Molecular detection of Toxoplasma gondii and Neospora caninum in seabirds collected along the coast of Santa Catarina, Brazil.

Toxoplasma gondii and Neospora caninum are two closely related protozoans that infect a wide range of animals, including birds. However, the occurrence of N. caninum and T. gondii in seabirds is unknown. Therefore, this study aimed to determine the presence of T. gondii and N. caninum DNA in tissue samples of seabirds. Tissue samples of the pectoral muscles, heart, and brain were collected from 47 birds along the coastline of Santa Catarina State, SC, Brazil. The DNA was extracted from the tissues and screened using nested-PCR (nPCR) targeting internal transcribed spacer 1 (ITS1). T. gondii DNA was detected in tissues from seven seabirds (7/47, 14.8%), kelp gull (Larus dominicanus) (5/21), and Manx shearwater (Puffinus puffinus) (2/8). N. caninum DNA was detected in tissues of nine seabirds (9/47, 19.1%), the kelp gull (L. dominicanus) (4/21), Manx shearwater (P. puffinus) (2/8), neotropic cormorant (Phalacrocorax brasilianus) (1/4), brown booby (Sula leucogaster) (1/5), and white-chinned petrel (Procellaria aequinoctialis) (1/1); however, no co-infection was observed. In conclusion, this study showed the circulation of N. caninum and T. gondii in seabirds along the coastline of Santa Catarina State. Further studies are required to clarify the role of these birds in the epidemiology of neosporosis and toxoplasmosis.

Detection of Toxoplasma gondii DNA in milk of dairy cows from southern Brazil.

Consumption of unpasteurized cow's milk may be a transmission route for some pathogenic microorganisms, but there is little information about the risk of Toxoplasma gondii infection. Blood and milk samples were collected in a paired and random fashion from 106 dairy cows and bulk-tank milk samples were also collected from each of the six farms, in southern Brazil. Serum anti-T.gondii antibodies (IgG) were detected by an indirect fluorescent antibody test (IFAT) with a cutoff point of 1:64. Nested PCR targeting the ITS1 was performed on milk samples to detect the Sarcocystidae family, confirmed to be T.gondii by Sanger sequencing. The occurrence of anti-T.gondii antibodies in the herds was 14.1%, (15/106) with seropositive cows in all herds. Antibody titers in positive samples ranged from 64 to 128. T.gondii DNA was detected in 2.8% (03/106) of the milk samples. The ITS1 sequences generated in this study were ON809793 - ON809794 and the sequencing revealed 98-100% identity with T. gondii DNA sequences deposited in GenBank. All cows PCR positive for T.gondii in milk were negative for IgG antibodies in serum, suggesting that naturally infected cows may shed T. gondii in milk in the acute phase of infection. The results of this study demonstrate that T. gondii DNA may be detected in raw cow's milk, so the potential risks of lactogenic infection should be considered. The presence of T. gondii DNA in milk does not confirm that the protozoa are viable and infective, and further investigations into the role of cow's milk in the epidemiology of toxoplasmosis are needed.

Blood parasites of passerines in the Brazilian Pampas and their implications for a potential population supplementation program for the endangered Yellow Cardinal (Gubernatrix cristata).

Espinilho savanna ("seasonal steppe savanna") is a unique vegetation formation of the Pampas biome that is found near the tri-border of Brazil, Uruguay, and Argentina. The Yellow Cardinal (Gubernatrix cristata) is a flagship species of this ecosystem, but it is classified as "critically endangered" in Brazil due to habitat loss and poaching for the illegal trade. Population supplementation through the release of individuals that were captive-bred or apprehended by authorities from the illegal trade has been considered as a conservation strategy for this species; however, the risk of pathogen introduction is a critical concern. We used microscopy and molecular methods to investigate the occurrence of blood parasites in wild passerines (n = 64, including three Yellow Cardinals) at Espinilho State Park, Rio Grande do Sul, Brazil, and in captive Yellow Cardinals (n = 30) at three facilities in Brazil. Haemosporidian parasites were detected in the blood smears of 10.9% of the wild passerines, comprising the morphospecies Haemoproteus erythrogravidus in Rufous-collared Sparrow (Zonotrichia capensis), H. quiscalus in Grayish Baywing (Agelaioides badius), and H. tyranni in Great Kiskadee (Pitangus sulphuratus); these are the southernmost records for these morphospecies and their first record for the Pampas biome. No haemosporidian parasites were detected in the blood smears of the Yellow Cardinals, wild or captive. Microfilariae were detected in the blood smears of 14.1% of the wild passerines, including all wild Yellow Cardinals, and in 43.3% of captive Yellow Cardinals. Trypanosoma sp. was detected in the blood smear of one captive Yellow Cardinal. Nested PCR and gene sequencing of the cyt-b gene of Haemoproteus/Plasmodium was used to test a subset of wild passerines and captive Yellow Cardinals, allowing for the molecular barcoding of H. quiscalus lineage AGEBAD04 and H. tyranni lineage PITSUL01; additionally, DNA identical to that of lineage PITSUL01 was detected in the blood of one captive Yellow Cardinal. This study provides valuable data to support the conservation management of the Yellow Cardinal and other threatened passerines from the Pampas and highlights the need for further studies on the epidemiology and pathology of filarioid worms and trypanosomes in passerines from this biome.

Molecular characterization of Sarcocystis spp. in seabirds from southern Brazil.

Sarcocystis spp. are cyst forming apicomplexan parasites that infect many vertebrates including birds. Sarcocystis spp. infection was investigated in tissue samples (pectoral muscles, heart, and brain) of 47 dead seabirds collected from the coastline of Santa Catarina State SC - Brazil, between August 2019 and March 2020. A portion of each tissue was fixed in 10% buffered formalin for histopathologic analysis while DNA was extracted from another portion and screened using nested-PCR targeting ITS1. Based on molecular analysis, Sarcocystis spp. were identified in 15/47 (31.9%) seabirds of five species, kelp gull (Larus dominicanus), manx shearwater (Puffinus puffinus), neotropic cormorant (Phalacrocorax brasilianus), brown booby (Sula leucogaster) and great skua (Stercorarius skua). Microscopically visible sarcocysts were observed only in the pectoral muscle of four seabirds 8.5% (4/47), while in one brown booby, sarcocysts were seen in both pectoral and cardiac muscles. Two types of sarcocysts, thin walled (≤1 µm) and thick-walled (≥ 2 µm) were identified. Based on ITS1 sequence comparison, S. halieti, S. falcatula and three not yet described Sarcocystis spp. were detected. Phylogenetically, S. falcatula isolates were classified as two distinct clusters. This is the first confirmation of S. halieti in seabird's species in South America and S. falcatula in birds of the order Charadriiformes. Further molecular studies are needed to understand the epidemiology of the Sarcocystis spp. infection and its impact on the health of seabirds.

Detection of Neospora spp. and Sarcocystis neurona in amniotic fluid and placentas from mares.

The closely related apicomplexa protozoa, Toxoplasma gondii, Neospora spp., and Sarcocystis neurona, have all been associated with neurological and reproductive diseases in horses. However, there is limited data regarding the presence of these three parasites in equine placental tissues and amniotic fluid. The aim of the present report was to investigate the presence of the DNA of T. gondii, Neospora spp. and S. neurona in placentas and amniotic fluid in mares. Anti-T. gondii, anti- S. neurona and anti- Neospora spp. antibody titers were evaluated in 31 mares in the final third of pregnancy by indirect fluorescent antibody test (IFAT). The presence of parasite DNA in placentas and amniotic fluid was evaluated by polymerase chain reaction (PCR), using two target loci (ITS1 and Nc5). No antibodies to were identified nor was any T. gondii DNA detected in any mare. Antibodies to Neospora spp. were found in 6 mares (19.35 %) and DNA from this protozoan was detected in four placentas (12.9 %) and in five amniotic fluid samples (16.6 %). Antibodies to Sarcocystis spp. were detected in nine mares and S. neurona DNA was found in only one placenta (3.23 %). Our results suggest that the transplacental route may be a potential source of Neospora caninum infection in mares. Further studies are needed to understand the role of transplacental transmission in the epidemiology of these protozoa.

Production and Functional Evaluation of Anti-Loxosceles Sera Raised by Immunizations of Rabbits with Mutated Recombinant Phospholipases-D.

Loxoscelism is the clinical condition triggered after the bite of spiders of the genus Loxosceles. The main species involved in accidents in South America are L. intermedia, L. laeta, and L. gaucho. The only specific treatment is the anti-Loxosceles serum produced with crude venoms. As phospholipases D (PLDs) trigger most of the effects observed in accidents, we developed and evaluated second-generation sera using mutated PLDs as antigens. Three isoforms of PLDs with site-directed mutations without biological activities were used for rabbit immunizations: D32A-E34A (L. gaucho), W230A (L. intermedia), and H12A-H47A (L. laeta). Sera were produced using crude venoms of three species of Loxosceles enriched with mutated recombinant PLDs (MIX) or using only mutated PLDs (REC). Immunizations stimulated the immune system from the second immunization with higher antibody production in the REC group. In vivo neutralization assays demonstrated that both sera reduced edema and dermonecrosis caused by Loxosceles intermedia crude venom. Follow-up of animals during the immunization protocols and in the neutralization assays demonstrated that the mutated proteins and the sera are safe. Results demonstrate the potential of using mutated recombinant PLDs in total or partial replacement of Loxosceles venoms in animal immunizations to produce anti-Loxosceles sera for treatments of Loxoscelism.

An abortion storm in dairy cattle associated with neosporosis in southern Brazil.

Between December 2016 and April 2017, a spate of abortions occurred in a closed dairy herd from the central eastern region of Paraná, Brazil, in which 75 cows aborted. To identify its cause, organ fragments were collected from an aborted fetus for histopathology, and the blood samples from a stillborn, 4 aborted fetuses, and 9 farm dogs for indirect fluorescent antibody technique (IFAT). These tests found multifocal non-suppurative encephalitis, periportal hepatitis, and multifocal lymphoplasmacytic myocarditis, and detected anti-Neospora antibodies in all aborted fetuses, and in 5 of the 9 dogs. DNA of Neospora caninum was detected in the brain tissue of an aborted fetus. Blood samples of 340 cows and 146 heifers showed 33.5% and 30.8% seropositivity, respectively. In this closed herd, the parasite was probably introduced by infected domesticated or wild carnivores inhabiting the farm, through the infective oocysts present in their stool.

Transplacental transmission of Neospora caninum to lambs in successive pregnancies of naturally infected sheep in Southern Brazil.

This study aims to report the detection of N. caninum DNA in a newborn lamb (1) with neurological signs and congenital neosporosis and in a stillborn lamb (2), both born from the same ewe in a herd of Southern Brazil. The lambs were born during different pregnancies of a Suffolk ewe seropositive to N. caninum and seronegative to T. gondii. Histopathological lesions were observed only in the central nervous system of the lambs. The newborn lamb (1) showed mild and focal gliosis in the frontal lobe. In the hippocampal region of the stillborn lamb (2), lymphoplasmacytic perivascular cuffs and N. caninum cysts were observed in the cytoplasm of neurons and confirmed by IHC. PCR was performed using brain samples to detect the protozoa N. caninum and Toxoplasma gondii. The infection with N. caninum was confirmed in the newborn lamb (1) by PCR and in the stillborn lamb (2) by histopathology, immunohistochemistry, and PCR tests.

Serosurvey of Toxoplasma gondii, Neospora caninum and Sarcocystis neurona in raptors and risk factor analysis.

Raptors are carnivorous birds with great hunting ability. Toxoplasma gondii, Neospora caninum and Sarcocystis spp. are intracellular Apicomplexan protozoans which infect a wide range of intermediate hosts, including birds. The aims of this study were to evaluate the serological reactivity of captive raptors serum to T. gondii, N. caninum and S. neurona antigens and identify possible risk factors associated with the infection. From August 2014 to September 2015, blood samples from 72 raptors were collected and serum samples were tested by immunofluorescence antibody test (IFAT). Antigen slides were prepared using tachyzoites of T. gondii and N. caninum and using merozoites of S. neurona. Serum samples were tested at the following cut-off dilutions: 1:16 for T. gondii and 1:50 for N. caninum and S. neurona. An anti-chicken IgY antibody conjugated with FITC was used as a secondary antibody at 1:50 dilution. Out of the 72 raptors serum tested by IFAT, 2.7% reacted to N. caninum, 8.3% to T. gondii and 11.1% to S. neurona antigens. The region in which the sample was collected, the reason the raptors were kept in captivity and diet were statistically associated with seropositivity to T. gondii and the use of the birds and diet were statistically associated with seropositivity to N. caninum and S. neurona (p ≤ 0.05). We highlight the occurrence of these protozoans in birds of prey and the importance of good hygiene and feeding management of these birds in captivity to reduce the risk of protozoal infections.

Molecular diagnosis of avian viruses in grassland passerines and captive yellow cardinals Gubernatrix cristata in Brazil

ABSTRACT: Avian influenza viruses (AIVs), Newcastle disease virus (NDV), West Nile virus (WNV), adenovirus (AV) and herpesvirus (HV) play an important role in the health of human and animal populations. However, knowledge of the prevalence of these viruses in wild birds is restricted to some groups (e.g. shorebirds) or regions worldwide. Information on grassland birds of South America, which is essential for their conservation, is scarce. The objectives of the present study were to evaluate occurrences of AIV, NDV, WNV, AV and HV for the first time in a bird community of a unique protected area in southern Brazil, which is home for the critically endangered yellow cardinal (Gubernatrix cristata), and captive yellow cardinals from fauna maintainers of the Brazilian Captive Program of the Yellow Cardinal. Passerine species of wild life were caught, identified and samples (swabs) were collected from the oropharynx and cloaca of 64 passerines of 26 species (including 3 yellow cardinals) and 30 yellow cardinals of captive, for molecular diagnosis. The samples were subjected to RNA and DNA extraction and the real-time polymerase chain reaction (RT-PCR) for AIV, NDV and WNV and nested PCR for AV and HV. One yellow cardinal of captive presented a positive result for AV, this result is important for planning, managing natural attributes and making decisions in relation to integrated conservation of threatened species. This is the first report of AV in yellow cardinal and epidemiological investigation of viruses in wild passerines of the Pampa biome, in Rio Grande do Sul, Brazil.

RESUMO: Os vírus da gripe aviária (VGA), vírus da doença de Newcastle (VDN), vírus do Nilo Ocidental (VNO), adenovírus (AV) e herpesvírus (HV) desempenham um papel importante na saúde das populações humana e animal. No entanto, o conhecimento da prevalência desses vírus em aves selvagens é restrito a alguns grupos (por exemplo, aves limícolas) ou regiões em todo o mundo. As informações sobre as aves campestres da América do Sul, essenciais para a sua conservação, são escassas. Os objetivos do presente estudo foram avaliar a ocorrência de VGA, VDN, VNO, AV e HV pela primeira vez em uma comunidade de aves de uma área única protegida no Sul do Brasil, que abriga o cardeal-amarelo (Gubernatrix cristata) criticamente ameaçado de extinção e em cardeais-amarelos de cativeiro dos mantenedores de fauna do Programa Brasileiro de Cativeiro do Cardeal-amarelo. Espécies de passeriformes silvestres foram capturadas, identificadas e amostras (swabs) foram coletadas da orofaringe e cloaca de 64 passeriformes de 26 espécies (incluindo 3 cardeais-amarelos) e 30 cardeais-amarelos de cativeiro, para diagnóstico molecular. As amostras foram submetidas à extração de RNA e DNA e à reação em cadeia da polimerase em tempo real (RT-PCR) para VGA, VDN e VNO e nested PCR para AV e HV. Um cardeal-amarelo de cativeiro apresentou resultado positivo para AV, este resultado é importante para o planejamento, manejo dos atributos naturais e tomada de decisões em relação à conservação integrada de espécies ameaçadas. Este é o primeiro relato de AV em cardeal-amarelo e de investigação epidemiológica de vírus em passeriformes silvestres do bioma Pampa, no Rio Grande do Sul, Brasil.

Molecular diagnosis of avian viruses in grassland passerines and captive yellow cardinals Gubernatrix cristata in Brazil / Diagnóstico molecular de vírus aviários em passeriformes campestres e cardeais-amarelos Gubernatrix cristata cativos no Brasil

Avian influenza viruses (AIVs), Newcastle disease virus (NDV), West Nile virus (WNV), adenovirus (AV) and herpesvirus (HV) play an important role in the health of human and animal populations. However, knowledge of the prevalence of these viruses in wild birds is restricted to some groups (e.g. shorebirds) or regions worldwide. Information on grassland birds of South America, which is essential for their conservation, is scarce. The objectives of the present study were to evaluate occurrences of AIV, NDV, WNV, AV and HV for the first time in a bird community of a unique protected area in southern Brazil, which is home for the critically endangered yellow cardinal (Gubernatrix cristata), and captive yellow cardinals from fauna maintainers of the Brazilian Captive Program of the Yellow Cardinal. Passerine species of wild life were caught, identified and samples (swabs) were collected from the oropharynx and cloaca of 64 passerines of 26 species (including 3 yellow cardinals) and 30 yellow cardinals of captive, for molecular diagnosis. The samples were subjected to RNA and DNA extraction and the real-time polymerase chain reaction (RT-PCR) for AIV, NDV and WNV and nested PCR for AV and HV. One yellow cardinal of captive presented a positive result for AV, this result is important for planning, managing natural attributes and making decisions in relation to integrated conservation of threatened species. This is the first report of AV in yellow cardinal and epidemiological investigation of viruses in wild passerines of the Pampa biome, in Rio Grande do Sul, Brazil.(AU)

Os vírus da gripe aviária (VGA), vírus da doença de Newcastle (VDN), vírus do Nilo Ocidental (VNO), adenovírus (AV) e herpesvírus (HV) desempenham um papel importante na saúde das populações humana e animal. No entanto, o conhecimento da prevalência desses vírus em aves selvagens é restrito a alguns grupos (por exemplo, aves limícolas) ou regiões em todo o mundo. As informações sobre as aves campestres da América do Sul, essenciais para a sua conservação, são escassas. Os objetivos do presente estudo foram avaliar a ocorrência de VGA, VDN, VNO, AV e HV pela primeira vez em uma comunidade de aves de uma área única protegida no Sul do Brasil, que abriga o cardeal-amarelo (Gubernatrix cristata) criticamente ameaçado de extinção e em cardeais-amarelos de cativeiro dos mantenedores de fauna do Programa Brasileiro de Cativeiro do Cardeal-amarelo. Espécies de passeriformes silvestres foram capturadas, identificadas e amostras (swabs) foram coletadas da orofaringe e cloaca de 64 passeriformes de 26 espécies (incluindo 3 cardeais-amarelos) e 30 cardeais-amarelos de cativeiro, para diagnóstico molecular. As amostras foram submetidas à extração de RNA e DNA e à reação em cadeia da polimerase em tempo real (RT-PCR) para VGA, VDN e VNO e nested PCR para AV e HV. Um cardeal-amarelo de cativeiro apresentou resultado positivo para AV, este resultado é importante para o planejamento, manejo dos atributos naturais e tomada de decisões em relação à conservação integrada de espécies ameaçadas. Este é o primeiro relato de AV em cardeal-amarelo e de investigação epidemiológica de vírus em passeriformes silvestres do bioma Pampa, no Rio Grande do Sul, Brasil.(AU)

Serosurvey of Toxoplasma gondii, Sarcocystis sp. and Neospora caninum in geese (Anser sp.) from urban parks and captivity.

Geese, ducks, mallards, and swans are birds of the order Anseriformes, which are found in the wild, in zoos and parks, and raised for meat consumption. Toxoplasma gondii, Sarcocystis sp., and Neospora caninum are protozoans of several species of animals. Wild and domestic birds can serve as intermediate hosts, disseminators and potential sources of infection of these protozoa to humans through contaminated meat. The aims of this study were: (i) to perform a serological survey of T. gondii, Sarcocystis sp. and N. caninum in geese (Anser sp.) from public parks and from captivity and (ii) to compare seroprevalence between these two locations. Antibodies were detected by Immunofluorescence antibody test using the serum of 149 geese. Antibodies to Sarcocystis sp., T. gondii, and N. caninum were detected in 28.18%, 18% and 0.67% of geese, respectively; 57% of geese from urban parks and 26.53% of geese from captivity were seropositive for at least one protozoa. The results indicate environmental contamination, particularly for the occurrence of antibodies against T. gondii - a zoonosis that causes toxoplasmosis and is transmitted through oocyte ingestion. This is the first serological survey of T. gondii, Sarcocystis sp. and N. caninum in geese from urban parks in Curitiba, Brazil.

HEMATOLOGICAL REFERENCE INTERVALS OF ENDANGERED CAPTIVE BLACK-FRONTED PIPING-GUANS (ABURRIA JACUTINGA) IN THE STATE OF PARANÁ, BRAZIL.

The black-fronted piping-guan (Aburria jacutinga) is and an endangered cracid species and little is known about its health. Hematology allows researchers to assess the health and physiological status of a population, but, to our knowledge, reference intervals are not available in the literature for A. jacutinga. Thus, the present study aimed to determine the hematological reference intervals of 31 captive black-fronted pipingguan adults in the state of Paraná, Brazil. Blood samples were collected from brachial vein and stored in heparinized tubes to be processed. The red blood cell (RBC) and white blood cell (WBC) counts were performed in a hemocytometer, and hemoglobin (HGB) concentration was obtained by the spectrophotometric cyanmethemoglobin method; packed cell volume (PCV) was determined by microhematocrit technique, and the mean corpuscular volume (MCV) and mean corpuscular hemoglobin concentration (MCHC) were calculated according to a previously established formula. The heterophil : lymphocyte ratio was calculated by dividing the number of heterophils by the number of lymphocytes. Determination of the 95% reference intervals and 90% confidence intervals were performed in accordance with American Society for Veterinary Clinical Pathology guidelines using Reference Value Advisor (version 2.0). The results obtained were as follows: RBC, 1.4-2.8 × 106/µl; PCV, 34.3-51.2%; HGB, 8.0-12.6 g/dl; MCV, 157.3-261.4 fl; MCHC, 22.0-26.7 g/dl; WBC, 2.3-33.1 × 103/µl; heterophils, 12.1-16.3%, 0.5-14.4 × 103/µl; lymphocytes, 19.8-69.0%, 1.4-16.4 × 103/µl; monocytes, 0.7-17.3%, 0.1-2.8×103/µl; eosinophils, 0.4-12.8%, 0.0-3.9 × 103/µl; basophils, 0.8-12.0%, 0.0-1.9 × 103/µl; and the heterophil : lymphocyte ratio, 0.2-3.0. The present study fills gaps in the literature on hematological values for A. jacutinga, which can be used to assess the health of the species populations. In addition, it contributed to establishing the health status of captive black-fronted piping-guans, indicating the possible welfare of the birds.

Serosurvey of Toxoplasma gondii, Sarcocystis sp. and Neospora caninum in geese (Anser sp.) from urban parks and captivity / Soroprevalência de Toxoplasma gondii, Sarcocystis sp. e Neospora caninum em gansos (Anser sp.) de parques urbanos e cativeiro

Abstract Geese, ducks, mallards, and swans are birds of the order Anseriformes, which are found in the wild, in zoos and parks, and raised for meat consumption. Toxoplasma gondii, Sarcocystis sp., and Neospora caninum are protozoans of several species of animals. Wild and domestic birds can serve as intermediate hosts, disseminators and potential sources of infection of these protozoa to humans through contaminated meat. The aims of this study were: (i) to perform a serological survey of T. gondii, Sarcocystis sp. and N. caninum in geese (Anser sp.) from public parks and from captivity and (ii) to compare seroprevalence between these two locations. Antibodies were detected by Immunofluorescence antibody test using the serum of 149 geese. Antibodies to Sarcocystis sp., T. gondii, and N. caninum were detected in 28.18%, 18% and 0.67% of geese, respectively; 57% of geese from urban parks and 26.53% of geese from captivity were seropositive for at least one protozoa. The results indicate environmental contamination, particularly for the occurrence of antibodies against T. gondii - a zoonosis that causes toxoplasmosis and is transmitted through oocyte ingestion. This is the first serological survey of T. gondii, Sarcocystis sp. and N. caninum in geese from urban parks in Curitiba, Brazil.

Resumo Gansos, patos, marrecos e cisnes são aves da ordem Anseriformes, encontrados em vida livre, zoológicos, parques e criados para consumo da carne. Toxoplasma gondii, Sarcocystis sp. e Neospora caninum são protozoários capazes de infectar diversas espécies animais. Aves domésticas e silvestres podem ser hospedeiras intermediárias e servir como disseminadoras e potenciais fontes de infecção para seres humanos por meio da carne. O objetivo do estudo foi 1) realizar a soroprevalência de T. gondii, Sarcocystis sp. e N. caninum em gansos (Anser sp.) provenientes de parques públicos e de um cativeiro e 2) comparar a soroprevalência entre os locais. Foi realizada sorologia de 149 Anser sp. pelo método da reação de imunofluorescência indireta. Anticorpos para Sarcocystis sp., T. gondii e N. caninum foram encontrados em 28,18%, 18%, e 0,67% dos animais, respectivamente; 57% dos gansos dos parques públicos e 26,53% dos animais cativos foram soropositivos para algum dos protozoários. A ocorrência de anticorpos para tais protozoários indica contaminação ambiental, ressaltando a alta prevalência de anticorpos para T. gondii, zoonose transmitida por ingestão dos oocistos. Sugere-se investigação da água e medidas ambientais para reduzir a contaminação dos animais e do ambiente. Este é o primeiro trabalho que avaliou sorologicamente gansos provenientes de parques urbanos de Curitiba, Paraná para T. gondii, Sarcocystis sp. e N. caninum.

Effects of silymarin on angiogenesis and oxidative stress in streptozotocin-induced diabetes in mice.

The present study evaluated the effects of acute treatment with silymarin, an extract that is obtained from Silybum marianum, on angiogenesis, oxidative stress, and inflammation in normoglycemic and diabetic mice. Diabetes was induced by streptozotocin (80 mg/kg, intraperitoneal) in male Swiss mice, 6 weeks of age. A polyether-polyurethane sponge was surgically implanted in the back of the mice as a model of healing in both diabetic and normoglycemic animals that were treated with oral silymarin or water for 10 days. The pancreas, liver, kidneys, blood, and sponges were collected and analyzed. Diabetes led to impairments of antioxidant defenses, reflected by a reduction of pancreatic superoxide dismutase and hepatic and renal catalase and an increase in pancreatic lipoperoxidation. An inflammatory process was observed in diabetic mice, reflected by an increase in pancreatic tumor necrosis factor α (TNF-α) and the infiltration of inflammatory cells in islets. The number of vessels was lower in the implanted sponges in diabetic mice. Silymarin treatment attenuated this damage, restoring antioxidant enzymes and reducing pancreatic TNF-α and inflammatory infiltration. However, silymarin treatment did not restore angiogenesis or glycemia. In conclusion, treatment with silymarin red uced oxidative stress and inflammation that were induced in the model of streptozotocin-induced diabetes in several organs, without apparent toxicity. Silymarin may be a promising drug for controlling diabetic complications.

Isolation of Neospora caninum from kidney and brain of a bovine foetus and molecular characterization in Brazil.

Bovine neosporosis has become a disease of international concern as it is among the main causes of abortion in cattle. Viable N. caninum has been isolated from brains of fetuses and neonatal calves, and there is no report of isolation of tachyzoites from kidney. Also, detailed information about the genetic diversity of N. caninum is scarce. N. caninum tachyzoites were isolated from the kidney and the brain of an aborted 4-month-old bovine foetus. The parasite was confirmed to be N. caninum by PCR. The tachyzoites of the new isolate, named BNC-PR4, were propagated in Vero cell cultures. Pathogenicity of the parasite was examined in BALB/c mice. Mice inoculated intraperitoneally with BNC-PR4 failed to yield clinical signs of disease and did not induce severe brain lesions, suggesting a bovine isolate with low virulence. The N. caninum-positive DNA sample was further analyzed by multilocus microsatellite (MS) genotyping for MS4, MS5, MS6A, MS6B, MS7, MS8, MS10, MS12, and MS21. Multilocus-microsatellite genotyping revealed a unique genetic profile that differed from previously reported isolates.

Survey of pathogens in threatened wild red-tailed Amazon parrot (Amazona brasiliensis) nestlings in Rasa Island, Brazil

ABSTRACT The red-tailed Amazon parrot (Amazona brasiliensis) is a threatened species of psittacine bird that inhabit coastal regions of Brazil. In view of the threat of this species, the aim of this study was to perform a health evaluation in wild nestlings in Rasa Island, determining the prevalence of enterobacteria and infectious agents according to type of nest. Blood samples were collected from 64 birds and evaluated for antibodies of Chlamydia psittaci by commercial dot-blot ELISA. Cloacal and oropharyngeal swabs samples were collected from 23 birds from artificial wooden nests, 15 birds from PVC nests and 2 birds from natural nests for microbiological analysis. Swab samples were collected from 58 parrots for C. psittaci detection by PCR and from 50 nestlings for Avian Influenza, Newcastle Disease and West Nile viruses' detection analysis by real-time RT-PCR. Ten bacterial genera and 17 species were identified, and the most prevalent were Escherichia coli and Klebsiella oxytoca. There was no influence of the type of nest in the nestlings' microbiota. All samples tested by ELISA and PCR were negative. There is currently insufficient information available about the health of A. brasiliensis and data of this study provide a reference point for future evaluations and aid in conservation plans.

Survey of pathogens in threatened wild red-tailed Amazon parrot (Amazona brasiliensis) nestlings in Rasa Island, Brazil.

The red-tailed Amazon parrot (Amazona brasiliensis) is a threatened species of psittacine bird that inhabit coastal regions of Brazil. In view of the threat of this species, the aim of this study was to perform a health evaluation in wild nestlings in Rasa Island, determining the prevalence of enterobacteria and infectious agents according to type of nest. Blood samples were collected from 64 birds and evaluated for antibodies of Chlamydia psittaci by commercial dot-blot ELISA. Cloacal and oropharyngeal swabs samples were collected from 23 birds from artificial wooden nests, 15 birds from PVC nests and 2 birds from natural nests for microbiological analysis. Swab samples were collected from 58 parrots for C. psittaci detection by PCR and from 50 nestlings for Avian Influenza, Newcastle Disease and West Nile viruses' detection analysis by real-time RT-PCR. Ten bacterial genera and 17 species were identified, and the most prevalent were Escherichia coli and Klebsiella oxytoca. There was no influence of the type of nest in the nestlings' microbiota. All samples tested by ELISA and PCR were negative. There is currently insufficient information available about the health of A. brasiliensis and data of this study provide a reference point for future evaluations and aid in conservation plans.

Hematologic and biochemical reference intervals of free-living Red-Tailed Amazon parrot (Amazona brasiliensis) nestlings on Rasa Island, Paraná, Brazil.

BACKGROUND: The Red-Tailed Amazon parrot (Amazona brasiliensis) is an endangered species of the Psittaciformes. There is little information about hematologic and biochemical variables of this species. OBJECTIVES: The purpose of this study was to determine hematologic and biochemical RIs for free-living A brasiliensis nestlings on Rasa Island, Paraná, Brazil, and to compare the results between sexes. METHODS: Thirty-seven parrots were taken from their nests and physically restrained for clinical examination and blood collection. The sex was diagnosed by PCR using the blood samples collected. Reference intervals were determined as recommended by the ASVCP guidelines in healthy nestlings. The difference between groups was analyzed using the Mann-Whitney test or Student's t-test. RESULTS: Sexing revealed 12 females and 25 males. The RIs for the measured variables were as follows: RBC 1.1-2.6 × 106 /µL, PCV 29.1-50.3%, HGB 7.2-12.9 g/dL, MCV 152-293 fL, MCHC 22.2-28.4 g/dL, WBC 4.9-28.5 × 103 /µL, 1.2-16 × 103 /µL, lymphocytes 2.4-18.7 × 103 /µL, monocytes 0.0-1.0 × 103 /µL, eosinophils 0.0-0.9 × 103 /µL, 0.0-1.3 × 103 /µL, heterophil:lymphocyte ratio 0.0-2.2, plasma total solids 2.1-3.7 g/dL, uric acid 0.5-2.0 mg/dL, glucose 184.9-284.3 mg/dL, AST 100.3-226.6 U/L, LDH 178.1-927.7 U/L, CK 149.8-1144.0 U/L, cholesterol 137.5-256.9 mg/dL, total protein 1.8-3.0 g/dL, calcium 7.0-8.6 mg/dL, and phosphorus 2.9-6.1 mg/dL. Increased concentrations of cholesterol (P < .05) were observed in females. DISCUSSION: This is the first study to establish hematologic and biochemical RIs for free-living A brasiliensis nestlings on Rasa Island. Hematologic and biochemical variables are important tools for evaluating the health status of free-living birds, and also support conservation planning for endangered species.

Laminar lesions in horses with systemic oxidative stress, committed by experimentally induced or naturally occurring gastrointestinal disorders / Lesões laminares em cavalos com estresse oxidativo sistêmico, acometidos por doenças gastrintestinais experimentalmente induzidas ou de ocorrência natural

Laminitis in horses can be associated with lesions in multiple organs secondary to sepsis. Twenty-one horses suffering from gastrointestinal disorders were used in the experiment; 7 horses with experimentally induced endotoxemia and intestinal ischaemia, and 14 horses suffering from naturally occurring colic syndrome. Tissue samples of lungs, liver, heart, brain, cerebellum and hoof laminar tissue were collected for histopathological and oxidative stress evaluation using nitrotyrosine and superoxide dismutase (SOD2) immunostaining. The horses were divided into two groups: the non-oxidative lesions group (NOLG), with 7 horses showing weak immunostaining in lungs, liver and kidney, and the oxidative lesions group (OLG), with 14 horses showing immunostaining indicating systemic oxidative stress in multiple organs. The horses from OLG showed increase of laminar lesions and SOD2 immunostaining in multiple organs when compared to the horses from the NOLG. No differences were found ln regard to laminar immunostaining by nitrotyrosine and SOD2 between experimental groups. It was concluded that systemic oxidative stress can be associated with the development of laminar lesions, and that the laminar tissue does not respond to oxidative stress with increase of SOD as occurs in other organs.(AU)

A laminite em equinos pode estar associada à lesão em múltiplos órgãos secundária a sepse. Foram utilizados 21 cavalos com afecções gastrintestinais, sendo sete com endotoxemia e isquemia intestinal induzidos experimentalmente, e 14 cavalos com síndrome cólica de origem natural. Amostras teciduais de pulmão, rim, fígado, coração, cérebro e cerebelo e de tecido laminar do casco foram coletadas para avaliação de lesão histopatológica e estresse oxidativo, pela imunomarcação de nitrotirosina e superóxido dismutase (SOD2). Os animais foram divididos em dois grupos: grupo sem lesão oxidativa (NOLG), com sete cavalos com fraca imunomarcação em pulmão, fígado e rim, e grupo lesão oxidativa (OLG), contendo 14 cavalos com imunomarcação indicando estresse oxidativo em múltiplos órgãos. Os cavalos do grupo OLG apresentaram aumento de lesões laminares e imunomarcação para SOD2 em múltiplos órgãos, quando comparados ao NOLG. Não houve diferença sobre a imunomarcação laminar para nitrotirosina e SOD2 entre os grupos experimentais. Conclui-se que o estresse oxidativo sistêmico está associado ao desenvolvimento de lesões laminares, e que o tecido laminar não responde ao estresse oxidativo com aumento de SOD como ocorre nos outros órgãos.(AU)