Negative seroprevalence for Toxoplasma gondii in free-living primates from Central Amazonia.

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.

The impact of dry ageing vacuum-packed pork on the viability of Toxoplasma gondii tissue cysts.

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 ±â€¯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.

Isolation of viable Toxoplasma gondii from organs and Brazilian commercial meat cuts of experimentally infected pigs.

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 × 103 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.

Typical Brazilian genotype of Toxoplasma gondii isolated from a horse destined for human consumption in Europe from a slaughterhouse.

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-SPECIFIC ANTIBODIES IN FREE-RANGING WHITE-LIPPED PECCARIES ( TAYASSU PECARI) FROM THE PERUVIAN AMAZON: DETECTION OF ANTIBODIES IN SERUM AND EVALUATION OF INDIRECT FLUORESCENT ANTIBODY TEST WITH HETEROLOGOUS SECONDARY ANTIBODY.

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.

ANTIBODIES AGAINST BRUCELLA ABORTUS AND LEPTOSPIRA SPP. IN CAPTIVE MAMMALS IN THE STATES OF PARá AND RIO GRANDE DO NORTE, BRAZIL.

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.

Isolation and biological and molecular characterization of Neospora caninum (NC-SP1) from a naturally infected adult asymptomatic cattle (Bos taurus) in the state of São Paulo, 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.

Serologic evidence of Toxoplasma gondii infection in wild birds and mammals from southeast Brazil.

In this study, serum samples of 53 wild animals from two different states from the southeast region of Brazil were analyzed for the presence of anti-Toxoplasma gondii antibodies by the modified agglutination test (MAT), with a cut-off of 1: 5 for birds and of 1: 25 for mammals. Out of the sampled animals, 27 were birds and 26 were mammals, and from this total, 83% (n = 44) were free-living animals. Antibodies were found in 13 mammals, from which 11 were free-living animals, and in five birds, all of which were free-living. In this study, T. gondii antibodies were detected in four bird species (crested seriema, Cariama cristata; buff-necked ibis, Theristicus caudatus; picazuro pigeon, Patagioenas picazuro; and burrowing owl, Athene cunicularia) and in a giant anteater (Myrmecophaga tridactyla) for the first time.

Chlamydia gallinacea in Brazilian backyard chicken farms.

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.

Molecular Investigation Confirms Myotis Genus Bats as Common Hosts of Polychromophilus in Brazil.

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.

Clinical, haematological and biochemical responses of sheep undergoing autologous blood transfusion.

BACKGROUND: This study aimed to evaluate the clinical, haematological and biochemical responses to autologous blood transfusion and the feasibility of this practice in sheep. Thus, we used eight male, 8 months old sheep, weighing on average 30 kg, from which 15 mL/kg of whole blood was collected and stored in CPDA-1 bags. Blood samples were refrigerated for 8 days and subsequently re-infused. The clinical, haematological and biochemical parameters were evaluated before blood collection and reinfusion, after 10 minutes of collection and reinfusion, after 3, 6, 12, 24, 48, 96 and 192 hours after collection and reinfusion. RESULTS: With respect to clinical parameters, we observed a decrease in heart rate after 24, 48 and 196 hours from reinfusion compared to basal values (p < 0.05). Haematological variables including globular volume and erythrocyte counts showed a significant decrease (p < 0.01) at all time points after collection and increased (p < 0.01) at all time points after reinfusion. There was a significant increase in total protein and calcium at all time points after reinfusion (p < 0.05). CONCLUSION: Autologous transfusion in sheep slightly altered the physiological, biochemical and haematological responses of sheep, indicating that the technique proposed is safe and can be applied in the clinical practice of this species. The 8 d period was not sufficient for complete recovery of the haematological parameters after blood collection.

Occurrence of Toxoplasma gondii and Neospora caninum antibodies and risk factors in domiciliated dogs of Manaus, Amazonas, Brazil.

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.

Dermocystidium sp. infection in farmed striped catfish Pangasianodon hypophthalmus farmed in Ceará state, Northeastern Brazil.

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.

Ticks and rickettsial exposure in lowland tapirs (Tapirus terrestris) of three Brazilian biomes.

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.

Occurrence of anti-Toxoplasma gondii and anti-Neospora caninum antibodies in pigs in the State of Pará, Brazil.

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.

Seroprevalence of Toxoplasma gondhi antibodies in captive wild mammals and birds in Brazil.

In this study, serum samples of 203 animals from different locations, from zoos and breeding facilities from the north and northeast regions of Brazil, were analyzed for the presence of anti-Toxoplasma gondii antibodies by the modified agglutination test (MAT) with a cutoff of 1:25. Of the sampled animals, 184 were adult mammals of both sexes and 19 were birds. Antibodies were found in 61 of 184 mammals, and no association between sex and age of the animals and the presence of T. gondii antibodies was observed (P < 0.05). Anti-T gondii antibodies were not found in birds. Toxoplasma gondii was detected in Brazilian tapir (Tapirus terrestris) for the first time.

Haliotrema aurigae (Yamaguti, 1968) Plaisance, Bouamer & Morand, 2004, in ornamental reef fish imported into Brazil.

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.

Parasitological diagnosis in food fish produced in a fish farm, in the Zona da Mata, Minas Gerais, southeastern Brazil.

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.

Scuticociliatosis caused by Uronema sp. in ten different ornamental aquarium reef fish in Brazil.

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.

First isolation and genotyping of Toxoplasma gondii in a free-living giant anteater (Myrmecophaga tridactyla) revealed a unique non-archetypal genotype.

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.