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Toxoplasma gondii is a coccidian that infects almost all warm-blooded animals, including birds. Rocas Atoll Biological Reserve, located in the northeast of Brazil, is the only atoll in the South Atlantic, and home to the largest population of seabirds in the western Atlantic. In this study the occurrence of T. gondii antibodies in seabirds from Rocas Atoll were determined. Birds were manually captured, blood samples were taken, and antibodies detected by the modified agglutination test (>5). In total, 267 birds of seven species belonging to three families (Sternidae, Fregatidae and Sulidae) and two orders (Charadriiformes and Suliformes) were sampled. Out of the 267 samples, 20 (7.3%) were seropositive: nine out of 48 brown noddies (Anous stolidus), one out of 26 black noddies (Anous minutus), three out of 20 magnificent frigatebirds (Fregata magnificens), five out of 95 sooty terns (Onychoprion fuscatus) and two out of 20 red-footed boobies (Sula sula). None of the 33 masked boobies (Sula dactylatra) and the 25 brown boobies (Sula leucogaster) were seropositive. The antibody titers were 5 (n=6), 10 (n=4), 20 (n=3), 40 (n=6) and 160 (n=1). Due to the uniqueness of this environment, monitoring the seabirds is suggested to maintaining this Conservation Unit.

In the aquaculture industry, biocides are routinely used to treat parasitosis in fish, and researchers are continually developing sustainable alternatives that can replace these harsh chemicals. In this context, the objective of this study was to evaluate the effectiveness of a new natural compound, BiokosTM, for the treatment against Epistylis sp. in Carassius auratus fish. The infestation was identified by the presence of whitish plaques on the integument of five animals, and the diagnosis was confirmed through skin scrapings. BiokosTM is a lipopeptide derived from the bacteria Pseudomonas that can destroy the functionality of the cell membrane of ciliated protozoa. The action of BiokosTM does not harm animals and the environment because the compound degrades into amino acids and fatty acids within days. A 0.15 m3 (150 L) aquarium was treated with an Ich-AwayTM water conditioner manufactured by the Danish company Sundew ApS, which has BiokosTM as the active ingredient. Six tablets were added to the water daily for two days, and new skin scrapings were performed. The fish were clinically well and no longer possessed lesions or parasites. The results obtained indicate that BiokosTM can be an innovative and more sustainable alternative for controlling epistyliasis in ornamental fish.

Leishmaniasis are zoonosis widely spread in Brazil, caused by the protozoan of the genus Leishmania, which includes several species. The disease manifests itself in the visceral or tegumentary form, and the main reservoir is the dogs. Manaus is the largest city in the Brazilian Amazon region, and despite the importance of the municipality, practically nothing is known about leishmaniosis in humans and animals. The objective of this study was to evaluate the occurrence and risk factors associated with the presence of Leishmania infantum in domiciliated dogs from Manaus. Molecular (polymerase chain reaction) and serological (immunofluorescent antibody test) methods were used as an indication of the circulation of the parasite. Blood samples for 154 domiciled dogs were obtained, and prevalence ratio and analysis of the variables were performed. Serum antibodies anti-Leishmania spp. were detected in 20.8% (95% CI: 14.4%-27.2%). Access to the street and zone of residence (P < 0.01) were associated with higher seropositivity. Molecular diagnosis for L. infantum detected positivity in 60 (39%) of the 154 (95% CI: 31.3%-46.7%) animals, and the variables street access, contact with dogs, and zone of residence were associated with higher frequencies of positivity (P < 0.05). Both serology and molecular diagnosis detected positive dogs in the municipality. This is the first description of the circulation of L. infantum infecting dogs in Manaus. As the municipality is classified as nonendemic, studies of isolation and characterization of the isolate must be done urgently., (© The Author(s) 2024. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

Avian chlamydiosis is a bacterial infectious disease of birds, considered until recently caused only by Chlamydia psittaci, that now includes the newly described species C. buteonis, C. avium, and C. gallinacea, associated with several avian hosts. Since its recognition as a species in 2014 and having chickens as one of its main hosts, C. gallinacea has already been described in backyard poultry on all continents. The present study aimed to survey by molecular techniques the presence and species of Chlamydia spp. in backyard chickens from three states of the southern region of Brazil (Paraná-PR, Santa Catarina-SC, and Rio Grande do Sul-RS). DNA extracted from cloacal swab samples were tested by polymerase chain reaction (PCR) for different species of Chlamydia, namely Chlamydiaceae (23 S rRNA gene), C. psittaci (ompA gene), C. avium (enoA gene) and C. gallinacea (gidA and enoA genes). The 16 S rRNA gene was used for sequencing and phylogenetic analysis. A total of 582 backyard chicken samples were collected and grouped in 238 pools, from 134 properties in 59 municipalities. Chlamydiaceae was detected in 25.2% (60/238) of the samples, in 38.8% (52/134) of the properties and in 66.1% (39/59) of the municipalities. None of the samples yielded positive PCR results for C. psittaci or C. avium. For C. gallinacea, the overall percentage was 16.3% (39/238) according to the results of gidA and enoA genes. Sequence analysis confirmed that the samples corresponded to C. gallinacea. This is the first report of C. gallinacea in Brazil., (© 2024. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.)

Background: There are few reports of tick-borne pathogens infecting dogs living in indigenous communities of Brazil. Herein, we aimed to molecularly detect vector-borne pathogens in dogs from two indigenous communities in the Brazilian Amazon. Materials and Methods: We surveyed 327 dogs raised in Amazon region at 2 distinct indigenous ethnicities for the molecular detection of tick-borne pathogens (114 from Tapirapé and 213 from Karajá indigenous ethnicity). Whole blood samples were subjected to PCR and sequencing for Ehrlichia , Babesia , and Hepatozoon . Univariate and multivariate analyses were used to investigate the factors affecting the pathogen infection patterns in dogs. Results: Among the 327 blood samples, 40 were positive for Ehrlichia canis (12.2%), 2 for Anaplasma platys (0.61%), and 204 were positive for Hepatozoon canis (66.5%). Binary Logistic Regression showed association between E. canis infection and ethnicity ( p  = 0.010) and tick attachment ( p  = 0.041). Karajá dogs were 3.4 times (95% CI 1.3-8.5) more likely to be positive for E. canis than Tapirapé dogs. Dogs with ticks were 2.5 times more likely (95% CI 1.0-7.6) to be positive for E. canis than dogs without ticks. Conclusions: Our survey expands the knowledge regarding the presence of vector-borne pathogens in dogs from indigenous communities in the Amazon region.

We investigated the biochemical profile of greater rheas ( Rhea americana ) in captivity and correlated these values according to the birds' sex. A total of 69 serum samples were collected from a breeding site in Mossoró, northeastern Brazil, and analyzed to quantify serum biochemical parameters (total protein, albumin, cholesterol, calcium, phosphorus, uric acid, urea, creatinine, ALP, AST, and CK). The birds had levels of urea, creatinine, total cholesterol, aspartate aminotransferase, calcium, and phosphorus similar to the values reported for ratite and ostrich species. By sex, females showed higher values ( p < 0.05) of calcium (3.5 mmol/L), total cholesterol (7.5 mmol/L), and uric acid (435.3 μmol/L) than males, which had 3.1 mmol/L, 3.8 mmol/L, and 390.7 μmol/L, respectively. This can be attributed to the difference in diet, the productive phase of females, or stress at the time of sampling. The data present a wide spectrum of biochemical results regarding the health of greater rheas, contributing to the veterinary clinical practice of this species.

Plasmodium spp. and some other blood parasites belonging to the order Haemosporida are the focus of many epidemiological studies worldwide. However, haemosporidian parasites from wild animals are largely neglected in scientific research. For example, Polychromophilus parasites, which are exclusive to bats, are described in Europe, Asia, Africa, and Oceania, but little is known about their presence and genetic diversity in the New World. In this study, 224 samples of bats from remaining fragments of the Atlantic Forest and Pantanal biomes, as well as urbanized areas in southern and southeastern Brazil, were analyzed for the presence of haemosporidian parasites by PCR of the mitochondrial gene that encodes cytochrome b ( cytb ). The PCR fragments of the positive samples were sequenced and analyzed by the Bayesian inference method to reconstruct the phylogenetic relationships between Polychromophilus parasites from bats in Brazil and other countries. Sequences from Brazilian lineages of Polychromophilus were recovered in a clade with sequences from Polychromophilus murinus and close to the one Polychromophilus sequence obtained in Panama, the only available sequence for the American continent. This clade was restricted to bats of the family Vespertilionidae and distinct from Polychromophilus melanipherus , a parasite species mainly found in bats of the family Miniopteridae. The detection of Polychromophilus and the genetic proximity to P. murinus were further confirmed with the amplification of two other genes ( clpc and asl ). We also found a Haemosporida parasite sequence in a sample of Noctilio albiventris collected in the Pantanal biome, which presents phylogenetic proximity with avian Haemoproteus sequences. Morphological and molecular studies are still needed to conclude and describe the Polychromophilus species in Brazilian Myotis bats in more detail and to confirm Haemoproteus parasites in bats. Nevertheless, these molecular results in Brazilian bats confirm the importance of studying these neglected genera.

This study aimed to search parasites in 333 ornamental fish from five Brazilian states (Ceará, Minas Gerais, São Paulo, Paraná and Santa Catarina). Fish were sent from eight farms located in the municipalities of Fortaleza, Patrocínio do Muriaé, São Francisco do Glória, Cascavel, Timbó, Iguape, Jacareí and Mairinque. All fish received anesthesia earlier to euthanasia procedures. After the search for parasites, it was verified that 70.6% (235/333) of fishes were infected by at least one type of parasite, being 12 types of parasites identified: monogeneans, digenean metacercariae, cestodes, nematodes, Lernaea cyprinacea, trichodinids, Piscinoodinium pillulare, Ichthyophthirius multifiliis, diplomonad flagellates, Ichthyobodo sp., Chilodonella sp., and Tetrahymena sp. The proportion of infected fishes among the farms is compared through statistical tests, besides, animal handling adopted in each farm is also discussed. The importance of ensuring fish health in order to make the ornamental freshwater fish industry economically viable and reduce losses in production is highlighted.

This study presents 35 negative serologies for antibodies anti-T. gondii in free-living primates from Central Amazonia. Our results suggest that these populations have not had contact with the parasite and, therefore, do not have antibodies. This was the first study surveying T. gondii in Cacajao, Callicebus, Pithecia, and Saguinus monkeys., (© 2022 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

The genus Dermocystidium infects a wide range of animals. The host infection often occurs through the ingestion of endospores. The diagnosis depends on wet mounts and histopathological analysis of the affected tissue. The aim of this study was to investigate the incidence of Dermocystidium sp. infection on the skin of farmed striped catfish (Pangasianodon hypophthalmus) from a fish farm located in Fortaleza, Ceará state, northeastern Brazil. From these observations, we determined that 100% of the analyzed animals were infected with Dermocystidium sp. The wet mount and histopathology of the fish lesions revealed spore-filled cysts between the dermis and epidermis, encapsulated by connective tissue. Owing to a lack of research on the parasite and its prevalence among different fish species in Brazil and the rest of the world, additional studies are required to understand their endemicity in fish farms of Brazil, and consequently develop better disease prevention methods and increase the overall productivity.

The presence of antibodies anti-Toxoplasma gondii and Neospora caninum have been described in dogs from virtually all Brazilian states, however in the state of Amazonas, there are few studies on these coccidia. In this study the occurrence of antibodies against T. gondii and N. caninum and risk factors were determined in domiciliated dogs of Manaus, AM. Blood samples were collected from 154 dogs and, during the harvest, a questionnaire was applied with questions related to the animals. The samples were analyzed, for the presence of anti-T. gondii and N. caninum antibodies, by indirect fluorescence antibody test, with cutoff of 16 and 50, respectively. Associations between the variables studied and the presence of antibodies were made by chi-square test, fisher's exact test or G test (p<0.05). Of the 154 samples, 19 (12.3% 95% CI = 7.1% - 17.5%) were reagents to T. gondii, and association (p <0.05) was observed between the presence of antibodies and contact with other dogs. The occurrence of dogs reactive to N. caninum was 1.9% (95% CI = 0.4% - 5.6%) with 3 of the 154 dogs positives, and no association (p>0.05) was observed between the presence of N. caninum antibodies, and the variables studied.

Species of the genus Leishmania parasitize mammals and have life cycles that alternate between vertebrate and invertebrate hosts. Most species develop in a hematophagous arthropod and infect a specific vertebrate host that may belong to diverse orders and families. Visceral leishmaniasis is a chronic zoonosis with a wide geographic distribution, affecting 350 million people globally, mostly in areas with a high risk of infection. In Brazil, this disease not only has a high incidence but is also expanding to new areas, both in urban centers and rural areas, including territories with tribal communities, due to increasing human intervention. The objective of this study was to perform cathepsin L-like gene-based molecular diagnosis of Leishmania infantum in the indigenous Tapirapé ethnic group in the state of Mato Grosso. From the 372 individuals assessed, only 0.8% (3/372) tested positive for L. infantum, all from the same village (Urubu Branco). Despite the small number of infected individuals, this study demonstrates the first human cases of Leishmania infantum infection in this population, suggesting the need for regular monitoring of visceral leishmaniasis in the area and leading to a broad discussion on the planning and implementation of public health measures for the indigenous population, while respecting their distinctive territories and culture.

Armadillos are specialist diggers and their burrows are used to find food, seek shelter and protect their pups. These burrows can also be shared with dozens of vertebrate and invertebrate species and; consequently, their parasites including the zoonotics. The aim of this study was to diagnose the presence of zoonotic parasites in four wild-caught armadillo species from two different Brazilian ecosystems, the Cerrado (Brazilian savanna) and the Pantanal (wetland). The investigated parasites and their correspondent diseases were: Toxoplasma gondii (toxoplasmosis), Trypanosoma cruzi (Chagas disease), Leishmania spp., (leishmaniasis), Paracoccidioides brasiliensis (Paracoccidioidomicosis) and Mycobacterium leprae (Hansen's disease). Forty-three free-living armadillos from Pantanal and seven road-killed armadillos from the Cerrado were sampled. Trypanosoma cruzi DTU TcIII were isolated from 2 out of 43 (4.65%) armadillos, including one of them also infected with Trypanosoma rangeli. Antibodies anti-T. gondii were detected in 13 out of 43 (30.2%) armadillos. All seven armadillos from Cerrado tested positive for P. brasiliensis DNA, in the lungs, spleen, liver fragments. Also, by molecular analysis, all 43 individuals were negative for M. leprae and Leishmania spp. Armadillos were infected by T. cruzi, T. rangeli, P. brasiliensis and presented seric antibodies to T. gondii, highlighting the importance of those armadillos could have in the epidemiology of zoonotic parasites., (© 2020 Wiley-VCH GmbH.)

This study evaluated ticks and rickettsial exposure in 220 free-ranging lowland tapirs, Tapirus terrestris, from 2006 to 2018 in selected areas of three major biomes of Brazil - Atlantic Forest, Pantanal, and Cerrado. Overall, a total of 5970 tick specimens representing the following nine species were collected from tapirs: Amblyomma brasiliense, Amblyomma coelebs, Amblyomma dubitatum, Amblyomma ovale, Amblyomma parvum, Amblyomma sculptum, Amblyomma triste, Haemaphysalis juxtakochi, and Rhipicephalus microplus. Amblyomma sculptum was the most prevalent and abundant tick species in all three biomes; however, mean intensity values for A. sculptum were significantly lower in Atlantic Forest than in the Pantanal or Cerrado, and at the same time, statistically similar among tapirs from Pantanal and Cerrado. Contrastingly, mean intensity values for A. coelebs were significantly higher in the Atlantic Forest than in the other biomes. The remaining tick species were collected in lower numbers, or were exclusive for one biome, e.g., A. brasiliense and H. juxtakochi only in the Atlantic Forest. A total of 177 blood sera (123 individuals plus 54 recaptures) were collected from tapirs and tested for the presence of reactive antibodies to six Rickettsia species by immunofluorescence assay. Overall, 69% (9/13), 49% (62/126), and 66% (25/38) tapir sera from Atlantic Forest, Pantanal and Cerrado, respectively, were seroreactive to at least one Rickettsia species, with no significant difference between the three areas. Although many tapir sera reacted simultaneously to ≥2 Rickettsia species, Rickettsia parkeri elicited highest % seroprevalence and endpoint titers, and was incriminated as the possible agent involved in a homologous reaction in tapirs from the three biomes, where A. ovale was previously found infected by R. parkeri. In fact, seroconversion to R. parkeri was demonstrated in five tapirs that were captured at least twice during the study. This study demonstrated that tapirs were found to be constantly infested by several tick species in the Atlantic Forest, Pantanal and Cerrado biomes; however, the richness of tick infestations was concordant to the tick species known to be established in each biome. Under natural conditions, lowland tapirs were shown to be exposed to tick-borne spotted fever group rickettsiae., (Copyright © 2021 Elsevier GmbH. All rights reserved.)

We investigated the occurrence of Toxoplasma gondii and Neospora caninum antibodies in pigs raised in the Northeast of Pará, Brazil. At Study I, convenience sampled 151 pigs at two slaughterhouses, with and without state inspection; and Study II, which assessed 159 pigs with probabilistic sampling from nine pig farms. Serological analysis was performed using indirect fluorescent antibody test for T. gondii and N. caninum with a cutoff of 64 and 50, respectively. Overall, 6.77% pigs were seropositive for T. gondii and 5.16% for N. caninum. In Study I, pigs slaughtered with and without state inspection presented similar occurrence for both coccidia (p>0.05). Study II found an association between N. caninum seropositivity and sludge discarded into the soil, feeding pigs with animal-based protein, subsistence system, and absence of nipple drinkers. No association was found for T. gondii. Pigs from Pará are a potential source of T. gondii infection to humans. To our best knowledge, this is the first study to report anti-N. caninum antibodies in the serum of pigs in Pará State, Brazilian Amazon.

Ixodes schulzei is an ixodid tick that parasitizes Cricetidae rodents, chiefly the South American water rat, Nectomys squamipes, in Brazil and Argentina. In the present study, we evaluated the life cycle of I. schulzei by exposing larvae and nymphs to feed on two rodent species, N. squamipes and Calomys callosus (large vesper mouse),while adult ticks were exposed to feed on N. squamipes. Off-host developmental periods were observed in an incubator at 27 °C, 95% relative humidity, and 0:24 (light:dark) regimen. Larvae and nymphs successfully fed on either C. callosus or N. squamipes. Mean larval and nymphal feeding periods were 8.8 and 8.7 days on N. squamipes and 8.5 and 9.7 days on C. callosus. The majority of engorged larvae (79.0-80.8%) and nymphs (67.0-86.0%) successfully molted to nymphs and adults, respectively. Mean premolt periods were 11.5-11.7 days for engorged larvae and 22.5-23.7 days for engorged nymphs. Only adult females emerged from engorged nymphs, regardless of host species, i.e., none of 120 engorged nymphs molted to male. Around 18% of the unfed females presented teratologies compatible with the metagynander type of gynandromorphism. Ixodes schulzei adult females successfully fed (mean feeding period, 9.4 days), oviposited, and presented high reproductive performance (high engorged weight, egg mass weight, and % egg mass hatching), in the absence of male ticks. Our results showed that I. schulzei successfully reproduces by parthenogenesis, and corroborate field data that indicate N. squamipes as the most important host for this tick species. The male of I. schulzei remains unknown.

This study identified the parasitic species in juvenile freshwater finfishes during the fattening stage, from a fish farm located in the Zona da Mata (MG), southeastern Brazil, and revealed both macro and microscopical lesions in fish gills. A total of 172 juvenile fishes of different species (Oreochromis niloticus, Ictalurus punctatus, Ctenopharyngodon idella, Cyprinus carpio, Astyanax bimaculatus and Brycon amazonicus) were transported to a laboratory in São Paulo city. The fish were sedated and then euthanized for parasitological analysis. All fish were infected by at least one parasite species. Ten different species of parasites were identified: Apiosoma sp., Epistylis sp., Ichthyobodo sp., trichodinids, Piscinoodinium pillulare, Ichthyophthirius multifiliis, Tetrahymena sp., monogeneans, Centrocestus formosanus metacercariae, and Dermocystidium sp. The best management practices and lack of sanitary control were also discussed.

Rickettsia amblyommatis has been reported infecting various tick species throughout the western hemisphere, including Amblyomma cajennense sensu stricto (s.s.). The present study aimed to evaluate whether R. amblyommatis can be maintained by transovarial transmission and transstadial passage in A. cajennense s.s. ticks, and whether these ticks are competent vectors of this rickettsial agent. We selected engorged females that were naturally infected or uninfected by R. amblyommatis, and reared their offspring (infected and control groups, respectively). Immature ticks were allowed to feed on guinea pigs, whereas adults fed on rabbits. All stages and individuals of each generation of the infected group yielded rickettsial DNA, which was not detected in any tick from the control group. No host from the control group seroconverted to R. amblyommatis. Among 16 guinea pigs and eight rabbits infested with R. amblyommatis-infected ticks, only one guinea pig seroconverted to R. amblyommatis. Some unfed adult ticks of the infected group were dissected, and DNA was extracted from their salivary glands and from their carcasses. DNA of R. amblyommatis was detected in all carcasses, but not in the salivary glands. Results of this study indicate very low vector competence of A. cajennense s.s. for R. amblyommatis. Previous studies reported domestic animals with high titers to R. amblyommatis in areas where these animals are naturally infested chiefly by A. cajennense s.s. ticks. One may consider that the vector competence of A. cajennense s.s. for R. amblyommatis could vary among tick populations and/or rickettsial strains.

The present study evaluated the viability of Toxoplasma gondii tissue cysts in dry-aged pork loins (m. longissimus) after 14, 21 and 28 days under controlled temperature (0 °C ± 1 °C). The pigs (n = 9) were orally inoculated with 3,000 T. gondii oocysts. The right loin of each pig was aged for a predetermined period, and the left loin was kept unprocessed as a control. Two experiments were performed. In Experiment 1, the loins of three pigs were aged for 14 days and then bioassayed in both cats and mice. In Experiment 2, the loins of six pigs were bioassayed only in mice, and the ageing periods were 14, 21, and 28 days. Toxoplasma gondii tissue cysts remained viable in loins aged up to 14 days, as confirmed by bioassays in cats and mice. Viable T. gondii was not recovered by bioassays in mice from loins that were aged for 21 or 28 days. These results demonstrate that T. gondii remained viable in vacuum-packed dry-aged pork loins for 14 days at controlled temperature but not for 21 days or longer., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The protozoan parasite Toxoplasma gondii can infect virtually all warm-blooded animals worldwide but little is known of its infection in the endangered giant anteaters (Myrmecophaga tridactyla). The present study found antibodies to T. gondii in 13 of 23 free-living M. tridactyla from the northwest region of São Paulo state, Brazil, by the Modified Agglutination Test (MAT, cut-off titer 1:25). Unfrozen tissues of seven M. tridactyla were bioassayed in mice and viable T. gondii (strain designated TgMytrBrSP1) isolated from one seropositive giant anteater. To our knowledge, this is a new host record for T. gondii. Genotyping using PCR-RFLP revealed the Brazilian clonal Type BrIII genotype, and a unique non-archetypal genotype was revealed by microsatellite analysis., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

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., (Copyright © 2019 Elsevier GmbH. All rights reserved.)

Scuticociliatosis, caused by an opportunistic ciliate protozoan, is responsible for significant economic losses in marine ornamental fish. This study reports the occurrence of Uronema spp., parasitizing ten species of marine reef fish at an ornamental fish wholesaler: Blue green damselfish (Chromis viridis), Vanderbilt's Chromis (Chromis vanderbilti), Pennant coralfish (Heniochus acuminatus), Threespot angelfish (Apolemichthys trimaculatus), Goldspotted angelfish (Apolemichthys xanthopunctatus), Sea goldie (Pseudanthias squamipinnis), Orchid dottyback (Pseudochromis fridmani), Threadfin butterflyfish (Chaetodon auriga), Vagabond butterflyfish (Chaetodon vagabundus), and Bluecheek butterflyfish (Chaetodon semilarvatus). Diseased fish showed disorders such as hemorrhages and ulcerative lesions on the body surface. Histopathological analysis of the muscle, liver, gut, kidney, spleen, gills, and stomach revealed hemorrhages and degeneration of muscle fiber, vacuolar degeneration of hepatocyte, inflammatory process and granuloma in the liver, atrophy of intestinal villi, inflammatory process and granuloma in the kidney, melanomacrophage centers, as well as inflammatory process in the spleen, epithelial cells hyperplasia and granuloma formation in the gills, and vacuolar degeneration and eosinophils in the stomach. Due to the severity of the disease, it is necessary to implement biosecurity measures with rapid and accurate diagnosis to minimize the risk of economic losses caused by Uronema spp.

More than 1.450 marine fish species are commercialized as ornamental fish and approximately 90% of these are wild-caught. Rainford's butterflyfish, Chaetodon rainfordi (McCulloch, 1923), is endemic to the western Pacific. This fish species belongs to one of the main ornamental fish families imported into Brazil. Haliotrema is a genus of the family Dactylogyridae. Species of this genus have very interesting host associations and geographic distributions. The aim of this study was to report on occurrences of exotic Haliotrema aurigae in diseased Rainford butterflyfish that were imported to Brazil. One Rainford's butterflyfish with suspected parasitic disease was euthanized and subjected to skin and gill scraping. The parasite specimens were mounted on semi-permanent slides with Hoyer's solution for morphological analysis of sclerotized structures. The hard-sclerotized structures of these specimens were compared and were identified as those of Haliotrema aurigae (Yamaguti, 1968). These results emphasize the importance of implementation of correct control measures at the country's borders, to avoid the introduction of exotic parasites. In addition, it can be emphasized that the easy adaptation of this parasite to new environments is a concern for ornamental fish farmers.

There is limited knowledge about tick diversity in the Amazon region. Here, we survey small terrestrial mammals for tick infestation at the Rio Pardo settlement, Amazonas State, Brazil. Sampling included rainy and dry seasons and four ecotones (primary forest, forest in regeneration, field crops and households). Each animal was inspected for ticks, which, if present, were placed in 70% alcohol and identified. Parasitological indexes were calculated and the presence/absence of ticks on hosts was tested for possible associations with independent variables (ecotone, host sex, host order, host family, host age and season). A total of 208 small mammals were captured, 47 individuals (10 species) in the primary forest, 124 (15 species) in the forest in regeneration, 11 (7 species) in the field crops, and 26 (4 species) in the households. A total of 14 small mammals were infested by ticks (overall prevalence: 6.7%; 95% CI: 3.72 - 11.04%), which consisted of 51 specimens that were identified into four species, as follows: Amblyomma humerale (32 nymphs); Ixodes luciae (6 females); Amblyomma coelebs (1 nymph); and Ornithodoros mimon (1 larva). In addition, 11 larvae were retained as Amblyomma spp. Only host order showed association (P = 0.002) with tick infestation, with marsupials 5.5 times more infested than rodents. Our record of O. mimon on D. marsupialis is the first on this host species, and the first record of a Argasidae tick in the Brazilian state of Amazonas. To the best of our knowledge, this is the first study that actively screened free-living terrestrial small mammals and provided data on prevalence, mean intensity and mean abundance of tick infestations in the Brazilian Amazonas state., (© 2024. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Toxoplasma gondii and Neospora caninum infections in dogs are predominantly manifest asymptomatic. However, these infections can also present highly varied and potentially severe clinical signs. This is due to the parasites' ability to replicate in a number of cell types within the host organism, with N. caninum exhibiting a particular tropism for the central and peripheral nervous systems, and T. gondii targeting the central nervous system and musculature. In clinical practice, toxoplasmosis and neosporosis are often considered to be closely related diseases, despite their distinct epidemiological, clinical, diagnostic, and therapeutic characteristics. The present review analyses the similarities and differences between these two protozoan infections, since an accurate and timely aetiological diagnosis is essential for establishing effective therapeutic protocols and control strategies., (© 2024. The Author(s).)

Cryptocaryon irritans is an obligate parasitic ciliate protozoan of wild and cultured marine fish. It causes white spot disease, and infections with this pathogen can cause significant losses for aquarists and commercial marine cultures worldwide. This study reports the occurrence of C. irritans parasitizing the ornamental reef fish, yellowtail tang, Zebrasoma xanthurum. Six days after being introduced to a new environment, 11 yellowtail tangs had white spots scattered across their bodies and fins. Suspicion of infection with C. irritans was evaluated by scraping the skin to confirm clinical diagnosis. After confirmation, the yellowtail tangs were transferred to a hospital aquarium and treated with the therapeutic agent Seachem Cupramine® for 15 days. During the treatment period, the copper concentration was monitored daily. At the end of the treatment, none of the yellowtail tangs showed clinical signs of white spots on their bodies, and skin scraping confirmed the yellowtail tangs were no longer infected. Subsequently, the yellowtail tangs were released for sale.

The present study evaluated the distribution and viability of Toxoplasma gondii tissue cysts in the organs and Brazilian commercial cuts of experimentally infected pigs. The pigs were infected with 3 × 10 3 oocysts of the T. gondii isolate TgCkBr57 (Type BrII). Mouse bioassays were performed on the brain, retina, tongue, diaphragm, and heart as well as the following muscle cuts: loin (longissimus), coppa (longissimus, spinalis dorsi, rhomboideus), tenderloin (psoas major), outside flat (biceps femoris), topside (semimembranosus), and top sirloin (gluteus medius). Toxoplasma gondii was isolated from the coppa, heart, diaphragm, and tongue of three pigs; from the tenderloin, outside flat, and brain of two pigs; and from the top sirloin and loin of one pig. Thus, the viability of T. gondii cysts was observed in all of the organs and cuts evaluated (except for the topside and retina), demonstrating the broad distribution of this parasite in pig organs and commercial meat cuts, and the importance of this species as a source of human infection.

Toxoplasmosis has been reported in many avian species, but little information is available from wild penguin populations. Leptospira can infects domestic and wild animals. Spheniscus magellanicus belong to the order Sphenisciformes, family Spheniscidae, and are colonial birds. These seabirds live in temperate waters along the Atlantic shores of South America, and their total population has been estimated to be 1,300,000 breeding pairs. Magdalena Island (Chile) hosts an important breeding colony but, over recent decades, a marked decline in the number of birds has been seen. The objective of this study was to determine occurrences of antibodies against T. gondii and Leptospira spp. in penguins (Spheniscus magellanicus) on Magdalena Island, from where no previous data on these agents were available. Serum samples were collected from 132 penguins on Magdalena Island. Antibodies against Toxoplasma gondii were detected using the modified agglutination test (Titer ≥20), and anti-Leptospira spp. antibodies were detected using the microscopic agglutination test (Titer ≥100). T. gondii antibodies were detected in 57 (43.18%) of the 132 serum samples, with titers that ranged from 20 to 320. None of the penguins in this study was reactive to anti-Leptospira spp. antibodies. This is the first report of T. gondii seropositivity in free-living Magellanic penguins in Chile., (Copyright © 2019 Elsevier B.V. All rights reserved.)

This study evaluated the species richness and seasonal dynamics of ticks and rickettsial agents infecting ticks in the largest natural Reserve of the Cerrado biome of Brazil, the Grande Sertão Veredas National Park. During 2012-2014, a total of 9531 host-seeking ticks were collected by dry ice traps and dragging, whereas 1563 ticks were collected from small mammals, and 1186 ticks from domestic animals. Overall, the following 12 tick species were identified: Amblyomma auricularium, Amblyomma dubitatum, Amblyomma naponense, Amblyomma ovale, Amblyomma parvum, Amblyomma sculptum, Amblyomma tigrinum, Amblyomma triste, Dermacentor nitens, Rhipicephalus microplus, Rhipicephalus sanguineus sensu lato, and Ornithodoros mimon. The three most abundant tick species, A. sculptum, A. parvum, and A. triste, are likely to develop one generation per year, with adults predominating between spring and autumn, and immature ticks during autumn-winter. Small mammals seem to be important hosts for immature stages of A. parvum, and A. triste, but not for A. sculptum. Molecular analyses revealed the presence of the human pathogen Rickettsia parkeri in 10% of the A. triste ticks, whereas two agents of unknown pathogenicity, Rickettsia bellii and 'Candidatus Rickettsia andeanae' were found in 7 and 5%, respectively, of the A. parvum ticks. A fourth rickettsial agent, Rickettsia amblyommatis, was found in a single A. sculptum tick. Several Vero cell-established isolates of R. parkeri and R. bellii were obtained from A. triste and A. parvum, respectively. Serological analyses of small mammals suggest that they have been infected by R. parkeri and R. bellii, possibly via natural infestations by A. triste and A. parvum, respectively. Because the Park has suffered low anthropic alterations, our results should provide baseline data that shall be used for future comparisons with other Cerrado areas with higher degree of anthropic changes., (Copyright © 2018 Elsevier GmbH. All rights reserved.)

The capsalid monogenean Neobenedenia melleni is known as a lethal pathogen for captured marine teleost ornamental fish, if left untreated. This study reports the occurrence of N. melleni parasitizing four species of ornamental reef fish imported into Brazil and maintained in quarantine: Arabian angelfish (Pomacanthus asfur ), yellowbar angelfish (Pomacanthus maculosus), regal angelfish (Pygoplites diacanthus), and bluecheek butterflyfish (Chaetodon semilarvatus). Ten days after the beginning of quarantine, some fish showed behavioral alterations, such as irritability, and corneal opacity, which were rapidly diagnosed to be caused by monogenean parasites by body surface scraping. The fish from the same batch were treated with two applications of 2 mg L-1 praziquantel each at an interval of four days. Seven days after the first treatment, the mucus surface of the fish was re-examined, which did not reveal the parasites presence being delivered for commercialization.

The main motivation for this study was to determine the occurrence of Toxoplasma gondii, a cosmopolitan widespread zoonotic parasite distribution that can infect a wide variety of mammals and birds, in Magellanic penguins (Spheniscus magellanicus) in Brazil. In recent decades there has been a significant increase in the number of penguins originating from Argentinian and Chilean Patagonia, where these birds are born, that arrive on the Brazilian coast, where many of them are stranded and rescued. Tissue samples were collected from 330 individuals surveyed from 2012-2015 at the Institute for Marine Animal Research and Rehabilitation (IPRAM) located in Cariacica, state of Espirito Santo, Brazil. Serum were collected from 145 animals surveyed in 2015 for the detection of anti-T. gondii antibodies using the Modified Agglutination Test (MAT ≥20) and 18 birds were positive, with titers of 20 (7 birds), 40 (9 birds) and 80 (2 birds). Mouse bioassay for the isolation of T. gondii was performed using tissues from 54 penguins that were also surveyed in 2015, but no isolates were obtained. DNA from tissue samples of 330 individuals was PCR amplified and sequenced to detect tissue cyst forming coccidians by using pan sarcocystids-directed primers (based on 18S rDNA). These samples were from animals surveyed in 2015 and from frozen stocked tissues from animals surveyed in the years 2012 and 2013. The positives were PCR amplified and sequenced with genus Sarcocystis-specific primers (based on internal transcribed spacer 1, RNA polymerase beta subunit coding gene, and cytochrome B coding gene) and with Sarcocystis falcatula/Sarcocystis neurona- specific primers (based on surface antigens SAG2, SAG3 and SAG4). Sixteen (3.0%) of pectoral muscle samples were positive by all the seven molecular markers and all the samples were identical to each other. Organisms close related to Sarcocystis falcatula were confirmed in all cases. This is the first report on molecular detection of infection by S. falcatula-related organisms and the first report of seropositivity for T. gondii in free-living Magellanic penguins in Brazil. Felids and didephid opossums are definitive hosts of T. gondii and S. falcatula, respectively. Where the penguins acquire the infective forms of the parasites shed by the terrestrial mammals remains to be elucidated., Competing Interests: The authors have declared that no competing interests exist.

Amblyomma aureolatum ticks are vectors of Rickettsia rickettsii, the etiologic agent of Rocky Mountain spotted fever in Brazil. Maintenance of R. rickettsii in nature depends on horizontal transmission along tick generations. Although such transmission is known to occur when uninfected and infected ticks feed simultaneously on susceptible animals (co-feeding systemic transmission), we investigated co-feeding nonsystemic transmission, which was based on R. rickettsii-infected and -uninfected A. aureolatum ticks feeding simultaneously on guinea pigs immune to R. rickettsii. Our acquisition and transmission infestations demonstrated that horizontal transmission of R. rickettsii by co-feeding ticks on immune hosts with no systemic infection did not occur when uninfected larvae fed distantly from infected nymphs but did occur in a few cases when uninfected larvae fed side-by-side with infected nymphs, suggesting that they shared the same feeding site. The co-feeding nonsystemic transmission type might have no epidemiologic importance for Rocky Mountain spotted fever.

Recent genetic population studies on Toxoplasma gondii in Brazil have shown large genetic variability. The objective of the present study was to isolate and genotypically characterize T. gondii from free-ranging and captive wild mammals and birds in Pernambuco state, Brazil. Fragments of heart, brain, skeletal muscle and diaphragm tissue from 71 birds and 34 mammals, which were either free-ranging or captive, were collected. Samples from 32 of these animals were subjected to bioassays in mice. Samples from the remaining 73 animals underwent biomolecular diagnosis, using PCR technique, targeting a repetitive DNA fragment of 529 bp in T. gondii. A non-virulent isolate (TgButstBrPE1) was obtained from a free-ranging striated heron (Butorides striata) and, based on primary samples, seven animals were found to be positive. The primary samples and the isolate obtained were subjected to PCR-RFLP using the markers SAG1, 5'3'SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, Apico and CS3. ToxoDB-RFLP genotype #13 from the striated heron isolate and Type BrIII genotype from a captive otter ( Lontra longicaudis) (PS-TgLonloBrPE1) were obtained. The present study describes the first isolation and genotypic characterization of T. gondii in free-ranging striated heron, and the first genotypic characterization of T. gondii in a captive otter.

Toxoplasmosis is a zoonotic disease caused by the protozoan Toxoplasma gondii. Infections occur via the ingestion of oocysts, consumption of cysts containing bradyzoites, and transplacental transmission of tachyzoites. Diversity in T. gondii strains may affect the outcome of clinical toxoplasmosis. The consumption of horse meat is a common practice in some parts of the world. The objectives of the present study were to isolate and genotype T. gondii from horses from an abattoir in the state of Rio Grande do Sul in southern Brazil that exports horse meat to Europe. Antibodies to T. gondii were found in 32.5% (13/40) of the horses using the modified agglutination test (MAT) with a cut-off of 1:25. Tissues from the 13 seropositive horses were bioassayed in mice, and one isolate, designated TgHorseBrRS1, was obtained. PCR-RFLP of the isolate revealed the ToxoDB-RFLP #228 genotype, a typical non-archetypal Brazilian genotype, and microsatellite analysis showed a unique non-archetypal genotype. This study showed that horses from Brazil can harbor viable T. gondii in their tissues, suggesting that recommendations to consumers should be made, especially in European countries where consumption of raw horse meat is common.

Neospora caninum is a coccidian parasite originally reported in dogs and widely prevalent in numerous species of wild and domestic animals and has as definitive hosts some species of canids. The white-lipped peccary (WLP) ( Tayassu pecari) is a Tayassuidae mammal, found from Mexico to south of Brazil and north of Argentina. It is a game species with great economic importance in the Peruvian Amazon. Blood samples from 101 WLPs were collected from near or within three different conservation reserves located in the southeastern region of the Peruvian Amazon. For the detection of antibodies against N. caninum, indirect fluorescent antibody tests (IFAT) were performed using collared peccary ( Pecari tajacu) and swine ( Sus scrofa domesticus) heterologous secondary antibodies. For both IFAT tests, the cutoff was 1:50. Positive samples were titrated by a two fold serial dilution. In addition to IFAT, samples were also analyzed using an immunoblotting test (IB) with anti-swine conjugate. To confirm the viability of the anti-swine conjugate, the results of these samples previously tested by a modified agglutination test (MAT) for Toxoplasma gondii were used as reference. From the total of 101 samples tested, 5 (4.9%) were N. caninum positive by the three tests and an extra sample was positive by both IFATs and negative in the IB. Comparing both IFATs and considering IB as the gold standard, the relative sensitivity of IFATs was 100%, the specificity was 98.9%, the positive predictive value was 83.3%, and the negative predictive value was 100%. The agreement between tests was characterized by a κ value of 0.904 (95% confidence interval, 0.717 to 1.0) and an SE of 0.095. This is the first report of N. caninum antibodies in free-ranging T. pecari, and swine and collared peccary conjugate can be used as a secondary antibody for detection of antibodies in Tayassu species.

Chickens are a host that is very resistant to the development of clinical toxoplasmosis. Free-range chickens have been used to indirectly track environmental contamination with Toxoplasma gondii oocysts because they feed on the ground. This study evaluated the genetic diversity of T. gondii isolates from free-range chickens from Florianópolis island in Santa Catarina state, southern Brazil. Sera from 21 chickens were tested for IgG anti-T. gondii antibodies using the modified agglutination test (MAT). Tissue homogenates from the 11 seropositive birds (MAT titres ≥5) were bioassayed in mice. The four obtained isolates (TgCkBrSC1-4) were genotyped using 11 PCR-RFLP markers and 15 microsatellite markers (MS). Four genotypes were identified, three of which are typical Brazilian genotypes (ToxoDB-RFLP #26 and #53 were previously reported and #278 is new), and the other is the rare clonal type I genotype. This type I isolate was considered a variant according to MS analysis, with two atypical alleles, which emphasizes the genetic diversity of the parasite in Brazil. The genetic variability of T. gondii in South America may be related to the high occurrence of severe ocular and congenital toxoplasmosis in humans in this region. High human seroprevalence and frequency of ocular toxoplasmosis are reported in southern Brazil, but there is limited information on the T. gondii strains that are circulating in this region, so more studies should be conducted to identify the strains in different hosts and in human toxoplasmosis cases., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Brazil has a large variety of wild animal species, but limited data are available on the occurrence of Brucella abortus and Leptospira spp. antibodies in these animals. Sera from 141 captive mammals belonging to 11 different species from the Northern and Northeastern regions of Brazil were screened. Antibodies against B. abortus and Leptospira spp. (24 live serovars) were investigated using the Rose Bengal plate and microscopic agglutination tests, respectively. Associations between the age, gender, and place of captivity were analyzed using the Pearson chi-square or the Fisher exact test. None of the animals were antibody positive for B. abortus. Among the animals tested, 11 (7.8%) were seropositive for Leptospira spp. These included one red brocket deer ( Mazama americana), two tufted capuchin ( Sapajus apella), seven agoutis ( Dasyprocta aguti), and one lowland paca ( Cuniculus paca). No association was observed between sex, age, and the occurrence of Leptospira spp. antibodies ( P > 0.05). However, an association was observed according to the place of captivity ( P = 0.046). From these 11 positive animals, six (54.5%) reacted to the serovars from the Icterohaemorraghiae serogroup, which is mainly responsible for the clinical cases of human leptospirosis in Brazil. To our knowledge, this is the first report of Leptospira spp. antibodies in M. americana and C. paca.

The importance of birds in the biological cycle of Neospora caninum is not clear. We report unsuccessful Neospora infection in chickens (Gallus gallus domesticus) using two isolates of N. caninum. In experiment #1, 30 White Leghorn chickens were orally inoculated with viable N. caninum oocysts (NC-SP1 isolate, 200 oocysts per bird) via the crop at 21days of age. Groups of three birds were euthanised at intervals of 7days (a total of 9weeks) and one group was challenged with the same oocyst dose at 37daysp.i. and observed for 11weeks. Blood samples were collected weekly, and sera were tested using IFAT. Chicken tissues were collected for PCR, quantitative PCR and immunohistochemistry. Two dogs approximately 45days of age were fed with tissues from chickens euthanised at 138 and 159daysp.i. The results indicated that the chickens were resistant to neosporosis as revealed by failure to seroconvert, to detect parasite DNA or N. caninum antigen by immunohistochemistry in inoculated bird tissues, and by no oocyst excretion by the dogs fed avian tissues. Similar results were obtained in experiment #2, in which 34 1-week-old chickens were each s.c. inoculated with 100,000 tachyzoites of the NcWTDMn1 isolate of N. caninum. The chickens were euthanised on days 7, 15, 22, 28, 36 and 60p.i. At necropsy, all tissues and serum from each bird were collected. All chickens remained asymptomatic, and N. caninum antigen was not detected by immunohistochemistry. Seven chickens euthanised at day 60p.i. demonstrated low (1:25 dilution) levels of antibodies by using the Neospora agglutination test. Two 12-week-old dogs fed tissues pooled from 10 inoculated chickens euthanised at day 60p.i. did not excrete N. caninum oocysts. This investigation indicates that chickens are resistant to experimental infection by N. caninum., (Copyright © 2017 Australian Society for Parasitology. Published by Elsevier Ltd. All rights reserved.)

Amblyomma ticks pose a significant public health threat due to their potential to transmit pathogens associated with rickettsial diseases. (E)-2-octenal, a compound found in donkeys (Equus asinus), exhibits strong repellent properties against Amblyomma sculptum nymphs under laboratory conditions. This study assessed the effectiveness of the (E)-2-octenal in wearable slow-release devices for personal human protection against Amblyomma ticks under natural conditions. Slow-release devices treated with (E)-2-octenal and untreated controls were prepared and tested on two volunteers walking through a tick-infested area in Goiania, Brazil. The experiment was conducted twice daily for three series of 10 days, with each volunteer wearing two devices attached to each leg, one on the ankle and one just above the thigh. Volunteers with control and treated devices exchanged them between rounds. Also, the daily release rate of (E)-2-octenal from the slow-release devices was determined in the laboratory, increasing significantly from 0.77 ± 0.14 µg/day on the first day to 9.93 ± 1.92 µg/day on the 4th day and remaining constant until the 16th day. A total of 5409 ticks were collected from both volunteers. Treated devices resulted in recovering fewer ticks (n = 1,666; 31%) compared to untreated devices (control: n = 3,743; 69%). (E)-2-octenal effectively repelled Amblyomma spp. larvae, A. sculptum adults, and exhibited pronounced repellency against A. dubitatum nymphs and adults. These findings suggest the potential of (E)-2-octenal delivered by wearable slow-release devices as a green-based repellent. Further improvements, however, are necessary to provide better protection for humans against A. sculptum and A. dubitatum in field conditions., (© 2024. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

During 2009-2012, wild animals were sampled in the Amazon biome of Brazil. Animal tissues and blood were tested by polymerase chain reaction (PCR) assays targeting DNA of the bacterial family Anaplasmataceae (genera Anaplasma, Ehrlichia, Wolbachia) and the genus Borrelia. Overall, 181 wild animals comprising 36 different species (2 reptiles, 5 birds, and 29 mammals) were sampled. All birds and reptiles were negative by all PCR assays, as well as all mammals for the Borrelia PCR assay. Anaplasmataceae agents were searched by PCR assays targeting two different genes, the ribosomal 16S rRNA gene and the protein-coding dsb gene. Three dsb closely related haplotypes were generated from 3 white-lipped peccaries (Tayassu pecari). In a phylogenetic analysis inferred from dsb partial sequences, these haplotypes grouped with previously reported Ehrlichia haplotypes from jaguar (Panthera onca) and horse from Brazil, suggesting that they could all represent a single species, yet to be properly characterized. A unique dsb haplotype was generated from a sloth (Bradypus tridactylus), and could also represent a different Ehrlichia species. All these dsb haplotypes formed a clade sister to the Ehrlichia ruminantium clade. Three distinct 16S rRNA gene haplotypes were generated from a wild guinea pig (Cavia sp.), a woolly mouse opossum (Micoureus demerarae), and two from robust capuchin monkeys (Sapajus sp.). In a phylogenetic analysis inferred from 16S rRNA gene partial sequence, these haplotypes grouped within the Wolbachia clade, and are likely to represent Wolbachia organisms that were infecting invertebrate metazoarians (e.g., filarids) associated with the sampled mammals. Two deer (Mazama americana) samples yielded two distinct 16S rRNA gene sequences, one identical to several sequences of Anaplasma bovis, and an unique sequence that grouped in a clade with different Anaplasma species. Our results indicate that a variety of genetically distinct Anaplasmataceae organisms, including potentially new Ehrlichia species, circulate under natural conditions in the Amazonian wildlife., (Copyright © 2017 Elsevier B.V. All rights reserved.)

This study was performed in Maranhão state, a transition area two Brazilian biomes, Amazon and Cerrado. During 2011-2013, 1,560 domestic dogs were sampled for collection of serum blood samples and ticks in eight counties (3 within the Amazon and 5 within the Cerrado). A total of 959 ticks were collected on 150 dogs (9.6%). Rhipicephalus sanguineus sensu lato (s.l.) was the most abundant tick (68% of all collected specimens), followed by Amblyomma cajennense sensu lato (s.l.) (12.9%), Amblyomma parvum (9.2%), and Amblyomma ovale (5.2%). Other less abundant species (<1%) were Amblyomma oblongoguttatum, Rhipicephalus microplus, Haemaphysalis juxtakochi, and Amblyomma rotundatum. Females of A. cajennense s.l. ticks were morphologically identified as A. cajennense sensu stricto (s.s.) or A. sculptum. Molecular analyses of 779 canine ticks revealed three Rickettsia species: Rickettsia amblyommatis in 1% (1/100) A. cajennense s.l., 'Candidatus Rickettsia andeanae' in 20.7% (12/58) A. parvum, Rickettsia bellii in 6.8% (3/44) A. ovale and 100% (1/1) A. rotundatum ticks. An additional collection of A. sculptum from horses in a Cerrado area, and A. cajennense s.s. from pigs in an Amazon area revealed R. amblyommatis infecting only the A. cajennense s.s. ticks. Serological analysis of the 1,560 canine blood samples revealed 12.6% canine seroreactivity to Rickettsia spp., with the highest specific seroreactivity rate (10.2%) for R. amblyommatis. Endpoint titers to R. amblyommatis were significantly higher than those for the other Rickettsia antigens, suggesting that most of the seroreactive dogs were exposed to R. amblyommatis-infected ticks. Highest canine seroreactivity rates per locality (13.1-30.8%) were found in Amazon biome, where A. cajennense s.s. predominated. Lowest seroreactivity rates (1.9-6.5%) were found in Cerrado localities that were further from the Amazon, where A. sculptum predominated. Multivariate analyses revealed that canine seroreactivity to Rickettsia spp. or R. amblyommatis was statistically associated with rural dogs, exposed to Amblyomma ticks.

During 2009-2012, wild animals were sampled in two areas within the Amazon biome of Brazil, in the states of Mato Grosso and Pará. Animal tissues and blood were molecularly tested for the presence of Piroplasmida (genera Babesia, Theileria, Cytauxzoon ) or Hepatozoon DNA. Overall, 181 wild animals comprising 36 different species (2 reptiles, 5 birds, and 29 mammals) were sampled. The following Piroplasmida agents were detected: Cytauxzoon felis in one ocelot ( Leopardus pardalis ), Theileria cervi in two red brocket deer ( Mazama americana ), Theileria spp. in three nine-banded-armadillos ( Dasypus novemcinctus ), one agouti ( Dasyprocta sp.), and four lowland pacas ( Cuniculus paca ), Babesia spp. in one common opossum ( Didelphis marsupialis ) and one white-lipped peccary ( Tayassu pecari ). The following Hepatozoon agents were detected: Hepatozoon sp. (possibly Hepatozoon caimani ) in three spectacled caimans ( Caiman crocodilus ), Hepatozoon felis in an ocelot ( Leopardus pardalis ), and Hepatozoon spp. in one scorpion mud turtle ( Kinosternon scorpioides ) and one lowland paca ( Cuniculus paca ). Phylogenetic analyses inferred by the 18S rRNA gene partial sequences supported these results, highlighting at least five novel Piroplasmida agents, and two novel Hepatozoon agents. This study screened the presence of tick-borne protozoa in a number of wildlife species from the Amazon for the first time. Our results indicate that a variety of genetically distinct Piroplasmida and Hepatozoon organisms circulate under natural conditions in the Amazonian wildlife.

Background: In Brazil, studies on animals and humans in mainland areas have shown that most strains of Toxoplasma gondii are pathogenic to mice and exhibit great genetic variability., Results: In this study, using a set of 11 PCR-RFLP and 15 microsatellite markers, we isolated and genetically characterised T. gondii strains from one cat and three rats on Fernando de Noronha Island. The cat had antibodies to T. gondii, which were revealed using a modified agglutination test (MAT, cut-off 1:25) and the seroprevalence among the 46 rodents was 15.2%. Viable T. gondii was isolated from one cat (TgCatBrFN1), two brown rats (TgRatnoBrFN1 and TgRatnoBrFN2) and one black rat (TgRatraBrFN1). Unlike the strains from mainland Brazil, these isolates were not pathogenic to outbred mice. The genotypes of these strains were compared with strains previously isolated on the island and in mainland Brazil. The analysis based on microsatellite data showed a limited genetic diversity of T. gondii on Fernando de Noronha Island with the majority of strains clustered into the following three groups: type II, III, and Caribbean 1., Conclusions: There was little variation among strains within the same group, suggesting that the majority of strains circulating on Fernando de Noronha are derived from only a few strains that were recently introduced to the island, likely from imported cats. Except for the strain belonging to the Caribbean 1 group that originates from northeast Brazil, there was little evidence that strains from the other groups were introduced to Fernando de Noronha via mainland Brazil.

The biological and genetic diversity of Neospora caninum is very limited because of availability of only a few viable isolates worldwide. This study describes the isolation and biological and molecular characterization of a new viable isolate of N. caninum (NC-SP1), from a cattle in Brazil. Approximately 400 g of brain from a naturally infected adult male cattle from an abattoir was fed to a 2-month-old dog. Neospora-like oocysts were observed on day 7 post-inoculation (PI) and the duration of oocyst shedding was 14 days. The DNA obtained from oocysts was characterized molecularly and the final sequence was 99% identical to homologous sequences of N. caninum available in GenBank®. For bioassay, gerbils (Meriones unguiculatus) were orally inoculated with 10 100 and 1000 oocysts; all gerbils remained clinically normal but developed N. caninum antibodies 14 days PI. Cell culture isolation was successful using the brain homogenate from one of the gerbils and tachyzoites were observed 24 days PI. Microsatellite genotyping revealed a unique genetic profile for this new reference isolate.

Introduction:: Leishmaniasis is endemic to the Northern, Northeastern, Central-Western, and Southeastern regions of Brazil. We aimed to assess the epidemiological situation of leishmaniasis in humans and dogs in indigenous villages located in the States of Mato Grosso and Tocantins using a serological survey conducted in May 2011., Methods:: Serum samples were collected from 470 humans and 327 dogs living in villages of the Urubu Branco and Tapirapé Karajá indigenous reserves. The samples were analyzed for the presence of Leishmania spp. antibodies using the indirect fluorescent antibody test (IFAT), enzyme-linked immunosorbent assay (ELISA) with a crude antigen (CA) and soluble antigen (SA), and Dual Path Platform (DPP®) immunoassay for canine visceral leishmaniasis., Results:: Of 470 human samples tested, two (0.4%) were positive using IFAT. Among 327 dog samples tested, 28 (8.6%) were positive using ELISA CA, five (1.5%) using ELISA SA, two (0.6%) using IFAT, and none using DPP® immunoassay with Leishmania infantum chagasi antigen. When Leishmania amazonensis antigen was used, 20 (6.1%) samples were positive using ELISA CA and four (1.2%) using IFAT., Conclusions:: There was a low prevalence of infection in the region, and significant differences among the main serological methods used for the diagnosis of leishmaniasis. These findings indicated that the detection of Leishmania spp. requires further study and improvement.

New World Nonhuman Primates are highly susceptible to clinical toxoplasmosis. Serum samples from 126 recently captured Leontopithecus chrysomelas, from an exotic and invasive population, were tested for Toxoplasma gondii antibodies by the modified agglutination test (MAT, cut-off 1:25); all were seronegative. The MAT is highly specific and is not species-specific. This is the first report of T. gondii survey in this tamarin in the wild. This result is consistent with prior reports that showed the high susceptibility of the species to infection by T. gondii usually with high mortality rates.

Although well established in dogs, Ehrlichia sp. infection has been scarcely reported in horses. The aim was to perform a comprehensive serological and molecular survey for the detection of Ehrlichia spp. in carthorses from Southern Brazil. Blood samples from 190 carthorses from Paraná State were sampled. Horses were also tested for Borrelia burgdorferi and Anaplasma phagocytophilum. Anti-Ehrlichia sp. antibodies were detected by a commercial rapid ELISA, and immunofluorescence antibody assays (IFA) with E. chaffeensis and E. canis as crude antigens. The molecular and phylogenetic analysis of Ehrlichia sp. was based on 16S rRNA and dsb genes. A total of 52 (27.4%), 4 (2.1%), and 3 (1.6%) horses were positive for Ehrlichia spp., Anaplasma spp. and Borrelia burgdorferi, respectively, by the commercial rapid ELISA. Thirty-eight (20.0%) and 37 (19.5%) horses showed anti-E. chaffeensis and anti-E. canis antibodies by IFA, respectively. One blood sample that also showed anti-E. chaffeensis antibodies was PCR positive for the 16S rRNA and dsb genes of Ehrlichia spp., showing an identity of>98.0% to the uncultured Ehrlichia sp. previously detected in Brazilian jaguars (Panthera onca). Anti-Ehrlichia sp. antibodies and Ehrlichia DNA were detected in carthorses from Southern Brazil, which may post public health concerns due to intimate contact with low-income owners. This is the first report of a natural infection of this bacteria in horses from South America. Clinical signs and the tick vector remain unknown., (Copyright © 2016. Published by Elsevier Ltd.)

The aim of this study was to determine the presence of antibodies to Neospora caninum and Toxoplasma gondii in serum samples of 187 dogs from 30 rural properties surrounding Córrego do Veado Biological Reserve, Espírito Santo State, Brazil. The Reserve is one of the last lowland Atlantic Forest remnants of the region, surrounded by agriculture farms and cattle pastures. The presence of IgG antibodies was determined by the Indirect Fluorescent Antibody Test for T. gondii (cut-off 1:16) and N. caninum (cut-off 1:50). Positive samples were diluted 2-fold until the last positive dilution. Antibodies to T. gondii were found in 77 (47.05%) dogs and antibodies to N. caninum in 22 dogs (11.76%) and one only dog was positive for both infections. No association between T. gondii and N. caninum infection and sex was observed (p>0.05). Control measures to prevent those infections in dogs that living surrounding the reserve and that had contact with wild animals are important to avoid the introduction of N. caninum in wild animals. This was the first study of frequency of occurrence of T. gondii and N. caninum in dogs from Espírito Santo, Brazil.

Toxoplasma gondii is a coccidian parasite that infects almost all warm-blooded animals, including birds. Abrolhos is an archipelago of five islands, located in the Atlantic Ocean, 56 nautical kilometers from the south coast of the state of Bahia, northeastern Brazil. Part of this archipelago is a National Marine Park, which is a conservation area protected by the Brazilian government. The objective of this study was to determine the occurrence of T. gondii antibodies in sera of seabird's species Sula spp. and Phaeton spp. from breeding colonies located in the Islands of Santa Bárbara and Redonda, Abrolhos's archipelago. Sera were tested by modified agglutination test, first screened at 1:5 dilution (cut-off point) and the positive samples were titrated at a two-fold serial dilution. Serum samples were obtained from 69 birds of four species: Sula dactylatra (23 birds), Sula leucogaster (19 birds), Phaeton aethereus (25 birds) and Phaeton lepturus (2 birds). Antibodies to T. gondii were found in 24 (34.8%) of 69 seabirds with titers that ranged from 5 to 640. Occurrence value in S. dactylatra was 34.8% (8/23), in S. leucogaster was 47.4% (9/19), in P. aethereus was 28% (7/25) and the 2 P. lepturus were negative. This is the first description of T. gondii antibodies in free ranging seabirds of the orders Suliformes and Phaethontiformes., (Copyright © 2016 Elsevier B.V. All rights reserved.)