0.840). However, receiver operating characteristic (ROC) curve analysis did not show significant diagnostic parameters. On the other hand, 12 out of 14 (85.7%) opossum samples of the IFA-positive group showed reactivity in ELISA, and this was significantly higher than of the IFA-negative group (0.7196 ± 0.440 vs. 0.2318 ± 0.098, respectively; 85.7% sensitivity, 100% specificity). Therefore, our results show that OmpA-pLMC has a potential to be used in immunodiagnostic assays to detect spotted fever group rickettsial infections.

Bacterial pathogens' screening in Brazilian chigger mites (Trombidiformes: Trombiculidae), with the first report of 'Candidatus Rickettsia colombianensi'-like in avian-associated chiggers.

Chiggers are larval ectoparasites of the Trombiculidae that can transmit pathogens to their hosts. In this study, chiggers collected from birds in Brazil were morphologically identified as Blankaartia sinnamaryi, Eutrombicula batatas, Eutrombicula daemoni, Eutrombicula goeldii, Eutrombicula tinami, and Parasecia gilbertoi. For these specimens, a beginning attempt at molecular identification were also provided, as well as, were genetically screened to detect bacterial pathogens. The species B. sinnamaryi and E. tinami were positive for Rickettsia felis-like and 'Candidatus Rickettsia colombianensi'-like, respectively. For the other agents (Anaplasmataceae, Borrelia spp. and Orientia tsutsugamushi), the tests were negative. This is the first report of 'Ca. R. colombianensi'-like and the second record of R. felis-like in chigger collected on birds from Brazil.

Haemosporidian parasites and incubation period influence plumage coloration in tanagers (Passeriformes: Thraupidae).

Birds are highly visually oriented and use plumage coloration as an important signalling trait in social communication. Hence, males and females may have different patterns of plumage coloration, a phenomenon known as sexual dichromatism. Because males tend to have more complex plumages, sexual dichromatism is usually attributed to female choice. However, plumage coloration is partly condition-dependent; therefore, other selective pressures affecting individuals' success may also drive the evolution of this trait. Here, we used tanagers as model organisms to study the relationships between dichromatism and plumage coloration complexity in tanagers with parasitism by haemosporidians, investment in reproduction and life-history traits. We screened blood samples from 2849 individual birds belonging to 52 tanager species to detect haemosporidian parasites. We used publicly available data for plumage coloration, bird phylogeny and life-history traits to run phylogenetic generalized least-square models of plumage dichromatism and complexity in male and female tanagers. We found that plumage dichromatism was more pronounced in bird species with a higher prevalence of haemosporidian parasites. Lastly, high plumage coloration complexity in female tanagers was associated with a longer incubation period. Our results indicate an association between haemosporidian parasites and plumage coloration suggesting that parasites impact mechanisms of sexual selection, increasing differences between the sexes, and social (non-sexual) selection, driving females to develop more complex coloration.

Host life-history traits predict haemosporidian parasite prevalence in tanagers (Aves: Thraupidae).

Vector-borne parasites are important ecological drivers influencing life-history evolution in birds by increasing host mortality or susceptibility to new diseases. Therefore, understanding why vulnerability to infection varies within a host clade is a crucial task for conservation biology and for understanding macroecological life-history patterns. Here, we studied the relationship of avian life-history traits and climate on the prevalence of Plasmodium and Parahaemoproteus parasites. We sampled 3569 individual birds belonging to 53 species of the family Thraupidae. Individuals were captured from 2007 to 2018 at 92 locations. We created 2 phylogenetic generalized least-squares models with Plasmodium and Parahaemoproteus prevalence as our response variables, and with the following predictor variables: climate PC1, climate PC2, body size, mixed-species flock participation, incubation period, migration, nest height, foraging height, forest cover, and diet. We found that Parahaemoproteus and Plasmodium prevalence was higher in species inhabiting open habitats. Tanager species with longer incubation periods had higher Parahaemoproteus prevalence as well, and we hypothesize that these longer incubation periods overlap with maximum vector abundances, resulting in a higher probability of infection among adult hosts during their incubation period and among chicks. Lastly, we found that Plasmodium prevalence was higher in species without migratory behaviour, with mixed-species flock participation, and with an omnivorous or animal-derived diet. We discuss the consequences of higher infection prevalence in relation to life-history traits in tanagers.

Higher infection probability of haemosporidian parasites in Blue-black Grassquits (Volatinia jacarina) inhabiting native vegetation across Brazil.

Human induced changes on landscape can alter the biotic and abiotic factors that influence the transmission of vector-borne parasites. To examine how infection rates of vector-transmitted parasites respond to changes on natural landscapes, we captured 330 Blue-black Grassquits (Volatinia jacarina) in Brazilian biomes and assessed the prevalence and diversity of avian haemosporidian parasites (Plasmodium and Haemoproteus) across avian host populations inhabiting environment under different disturbance and climatic conditions. Overall prevalence in Blue-black Grassquits was low (11%) and infection rates exhibited considerable spatial variation, ranging from zero to 39%. Based on genetic divergence of cytochrome b gene, we found two lineages of Haemoproteus (Parahaemoproteus) and 10 of Plasmodium. We showed that Blue-black Grassquit populations inhabiting sites with higher proportion of native vegetation cover were more infected across Brazil. Other landscape metrics (number of water bodies and distance to urban areas) and climatic condition (temperature and precipitation) known to influence vector activity and promote avian malaria transmission did not explain infection probability in Blue-black Grassquit populations. Moreover, breeding season did not explain prevalence across avian host populations. Our findings suggest that avian haemosporidian prevalence and diversity in Blue-black Grassquit populations are determined by recent anthropogenic changes in vegetation cover that may alter microclimate, thus influencing vector activity and parasite transmission.

Ticks biting humans in the Brazilian savannah: Attachment sites and exposure risk in relation to species, life stage and season.

Information about human tick bites in Brazil is mostly anecdotal. Published information is typically restricted to single tick infestation episodes and does not address human exposure occurring on a daily basis in natural, rural, or green urban areas. We present a comprehensive 2-yr study on human parasitism during a broad survey on ticks within a Brazilian savannah reserve. Overall, 439 tick bites were recorded from the following species: Amblyomma sculptum (n = 331 bites), Amblyomma parvum (n = 64), Amblyomma auricularium (n = 3), Rhipicephalus microplus (n = 2), Dermacentor nitens (n = 1) and 38 Amblyomma spp. ticks. Nymphs accounted for most tick bites (n = 292, 66.5 % of all bites) and these were overwhelmingly A. sculptum (92.8 % of nymphal bites). The main adult tick biting humans was A. parvum (n = 61, 50 % of all adult tick bites) followed closely by A. sculptum (n = 58, 47.5 % of all adult tick bites). Winter was the season with the highest percentage of tick bites (39.7 % of all bites), followed by spring (32.4 %); summer (18.9 %) and autumn (9 %). Amblyomma sculptum adult bites peaked in summer whereas nymphal bites occurred primarily in winter and spring. Amblyomma parvum adult bites peaked in spring and summer. The most common tick attachment sites included the waist, legs and belly, but A. parvum adults were recorded from the head of humans as well. A noteworthy observation was the lack of human parasitism by Amblyomma triste, the third most prevalent species in the environment. This tick species is a frequent human biter in both Uruguay and Argentina where it transmits a pathogen, Rickettsia parkeri, to humans.

First record of haemosporidian parasites infecting swifts (Aves: Apodidae).

Avian malaria is a widespread infection caused by parasites from the Order Haemosporida. Indeed, Neotropical swifts are interesting models for host-parasite coevolution studies due to their unique life history that may allow them to escape parasitism. Considering this, we evaluated haemosporidians from Neotropical swifts. We collected blood and prepared smears from 277 individuals from waterfalls in Brazil. Despite low parasitemia or absence of parasites detected by microscopy haemosporidian infection was detected in 44 swifts (15.8%) using a screening PCR that amplifies a 154-nucleotide segment of ribosomal RNA coding sequence within the mitochondrial DNA (mtDNA) of Plasmodium and Haemoproteus. Although previous studies reported absence of these parasites in European and North American swifts our data suggest that Neotropical swifts are susceptible to haemosporidians. Further studies will add information about evolutionary and ecological aspects of avian haemosporidia in Neotropical Cypseloidinae swifts.

Ticks and Rickettsia on anteaters from Southeast and Central-West Brazil.

The giant anteater (Myrmecophaga tridactyla) and the collared anteater (Tamandua tetradactyla) are widespread in Brazil and found in all Brazilian biomes. These hosts frequently use domestic animal environments such as pastures, where tick and related microorganism interchange may occur between hosts. Reports of tick infestations of these animals are scattered and refer to small samples and/or are geographically restricted. We herein present data on a wide geographic distribution of ticks and their Rickettsia collected from 72 giant and 30 collared anteaters, mostly road killed, over a period of 18 years, from Southeast and Central-West Brazil encompassing four States and 46 Municipalities. Overall nine tick species (Amblyomma auricularium, A. calcaratum, A. nodosum, A. ovale, A. parvum, A. sculptum, A. triste, Rhipicephalus microplus and R. sanguineus sensu lato) were collected from anteaters. Amblyomma sculptum, A. nodosum, and A. calcaratum were the most prevalent corresponding to, respectively, 48.8%, 39.3% and 2.7% of all ticks (n = 1775). However, A. nodosum tick numbers on collared anteaters were significantly higher (P < 0.001) than those on giant anteaters. At the same time, an abundance of A. sculptum adults on giant anteaters was significantly higher (Z = 2.875; P = 0.004) than that of A. nodosum and only eight A. sculptum nymphs were found on collared anteaters. DNA samples from 20 ticks from nine different animals yielded a visible amplicon in PCR targeting gltA. The PCR products targeting spotted-fever Rickettsia gene (ompA) from five adults of A. nodosum were sequenced and were shown to be 100% identical to Rickettsia parkeri strain NOD (MF737635.1). The product of one nymph and one adult of A. sculptum yielded a sequence 99% identical to R. parkeri strain NOD. Further, Rickettsia bellii genes were found in three A. nodosum adults. Ecological, behavioral and anatomical traits of anteaters are discussed to explain reported tick infestations and Rickettsia DNA found.

Ticks (Acari: Ixodidae) in the Serra da Canastra National Park in Minas Gerais, Brazil: species, abundance, ecological and seasonal aspects with notes on rickettsial infection.

The Cerrado Biome is the second largest in Brazil covering roughly 2 million km2, with varying features throughout its area. The Biome is endangered but it is also source of animal species for rural, green urban and degraded rainforest areas. Ticks are among Cerrado species that establish at anthropogenic sites and although information about them is steadily increasing, several features are unknown. We herein report tick species, abundance and some ecological relationships within natural areas of the Cerrado at higher altitudes (800-1500 m) within and around Serra da Canastra National Park, in Minas Gerais State Brazil. In total of 1196 ticks were collected in the environment along 10 campaigns held in 3 years (2007-2009). Amblyomma sculptum was the most numerous species followed by Amblyomma dubitatum and Amblyomma brasiliense. Distribution of these species was very uneven and an established population of A. brasiliense in the Cerrado is reported for the first time. Other tick species (Amblyomma ovale, Amblyomma nodosum, Amblyomma parvum, Ixodes schulzei and Haemaphysalis leporispalustris) were found in lesser numbers. Domestic animals displayed tick infestations of both rural and urban origin as well as from natural areas (Rhipicephalus sanguineus sensu lato, Rhipicephalus microplus, Dermacentor nitens, A. sculptum, A. ovale, Amblyomma tigrinum, Argas miniatus). Amblyomma sculptum had the widest domestic host spectrum among all tick species. DNA of only one Rickettsia species, R. bellii, was found in an A. dubitatum tick. Several biological and ecological features of ticks of the studied areas are discussed.

Ticks (Acari: Ixodidae) on swifts (Apodiformes: Apodidae) in Minas Gerais, southeastern Brazil.

Brazil harbors five species of Cypseloidinae swifts. Those from Streptoprocne and Cypseloides genera have a very distinct ecology. They shelter at night and build nests in moist cliffs by waterfalls. Information about tick infestation of these birds is virtually non-existent and restricted to the description of a new species, Ixodes paranaensis, in Streptoprocne biscutata in Paraná State and another record of this species in Streptoprocne zonaris in Minas Gerais State. We herein report tick infestation of swifts at eight waterfalls in the Cerrado biome of Minas Gerais State, southeastern Brazil. Swifts were captured during six campaigns from November 2008 to April 2013. Overall, 584 swifts were captured (527 C. senex, four C. fumigatus and 53 S. zonaris). Four birds were tick infested (prevalence of 0.7 %). Three individuals of C. senex hosted one tick each; a nymph of I. paranaensis, a female of I. paranaensis and a nymph of Amblyomma cajennense. One S. zonaris hosted an I. paranaensis nymph and an Ornithodoros sp. larva (Argasidae).

Ticks on birds from Cerrado forest patches along the Uberabinha river in the Triângulo Mineiro region of Minas Gerais, Brazil / Carrapatos em aves em matas de cerrado ao longo do rio Uberabinha na região do Triângulo Mineiro de Minas Gerais, Brasil

We herein describe ticks parasitizing birds in forest fragments along the Uberabinha River, a major watercourse that cuts through patches of remnants of Brazilian savannah in Uberlândia, Minas Gerais, Brazil. Overall 352 birds from 62 species, overwhelmingly, Passeriformes, were captured with mist-nets. The most abundant bird species was Basileuterus hypoleucus (n=36), followed by Lanio penicillata (n=24) and Thalurania furcata (n=23). Thirty one birds, all Passeriformes, were found infested with 56 ticks from which 12 were larvae and 44 nymphs, all from the Amblyomma genus. Highest infestation prevalence was found on Taraba major (66.6%), Thamnophilus pelzeni (60%) and Saltator maximus (50%). The mean intensity of tick infestation was low (1.8 tick per infested bird) with most of the parasites located on the neck (60%) of birds, followed by the head (20%). All larvae were attached to the skin around the eyes of birds. Amblyomma nodosum was the most numerous tick species found attached to birds (n=23 nymphs, 52.3% of nymphs) followed by Amblyomma longirostre (n=5, 11.4% of nymphs). Ecological relationships are discussed.

Descrevemos aqui carrapatos que parasitam aves em fragmentos florestais ao longo do rio Uberabinha, o principal curso de água que corta manchas de remanescentes de Cerrado em Uberlândia, Minas Gerais, Brasil. Ao todo, foram capturadas, com redes de neblina, 352 aves, de 62 espécies, em sua maioria, Passeriformes. As espécies de aves mais abundantes foram Basileuterus hypoleucus com 36 indivíduos, seguido de Lanio penicillata (n=24) e Thalurania furcata (n=23). Trinta e uma aves, todas Passeriformes, estavam infestadas com 56 carrapatos, sendo 12 larvas e 44 ninfas, todos do gênero Amblyomma. A prevalência de infestação mais elevada foi encontrada em Taraba major (66,6%), Thamnophilus pelzeni (60%) e Saltator maximus (50%). A intensidade média de infestação por carrapatos foi baixa (1,8 carrapato por ave infestada) e a maior parte dos parasitas recolhidos estava localizada no pescoço (60%) das aves, seguido pela cabeça (20%). Todas as larvas estavam fixadas à pele ao redor dos olhos das aves. Amblyomma nodosum foi a espécie de carrapato mais numerosa encontrada nas aves (n=23 ninfas, 52,3% das ninfas), seguido de Amblyomma longirostre (n=5, 11,4% das ninfas). Relações ecológicas são discutidas.

Ticks on birds in a forest fragment of Brazilian cerrado (savanna) in the municipality of Uberlândia, State of Minas Gerais, Brazil.

This is a report of tick species, parasite prevalence and infestation intensity of birds in a forest fragment (18° 56' 57'' S and 48° 12' 14'' W) within the Brazilian cerrado (savanna), in the municipality of Uberlândia, State of Minas Gerais, Brazil. A total of 162 birds from 26 species were captured. One adult tick, 296 larvae and 67 nymphs were found on passerine birds. Of these, it was identified 31 larvae and 27 nymphs of Amblyomma longirostre, 17 nymphs of A. nodosum, one A. cajennense larvae and one male of Rhipicephalus sanguineus. All other ticks were identified as Amblyomma sp. larvae (n = 264) or nymphs (n = 26). Overall tick infestation intensity and prevalence were 4.32 ticks/infested bird and 52%, respectively. Sampling of host-seeking ticks on the ground within the forest during a two-year period showed only five Amblyomma sp. nymphs and one adult male of A. nodosum whereas a search for ticks on domestic animals (cattle, horses and dogs) found Rhipicephalus (Boophilus) microplus ticks. Although identification was possible in only 27% of bird ticks there seemed to be no correlation between environmental and domestic animal and bird infestation. It can be assumed that bird infestation may occur above the ground or at specific sites not sampled.

Ticks on birds in a forest fragment of Brazilian cerrado (savanna) in the municipality of Uberlândia, State of Minas Gerais, Brazil / Carrapatos de aves em um fragmento florestal de Cerrado, município de Uberlândia, Estado de Minas Gerais, Brasil

This is a report of ticks species, parasite prevalence and infestation intensity of birds in a forest fragment (18º 56' 57" S and 48º 12' 14" W) within the Brazilian cerrado (savanna), in the municipality of Uberlândia, State of Minas Gerais, Brazil. A total of 162 birds from 26 species were captured. One adult tick, 296 larvae and 67 nymphs were found on passerine birds. Of these, it was identified 31 larvae and 27 nymphs of Amblyomma longirostre, 17 nymphs of A. nodosum, one A. cajennense larvae and one male of Rhipicephalus sanguineus. All other ticks were identified as Amblyomma sp. larvae (n = 264) or nymphs (n = 26). Overall tick infestation intensity and prevalence were 4.32 ticks/infested bird and 52 percent, respectively. Sampling of host-seeking ticks on the ground within the forest during a two-year period showed only five Amblyomma sp. nymphs and one adult male of A. nodosum whereas a search for ticks on domestic animals (cattle, horses and dogs) found Rhipicephalus (Boophilus) microplus ticks. Although identification was possible in only 27 percent of bird ticks there seemed to be no correlation between environmental and domestic animal and bird infestation. It can be assumed that bird infestation may occur above the ground or at specific sites not sampled.

Neste trabalho, são apresentadas as espécies de carrapatos em aves silvestres, sua prevalência e a intensidade de infestação em um fragmento florestal (18º 56' 57" S e 48º 12' 14" W) do Cerrado, no município de Uberlândia, Estado de Minas Gerais. Foram capturadas 162 aves de 26 espécies, mas apenas as aves Passeriformes estavam infestadas. Nestas, foram encontrados um carrapato adulto, 67 ninfas e 296 larvas. Dentre os carrapatos foi possível a identificação de 31 larvas e 27 ninfas de Amblyomma longirostre, 17 ninfas de A. nodosum, uma larva de A. cajennense e um Rhipicephalus sanguineus macho. Todos os outros carrapatos foram classificados como larvas (n = 264) ou ninfas (n = 26) de Amblyomma sp. A intensidade de infestação e a prevalência de carrapatos foram de 4,32 carrapatos/ave infestada e 52 por cento, respectivamente. Na pesquisa de carrapatos em vida livre sobre a vegetação do fragmento florestal, foram capturados apenas cinco ninfas de Amblyomma sp. e um macho adulto de A. nodosum, por um período de dois anos. Carrapatos da espécie Rhipicephalus (Boophilus) microplus foram encontrados parasitando bovinos, cavalos e cães domésticos da região. Mesmo considerando a identificação de apenas 27 por cento dos carrapatos das aves, parece não haver correlação entre infestação ambiental, nos animais domésticos, com a infestação das aves. Pode-se supor que a infestação das aves ocorra acima do solo ou em locais específicos não pesquisados neste trabalho.