Trypanosoma cruzi infection in captive Neotropical primates in the Brazilian Amazon.

The aim of this study was to detect the infection by Trypanosoma cruzi in captive Neotropical primates in the Brazilian Amazon. From February 2013 to July 2014, 112 blood samples were collected from Neotropical primates from the Amazonas, Amapá, and Pará States, north of Brazil. The subjects belonged to the families Cebidae (N = 59), Atelidae (N = 41), Callitrichidae (N = 5), Pitheciidae (N = 4), and Aotidae (N = 3). Blood smears also were examined for the presence of trypomastigotes by optical microscopy. For the detection of T. cruzi DNA, a Nested-PCR with primers TCZ1/TCZ2 and TCZ3/TCZ4 was performed. T. cruzi DNA was detected in 12.5% (14/112) of Neotropical primates examined. Positive samples were detected in 16%, 12.5%, and 11.11% of the different species of primates sampled from the Amapá, Pará, and Amazonas states, respectively. The analysis of the blood smears did not reveal trypomastigote forms of T. cruzi. In conclusion, Neotropical primates kept in captivity were infected by T. cruzi in the studied areas. We recommend that a health management protocol be put into place to prevent the transmission of infectious agents among captive populations, captive and wild populations, and between NHPs and the technicians who handle these animals.

Phylogenetic congruence of Plasmodium spp. and wild ungulate hosts in the Peruvian Amazon.

Malaria parasites are known to infect a variety of vertebrate hosts, including ungulates. However, ungulates of Amazonia have not been investigated. We report for the first time, the presence of parasite lineages closely related to Plasmodium odocoilei clade 1 and clade 2 in free-ranging South American red-brocket deer (Mazama americana; 44.4%, 4/9) and gray-brocket deer (Mazama nemorivaga; 50.0%, 1/2). We performed PCR-based analysis of blood samples from 47 ungulates of five different species collected during subsistence hunting by an indigenous community in the Peruvian Amazon. We detected Plasmodium malariae/brasilianum lineage in a sample from red-brocket deer. However, no parasite DNA was detected in collared peccary (Pecari tajacu; 0.0%, 0/10), white-lipped peccary (Tayassu pecari; 0.0%, 0/15), and tapir (Tapirus terrestris; 0.0%, 0/11). Concordant phylogenetic analyses suggested a possible co-evolutionary relationship between the Plasmodium lineages found in American deer and their hosts.

Detection of Pathogenic Leptospira in Captive Chelonians (Kinosternon scorpioides-Linnaeus, 1766) in the Brazilian Amazon.

Leptospirosis is a zoonosis of great importance for One Health. In this context, the Amazonian biome may harbor numerous hosts for Leptospira spp. that contribute to the maintenance of the pathogen in the environment. Some reptiles, such as chelonians, have been little studied in terms of their involvement with leptospires. The objective of this study was to detect Leptospira spp. DNA in Kinosternon scorpioides turtles kept in captivity in a region of the Brazilian Amazon. A total of 147 samples of blood (n = 40), cloacal fluid (n = 27), cloacal lavage (n = 40), and stomach (n = 40) were collected from 40 chelonians. After DNA extraction, the samples were subjected to amplification of a 331 base pair product of the 16S rRNA gene using the Lep1 and Lep2 primers. PCR products were Sanger sequenced, assembled, and subjected to online blast search and phylogenetic analysis. Of the animals tested, 40% (16/40, 95% confidence interval [CI]: 25-55) had at least one or two samples positive for Leptospira spp. Considering the total number of samples collected, 12.93% (19/147) were positive, being blood clots (27.5%; 11/40), followed by cloacal washings (10%; 4/40), cloacal fluid (11.11%; 3/27) and gastric washings (2.5%; 1/40). Of these, 11 samples were sequenced and showed 99% to 100% identity with Leptospira interrogans sequences, which was confirmed by phylogenetic analysis. This is the first study to detect pathogenic Leptospira DNA in chelonians in a region of the Brazilian Amazon. It has been concluded that K. scorpioides turtles in captivity have been exposed to pathogenic Leptospira.

Tick and flea infestations in captive Tapirus terrestris and Tapirus kabomani (Perissodactyla: Tapiridae) in the Brazilian Amazon.

The present study aimed to identify tick and flea species infesting captive tapirs (Tapirus terrestris and Tapirus kabomani) in the Brazilian Amazon. Ectoparasites were collected from tapirs chemically restrained in Zoo-botanical Parks, breeding facilities, conservationists and, environmental compensation areas in the states of Amapá, Amazonas, and Pará. After collection, the tick and flea specimens were placed in plastic pots containing isopropanol and identified according to dichotomous keys. Ectoparasite infestations were observed in 55% (18/33) tapirs, of which 61% (11/18) were single infestations with ticks or fleas, and 39% (7/18) were mixed infestations with different species of ticks and/or fleas. In total, 227 ticks (15 larvae, 107 nymphs, 38 females and 67 males) and 14 fleas (eight females and six males) were collected, identifying four tick species (Amblyomma cajennense sensu stricto , Amblyomma naponense, Amblyomma oblongoguttatum, and Amblyomma pacae) and three flea species and/or subspecies (Ctenocephalides felis felis, Rhopalopsyllus australis australis, and Tunga penetrans). In conclusion, infestations with Ixodidae ticks and Rhopalopsyllidae, Pulicidae and Tungidae fleas were found in captive tapirs in the Brazilian Amazon, and A. cajennense s.s. was the most frequent species. Additionally, the present study reports new associations between A. pacae and R. a. australis with T. terrestris and between A. cajennense s.s. with T. kabomani.

EFFECTS OF DETOMIDINE AND DEXMEDETOMIDINE ON PHYSIOLOGICAL PARAMETERS AND ANESTHETIC RECOVERY IN TAPIRUS TERRESTRIS ANESTHETIZED WITH KETAMINE, GLYCERYL GUAIACOL ETHER, AND MIDAZOLAM.

The aim of this study was to compare the effects of two anesthetic induction protocols for long procedures carried out in the field in Tapiridae. Sixteen tapirs were divided into two groups (n=8) receiving either detomidine (DET) or dexmedetomidine (DEX) for anesthetic induction. All animals were anesthetized by intramuscular administration of a combination of ketamine (1.5 mg/kg), midazolam (0.2 mg/kg), plus either DET (0.04 mg/kg) or DEX (0.007 mg/kg). Anesthetic maintenance was by continuous infusion of ketamine, midazolam, and glyceryl guaiacol ether at 2 mg/kg per hour, 0.1 mg/kg per hour, and 100 mg/kg per hour, respectively). The animals were kept anesthetized for a total of 50 min to allow physical examination and collection of biological material as part of a research program, and physiological variables (heart rate [HR], respiratory rate, oxyhemoglobin saturation [SpO2], rectal temperature [RT], mean arterial pressure [MAP], blood glucose [GLI], and cortisol) and electrocardiogram were recorded during anesthesia. Anesthetic recovery was monitored by two researchers who were not informed of the induction protocol group. The recorded results were statistically evaluated. In both groups there was an initial increase in MAP, which subsequently decreased; RT gradually decreased during anesthesia; HR and GLI increased throughout the procedure; SpO2 was below normal throughout the procedure. Cortisol levels were significantly higher in the DEX group than in the DET group. Also, the animals in the DEX group had a longer recovery time than those in the DET group. On the basis of the results, we conclude that the combination of alpha-2 agonists and midazolam, ketamine, and glyceryl guaicol ether is an appropriate protocol for the anesthesia of tapirs in the field. However, in moderately extended procedures oxygen supplementation is recommended. Additionally, DEX resulted in fewer cardiovascular effects and longer-lasting sedation than DET.

Molecular characterization of Trypanosoma cruzi DTUs of the triatomine species in a Chagas disease endemic area.

Trypanosoma cruzi is the etiological agent of Chagas disease, a neglected tropical infection with great public health importance. This protozoan has triatomine insects as vector but may also be transmitted through blood transfusion, organ transplants, ingestion of contaminated food, or congenitally. It has a heterogeneous population classified into Discrete Typing Units (DTUs), TcI-TcVI and TcBat. The aim of this study was to molecularly characterize the DTUs of T. cruzi in triatomines from a Chagas disease endemic area in Northeastern Brazil. Triatomines were collected and the gut content was microscopically analyzed to investigate the presence of trypanosomatid flagellates. In addition, digestive tracts of some specimens were dissected and molecularly analyzed through PCR for Trypanosoma spp. and sequencing. PCR positive samples were further submitted to a multiplex PCR for DTUs of T. cruzi. A total of 117 triatomines were collected, 93.16% being in intradomicile and 6.84% in peridomicile environments. Insects were identified as Panstrongylus lutzi (37.60%), Triatoma pseudomaculata (26.50%), Triatoma brasiliensis (23.08%) and Panstrongylus megistus (12.82%). The specimens herein analyzed presented infection rates by T. cruzi of 5.49% and 12.09% in parasitological and molecular examinations, respectively. Multiplex PCR screening revealed 70.59% of the TcI genotype, detected in all triatomine species identified in this study and 29.41% of the DTU TcIII/TcIV detected in P. megistus and P. lutzi. T. cruzi infect triatomines in intradomicile and peridomicile environments, which brings attention to the risk of human infections and to the importance of the implementation of surveillance and entomological control actions.

Trypanosoma cruzi infection in triatomines (Hemiptera: Reduviidae) from rural areas of the state of Pará, Brazil.

The present study aimed to identify the triatomine species and evaluate Trypanosoma cruzi infection in insects captured in endemic areas of Pará State, Brazil. Triatomines were captured in nine rural communities in the municipality of São Domingos do Capim in August, September and December 2014 using active searches and Malaise and Noireau traps. Additionally, from 2014 to 2018, residents and community health agents submitted captured triatomines to the study team. The analysis of T. cruzi infection in the insects was performed by direct parasitological examination and nested-PCR. A total of 225 triatomines were captured and identified: Rhodnius robustus (n = 111), Rhodnius pictipes (n = 54), Panstrongylus geniculatus (n = 44), Eratyrus mucronatus (n = 11), Panstrongylus lignarius (n = 4), and Panstrongylus rufotuberculatus (n = 1). Direct parasitological examination was performed in 27 living triatomines R. robustus (n = 14), P. geniculatus (n = 7) and R. pictipes (n = 6) and metacyclic trypomastigote forms similar to those of T. cruzi were observed in 66.6% (18/27) samples. Of 174 samples analysed by nested-PCR, 81.6% were positive for T. cruzi DNA: R. robustus (84.7%; 72/85), R. pictipes (84.1%; 37/44), P. geniculatus (69.4%; 25/36), P. lignarius (100%; 4/4), E. mucronatus (75%; 3/4) and P. rufotuberculatus (100%; 1/1). R. robustus, R. pictipes and P. geniculatus were the main vectors of T. cruzi in the studied areas; however, the detection of infections in P. lignarius, E. mucronatus and P. rufotuberculatus indicated that these species can also act as potential vectors of T. cruzi in the study areas.

Molecular investigation of Toxoplasma gondii in oysters (Crassostrea spp.) sold on beaches in the State of Pará, Brazil.

The aim of this study was to detect Toxoplasma gondii DNA in oysters (Crassostrea spp.) sold on seven beaches in the State of Pará, Brazil. According to the National Program for Hygiene and Sanitary Control of Bivalve Mollusks, 100 g of the edible part of mollusks is required to analyze contaminating microorganisms. In this study, 12 oysters were assumed to be equivalent to 100 g of edible parts when preparing each pooled sample. In total, 360 oysters were purchased from 30 vendors. From groups of 12 oysters purchased per vendor, 60 pooled samples were obtained, comprising 30 gill tissues and 30 gastrointestinal tracts. For molecular analysis, nested-PCR was conducted to amplify a 155-base-pair product of the B1 gene from T. gondii. All analyzed samples were negative for T. gondii. Our findings indicate that the oyster samples sold on the beaches in the State of Pará were not contaminated by T. gondii.

Molecular detection of Histoplasma capsulatum in bats of the Amazon biome in Pará state, Brazil.

Histoplasma capsulatum, the fungus causing histoplasmosis, has a strong impact on public health. Histoplasmosis is one of the most prevalent systemic mycoses in the Americas and occurs in several mammalian species. Bats are important in the epidemiological cycle of histoplasmosis because they disseminate the fungus throughout the environment. The aim of the present study was to investigate natural H. capsulatum infection in bats located in forested areas, which have undergone anthropogenic perturbations, as well as in the urban areas of the state of Pará. Twenty-two species of bats were captured in 18 municipalities of Pará; the samples obtained from these animals were subjected to nested PCR for amplification of H. capsulatum DNA. The HCI/HCII and HCIII/HCIV primers were used, and the final 210-pb fragment was amplified. Of the 100 bats analysed, two were confirmed to be positive for H. capsulatum. Samples amplified by nested PCR were sequenced and found to share identity and have 100% match with H. capsulatum DNA. H. capsulatum was detected in the area of study: the state of Pará has a wide diversity of bat species, and the region under investigation is situated in the north of the state, which suffers the most severe environmental and climatic changes. Therefore, it is essential to elucidate the distribution of H. capsulatum hosts in this region to facilitate the implementation of effective disease surveillance.

Chagas disease in urban and peri­urban environment in the Amazon: Sentinel hosts, vectors, and the environment.

Chagas disease is an anthropozoonosis, caused by a flagellated protozoan, Trypanosoma cruzi, in which the enzootic cycle occurs between mammals and triatomines. Two dogs with a history of sudden death were necropsied at the Federal University of Pará (UFPA). One dog had a pale area in the myocardium, which on histopathological examination showed a T. cruzi amastigote nest; immunohistochemistry (IHC) analysis characterized it as acute Chagas disease (ACD). The second dog showed no macroscopic changes. Microscopically, a few cardiomyocytes were replaced by adipocytes, and IHC result was negative for T. cruzi. However, results of polymerase chain reaction (PCR) of the cardiac tissue of both dogs was positive for T. cruzi DNA. After that, an epidemiological study was conducted in the region. For this study, we selected four areas in Castanhal. One of the four areas (Area 1) is where one of the dogs lived. The other three areas were chosen because they were recently deforested for housing. Blood samples were collected from dogs, cats, wild small mammals (marsupials and rodents), and the digestive tract of triatomines. Nested PCR was performed on all the blood samples and the triatomine digestive tracts. In Area 1, T. cruzi DNA was detected in 50% (12/24) of the tested dogs, in the only tested cat (1/1), 50% (1/2) of the tested marsupials (Didelphis marsupials), and 100% of the captured triatomines (Rhodnius pictipes) (2/2). In Area 2, T. cruzi DNA was not detected in any of the 11 (0/11) dogs and two marsupials tested (0/2), and no triatomines were found in this area. In Area 3, T. cruzi DNA was detected in 42.25% (30/71) of the dogs, in 66,6% (2/3) of the cats, the only captured marsupial (D. marsupialis) (1/1), and all three triatomines (3/3) (R. pictipes) tested. In Area 4, the two dogs tested were negative (0/2), 25% (1/4) of the captured marsupials (D. marsupialis) was positive, and no triatomine was captured in this area. The data demonstrate the importance of detecting T. cruzi in dogs, cats, small rodents, and marsupials in the Amazon metropolitan areas, where ecotopes carry reservoirs and vectors capable of participating in the Chagas disease cycle. The proximity between humans and T. cruzi vectors in these places might contribute to increased disease transmission risk and maintenance of agents. It was noted that high-standard condominiums, previously thought to reduce the risk for this disease, presented a new epidemiological risk. The presence of T. cruzi DNA in a dog who, a year earlier had tested negative, when another dog in the same house died of ACD, shows that the transmission cycle is present and active, with a high possibility of disease transmission to animals and humans.

First report of natural infection by Trypanosoma cruzi in secretions of the scent glands and myocardium of Philander opossum (Marsupialia: Didelphidae): Parasitological and clinicopathological findings.

Trypanosoma cruzi is the etiologic agent of American trypanosomiasis and can infect humans and different species of domestic and wild animals. The marsupials are important wild reservoirs of T. cruzi, aiding in the maintenance of this agent in sylvatic and peri-domestic environments. The objective of this study was to report the parasitological and clinicopathological findings of a natural infection by T. cruzi in one specimen of Philander opossum that originated from the Brazilian Amazon. The animal was captured in a forest fragment near a rural community with reports of human Chagas disease. T. cruzi infection was diagnosed by blood smear examinations, blood culture, scent glands secretion culture, histopathological examination, and nested-PCR. Positive samples were subjected to PCR to characterize the discrete typing units (DTUs) of T. cruzi. Characteristic trypomastigotes of T. cruzi were observed in the blood smear, and spheromastigotes, epimastigotes, and trypomastigotes were visualized in the cultures. Non-suppurative myocarditis associated with amastigote clusters was the principal histopathological finding. DNA from T. cruzi was detected in samples of blood, blood cultures, scent glands secretion cultures, cardiac muscles, and the spleen. The TcI and the TcII/V/VI group DTUs were detected in blood culture and scent glands secretion cultures. Infection by T. cruzi can cause myocarditis in P. opossum and DTUs TcI and TcII/V/VI group mixed infection can be detected in the acute phase. P. opossum can be a source of infection for triatomine vectors and has the potential source for direct transmission of T. cruzi by secretions from the scent glands. These data are important to improve the understanding of the complex enzootic transmission cycle of T. cruzi in the Brazilian Amazon.

Natural Theileria equi infection in captive Tapirus terrestris (Perissodactyla: Tapiridae) in the Brazilian Amazon.

The objective of this study was to detect natural Theileria equi infection in captive tapirs (Tapirus terrestris) in the Brazilian Amazon. Samples from 19 captive tapirs were collected from zoological and botanical gardens and conservation parks in the Pará (n = 18) and Amazonas (n = 1) states. Whole-blood samples were collected for subsequent screening of T. equi DNA by PCR using the BEC-UF2 and EQUI-R primer set. Microscopic analyses of blood smears revealed T. equi trophozoites in 37% (7/19) of the animals examined, and T. equi DNA was detected in 58% (11/19) of the blood samples analyzed. Sequencing of amplified PCR products revealed an identity with T. equi isolates obtained from horses and waterbuck available in GenBank. In conclusion, T. equi infection occurs in captive tapirs in the Brazilian Amazon, and these mammals could potentially act as reservoirs.

Serological prevalence of Toxoplasma gondii infection in cats (Belém, Pará, Brazil).

We evaluated the prevalence of anti-Toxoplasma gondii antibodies in the serum samples collected from domestic cats in Belém, Pará, Brazil. We also correlated the presence of T. gondii antibodies with environmental variables and cat-owner habits. Four-hundred and forty-seven serum samples from domestic cats were analyzed. The sera were tested using an indirect immunofluorescence assay. Among the animals analyzed, 21.92% (98/447) were seropositive. A statistically significant association was found in relation to age and serology among the animals over 1 year old (p<0.01): in the group up to 1 year old, 12.82% (20/156) of the animals were positive, and in the group over 1 year old, 26.80% (78/291) were positive. Our results show that the cats in Belém, Pará region have anti-T. gondii antibodies, and their owners are not aware of toxoplasmosis or how to prevent its transmission.

Detection of Toxoplasma gondii in Crassostrea spp. oysters cultured in an estuarine region in eastern Amazon.

The aim of the present study was to detect DNA of Toxoplasma gondii in Crassostrea spp. oysters cultured in the state of Pará, Brazil. A total of 400 oysters were directly collected from a fixed rack system. Gills, gastrointestinal tract (GIT) and intervalvular liquid were separated and grouped into pool samples of 10 animals, resulting in 40 samples each of gills, GIT and intervalvular liquid. DNA extraction was performed using a commercial kit, and T. gondii DNA was detected by nested PCR using the primers Toxo3 and Toxo4, which produced an amplification product of 155 bp of the T. gondii gene B1. Nucleotide sequencing was performed for positive samples, and the obtained sequences were identified by comparison with sequences in GenBank. The DNA of T. gondii was detected in 5.8% (7/120) of the pool samples, of which 7.5% (3/40) was in the GIT, 5% (2/40) in the gills, and 5% (2/40) was in the intervalvular liquid. The obtained sequences presented 100% identity and overlap with T. gondii DNA sequences. This is the report of detection of T. gondii DNA in oysters from genus Crassostrea spp. originating from the state of Pará, eastern Amazon.

Plasmodium vivax and Plasmodium falciparum infection in Neotropical primates in the western Amazon, Brazil.

The Brazilian Amazon is endemic for malaria and natural infections by Plasmodium spp. have been detected in Neotropical primates. Despite the diversity of primate species in the region, studies on infections by these agents are limited. The aim of the present study was to investigate the frequency of infection by Plasmodium vivax and P. falciparum in free-born primates that were kept in captivity, in the western Amazon, Brazil. Blood samples were collected from 98 Neotropical primates. Detection of P. vivax and P. falciparum DNA was performed using a semi-nested PCR, and the amplified products were sequenced. Plasmodium spp. DNA was detected in 6.12% (6/98) of the primates. P. vivax, and P. falciparum DNA was detected in 2.04% (2/98) and 4.08% (4/98) of these mammals, respectively. Sequencing and phylogenetic analysis confirmed the results obtained from the semi-nested PCR. The presence of infected non-human primates (NHP) can be auxiliary in the maintenance of P. falciparum and P. vivax and may have implications for the malaria surveillance and control in the Brazilian Amazon. It is necessary to structure an efficient surveillance system for the aetiological agents of malaria that infect NHP and humans to reduce the risk of Plasmodium spp. introduction into new areas, to protect all susceptible species.

F200Y polymorphism of the ß-tubulin isotype 1 gene in Haemonchus contortus and sheep flock management practices related to anthelmintic resistance in eastern Amazon.

The objective of the present study was to determine the frequency of the F200Y polymorphism in the ß-tubulin isotype 1 gene of Haemonchus contortus from various sheep flocks in eastern Amazon, and to identify management practices that may favor the emergence of resistance to anthelmintic drugs in the same area. In total, 305 specimens of H. contortus were collected from sheep at 12 farms located in the state of Pará. An allele-specific PCR was performed to detect the F200Y polymorphism, and questionnaires were used to obtain information about the farms and flocks. All genotypes were detected as follows: 31% of the parasites were RR, 37% of the parasites were SR, and 32% were SS. The completed questionnaires revealed that all farms employed semi-intensive farming systems, performed suppressive anthelmintic treatment, and based their choice of drug on cost and availability rather than on any knowledge regarding drugs that remained effective on their property. It can thus be concluded that the SNP in codon 200 of the ß-tubulin isotype 1 gene is present in the H. contortus populations from eastern Amazon, and that a series of management practices that favor the emergence of anthelmintic resistance are employed on these farms.

Geographical distribution of Amblyomma cajennense (sensu lato) ticks (Parasitiformes: Ixodidae) in Brazil, with description of the nymph of A. cajennense (sensu stricto).

BACKGROUND: Until recently, Amblyomma cajennense (Fabricius, 1787) was considered to represent a single tick species in the New World. Recent studies have split this taxon into six species. While the A. cajennense species complex or A. cajennense (sensu lato) (s.l.) is currently represented by two species in Brazil, A. cajennense (sensu stricto) (s.s.) and Amblyomma sculptum Berlese, 1888, their geographical distribution is poorly known. METHODS: The distribution of the A. cajennense (s.l.) in Brazil was determined by morphological examination of all lots of A. cajennense (s.l.) in two large tick collections of Brazil, and by collecting new material during three field expeditions in the possible transition areas between the distribution ranges of A. cajennense (s.s.) and A. sculptum. Phylogenetic analysis inferred from the ITS2 rRNA gene was used to validate morphological results. Morphological description of the nymphal stage of A. cajennense (s.s.) is provided based on laboratory-reared specimens. RESULTS: From the tick collections, a total 12,512 adult ticks were examined and identified as 312 A. cajennense (s.s.), 6,252 A. sculptum and 5,948 A. cajennense (s.l.). A total of 1,746 ticks from 77 localities were collected during field expeditions, and were identified as 249 A. cajennense (s.s.), 443 A. sculptum, and 1,054 A. cajennense (s.l.) [these A. cajennense (s.l.) ticks were considered to be males of either A. cajennense (s.s.) or A. sculptum]. At least 23 localities contained the presence of both A. cajennense (s.s.) and A. sculptum in sympatry. DNA sequences of the ITS2 gene of 50 ticks from 30 localities confirmed the results of the morphological analyses. The nymph of A. cajennense (s.s.) is morphologically very similar to A. sculptum. CONCLUSION: Our results confirmed that A. cajennense (s.l.) is currently represented in Brazil by only two species, A. cajennense (s.s.) and A. sculptum. While these species have distinct distribution areas in the country, they are found in sympatry in some transition areas. The current distribution of A. cajennense (s.l.) has important implications to public health, since in Brazil A. sculptum is the most important vector of the bacterium Rickettsia rickettsii, the etiological agent of Brazilian spotted fever.

Morphological description of the nymphal stage of Amblyomma geayi and new nymphal records of Amblyomma parkeri.

The external morphology of the nymph of Amblyomma geayi Neumann is described by optical and scanning electron microscopy. Unfed nymphs were obtained from an engorged A. geayi female, which had been collected on a sloth (Bradypus variegatus) from Belém municipality, State of Pará, northern Brazil, and was kept under laboratory conditions. With the present description, we propose a modification of a taxonomic key published in 2010 for the Amblyomma nymphs that occur in Brazil, through the inclusion of A. geayi. The nymph of A. geayi is morphologically very similar to the nymph of Amblyomma parkeri Fonseca and Aragão, with only slight morphological differences related to scutal surface and punctuations (more shagreened and less punctuated in A. geayi). These 2 nymphs differ from all other known Amblyomma nymphs from Brazil by the combination of auriculae present as small posterolateral rounded projections, eyes located at the level of the scutal midlength, and a rounded hypostome. These nymphal similarities as well the morphology of the adult stage corroborate previous studies that showed that A. geayi and A. parkeri are genetically closely related. Unpublished host records of the nymphs of both A. geayi and A. parkeri are provided. Established populations of A. geayi and A. parkeri seem to be geographically separated, since all confirmed records of A. geayi are from the northern half of South America (mainly the Amazonian region) and Central America, whereas all known records of A. parkeri are from the Atlantic rainforest biome in northeastern, southeastern, and southern Brazil.

Infestation by Haematopinus quadripertusus on cattle in São Domingos do Capim, state of Pará, Brazil.

Severe infestation with lice was observed on crossbred cattle (Bos taurus indicus ×Bos taurus taurus) in the municipality of São Domingos do Capim, state of Pará, Brazil. Sixty-five animals were inspected and the lice were manually collected, preserved in 70% alcohol and taken to the Animal Parasitology Laboratory, School of Veterinary Medicine, Federal University of Pará, Brazil, for identification. The adult lice were identified as Haematopinus quadripertusus, and all the cattle examined were infested by at least one development stage of this ectoparasite. The specimens collected were located only on the tail in 80% (52/65) of the cattle, while they were around the eyes as well as on the ears and tail in 20% (13/65). Nits, nymphs and adults of the parasite were respectively collected from 98.46% (64/65), 38.46% (25/65) and 23.08% (15/65) of the animals examined. This is the first report of bovine pediculosis caused by H. quadripertusus in the state of Pará, Brazil. Further studies should be conducted to determine the occurrence pattern of this species in Brazil and its importance to livestock production.

Leishmania infantum infection in dogs from maroon communities in the Eastern Amazon / Infecção por Leishmania infantum em cães de comunidades quilombolas na Amazônia Oriental

ABSTRACT: This study was designed to detect L. infantum infection in dogs and to evaluate the factors associated with canine visceral leishmaniasis in the maroon communities of Menino Jesus de Petimandeua and Itaboca in the municipality of Inhangapi, Pará, Brazil. Whole blood and intact skin samples were collected from 143 dogs, and a questionnaire was applied. L. infantum DNA was detected by polymerase chain reaction (PCR) using primers RV1 and RV2. Collection sites were georeferenced to obtain a spatial distribution of the residences visited and infected dogs. L. infantum DNA was detected in 8.4% (12/143) of the skin samples and in 1.4% (2/143) of the blood samples. On the risk map, three clusters were observed in Itaboca and one was observed in Menino Jesus de Petimandeua. We observed that most of the inhabitants in these maroon communities live close to forested areas and do not use protection against insect vectors. The presence of canine reservoirs of L. infantum associated to environment characteristics (preserved forests and deforested areas) and habits of dog owners (living near forested areas and not using any protection against insects) may favor the transmission of L. infantum in the studied areas.

RESUMO: O presente estudo objetivou detectar a infecção por Leishmania infantum em cães e avaliar os fatores associados com a leishmaniose visceral canina nas comunidades quilombolas Menino Jesus de Petimandeua e Itaboca, município de Inhangapi, Pará, Brasil. Foram coletadas amostras de sangue e de pele íntegra de 143 cães, sendo aplicado um questionário. A detecção do DNA de L. infantum foi realizada através da PCR com os iniciadores RV1 e RV2. Os locais de coleta foram georreferenciados para realizar a distribuição e análise espacial das residências visitadas e dos cães infectados. DNA de L. infantum foi detectado em 8,4% (12/143) das amostras de pele dos cães e em 1,4% (2/143) das amostras de sangue. No mapa de risco, foram observados três aglomerados em Itaboca e um em Menino Jesus de Petimandeua. Pôde-se constatar que a maioria dos moradores das comunidades quilombolas reside em áreas próximas de mata e não utilizam proteção contra insetos vetores. A presença de reservatórios caninos de L. infantum associada com características ambientais (floresta preservada e áreas de desmatamento) e os hábitos dos proprietários dos cães (que vivem perto de áreas de floresta e não usam qualquer tipo de proteção contra insetos) podem favorecer a transmissão de L. infantum nas áreas estudadas.