Molecular detection of Babesia spp. and Rickettsia spp. in coatis (Nasua nasua) and associated ticks from midwestern Brazil.

Procyonids are reservoirs of many zoonotic infectious diseases, including tick-borne pathogens. The role of coatis (Nasua nasua) in the epidemiology of piroplasmids and Rickettsia has not been fully addressed in Brazil. To molecularly study these agents in coatis and associated ticks, animals were sampled in two urban areas in Midwestern Brazil. Blood (n = 163) and tick (n = 248) DNA samples were screened by PCR assays targeting the 18S rRNA and gltA genes of piroplasmids and Rickettsia spp., respectively. Positive samples were further molecularly tested targeting cox-1, cox-3, ß-tubulin, cytB, and hsp70 (piroplasmid) and ompA, ompB, and htrA 17-kDa (Rickettsia spp.) genes, sequenced and phylogenetically analyzed. All coatis' blood samples were negative for piroplasmids, whereas five pools of ticks (2%) were positive for two different sequences of Babesia spp.. The first from Amblyomma sculptum nymphs was close (i.e., ≥ 99% nucleotide identity) to a Babesia sp. previously found in capybaras (Hydrochoerus hydrochaeris); the second from Amblyomma dubitatum nymphs and Amblyomma spp. larvae was identical (100% nucleotide identity) to a Babesia sp. detected in opossums (Didelphis albiventris) and associated ticks. Four samples (0.8%) were positive by PCR to two different Rickettsia spp. sequences, being the first from Amblyomma sp. larva identical to Rickettsia belli and the second from A. dubitatum nymph identical to Rickettsia species from Spotted Fever Group (SFG). The detection of piroplasmids and SFG Rickettsia sp. highlights the importance of Amblyomma spp. in the maintenance of tick-borne agents in urban parks where humans and wild and domestic animals are living in sympatry.

Longitudinal dynamics and health impact of Hepatozoon procyonis (Apicomplexa: Hepatozoidae) on naturally infected ring-tailed coatis Nasua nasua (Carnivora: Procyonidae) from Midwestern Brazil.

This study aimed to morphologically and molecularly detect Hepatozoon procyonis in ring-tailed coatis' (Nasua nasua) blood and associated ticks from central-western Brazil, Campo Grande, Mato Grosso do Sul state and also evaluate the impact of the protozoa in blood parameters and coati´s health. Samplings were performed in a conservation area Parque Estadual do Prosa (PEP) and in a Brazilian Air Force Private Area namely Vila da Base Aérea (VBA), between March 2018 and April 2019. We collected 165 blood samples, 61 from recaptured coatis. Peripheral blood smears were stained with Romanovsky-type stain for H. procyonis parasitemia assessment. DNA extracted from blood samples and ticks (Amblyomma spp.) were submitted to a nested PCR (nPCR) assay based on the 18S rRNA gene for Hepatozoon spp. Out of 104 individuals sampled, 80 (77%) were positive for H. procyonis in at least one capture. Overall, 67/165 (40.6%) blood smears showed H. procyonis gametocytes (PEP: 41/63 - 65%; VBA: 26/102 - 25.5%). Parasitemia based on 500 assessed leucocytes ranged from 1 (0.2%) to 50 (10%) and 1 (0.2%) to 25 (5%), from animals sampled in PEP and VBA, respectively. Fluctuation on the parasitemia was observed during recaptures. nPCR results showed higher positivity when compared to blood smears, i.e. 112/165 (68%) positive blood samples [PEP: 41/63 (65%), VBA: 26/102 (25.5%)]. In total, 63/248 (25.4%) tick DNA samples were positive at nPCR for Hepatozoon sp., including 32/87 (37%) pools (1 to 10 larvae) of Amblyomma larvae, 21/105 (20%) pools (1 to 5 nymphs) of Amblyomma sculptum nymphs, 9/43 (21%) pools (1 to 5 nymphs) of Amblyomma dubitatumnymphs, and 1/12 (8%) A. sculptum adult female. The partial 18S rRNA sequence from one coati's blood sample and one representative of each positive tick species randomly selected from each area for sequencing (1,000 bp) showed 100% identity with sequences of H. procyonis from GenBank previously detected in coatis. Regarding H. procyonis infection, no statistical differences were obtained when comparing males vs. females (p-value 0.67), immature animals vs. adults (p-value 0.31), rainy vs. dry season (p-value 0.51) and sampling location (p-value 0.42). No noticeable alteration in blood parameters or heath status was observed in parasite animals. H. procyonis circulates in a high prevalence in coatis from central-western Brazil. Parasitemia fluctuates among different coatis' recaptures and apparently the infection has no influence in coatis' hematological and clinical parameters.

Batmanacarus robini, n. gen., n. sp. (Trombidiformes: Trombiculidae) collected parasitizing a ghost-faced bat (Chiroptera: Mormoopidae) from a cave in Trinidad and Tobago.

Trinidad and Tobago, a neotropical country, has 38 reported chigger species. Of these species, 18 were parasitizing bats. Here, we describe a new genus and species parasitizing a ghost-faced bat in this country.

First record of Eutrombicula tinami (Oudemans, 1910) (Trombidiformes: Trombiculidae) parasitizing a cat in Brazil.

Chigger mites are ectoparasites of terrestrial tetrapods and can cause dermatitis in the host, known as trombiculiasis. In Brazil, there are 73 species of chiggers; however, cats never have recorded as a host in this country. Here, we report the first record of chiggers parasitizing a domestic cat in Brazil; and a new locality for Eutrombicula tinami (Oudemans 1910) in the Rio Grande do Sul state, Brazil.

Anaplasmataceae agents among wild mammals and ectoparasites in Brazil.

Anaplasmataceae agents comprise obligate intracellular bacteria that can cause disease in humans and animals. Between August 2013 and March 2015, 31 Nasua nasua (coati), 78 Cerdocyon thous (crab-eating fox), seven Leopardus pardalis (ocelot), 110 wild rodents, 30 marsupials, and 42 dogs were sampled in the Pantanal wetland, Brazil. In addition, ectoparasites found parasitizing the animals were collected and identified. The present work aimed to investigate the occurrence of Anaplasmataceae agents in wild mammals, domestic dogs and ectoparasites, by molecular and serological techniques. Overall, 14 (17·9%) C. thous, seven (16·6%) dogs and one (3·2%) N. nasua were seroreactive to Ehrlichia canis. Nine dogs, two C. thous, one N. nasua, eight wild rodents, five marsupials, eight Amblyomma sculptum, four Amblyomma parvum, 13 A. sculptum nymphal pools, two Amblyomma larvae pools and one Polygenis (Polygenis) bohlsi bohlsi flea pool were positive for Ehrlichia spp. closely related to E. canis. Seven N. nasua, two dogs, one C. thous, one L. pardalis, four wild rodents, three marsupials, 15 A. sculptum, two Amblyomma ovale, two A. parvum and one Amblyomma spp. larval pools were positive for Anaplasma spp. closely related to A. phagocytophilum or A. bovis. The present study provided evidence that wild animals from Brazilian Pantanal are exposed to Anaplasmataceae agents.

Description of Blankaartia shatrovi n. sp. (Acari: Trombiculidae) From Brazil.

The chigger mite genus Blankaartia includes 28 known species, of which 10 are distributed in the Nearctic and Neotropical regions. These species preferentially parasitize birds, but occasionally they can also be found on rodents, bats, and reptiles, showing low host selectivity. In the present study, we report the presence of this genus in Brazil for the first time, including the first report of Blankaartia sinnamaryi (Floch and Fauran) and the description of a new species of Blankaartia collected from birds (Order Passeriformes).

Ectoparasites of small-mammals: determinants of community structure in South American savannah.

This study aimed to assess the contribution of hosts characteristics (rodents and marsupials) in the organization of ectoparasite communities present in woodland patches in western central Brazil. We verified the effect of host species, sex, body mass and vertical strata in addition to the role of seasonality on the ectoparasite composition, richness and abundance. The total sampling effort was 22 032 trap-nights equally distributed in 54 woodland patches. Variance partition and principal coordinate analysis were used to verify the existence of significant relationships between response variables and predictors. As expected, host species was the most important variable in ectoparasite community assembly. The composition, richness and abundance of mites and lice were highly influenced by host species, although higher for mites than for lice. Host body mass had a determining role on the richness and abundance of tick species. Vertical stratification and seasonality had weak influence, while the sex of the host had no influence on the organization of these communities. The results are closely related to the evolutionary characteristics of the species involved, as well as with local environmental characteristics of the study area.

Life Cycle of Amblyomma romitii (Acari: Ixodidae) Under Laboratory Conditions.

The life cycle of Amblyomma romitii Tonelli-Rondelli, 1939 is reported for the first time, using rabbits (Oryctolagus cuniculus) for larvae and capybara (Hydrochoerus hydrochaeris) for nymphs and adults, as experimental hosts. Developmental periods of free-living stages were observed in an incubator at 27 ± 1°C, 80 ± 10% relative humidity (RH), and 24-h darkness. The life cycle of A. romitii in the laboratory could be completed in an average period of 216.4 d. The overall sex ratio (M:F) was 1:1.4. The results showed that rabbits are quite suitable as experimental hosts for the larval stages of A. romitii, while capybaras are suitable experimental hosts for nymphs and adults.

Technique for restoration of mite (Acari) preparations in deteriorated Hoyer's medium.

The Acari Collection of Instituto Butantan (IBSP), São Paulo, Brazil, includes many types and other identified mite specimens that were mounted in Hoyer's medium, mainly in the first part of last century. An effort to restore degraded preparations was initiated in 1996. In this process, an improved technique was developed, allowing the adequate cleaning of specimens mounted up to 50-70 years before. Types and other identified specimens of Trombidiformes (Harpirhynchidae and Trombiculidae), Sarcoptiformes (Acaridae, Atopomelidae, Listrophoridae, and Psoroptidae) and Mesostigmata (Dermanyssidae, Ixodorhynchidae, Laelapidae, Macronyssidae, and Spinturnicidae) deposited at IBSP Collection have been satisfactorily restored.

Occurrence of Ornithodoros brasiliensis Aragão (Acari: Argasidae) in São Francisco de Paula, RS, Southern Brazil.

There have been no reports of the endemic Ornithodoros brasiliensis (Aragão) in Rio Grande do Sul, southern Brazil, since the 1950s. In January 2007, 21 O. brasiliensis ticks were collected in a rural area named "Cruzinha" in the municipality of São Francisco de Paula, RS, and another population was sampled later that year (October) in Vargem do Cedro, another rural area of São Francisco de Paula, following reports of human parasitism by ticks. The reappearance of this tick is a reason for concern in terms of public health.

Ticks (Ixodidae) on humans in South America.

Twenty eight species of Ixodidae have been found on man in South America (21 Amblyomma, 1 Boophilus, 2 Dermacentor, 2 Haemaphysalis, 1 Ixodes and 1 Rhipicephalus species). Most of them are rarely found on man. However, three species frequently parasitize humans in restricted areas of Argentina (A. neumanni reported from 46 localities), Uruguay (A. triste from 21 sites) and Argentina-Brazil (A. parvum from 27 localities). The most widespread ticks are A. cajennense (134 localities in Argentina, Bolivia, Brazil, Colombia, Ecuador, French Guiana, Guyana, Paraguay, Suriname and Venezuela), A. ovale (37 localities in Argentina, Brazil, Ecuador, French Guiana, Guyana, Paraguay, Suriname and Venezuela) and A. oblongoguttatum (28 sites in Brazil, Colombia, French Guiana, Guyana, Suriname and Venezuela). Amblyomma aureolatum (18 localities in Argentina, Brazil, French Guiana and Paraguay), A. cajennense, and A. triste are vectors of rickettsioses to man in South America. A better understanding of the respective roles of these and other tick species in transmitting pathogens to humans will require further local investigations. Amblyomma ticks should be the main subjects of these studies followed by species of Boophilus, Dermacentor, Haemaphysalis and Rhipicephalus species. In contrast with North America, Europe and Asia, ticks of the genus Ixodes do not appear to be major players in transmitting diseases to human. Indeed, there is only one record of an Ixodes collected while feeding on man for all South America.

Experimental infestation with the immatures of Amblyomma dissimile Koch, 1844 (Acari: Ixodidae) on Tropidurus torquatus (Lacertilia: Iguanidae) and Oryctolagus cuniculus

Larvas provenientes de duas fêmeas de Amblyomma dissimile Koch, 1844 (Acari: Ixodidae), naturalmente ingurgitadas em uma iguana (Iguana iguana) e provenientes do Estado do Mato Grosso, foram utilizadas na infestação experimental de lagartos da espécie Tropidurus torquatus e coelhos domésticos. As larvas alimentadas em ambos os hospedeiros realizaram ecdise para ninfas. As ninfas apenas ingurgitaram no lagarto e mudaram para machos e fêmeas. Este é o primeiro registro do parasitismo de larvas e ninfas de A. dissimile em T. torquatus e de larvas em coelhos.

Amblyomma aureolatum (Pallas, 1772) and Amblyomma ovale Koch, 1844 (Acari: Ixodidae): hosts, distribution and 16S rDNA sequences.

DNA sequences of Amblyomma aureolatum (Pallas, 1772) and Amblyomma ovale Koch, 1844 were obtained to determine genetic differences between these tick species. Collections of these species are discussed in relation to distribution and hosts. Seven ticks collections (four from Brazil, one from Argentina, one from Uruguay and one from USA) house a total of 1272 A. aureolatum (224 males, 251 females, 223 nymphs and 574 larvae) and 1164 A. ovale (535 males, 556 females, 66 nymphs and 7 larvae). The length of the sequenced mitochondrial 16S rRNA gene fragment for A. aureolatum was 370bp and for A. ovale was 373bp. The DNA sequence analysis showed a 13.1% difference between the two species. Apart from one male A. ovale found on a toad, all adult ticks were found on mammals. The majority of adult specimens of both tick species were removed from Carnivora (96.1 and 84.3% of A. aureolatum and A. ovale, respectively), especially from dogs (53.1% of A. aureolatum, and 46.4% of A. ovale). Collections on wild Canidae were higher for A. aureolatum (23.3%) than for A. ovale (7.1%). On the other hand, collections of A. ovale adults on wild Felidae were higher (18.3%) than findings of A. aureolatum (9.2%). The contribution of other mammalian orders as hosts for adults of A. aureolatum and A. ovale was irrelevant, with the exception of Perissodactyla because Tapiridae contributed with 13.0% of the total number of A. ovale adults. Adults of both tick species have been found occasionally on domestic hosts (apart of the dog) and humans. Most immature stages of A. aureolatum were found on Passeriformes birds, while rodents and carnivores were the most common hosts for nymphs and larvae of A. ovale. A. aureolatum has been found restricted to the Neotropical region, covering the eastern area of South America from Uruguay to Surinam, including northeastern Argentina, eastern Paraguay, southeastern Brazil and French Guiana. A. ovale showed a distribution that covers the Neotropical region from central-northern Argentina throughout the Neotropics into the Nearctic region of Mexico with a few records from the USA, also with collection sites in Paraguay, Bolivia, most Brazilian states, Peru, Ecuador, French Guiana, Surinam, Guyana, Trinidad & Tobago, Venezuela, Colombia, Panama, Costa Rica, Nicaragua, Belize, Guatemala and several states of Mexico.

Ixodes (Haemixodes) longiscutatum Boero (new status) and I. (H.) uruguayensis Kohls & Clifford, a new synonym of I. (H.) longiscutatum (Acari: Ixodidae).

Females of Ixodes (Haemixodes) uruguayensis Kohls & Clifford, 1967, a species whose adults were unknown until the present, were obtained in the laboratory from engorged nymphs collected on rodents (Scapteromys tumidus and Oxymycterus nasutus) in the counties of Maldonado and San José, Uruguay. Morphological characters of these females were identical to those given in the description of the female of Ixodes longiscutatum Boero, 1944. I. uruguayensis is, thus, relegated to a junior subjective synonym of I. longiscutatum. However, because of the unique morphological characters of the immature stages, the validity of the subgenus Haemixodes Kohls & Clifford, 1967 is not questioned. Therefore, the new status of Ixodes (Haemixodes) uruguayensis Kohls & Clifford, 1967 is Ixodes (Haemixodes) longiscutatum Boero, 1944.

Oviposition and eclosion periods of Ixodes didelphidis Fonseca and Aragão, 1951 (Acari: Ixodidae) under laboratory conditions.

Oviposition and eclosion periods for Ixodes didelphidis were observed under two temperatures (25 degrees C and 27 degrees C) and 90-95% humidity. Although there was a significant increase in the eclosion period (p<0.05) and a tendency to increase the oviposition period at 25 degrees C, there was neither significant differences in the interval (days), until maximum peak of eclosion nor in the number of emerging larvae during the peak nor the total number of emerged larvae. These temperature values are not critical for embryological development of the species. Because at 27 degrees C and under high humidity the oviposition and eclosion periods are shorter, and the percentage of emerged larvae is higher, we consider this to be the ideal temperature for laboratory studies.

Parasitism by Ixodes didelphidis and I. loricatus (Acari: Ixodidae) on small wild mammals from an Atlatic Forest in the State of Sao Paulo, Brazil.

From January 1995 to June 1996, ticks were studied in a fragment of Atlantic Forest in a residential area in the city of Itapevi, State of Sao Paulo, Brazil. Cases of human Lyme disease-like illness were registered in this area during the spring of 1992. The monthly relative density of ticks was determined and the influence of seasonal climatic conditions was evaluated as well as the relationship between ticks and hosts. Ticks (n = 88) were collected from small mammals captured (n = 134) in monthly trappings where the traps were operated for five consecutive days. Immature ticks (n = 42, 47.7%) were identified as Ixodes spp. The adult ticks (n = 46, 52.3%) were identified as Ixodes didelphidis Fonseca and Aragão (n = 19, 21.6%), I. loricatus Neumann (n = 26, 29.5%) and Amblyomma cajennense (F.) (n = 1, 1.1%). The monthly correlations between tick numbers and meteorological data were not significant. The correlation between the different stages of ticks and the two seasons (cold-dry and warm-rainy) indicated that both immature and adult ticks showed a seasonal pattern of abundance. The highest numbers of immature ticks were observed on rodents, during the cold-dry season (from April to September) with a peak in August 1995. Adults ticks were predominant during the warm-rainy season (from October to March) with a peak in January 1995. Adult ticks were only collected on marsupials.

Parasitism of Ixodes (Multidentatus) auritulus Neumann (Acari: Ixodidae) on birds from the city of Curitiba, State of Paraná, Southern Brazil.

The tick-bird relationship of 56 specimens of birds (Passeriformes and Columbiformes) collected in the city of Curitiba, State of Paraná, between 1990 and 1995, among which 102 specimens of Ixodes (Multidentatus) auritulus were found and analyzed. New host records were also produced including the first report of I. auritulus on a Columbiformes bird in Brazil.

Características biológicas da fase näo parasitária do Ixodes amarali (Acari: Ixodidae) em gambá (Didelphis sp.) no Estado do Rio de Janeiro / Biological characteristics of the non-parasitic phase of Ixodes amarali (Acari: Ixodidae) from opossum (Didelphis sp.) in the State of Rio de Janeiro, Brazil

Ixodes amarali was reported for the second time in the State of Rio de Janeiro, southeastern Brazil, from an engorged female obtained from a naturally infested opossum (Didelphis sp.). Biological characteristics of de non-parasitic phase were studied, under controlled conditions (temperature of 27ñ1§C, relative humidity higther than 80 per cent and scotophase)

Interrelationship between ectoparasites and wild rodents from Tijucas do Sul, state of Paraná, Brazil.

Sixteen species of ectoparasites were collected from 50 wild rodents, from August 1990 to August 1991, in an area of Araucaria augustifolia forest, in the municipality of Tijucas do Sul, State of Paraná, Brazil. Ectoparasites infested 98% of the rodents, with the highest indices of infestation found in the drycool season. Species that occurred in single or multiple infestations were recorded. Ectoparasite/host associations were significant (p < 0.01) for Gigantolaelaps wolffsohni/Oryzomys nigripes, Polygenis pradoi/Oxymycterus sp. and Amblyopinus sp./Oxymycterus sp. The following represent new host records: Polygenis (Polygenis) tripus from Akodon serrensis and Hoplopleura sciuricola from Sciurus aestuans. New geographic records are given for two species of flea and one sucking lice.