Alexandre Munhoz | Universidade Estadual de Santa Cruz (original) (raw)
Papers by Alexandre Munhoz
Revista Brasileira de Parasitologia Veterinária, 2011
The aim of this study was to determine the presence of species of the genus Eimeria species in na... more The aim of this study was to determine the presence of species of the genus Eimeria species in naturally infected bovines in Southern Bahia, Northeast Brazil. The study population comprised 117 Zebu crossbred cattle that belonged to 10 dairy herds with extensive or semi-extensive production systems. The modified Gordon and Whitlock technique was used to determine positive samples and number of oocysts per gram of feces. Statistical analyses were performed using the chi-square test with Yates correction and a 95% confidence interval. Thirty-nine cattle (33.33%) were positive, and ten different species were identified in infected animals: E. bovis (24.79%); E. canadensis (8.55%); E. zuernii (6.83%); E. ellipsoidalis (5.99%); E. cylindrica (3.42%); E. auburnensis (3.42%); E. brasiliensis (2.56%); E. bukidnonensis (1.71%); E. alabamensis (0.85%), and E. subspherica (0.85%). Higher parasitism was observed in animals up to one year of age (p = 0.005), but no animal presented clinical signs of the disease. As the presence of clinical eimeriosis was not evidenced and all animals were Zebu crossbred cattle from extensive or semi-extensive production systems, further studies should be conducted to investigate the effects of these factors on disease development.
Revista Brasileira de Parasitologia Veterinária, 2015
Ehrlichiosis is a zoonotic disease that is caused by bacteria of the genus Ehrlichia. The aims of... more Ehrlichiosis is a zoonotic disease that is caused by bacteria of the genus Ehrlichia. The aims of this study were to detect the presence of Ehrlichia spp. in the blood of dogs in Ituberá, Bahia, and to compare the sensitivities and specificities of blood smear, serological, and molecular examinations. Furthermore, this study identified factors associated with exposure to the agent in dogs in this locality. Blood samples were collected from 379 dogs and submitted for indirect immunofluorescent assay and polymerase chain reaction testing for the detection of Ehrlichia spp. antibodies and DNA, respectively. Additionally, a peripheral blood smear was obtained from the ear tip for parasite identification. Of the 379 animals, 12.4%, 32.7%, and 25.6% were identified as positive on the blood smear, serological, and molecular tests, respectively. The dogs positive in one of the three techniques were considered exposed (46.9%). Younger dogs and rural habitat were protective factors and presence of ticks and contact with other dogs were the risk factors associated with exposure to the agent. It was concluded that dogs of Ituberá have high positivity for Ehrlichia spp. and that the diagnostic methods used for detection are complementary.
Acta Tropica, 2015
Leishmaniosis is a zoonosis caused by protozoa of the genus Leishmania. American cutaneous leishm... more Leishmaniosis is a zoonosis caused by protozoa of the genus Leishmania. American cutaneous leishmaniosis (ACL) is mainly caused by the species L. amazonensis and L. braziliensis, and American visceral leishmaniosis (AVL) is caused by L. infantum chagasi. In addition to their proven roles as reservoirs of AVL, dogs are also suspected by researchers to be reservoirs of ACL due to reports of this infection in domestic environments and of infected dogs in endemic areas. The aim of this study was to detect Leishmania sp. infection in dogs from Vila Operária, Buerarema, Bahia, using parasitological tests, indirect immunofluorescent assay (IFA) and polymerase chain reaction (PCR). Furthermore, this study also aimed to identify risk factors associated with illness in dogs in this locality by conducting an epidemiological survey. For this purpose, 292 dogs were clinically evaluated for the presence of skin lesions, and the dogs that showed these changes were submitted to scarification injury to enable preparation of slides for microscopic study of amastigotes. Subsequently, the dogs underwent blood sampling for serological (IFA) and molecular (PCR) tests. Additionally, the owners of the dogs answered an epidemiological questionnaire to facilitate the identification of risk factors for exposure of dogs to pathogens of ACL. Of the 292 dogs studied, 13 (4.5%) had lesions suggestive of ACL, but with a negative parasitological examination and 147 (50.3%) were seropositive according to the IFA. Of the 273 dogs studied using PCR test, 10 (3.66%) were positive for L. braziliensis, and all samples were negative for L. infantum chagasi. Wastelands in the peridomicile and the presence of light in the household were risk factors associated with ACL. The results show that Vila Operária has asymptomatic dogs with ACL and that the detection sensitivity of the IFA was higher than that of PCR for the infected dogs.
The aim of this study was to characterize the role of quails (Coturnix coturnix japonica) as inte... more The aim of this study was to characterize the role of quails (Coturnix coturnix japonica) as intermediate hosts for Neospora caninum. Fifty-eight 20-day old quails were individually identified with numbered rings and kept in cages adequate for the species. The birds were fed commercial feed and distilled water ad libitum. They were weighed daily during the first month and every seven days from then on. The birds were randomly distributed into 3 groups; 40 quails (group A) and 8 quails (group B) received 3.5 × 10 6 and 5 × 10 6 N. caninum tachyzoites, respectively, and 10 quails (group C) received placebo. Four quails from group A and one from group C had their blood collected and were euthanized on the 1st, 3rd, 5th, 7th, 14th, 21th, 30th, 60th, 120th day after infection (DAI), and 4 quails from group B were euthanized on days 60th and 120th DAI. Following euthanasia, the birds were submitted to necropsy, the organs were weighed and fragments were collected for histopathology, immunohistochemistry (IHC) and PCR. On the 60th DAI, two dogs were fed tissue obtained from group A quails, and one dog was fed quail tissues from group B. The dogs' blood was collected weekly for serology, and their feces were collected daily for 33 days for microscopy and molecular identification of oocysts. Student's t-test was used to compare the weights of the collected organs and the quail and cloacal temperature. No quail presented any clinical signs or died. Splenomegaly and hepatomegaly were the primary necropsy findings during the first week of infection (p < 0.05). Positive serology, immunoreactivity through IHC and molecular identification of the parasite, were observed, especially during the first two weeks of infection. No dogs eliminated the oocysts or seroconverted. The infected quails gained more weight than the control quails. The results demonstrated that quails are resistant to infection with N. caninum tachyzoites under the conditions of this study.
Pesquisa Veterinária Brasileira, 2009
RESUMO.-[Soropositividade para Neospora caninum e raças bovinas no vale do Paraíba sulfluminense,... more RESUMO.-[Soropositividade para Neospora caninum e raças bovinas no vale do Paraíba sulfluminense, Estado do Rio de Janeiro.] A realização deste estudo teve como objetivo verificar a ocorrência diferenças na distribuição da infecção por Neospora caninum em bovinos leiteiros da raça Holandesa Preta e Branca e mestiços na Mesorregião Fluminense a maior bacia leiteira do Rio de Janeiro. De um total de 2.491 vacas foram coleta-das amostras de sangue de 563 de forma não randomizada, pertencentes a 57 propriedades localizadas nos municípios de Resende e Rio Claro. O tamanho da amostra por propriedade foi proporcional ao número de animais alvo da mesma. O abortamento ou distúrbios reprodutivos não foram utilizados como critérios de seleção. Foi verificada associação altamente significativa entre a soropositividade e o padrão racial (p=0,006). Ao conduzir as análises entre apenas dois padrões verificou-se os seguintes resultados: HPB versus zebuínos (p=0,0028) com 2,65 mais chances de sorpositividade em bovinos HPB; HPB versus (zebuínos + Gir x Holandês) p=0,01, com 2,23 mais chances de sopositividade também na raça HPB. Não houve diferença significativa entre HPB versus Gir x Holandês (p=0,08) e entre os zebuínos e Gir x Holandês (p=0,11). Este estudo suporta a hipótese que existe uma estreita relação entre a presença da infecção e a 1 INDEX TERMS: Neospora caninum, association, cattle breed. Pesq. Vet. Bras. 29(1):29-32, janeiro 2009 Alexandre D. Munhoz et al. 30 raça européia Holandesa Preta e Branca, na região e condições em que foi desenvolvido o presente estudo. TERMOS DE INDEXAÇÃO: Neospora caninum, associação, padrões bovinos.
Parasitología latinoamericana, 2002
To determine the hepatic profiles due to Toxoplasma gondii infection in rabbits, 15 animals were ... more To determine the hepatic profiles due to Toxoplasma gondii infection in rabbits, 15 animals were divided in three groups of five animals each. The first and second groups were inoculated subcutaneously with 10 6 and 10 3 tachyzoites of T. gondii respectively, while the third was left as control. Blood samples were taken on days 0, 4, 8, e 11 after inoculation (DAI). Livers of each rabbit was analyzed in other to observed lesions. Hepathomegaly was observed in all of the infected groups, independently of the dose used, where was observed a significant increase of AST e ALT at 8 th DAI. Tachyzoites were observed in all infected animals, and were reisolated in 90% of livers bioassed in mice. Histopathology was consisted by diffuse infiltration inflammatory, consisted mainly by lymphocytes at sinusoids e portal space. The increase of AST and ALT during acute toxoplasmosis in rabbits was justified by presence of tachyzoites in tissue smears associated with gross and histopathologic lesions.
Veterinary parasitology, Jan 30, 2012
From August 2006 to 2008, 411 dogs in northeastern Brazil were evaluated for seropositivity to Ne... more From August 2006 to 2008, 411 dogs in northeastern Brazil were evaluated for seropositivity to Neospora caninum. The dogs were clinically examined, and their owners were interviewed about the conditions in which the animals were maintained in order to assess the factors associated with infection by this parasite. A serum sample was taken from each dog for serological examination in an indirect fluorescent antibody test for N. caninum. The Yates' Chi-square test or Fisher's exact test was used to select the variables for the multivariate logistic regression model. Seropositivity was detected in 9.26% of dogs. The seropositivity rates of dogs from different environments were 2.6% (4/156) in urban areas, 13.1% (28/214) in peri-urban areas, and 14.6% (6/41) in rural areas. Factors associated with seropositivity for N. caninum were the following: contact with other dogs, access to food outside the home and residing in the peri-urban or rural environments (p<0.05). Results of...
Breast Cancer Research and Treatment, 2011
We examined the impact of surgical treatments (breast-conserving surgery [BCS], mastectomy alone,... more We examined the impact of surgical treatments (breast-conserving surgery [BCS], mastectomy alone, mastectomy with reconstruction) and surgical side-effects severity on early stage (0-IIA) breast cancer patients' body image over time. We interviewed patients at 4-6 weeks (T1), six (T2), 12 (T3), and 24 months (T4) following definitive surgical treatment. We examined longitudinal relationships among body image problems, surgery type, and surgical side-effects severity using the Generalized Estimating Equation approach, controlling for demographic, clinical, and psychosocial factors. We compared regression coefficients of surgery type from two models, one with and one without surgical side-effects severity. Of 549 patients enrolled (mean age 58; 75% White; 65% BCS, 12% mastectomy, 23% mastectomy with reconstruction), 514 (94%) completed all four interviews. In the model without surgical side-effects severity, patients who underwent mastectomy with reconstruction reported poorer body image than patients who underwent BCS at T1-T3 (each P < 0.02), but not at T4. At T2, patients who underwent mastectomy with reconstruction also reported poorer body image than patients who underwent mastectomy alone (P = 0.0106). Adjusting for surgical side-effects severity, body image scores did not differ significantly between patients with BCS and mastectomy with reconstruction at any interview; however, patients who underwent mastectomy alone had better body image at T2 than patients who underwent mastectomy with reconstruction (P = 0.011). The impact of surgery type on body image within the first year of definitive surgical treatment was explained by surgical side-effects severity. After 2 years, body image problems did not differ significantly by surgery type.
Revista brasileira de parasitologia veterinária = Brazilian journal of veterinary parasitology : Órgão Oficial do Colégio Brasileiro de Parasitologia Veterinária
The aim of this study was to evaluate parasitism kinetics and tissue lesions in the first week of... more The aim of this study was to evaluate parasitism kinetics and tissue lesions in the first week of infection by Neospora caninum in dogs fed Gallus gallus chorioallantoic membranes (CMs) previously infected in ovo. Five two-month-old pups were used. Each dog was given five CMs that were previously infected with N. caninum via the oral route. Four animals were euthanized in the first week of infection. All four dogs had their stools examined one week prior to and up to the day they were euthanized. The stools of the uneuthanized dog were collected for 30 days. After euthanasia, organ sections were utilized for histopathology, immunohistochemistry, indirect immunofluorescent tissue reactions, PCR and real-time PCR to detect parasites. Necropsy revealed that the small and large intestines, spleen, and lungs were affected. No oocysts or N. caninum DNA were identified in the stool samples. Real-time PCR was the most sensitive technique used to detect the protozoa in tissues, which were id...
Revista Brasileira de Parasitologia Veterinária, 2011
The aim of this study was to determine the presence of species of the genus Eimeria species in na... more The aim of this study was to determine the presence of species of the genus Eimeria species in naturally infected bovines in Southern Bahia, Northeast Brazil. The study population comprised 117 Zebu crossbred cattle that belonged to 10 dairy herds with extensive or semi-extensive production systems. The modified Gordon and Whitlock technique was used to determine positive samples and number of oocysts per gram of feces. Statistical analyses were performed using the chi-square test with Yates correction and a 95% confidence interval. Thirty-nine cattle (33.33%) were positive, and ten different species were identified in infected animals: E. bovis (24.79%); E. canadensis (8.55%); E. zuernii (6.83%); E. ellipsoidalis (5.99%); E. cylindrica (3.42%); E. auburnensis (3.42%); E. brasiliensis (2.56%); E. bukidnonensis (1.71%); E. alabamensis (0.85%), and E. subspherica (0.85%). Higher parasitism was observed in animals up to one year of age (p = 0.005), but no animal presented clinical signs of the disease. As the presence of clinical eimeriosis was not evidenced and all animals were Zebu crossbred cattle from extensive or semi-extensive production systems, further studies should be conducted to investigate the effects of these factors on disease development.
Revista Brasileira de Parasitologia Veterinária, 2015
Ehrlichiosis is a zoonotic disease that is caused by bacteria of the genus Ehrlichia. The aims of... more Ehrlichiosis is a zoonotic disease that is caused by bacteria of the genus Ehrlichia. The aims of this study were to detect the presence of Ehrlichia spp. in the blood of dogs in Ituberá, Bahia, and to compare the sensitivities and specificities of blood smear, serological, and molecular examinations. Furthermore, this study identified factors associated with exposure to the agent in dogs in this locality. Blood samples were collected from 379 dogs and submitted for indirect immunofluorescent assay and polymerase chain reaction testing for the detection of Ehrlichia spp. antibodies and DNA, respectively. Additionally, a peripheral blood smear was obtained from the ear tip for parasite identification. Of the 379 animals, 12.4%, 32.7%, and 25.6% were identified as positive on the blood smear, serological, and molecular tests, respectively. The dogs positive in one of the three techniques were considered exposed (46.9%). Younger dogs and rural habitat were protective factors and presence of ticks and contact with other dogs were the risk factors associated with exposure to the agent. It was concluded that dogs of Ituberá have high positivity for Ehrlichia spp. and that the diagnostic methods used for detection are complementary.
Acta Tropica, 2015
Leishmaniosis is a zoonosis caused by protozoa of the genus Leishmania. American cutaneous leishm... more Leishmaniosis is a zoonosis caused by protozoa of the genus Leishmania. American cutaneous leishmaniosis (ACL) is mainly caused by the species L. amazonensis and L. braziliensis, and American visceral leishmaniosis (AVL) is caused by L. infantum chagasi. In addition to their proven roles as reservoirs of AVL, dogs are also suspected by researchers to be reservoirs of ACL due to reports of this infection in domestic environments and of infected dogs in endemic areas. The aim of this study was to detect Leishmania sp. infection in dogs from Vila Operária, Buerarema, Bahia, using parasitological tests, indirect immunofluorescent assay (IFA) and polymerase chain reaction (PCR). Furthermore, this study also aimed to identify risk factors associated with illness in dogs in this locality by conducting an epidemiological survey. For this purpose, 292 dogs were clinically evaluated for the presence of skin lesions, and the dogs that showed these changes were submitted to scarification injury to enable preparation of slides for microscopic study of amastigotes. Subsequently, the dogs underwent blood sampling for serological (IFA) and molecular (PCR) tests. Additionally, the owners of the dogs answered an epidemiological questionnaire to facilitate the identification of risk factors for exposure of dogs to pathogens of ACL. Of the 292 dogs studied, 13 (4.5%) had lesions suggestive of ACL, but with a negative parasitological examination and 147 (50.3%) were seropositive according to the IFA. Of the 273 dogs studied using PCR test, 10 (3.66%) were positive for L. braziliensis, and all samples were negative for L. infantum chagasi. Wastelands in the peridomicile and the presence of light in the household were risk factors associated with ACL. The results show that Vila Operária has asymptomatic dogs with ACL and that the detection sensitivity of the IFA was higher than that of PCR for the infected dogs.
The aim of this study was to characterize the role of quails (Coturnix coturnix japonica) as inte... more The aim of this study was to characterize the role of quails (Coturnix coturnix japonica) as intermediate hosts for Neospora caninum. Fifty-eight 20-day old quails were individually identified with numbered rings and kept in cages adequate for the species. The birds were fed commercial feed and distilled water ad libitum. They were weighed daily during the first month and every seven days from then on. The birds were randomly distributed into 3 groups; 40 quails (group A) and 8 quails (group B) received 3.5 × 10 6 and 5 × 10 6 N. caninum tachyzoites, respectively, and 10 quails (group C) received placebo. Four quails from group A and one from group C had their blood collected and were euthanized on the 1st, 3rd, 5th, 7th, 14th, 21th, 30th, 60th, 120th day after infection (DAI), and 4 quails from group B were euthanized on days 60th and 120th DAI. Following euthanasia, the birds were submitted to necropsy, the organs were weighed and fragments were collected for histopathology, immunohistochemistry (IHC) and PCR. On the 60th DAI, two dogs were fed tissue obtained from group A quails, and one dog was fed quail tissues from group B. The dogs' blood was collected weekly for serology, and their feces were collected daily for 33 days for microscopy and molecular identification of oocysts. Student's t-test was used to compare the weights of the collected organs and the quail and cloacal temperature. No quail presented any clinical signs or died. Splenomegaly and hepatomegaly were the primary necropsy findings during the first week of infection (p < 0.05). Positive serology, immunoreactivity through IHC and molecular identification of the parasite, were observed, especially during the first two weeks of infection. No dogs eliminated the oocysts or seroconverted. The infected quails gained more weight than the control quails. The results demonstrated that quails are resistant to infection with N. caninum tachyzoites under the conditions of this study.
Pesquisa Veterinária Brasileira, 2009
RESUMO.-[Soropositividade para Neospora caninum e raças bovinas no vale do Paraíba sulfluminense,... more RESUMO.-[Soropositividade para Neospora caninum e raças bovinas no vale do Paraíba sulfluminense, Estado do Rio de Janeiro.] A realização deste estudo teve como objetivo verificar a ocorrência diferenças na distribuição da infecção por Neospora caninum em bovinos leiteiros da raça Holandesa Preta e Branca e mestiços na Mesorregião Fluminense a maior bacia leiteira do Rio de Janeiro. De um total de 2.491 vacas foram coleta-das amostras de sangue de 563 de forma não randomizada, pertencentes a 57 propriedades localizadas nos municípios de Resende e Rio Claro. O tamanho da amostra por propriedade foi proporcional ao número de animais alvo da mesma. O abortamento ou distúrbios reprodutivos não foram utilizados como critérios de seleção. Foi verificada associação altamente significativa entre a soropositividade e o padrão racial (p=0,006). Ao conduzir as análises entre apenas dois padrões verificou-se os seguintes resultados: HPB versus zebuínos (p=0,0028) com 2,65 mais chances de sorpositividade em bovinos HPB; HPB versus (zebuínos + Gir x Holandês) p=0,01, com 2,23 mais chances de sopositividade também na raça HPB. Não houve diferença significativa entre HPB versus Gir x Holandês (p=0,08) e entre os zebuínos e Gir x Holandês (p=0,11). Este estudo suporta a hipótese que existe uma estreita relação entre a presença da infecção e a 1 INDEX TERMS: Neospora caninum, association, cattle breed. Pesq. Vet. Bras. 29(1):29-32, janeiro 2009 Alexandre D. Munhoz et al. 30 raça européia Holandesa Preta e Branca, na região e condições em que foi desenvolvido o presente estudo. TERMOS DE INDEXAÇÃO: Neospora caninum, associação, padrões bovinos.
Parasitología latinoamericana, 2002
To determine the hepatic profiles due to Toxoplasma gondii infection in rabbits, 15 animals were ... more To determine the hepatic profiles due to Toxoplasma gondii infection in rabbits, 15 animals were divided in three groups of five animals each. The first and second groups were inoculated subcutaneously with 10 6 and 10 3 tachyzoites of T. gondii respectively, while the third was left as control. Blood samples were taken on days 0, 4, 8, e 11 after inoculation (DAI). Livers of each rabbit was analyzed in other to observed lesions. Hepathomegaly was observed in all of the infected groups, independently of the dose used, where was observed a significant increase of AST e ALT at 8 th DAI. Tachyzoites were observed in all infected animals, and were reisolated in 90% of livers bioassed in mice. Histopathology was consisted by diffuse infiltration inflammatory, consisted mainly by lymphocytes at sinusoids e portal space. The increase of AST and ALT during acute toxoplasmosis in rabbits was justified by presence of tachyzoites in tissue smears associated with gross and histopathologic lesions.
Veterinary parasitology, Jan 30, 2012
From August 2006 to 2008, 411 dogs in northeastern Brazil were evaluated for seropositivity to Ne... more From August 2006 to 2008, 411 dogs in northeastern Brazil were evaluated for seropositivity to Neospora caninum. The dogs were clinically examined, and their owners were interviewed about the conditions in which the animals were maintained in order to assess the factors associated with infection by this parasite. A serum sample was taken from each dog for serological examination in an indirect fluorescent antibody test for N. caninum. The Yates' Chi-square test or Fisher's exact test was used to select the variables for the multivariate logistic regression model. Seropositivity was detected in 9.26% of dogs. The seropositivity rates of dogs from different environments were 2.6% (4/156) in urban areas, 13.1% (28/214) in peri-urban areas, and 14.6% (6/41) in rural areas. Factors associated with seropositivity for N. caninum were the following: contact with other dogs, access to food outside the home and residing in the peri-urban or rural environments (p<0.05). Results of...
Breast Cancer Research and Treatment, 2011
We examined the impact of surgical treatments (breast-conserving surgery [BCS], mastectomy alone,... more We examined the impact of surgical treatments (breast-conserving surgery [BCS], mastectomy alone, mastectomy with reconstruction) and surgical side-effects severity on early stage (0-IIA) breast cancer patients' body image over time. We interviewed patients at 4-6 weeks (T1), six (T2), 12 (T3), and 24 months (T4) following definitive surgical treatment. We examined longitudinal relationships among body image problems, surgery type, and surgical side-effects severity using the Generalized Estimating Equation approach, controlling for demographic, clinical, and psychosocial factors. We compared regression coefficients of surgery type from two models, one with and one without surgical side-effects severity. Of 549 patients enrolled (mean age 58; 75% White; 65% BCS, 12% mastectomy, 23% mastectomy with reconstruction), 514 (94%) completed all four interviews. In the model without surgical side-effects severity, patients who underwent mastectomy with reconstruction reported poorer body image than patients who underwent BCS at T1-T3 (each P < 0.02), but not at T4. At T2, patients who underwent mastectomy with reconstruction also reported poorer body image than patients who underwent mastectomy alone (P = 0.0106). Adjusting for surgical side-effects severity, body image scores did not differ significantly between patients with BCS and mastectomy with reconstruction at any interview; however, patients who underwent mastectomy alone had better body image at T2 than patients who underwent mastectomy with reconstruction (P = 0.011). The impact of surgery type on body image within the first year of definitive surgical treatment was explained by surgical side-effects severity. After 2 years, body image problems did not differ significantly by surgery type.
Revista brasileira de parasitologia veterinária = Brazilian journal of veterinary parasitology : Órgão Oficial do Colégio Brasileiro de Parasitologia Veterinária
The aim of this study was to evaluate parasitism kinetics and tissue lesions in the first week of... more The aim of this study was to evaluate parasitism kinetics and tissue lesions in the first week of infection by Neospora caninum in dogs fed Gallus gallus chorioallantoic membranes (CMs) previously infected in ovo. Five two-month-old pups were used. Each dog was given five CMs that were previously infected with N. caninum via the oral route. Four animals were euthanized in the first week of infection. All four dogs had their stools examined one week prior to and up to the day they were euthanized. The stools of the uneuthanized dog were collected for 30 days. After euthanasia, organ sections were utilized for histopathology, immunohistochemistry, indirect immunofluorescent tissue reactions, PCR and real-time PCR to detect parasites. Necropsy revealed that the small and large intestines, spleen, and lungs were affected. No oocysts or N. caninum DNA were identified in the stool samples. Real-time PCR was the most sensitive technique used to detect the protozoa in tissues, which were id...