RESUMO

Neospora caninum is an obligate intracellular parasite that can infect many domestic and wild animals, including birds. These animals are important sources for monitoring of environmental contamination, as they could become infected through sporulated oocysts; however, the real role of birds in the biological cycle of N. caninum remains uncertain. This study aimed to determine the prevalence of anti-N. caninum antibodies, evaluate associated factors, detect the parasite by molecular testing of free-range chickens from Brazil, and evaluate different techniques for its serological diagnosis. Blood samples of 366 chickens from 25 farms were collected for serological assays. The indirect fluorescent antibody test (IFAT) and the indirect enzyme-linked immunosorbent assay (ELISA) were used to detect anti-N. caninum antibodies. Chickens that tested seropositive by IFAT had their brain tissues and a pool of organs (heart, lung, and liver) submitted to PCR for molecular detection of the parasite. Out of 366 chickens, 65 (17.8%) and 163 (44.6%) were seropositive by IFAT and ELISA, respectively. Brain tissues (n=60) and the pools of organs (n=65) were negative in the PCR. Our results showed a high prevalence of antibodies in free-range chickens and that IFAT is the more sensitive technique for the detection of anti-N. caninum antibodies.

Assuntos

RESUMO

Neospora caninum is an obligate intracellular parasite that can infect many domestic and wild animals, including birds. These animals are important sources for monitoring of environmental contamination, as they could become infected through sporulated oocysts; however, the real role of birds in the biological cycle of N. caninum remains uncertain. This study aimed to determine the prevalence of anti-N. caninum antibodies, evaluate associated factors, detect the parasite by molecular testing of free-range chickens from Brazil, and evaluate different techniques for its serological diagnosis. Blood samples of 366 chickens from 25 farms were collected for serological assays. The indirect fluorescent antibody test (IFAT) and the indirect enzyme-linked immunosorbent assay (ELISA) were used to detect anti-N. caninum antibodies. Chickens that tested seropositive by IFAT had their brain tissues and a pool of organs (heart, lung, and liver) submitted to PCR for molecular detection of the parasite. Out of 366 chickens, 65 (17.8%) and 163 (44.6%) were seropositive by IFAT and ELISA, respectively. Brain tissues (n=60) and the pools of organs (n=65) were negative in the PCR. Our results showed a high prevalence of antibodies in free-range chickens and that IFAT is the more sensitive technique for the detection of anti-N. caninum antibodies.(AU)

Neospora caninum é um parasita intracelular obrigatório que pode infectar vários animais domésticos e silvestres, incluindo as aves. Esses animais são importantes fontes para o monitoramento da contaminação ambiental, uma vez que eles podem se infectar por meio da ingestão de oocistos. Porém, o real papel das aves no ciclo biológico do N. caninum ainda é incerto. Este estudo teve como objetivos determinar a prevalência de anticorpos anti-N. caninum, avaliar fatores associados, detectar o parasita por meio de teste molecular em galinhas de vida livre do Brasil e avaliar diferentes técnicas para o diagnóstico sorológico. Amostras de sangue de 366 galinhas de 25 fazendas foram coletadas para testes sorológicos. A reação de imunofluorescência indireta (RIFI) e o ensaio de imunoabsorção enzimática (ELISA) foram utilizados para detectar anticorpos anti-N. caninum. As amostras de tecidos (cérebro, coração, pulmão e fígado) de galinhas soropositivas na RIFI foram coletadas e submetidas à PCR para detecção molecular do parasita. Das 366 galinhas, 65 (17,8%) e 163 (44,6%) foram soropositivas pela RIFI e ELISA, respectivamente. Nenhuma amostra foi positiva na PCR. Esses resultados demonstram uma elevada prevalência de anticorpos em galinhas de vida livre e que a RIFI é uma técnica mais sensível para a detecção de anticorpos anti-N. caninum.(AU)

Assuntos

RESUMO

Abstract Neospora caninum is an obligate intracellular parasite that can infect many domestic and wild animals, including birds. These animals are important sources for monitoring of environmental contamination, as they could become infected through sporulated oocysts; however, the real role of birds in the biological cycle of N. caninum remains uncertain. This study aimed to determine the prevalence of anti-N. caninum antibodies, evaluate associated factors, detect the parasite by molecular testing of free-range chickens from Brazil, and evaluate different techniques for its serological diagnosis. Blood samples of 366 chickens from 25 farms were collected for serological assays. The indirect fluorescent antibody test (IFAT) and the indirect enzyme-linked immunosorbent assay (ELISA) were used to detect anti-N. caninum antibodies. Chickens that tested seropositive by IFAT had their brain tissues and a pool of organs (heart, lung, and liver) submitted to PCR for molecular detection of the parasite. Out of 366 chickens, 65 (17.8%) and 163 (44.6%) were seropositive by IFAT and ELISA, respectively. Brain tissues (n=60) and the pools of organs (n=65) were negative in the PCR. Our results showed a high prevalence of antibodies in free-range chickens and that IFAT is the more sensitive technique for the detection of anti-N. caninum antibodies.

Resumo Neospora caninum é um parasita intracelular obrigatório que pode infectar vários animais domésticos e silvestres, incluindo as aves. Esses animais são importantes fontes para o monitoramento da contaminação ambiental, uma vez que eles podem se infectar por meio da ingestão de oocistos. Porém, o real papel das aves no ciclo biológico do N. caninum ainda é incerto. Este estudo teve como objetivos determinar a prevalência de anticorpos anti-N. caninum, avaliar fatores associados, detectar o parasita por meio de teste molecular em galinhas de vida livre do Brasil e avaliar diferentes técnicas para o diagnóstico sorológico. Amostras de sangue de 366 galinhas de 25 fazendas foram coletadas para testes sorológicos. A reação de imunofluorescência indireta (RIFI) e o ensaio de imunoabsorção enzimática (ELISA) foram utilizados para detectar anticorpos anti-N. caninum. As amostras de tecidos (cérebro, coração, pulmão e fígado) de galinhas soropositivas na RIFI foram coletadas e submetidas à PCR para detecção molecular do parasita. Das 366 galinhas, 65 (17,8%) e 163 (44,6%) foram soropositivas pela RIFI e ELISA, respectivamente. Nenhuma amostra foi positiva na PCR. Esses resultados demonstram uma elevada prevalência de anticorpos em galinhas de vida livre e que a RIFI é uma técnica mais sensível para a detecção de anticorpos anti-N. caninum.

Assuntos

RESUMO

Toxoplasma gondii is an intracellular parasite that can infect all warm-blooded animals including humans. Recent studies showed that T. gondii strains from South America are genetically diverse. The present work aimed to determine T. gondii prevalence in free-ranging chicken in northwest Parana state in Brazil by two serological tests, to isolate the parasites from seropositive chickens and to genotype the isolates. Antibodies to T. gondii in 386 serum samples from 24 farms were investigated by immunofluorescence antibody assay (IFA) and modified agglutination test (MAT). Samples having titers ≥ 16 were considered positive for both tests. Among the 386 serum samples, 102 (26.4%) were positive for IFA, 64 (16.6%) were positive for MAT, 47 (12.2%) were positive in both tests, and 119 (30.8%) were positive in at least one of the two tests. Brain and pool of heart, lung, and liver from the 119 seropositive chickens were used for mouse bioassay to isolate the parasites. Thirty eight (31.9%) of these seropositive chickens were considered positives in mouse bioassay and 18 isolates were obtained. The isolates were characterized by 10 PCR-RFLP genetic markers including SAG1, SAG2 (5'-3'SAG2, alt.SAG2), SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico. Results of genotyping were compared with the genotypes in ToxoDB database. It revealed ten genotypes, including ToxoDB PCR-RFLP genotypes #6 (n = 2), #19 (n = 1), #21 (n = 2), #111 (n = 2), #152 (n = 1), and #175 (n = 1) and four new types not described before. Our results confirmed a high genetic diversity of this parasite in southern Brazil and also showed that the use of two serological tests in combination can improve the chance of T. gondii isolation. More studies should be taken to determine the zoonotic potential of chickens in the transmission of T. gondii.

Assuntos

RESUMO

Rearing free-range chicken is based on grazing feeding patterns, and these animals could be potential environmental contaminants of Cryptosporidium oocysts for humans and other animals. Therefore, the present study aimed to evaluate the molecular prevalence of Cryptosporidium spp. in free-range chickens from Brazil. A total of 351 fecal samples from chickens were examined from 20 farms. For detection of Cryptosporidium spp., 18S rRNA gene fragments were amplified using a nested PCR reaction. Positive samples were sent for sequencing. The overall prevalence of Cryptosporidium was 25.6% (95% CI = 21.2% - 30.6%). Sequencing of the amplified fragments allowed for the identification of three species: C. meleagridis in 57 (62.6%), C. baileyi in 15 (16.4%), C. parvum in 3 (3.2%) samples, and a new Cryptosporidium genotype (C. genotype BrPR1) in 3 (3.2%) samples. Cryptosporidium genotype BrPR1 has not yet been classified as a species, and its host spectrum is not known. Cryptosporidium, including zoonotic species, exists at a high prevalence in free-range chickens within the region studied.(AU)

A criação de galinhas no estilo colonial/caipira é baseada em padrões de alimentação de pastagem, o que as torna potenciais contaminantes ambientais de oocistos de Cryptosporidium para humanos e outros animais. Portanto, o presente estudo teve como objetivo avaliar a prevalência molecular de Cryptosporidium spp. em galinhas criadas em sistema colonial/caipira. Um total de 351 amostras de fezes de frangos foram examinadas em 20 fazendas. Para a detecção de Cryptosporidium spp., os fragmentos do gene rRNA 18S foram amplificados utilizando-se a reação de nested-PCR. A prevalência global de Cryposporidium foi de 25,6% (IC 95% = 21,2% - 30,6%). O sequenciamento dos fragmentos amplificados permitiu a identificação de três espécies que infectam aves: C. meleagridis em 57 (62,6%), C. baileyi em 15 (16,4%), C. parvum em 3 (3,2%) amostras, bem como, um novo genótipo de Cryptosporidium (C. genótipo BrPR1) foi identificado em 3 (3,2%) amostras. Cryptosporidium genotipo BrPR1 não foi ainda classificado como uma espécie, e seu espectro de hospedeiros é desconhecido. O presente trabalho permitiu concluir que Cryptosporidium, incluindo espécies zoonóticas, existe com alta prevalência em galinhas criadas em sistema colonial/caipira na região estudada.(AU)

Assuntos

RESUMO

Abstract Rearing free-range chicken is based on grazing feeding patterns, and these animals could be potential environmental contaminants of Cryptosporidium oocysts for humans and other animals. Therefore, the present study aimed to evaluate the molecular prevalence of Cryptosporidium spp. in free-range chickens from Brazil. A total of 351 fecal samples from chickens were examined from 20 farms. For detection of Cryptosporidium spp., 18S rRNA gene fragments were amplified using a nested PCR reaction. Positive samples were sent for sequencing. The overall prevalence of Cryptosporidium was 25.6% (95% CI = 21.2% - 30.6%). Sequencing of the amplified fragments allowed for the identification of three species: C. meleagridis in 57 (62.6%), C. baileyi in 15 (16.4%), C. parvum in 3 (3.2%) samples, and a new Cryptosporidium genotype (C. genotype BrPR1) in 3 (3.2%) samples. Cryptosporidium genotype BrPR1 has not yet been classified as a species, and its host spectrum is not known. Cryptosporidium, including zoonotic species, exists at a high prevalence in free-range chickens within the region studied.

Resumo A criação de galinhas no estilo colonial/caipira é baseada em padrões de alimentação de pastagem, o que as torna potenciais contaminantes ambientais de oocistos de Cryptosporidium para humanos e outros animais. Portanto, o presente estudo teve como objetivo avaliar a prevalência molecular de Cryptosporidium spp. em galinhas criadas em sistema colonial/caipira. Um total de 351 amostras de fezes de frangos foram examinadas em 20 fazendas. Para a detecção de Cryptosporidium spp., os fragmentos do gene rRNA 18S foram amplificados utilizando-se a reação de nested-PCR. A prevalência global de Cryposporidium foi de 25,6% (IC 95% = 21,2% - 30,6%). O sequenciamento dos fragmentos amplificados permitiu a identificação de três espécies que infectam aves: C. meleagridis em 57 (62,6%), C. baileyi em 15 (16,4%), C. parvum em 3 (3,2%) amostras, bem como, um novo genótipo de Cryptosporidium (C. genótipo BrPR1) foi identificado em 3 (3,2%) amostras. Cryptosporidium genotipo BrPR1 não foi ainda classificado como uma espécie, e seu espectro de hospedeiros é desconhecido. O presente trabalho permitiu concluir que Cryptosporidium, incluindo espécies zoonóticas, existe com alta prevalência em galinhas criadas em sistema colonial/caipira na região estudada.

Assuntos

RESUMO

Rearing free-range chicken is based on grazing feeding patterns, and these animals could be potential environmental contaminants of Cryptosporidium oocysts for humans and other animals. Therefore, the present study aimed to evaluate the molecular prevalence of Cryptosporidium spp. in free-range chickens from Brazil. A total of 351 fecal samples from chickens were examined from 20 farms. For detection of Cryptosporidium spp., 18S rRNA gene fragments were amplified using a nested PCR reaction. Positive samples were sent for sequencing. The overall prevalence of Cryptosporidium was 25.6% (95% CI = 21.2% - 30.6%). Sequencing of the amplified fragments allowed for the identification of three species: C. meleagridis in 57 (62.6%), C. baileyi in 15 (16.4%), C. parvum in 3 (3.2%) samples, and a new Cryptosporidium genotype (C. genotype BrPR1) in 3 (3.2%) samples. Cryptosporidium genotype BrPR1 has not yet been classified as a species, and its host spectrum is not known. Cryptosporidium, including zoonotic species, exists at a high prevalence in free-range chickens within the region studied.

Assuntos

RESUMO

The effects of managerial systems on the occurrence of gastrointestinal helminths in Free-Range Chickens (FRCs) from northern Paraná, Brazil were investigated. The most predominant (23.3%; 61/262) cestode observed was Raillietina cesticillus; Heterakis gallinarum (71.4%; 187/262) and Ascaridia galli (45%; 118/262) were the predominant nematodes; Postharmostomum commutatum was the only trematode observed in 2.7% (7/262) of FRCs. The most elevated parasitic burdens were associated with Heterakis gallinarum, Ascaridia galli, and Raillietina cesticillus. Significant (p ? 0.05) associations were observed when the effects of the types of bedding, soil type, and fence restriction of FRCs were considered relative to the possibility of helminthiasis. The type of bedding, the length of the sanitary break, and the presence of shading significantly (p ? 0.05) influenced the possibility of FRCs being infected by H. gallinarum. Most parameters evaluated were significantly associated with infection of FRCs by A. galli. These findings suggest that FRCs from northern Paraná are infected by a wide-range of gastrointestinal helminths, but more frequently by R. cesticillus, H. gallinarum, and A. galli. Moreover, the type of floor bedding, the soil type, and the usage of fences in the management of FRCs is directly related to gastrointestinal helminthiasis.(AU)

Foram investigados os efeitos dos sistemas de manejo sobre a ocorrência de helmintos gastrintestinais em galinhas caipiras (GCs) do norte do Paraná, Brasil. O cestódeo predominante (23,3%; 61/262) foi Raillietina cesticillus; entre os nematódeos Heterakis gallinarum (71,4%, 187/262) e Ascaridia galli (45%, 118/262); Postharmostomum commutatum foi o único trematódeo encontrado em 2,7% (7/262) das GCs. As cargas parasitárias mais elevadas foram associadas à Heterakis gallinarum, Ascaridia galli e Raillietina cesticillus. Associações significantes (p ? 0,05) quanto às helmintíases foram observadas em relação ao tipo de cama, ao tipo de solo e à presença de restrição das GCs em áreas cercadas. O tipo de cama, o tempo do vazio sanitário e a presença de sombreamento influenciaram significativamente (p ? 0,05) quanto à infecção por H. gallinarum. A maioria dos parâmetros avaliados foram significativamente associados à infecção das CGs por A. galli. Estes resultados sugerem que GCs do norte do Paraná são infectadas por uma ampla variedade de helmintos gastrintestinais, porém mais frequentemente por R. cesticillus, H. gallinarum e A. galli. Além disso, o tipo de cama de forração, o tipo de solo e o uso de cercas no manejo das GCs são variáveis que estão diretamente relacionados à presença de helmintíases gastrintestinais.(AU)

Assuntos

RESUMO

The effects of managerial systems on the occurrence of gastrointestinal helminths in Free-Range Chickens (FRCs) from northern Paraná, Brazil were investigated. The most predominant (23.3%; 61/262) cestode observed was Raillietina cesticillus; Heterakis gallinarum (71.4%; 187/262) and Ascaridia galli (45%; 118/262) were the predominant nematodes; Postharmostomum commutatum was the only trematode observed in 2.7% (7/262) of FRCs. The most elevated parasitic burdens were associated with Heterakis gallinarum, Ascaridia galli, and Raillietina cesticillus. Significant (p ? 0.05) associations were observed when the effects of the types of bedding, soil type, and fence restriction of FRCs were considered relative to the possibility of helminthiasis. The type of bedding, the length of the sanitary break, and the presence of shading significantly (p ? 0.05) influenced the possibility of FRCs being infected by H. gallinarum. Most parameters evaluated were significantly associated with infection of FRCs by A. galli. These findings suggest that FRCs from northern Paraná are infected by a wide-range of gastrointestinal helminths, but more frequently by R. cesticillus, H. gallinarum, and A. galli. Moreover, the type of floor bedding, the soil type, and the usage of fences in the management of FRCs is directly related to gastrointestinal helminthiasis.

Foram investigados os efeitos dos sistemas de manejo sobre a ocorrência de helmintos gastrintestinais em galinhas caipiras (GCs) do norte do Paraná, Brasil. O cestódeo predominante (23,3%; 61/262) foi Raillietina cesticillus; entre os nematódeos Heterakis gallinarum (71,4%, 187/262) e Ascaridia galli (45%, 118/262); Postharmostomum commutatum foi o único trematódeo encontrado em 2,7% (7/262) das GCs. As cargas parasitárias mais elevadas foram associadas à Heterakis gallinarum, Ascaridia galli e Raillietina cesticillus. Associações significantes (p ? 0,05) quanto às helmintíases foram observadas em relação ao tipo de cama, ao tipo de solo e à presença de restrição das GCs em áreas cercadas. O tipo de cama, o tempo do vazio sanitário e a presença de sombreamento influenciaram significativamente (p ? 0,05) quanto à infecção por H. gallinarum. A maioria dos parâmetros avaliados foram significativamente associados à infecção das CGs por A. galli. Estes resultados sugerem que GCs do norte do Paraná são infectadas por uma ampla variedade de helmintos gastrintestinais, porém mais frequentemente por R. cesticillus, H. gallinarum e A. galli. Além disso, o tipo de cama de forração, o tipo de solo e o uso de cercas no manejo das GCs são variáveis que estão diretamente relacionados à presença de helmintíases gastrintestinais.

Assuntos