Rabies Virus-Neutralizing Antibodies in Free-Ranging Invasive Wild Boars (Sus scrofa) from Brazil.

Rabies, one of the most lethal global zoonoses, affects all mammals. It remains circulating worldwide in sylvatic cycles through terrestrial and airborne reservoirs, and in Brazil, bats are currently the main reservoirs and source of transmission. Wild boars, an important invasive alien species in Brazil, are a proven food source for hematophagous bats and may participate in the Brazilian sylvatic cycle of rabies. We evaluated the presence of this pathogen in hunted wild boars from the São Paulo state using histopathology, the direct fluorescent antibody test (DFA), viral isolation in cell culture (VICC), the rapid fluorescent focus inhibition test (RFFIT), and quantitative reverse transcription polymerase chain reaction (RT-qPCR). The results of histopathological, DFA, VICC, and RT-qPCR analysis were negative for all samples; seven serum samples tested positive in the RFFIT, and titers ranged from 0.13 IU/mL to 0.5 IU/mL. The presence of rabies virus-neutralizing antibodies in the studied wild boars suggests the circulation of the virus in these animals. Educative actions directed at hunters should include information on the prevention of this important zoonosis.

Integrative taxonomy of Metastrongylus spp. in wild boars from Brazil.

BACKGROUND: Wild boars (Sus scrofa) may cause substantial damage to crops and can spread zoonotic parasites to domestic animals, posing a risk to health and animal production. Metastrongylus spp. can negatively affect the wild boar population, increasing piglet mortality. In addition to that, studies with Metastrongylus genetic characterization are still scarce in Brazil. The present study aims to characterize Metastrongylus spp. from wild boars hunted in the states of São Paulo, Paraná, and Rio Grande do Sul, Brazil, using traditional morphological description and DNA sequences in an integrative taxonomic approach. METHODS: After nematode collection from 58 wild boars, the parasites were morphologically identified and genetically characterized by the amplification of 18S ribosomal DNA (rDNA), 28S rDNA, internal transcribed spacer (ITS) region, and cox-1 mitochondrial DNA (mtDNA). Descriptors of infection were determined and Pearson's Chi-square test was applied to compare the prevalence of infections among the identified parasite species, host age group (juveniles and adults), and sex. The Mann-Whitney U test was performed to compare the mean intensity between the age groups and sex. RESULTS: Metastrongylus salmi, Metastrongylus apri, and Metastrongylus pudendotectus were identified in 77.6% (45/58) of the necropsied wild boars. Metastrongylus salmi was the most prevalent and abundant species (70.7%, 11.1), followed by M. pudendotectus (18.9%, 4.3) and M. apri (17.2%, 2.2). Metastrongylus pudendotectus showed the highest mean intensity and range (25.2, 1-93), followed by M. salmi (15.7, 1-58) and M. apri (12.6, 3-27). We found a significantly higher prevalence of Metastrongylus spp. and M. salmi in adult wild boars, probably associated with a more prolonged time of exposure to intermediate host species. The phylogenetic analysis revealed that ITS2 region and cox-1 mtDNA are the most suitable genetic markers for Metastrongylus species characterization. Genetic variability between M. apri and M. salmi isolates was verified. CONCLUSIONS: We expand the knowledge about the Metastrongylus community in the non-captive wild boar population from Brazil as well as the importance of this exotic species in the maintenance of Metastrongylus spp. in its areas of occurrence. The novel genetic sequences obtained may help further studies to understand the genetic diversity in other nematode populations from Brazil and other countries.

Epidemiology and Integrative Taxonomy of Helminths of Invasive Wild Boars, Brazil.

Wild boars (Sus scrofa) are a significant invasive species in Brazil. We evaluated the helminth diversity of 96 wild boars in São Paulo state. Helminth infection descriptors were calculated, the species were identified and their 18S, 28S rDNA and internal transcribed spacer (ITS) regions were amplified for phylogenetic analyses. Ascarops strongylina, Strongyloides ransomi, Globocephalus urosubulatus, Oesophagostomum dentatum, Trichuris suis, Metastrongylus salmi, Metastrongylus pudendotecus, Ascaris suum and Stephanurus dentatus and Macracanthorhynchus hirudinaceus were identified. Globocephalus urosubulatus had the highest prevalence and mean abundance, and most animals had mixed infections with three parasite species. There was no association between parasite intensity and prevalence and host sex and body condition index (p > 0.05). Novel DNA sequences were obtained from G. urosubulatus, A. strongylina, and S. dentatus. This is the first study on the helmint diversity of non-captive wild boars in Brazil, and the first report of the occurrence of M. hirudinaceus, G. urosubulatus and S. dentatus in Brazilian wild boars. Non-captive wild boars of São Paulo State did not act as capture hosts for native helminth species but maintained their typical parasites, common to domestic pigs. They may act as parasite dispersers for low-tech subsistence pig farming and for native Tayassuidae.

Filarids (Spirurida: Onchocercidae) in wild carnivores and domestic dogs from the Brazilian Atlantic forest.

Onchocercidae nematodes are heteroxenous parasites with worldwide distribution, and some of the species associated to animals may present zoonotic potential. Climatic changes and anthropic influences on the environment may result in vectors' proliferation, facilitating the spillover to humans and/or non-typical animal hosts. The Iguaçu National Park (PARNA Iguaçu), one of the most important Brazilian natural remanescents of Atlantic rainforest, is strongly affected by human activities such as tourism and agriculture. The complexity of this area is especially characterized by the close nexus between the rich wildlife, humans, and domestic animals, especially domestic dogs. Based on this, this research aimed to diagnose the Onchocercidae nematodes in wild carnivores and domestic dogs in the PARNA Iguaçu and the surrounding areas. For this, we collected 162 samples of seven species of wild carnivores and 225 samples of domestic dogs. The presence of microfilariae in the blood samples was diagnosed by the modified Knott's test and molecular screening, and the specific identification was based on sequencing of the myoHC and hsp70 genes. Microfilariae were detected only in ring-tailed coatis, in which we found five species: Mansonella sp. 1, Mansonela sp. 2, Onchocercidade gen. sp. 1, Onchocercidade gen. sp. 2, and Dirofilaria immitis. The morphological analysis supported the molecular findings. The domestic dogs were parasitized by Acanthocheilonema reconditum, representing a new locality record for this species. Phylogenetic analysis showed high genetic similarity among the four undetermined species and Mansonella spp., Brugia spp., and Wuchereria bancrofti. The presence of D. immitis in ring-tailed coatis may be result of spillover from dogs, even though the parasite was not diagnosed in the sampled dogs. The presence of several undetermined Onchocercidae species indicates the necessity of continuous investigations on wild and domestic animals from Neotropical area, especially considering the growing anthropic influence on forest remnants.

Toxoplasma gondii infection in wild boars (Sus scrofa) from the State of São Paulo, Brazil: Serology, molecular characterization, and hunter's perception on toxoplasmosis.

The consumption of wild boar meat, common in many countries, became popular in Brazil after the hunting of these animals was authorized in 2013. The meat of these animals is often consumed by hunters and their social groups, and their offal is occasionally used as supplemental food in the diet of hunting dogs. Given the high frequency of foodborne diseases related to wild boar meat consumption in other countries, including toxoplasmosis, knowledge on these diseases is essential for risk assessment and elaboration of education campaigns for the exposed public. Thus, this study aimed diagnosing, isolating, and genotyping Toxoplasma gondii in hunted wild boars. For that, we obtained samples of serum and tissues (brain, tongue, diaphragm, and heart) from 26 wild boar hunted in three areas in São Paulo State, Brazil, based on convenience sampling strategy. The serum samples were submitted to the indirect immunofluorescence reaction test (IFAT) test while the tissue samples (n = 22) were used to perform a bioassay in mice to isolate the parasite. The isolated samples were genetically characterized by PCR-RFLP with SAG1, 5' and 3' SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico markers. Questionnaires were also formulated and applied to wildlife hunters to assess knowledge about toxoplasmosis. The seroprevalence of T. gondii was 76.9% (20/26), with titers ranging from 16 to 1024. Viable parasites accounted for 4.5% (1/22) of the samples. The ToxoDB #6 genotype of TgJava1 alone was detected. Most interviewed hunters, 84.2% (16/19) consume game meat and a few of them (15.7%; 3/19) prefer undercooked meat. Also, 15.7% (3/19) of the hunters reported supplementing their hunting dogs' diet with wild boar meat and/or offal. As antibodies to T. gondii were detected in 76.9% (20/26) of the studied wild boars, we concluded that infection by T. gondii is frequent in wild boars used for human and animal consumption in the studied areas. Although genotype #6 is commonly found in Brazil in domestic animals, wild animals, and humans, causing everything from mild clinical symptoms to death, this study found, for the first time, the detection of this genotype in wild boars. These results also reaffirm the importance of these animals as a possible source of T. gondii infection for humans and domestic animals.

Eurytrema coelomaticum natural infection in small ruminants: a neglected condition.

Pancreatic eurytrematosis (PE) is an under diagnosed and neglected parasitosis in goats and sheep in the Americas. Clinical and pathological features of PE are not well defined in small ruminants worldwide. Natural cases of PE in small ruminants were detected in the Federal District, Brazil. A survey of necropsy records, including epidemiological and clinicopathological data, in goats and sheep was conducted. Most cases of PE occurred during the rainy season in adult females, with an incidence of 12.9% in goats and 0.8% in sheep. Clinical signs varied from asymptomatic infections to anorexia, lethargy, weakness, marked weight loss and death in some goats. Overall, most cases of PE in goats and sheep were incidental necropsy findings with minor pancreatic lesions. Three goats, however, showed severe chronic pancreatitis, dilation of major pancreatic ducts with numerous trematodes present and marked abdominal fat necrosis. Morphological and molecular characterization of flukes detected Eurytrema coelomaticum. Our findings shed light on the prevalence of E. coelomaticum infections in small ruminants in the region and highlight the possibility of severe and lethal cases in goats. PE must be further investigated in small ruminant populations in relevant livestock production regions of the Americas.

Oncicola luehei in a wild crab-eating raccoon (Procyon cancrivorus) from the Brazilian Cerrado Savanna

The occurrence of Oncicola luehei is reported in a road killed crab-eating raccoon (Procyon cancrivorus) near the municipality of Uberlândia, Minas Gerais State. The animal was collected as part of a study that monitors wildlife road killing in the Triângulo Mineiro region. In necropsy, a single male acanthocephalan was recovered from the large intestine. The parasite was wrinkled, whitish in color, with a total body length of 15.88mm, globular proboscis (0.71 x 0.81mm) armed with 36 spiraled hooks, long lemniscus (7.30 x 0.81 mm) surpassing the anterior testis. The testes were ellipsoid in shape, disposed in tandem, the anterior measuring 1.44 x 0.53mm and the posterior 1.5 x 0.50mm. At the posterior part of the body, eight cement glands arranged in two rows of four, measuring 0.38 x 0.46 mm each. Based on this, the parasite was classified as Oncicola luehei. This study represents a new host and locality records for the parasite.

Removal of total coliforms, thermotolerant coliforms, and helminth eggs in Swine production wastewater treated in anaerobic and aerobic reactors.

The present work evaluated the performance of two treatment systems in reducing indicators of biological contamination in swine production wastewater. System I consisted of two upflow anaerobic sludge blanket (UASB) reactors, with 510 and 209 L in volume, being serially arranged. System II consisted of a UASB reactor, anaerobic filter, trickling filter, and decanter, being also organized in series, with volumes of 300, 190, 250, and 150 L, respectively. Hydraulic retention times (HRT) applied in the first UASB reactors were 40, 30, 20, and 11 h in systems I and II. The average removal efficiencies of total and thermotolerant coliforms in system I were 92.92% to 99.50% and 94.29% to 99.56%, respectively, and increased in system II to 99.45% to 99.91% and 99.52% to 99.93%, respectively. Average removal rates of helminth eggs in system I were 96.44% to 99.11%, reaching 100% as in system II. In reactor sludge, the counts of total and thermotolerant coliforms ranged between 10(5) and 10(9) MPN (100 mL)(-1), while helminth eggs ranged from 0.86 to 9.27 eggs g(-1) TS.

Morphology of eggs of Dioctophyme renale Goeze, 1782 (Nematoda: Dioctophymatidae) and influences of temperature on development of first-stage larvae in the eggs.

This study aims to provide information on morphological data of D. renale eggs, as well as on first-stage larvae development into eggs kept at different temperatures. Eggs were obtained by centrifugation of infected dog urine, placed in Petri dishes, and stored in BOD chamber for a 90-day period. Each treatment (GI--15 degrees C, GII--20 degrees C, and GIII--26 degrees C) was repeated five times. Eggs showed average measures of 67.23 x 42.78 microm, and the mean incubation time was inversely proportional to the incubation temperature. Larvae motility was observed one week after being observed in eggs.