PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

ABSTRACT.- Silva Júnior A., Castro L.A., Chiarelli Neto O., Silva F.M.F., Vidigal P.M.P., Moraes M.P. & Almeida M.R. 2009. Development and evaluation of a recombinant DNA vaccine candidate expressing porcine circovirus 2 structural protein. Pesquisa Veterinária Brasileira 29(1):76-82. Laboratório de Infectologia Molecular Animal, Instituto de Biotecnologia Aplicada à Agropecuária, Universidade Federal de Viçosa, Av. PH Rolfs s/n, Campus Universitário, Viçosa, MG 36570-000, Brazil. E-mail: marcia@ufv.br Porcine circovirus 2 (PCV2) is generally associated with the porcine circovirosis syndrome, which is considered an important disease of swine and has potentially serious economic impact on the swine industry worldwide. This article describes the construction of a recombinant plasmid expressing the PCV2 structural protein and the evaluation of cellular and humoral immune responses produced by this recombinant vaccine in BALB/c mice. The vaccine candidate was obtained and analyzed in vivo, in an effort to determine the ability to induce a specific immune response in mice. DNA was extracted from a Brazilian PCV2 isolate and the gene coding for Cap protein was amplified by PCR and inserted into an expression plasmid. Groups of BALB/c mice were inoculated intra-muscularly and intradermally in a 15-day interval, with 100 µg and 50 µg of the vaccine construct, respectively. Another group was inoculated intramuscularly with 100 µg of empty plasmid, corresponding to the control group. Seroconversion and cellular response in BALB/c mice were compared and used for vaccine evaluation. Seroconversion was analyzed by ELISA. After a series of 3 immunizations the spleen cells of the immunized animals were used to perform lymphocyte proliferation assays. Seroconversion to PCV2 was detected by ELISA in the animals inoculated with the vaccine construct when compared with control groups. Lymphocyte proliferation assays showed a stronger cell proliferation in the inoculated animals compared with the control group. Thus, the vaccine candidate construct demonstrated to be able to induce both humoral and cellular responses in inoculated mice.

• PCV2 DNA vaccine porcine circovirosis

Abstract in Portuguese:

ABSTRACT.- Silva Júnior A., Castro L.A., Chiarelli Neto O., Silva F.M.F., Vidigal P.M.P., Moraes M.P. & Almeida M.R. 2009. Development and evaluation of a recombinant DNA vaccine candidate expressing porcine circovirus 2 structural protein. Pesquisa Veterinária Brasileira 29(1):76-82. Laboratório de Infectologia Molecular Animal, Instituto de Biotecnologia Aplicada à Agropecuária, Universidade Federal de Viçosa, Av. PH Rolfs s/n, Campus Universitário, Viçosa, MG 36570-000, Brazil. E-mail: marcia@ufv.br Porcine circovirus 2 (PCV2) is generally associated with the porcine circovirosis syndrome, which is considered an important disease of swine and has potentially serious economic impact on the swine industry worldwide. This article describes the construction of a recombinant plasmid expressing the PCV2 structural protein and the evaluation of cellular and humoral immune responses produced by this recombinant vaccine in BALB/c mice. The vaccine candidate was obtained and analyzed in vivo, in an effort to determine the ability to induce a specific immune response in mice. DNA was extracted from a Brazilian PCV2 isolate and the gene coding for Cap protein was amplified by PCR and inserted into an expression plasmid. Groups of BALB/c mice were inoculated intra-muscularly and intradermally in a 15-day interval, with 100 µg and 50 µg of the vaccine construct, respectively. Another group was inoculated intramuscularly with 100 µg of empty plasmid, corresponding to the control group. Seroconversion and cellular response in BALB/c mice were compared and used for vaccine evaluation. Seroconversion was analyzed by ELISA. After a series of 3 immunizations the spleen cells of the immunized animals were used to perform lymphocyte proliferation assays. Seroconversion to PCV2 was detected by ELISA in the animals inoculated with the vaccine construct when compared with control groups. Lymphocyte proliferation assays showed a stronger cell proliferation in the inoculated animals compared with the control group. Thus, the vaccine candidate construct demonstrated to be able to induce both humoral and cellular responses in inoculated mice.

Abstract in English:

ABSTRACT.- Furlan F.H., Lucioli J., Borelli V., Fonteque J.H., Stolf L., Traverso S.D. & Gava A. 2009. [Poisoning by closantel in sheep and goats in the State of Santa Catarina, Brazil.] Intoxicação por closantel em ovinos e caprinos no Estado de Santa Catarina. Pesquisa Veterinária Brasileira 29(1):89-93. Departamento de Medicina Veterinária, Centro de Ciências Agroveterinárias, Universidade do Estado de Santa Catarina, Lages, SC 88520-000, Brazil. E-mail: a2ag@cav.udesc.br Two outbreaks of closantel overdosage in sheep and goat flocks are described. In the first outbreak 12 sheep were affected, 5 of them showed blindness, three sheep died (Sheep 1-3) and two were euthanized 6 months after the onset of clinical manifestation (Sheep 4 and 5). In the second outbreak 26 goats were affected, from which six survived despite blindness and one was euthanized. Clinically the animals showed depression, ataxia, motor incoordination, decreased or absent pupil reflexes. In some animals this clinical picture developed to bilateral blindness, with no reaction to threat and bilateral irresponsive midriasis. In the ophthalmic examination retinal vessel atrophy and hyperreflexia were observed. The histological examination showed myelin edema leading to status spongiosus in the central nervous system and compressive neuropathy of the optic nerve, associated with retinal degeneration and/or atrophy. This report aims to describe the epidemiologic, clinic and pathologic aspects of closantel overdosage in sheep and goats.

• Closantel sheep goats status spongiosus retinal degeneration

Abstract in Portuguese:

ABSTRACT.- Furlan F.H., Lucioli J., Borelli V., Fonteque J.H., Stolf L., Traverso S.D. & Gava A. 2009. [Poisoning by closantel in sheep and goats in the State of Santa Catarina, Brazil.] Intoxicação por closantel em ovinos e caprinos no Estado de Santa Catarina. Pesquisa Veterinária Brasileira 29(1):89-93. Departamento de Medicina Veterinária, Centro de Ciências Agroveterinárias, Universidade do Estado de Santa Catarina, Lages, SC 88520-000, Brazil. E-mail: a2ag@cav.udesc.br Two outbreaks of closantel overdosage in sheep and goat flocks are described. In the first outbreak 12 sheep were affected, 5 of them showed blindness, three sheep died (Sheep 1-3) and two were euthanized 6 months after the onset of clinical manifestation (Sheep 4 and 5). In the second outbreak 26 goats were affected, from which six survived despite blindness and one was euthanized. Clinically the animals showed depression, ataxia, motor incoordination, decreased or absent pupil reflexes. In some animals this clinical picture developed to bilateral blindness, with no reaction to threat and bilateral irresponsive midriasis. In the ophthalmic examination retinal vessel atrophy and hyperreflexia were observed. The histological examination showed myelin edema leading to status spongiosus in the central nervous system and compressive neuropathy of the optic nerve, associated with retinal degeneration and/or atrophy. This report aims to describe the epidemiologic, clinic and pathologic aspects of closantel overdosage in sheep and goats.

Abstract in English:

ABSTRACT. Riet-Correa F., Haraguchi M., Dantas A.F., Burakovas R.G., Yokosuka A., Mimaki Y., Medeiros R.M.T. & Matos P.F. 2009. Sheep poisoning by Panicum dichotomiflorum in northeastern Brazil. Pesquisa Veterinária Brasileira 29(1):94-98. Hospital Veterinário, Centro de Saúde e Tecnologia Rural, Universidades Federal de Campina Grande, Patos, PB 58700-000, Brazil. E-mail: franklin.riet@pq.cnpq.br Different species of Panicum, including P. dichotomiflorum, have been reported as a cause of photosensitization in sheep, horses, cattle and goats. An outbreak of hepatogenous photosensitization occurred in 3 flocks of hair sheep in the Brazilian semiarid region. Eighty one out of 365 sheep were affected and 39 died. The main affected animals were nursing lambs and sheep younger than one year old. Donkeys, goats and cattle grazing in the same pasture were not affected. Clinical signs were edema of the head, followed by dermatitis, mainly in the face, ears, and croup, ocular discharge, corneal opacity with blindness, and redness of the coronary band and hoof. At necropsy of one affected lamb the liver was yellowish. Upon histologic examination scattered necrotic hepatocytes were observed in the liver and focal areas of necrosis of myocytes appeared in the heart. Samples of P. dicotomiflorum were analyzed by TLC and those containing saponins were isolated by HPLC using RP-C18 column and eluted with a mixture of MeOH and H2O. The isolated compounds were submitted to 1H and 13C NMR spectroscopy. Reactions were positive to furostanol saponins with the same Rf of the standard protodioscin (0.21) and methylprotodioscin (0.32). The spectroscopic results indicated a mixture of (25R)- and (25S)-protodioscin isomers in a proportion of 3:1, and methylprotodioscin.

• Lithogenic steroidal saponins Panicum dichotomiflorum photosensitization protodioscin

Abstract in Portuguese:

ABSTRACT. Riet-Correa F., Haraguchi M., Dantas A.F., Burakovas R.G., Yokosuka A., Mimaki Y., Medeiros R.M.T. & Matos P.F. 2009. Sheep poisoning by Panicum dichotomiflorum in northeastern Brazil. Pesquisa Veterinária Brasileira 29(1):94-98. Hospital Veterinário, Centro de Saúde e Tecnologia Rural, Universidades Federal de Campina Grande, Patos, PB 58700-000, Brazil. E-mail: franklin.riet@pq.cnpq.br Different species of Panicum, including P. dichotomiflorum, have been reported as a cause of photosensitization in sheep, horses, cattle and goats. An outbreak of hepatogenous photosensitization occurred in 3 flocks of hair sheep in the Brazilian semiarid region. Eighty one out of 365 sheep were affected and 39 died. The main affected animals were nursing lambs and sheep younger than one year old. Donkeys, goats and cattle grazing in the same pasture were not affected. Clinical signs were edema of the head, followed by dermatitis, mainly in the face, ears, and croup, ocular discharge, corneal opacity with blindness, and redness of the coronary band and hoof. At necropsy of one affected lamb the liver was yellowish. Upon histologic examination scattered necrotic hepatocytes were observed in the liver and focal areas of necrosis of myocytes appeared in the heart. Samples of P. dicotomiflorum were analyzed by TLC and those containing saponins were isolated by HPLC using RP-C18 column and eluted with a mixture of MeOH and H2O. The isolated compounds were submitted to 1H and 13C NMR spectroscopy. Reactions were positive to furostanol saponins with the same Rf of the standard protodioscin (0.21) and methylprotodioscin (0.32). The spectroscopic results indicated a mixture of (25R)- and (25S)-protodioscin isomers in a proportion of 3:1, and methylprotodioscin.

Abstract in English:

ABSTRACT.- Bezerra F.S.B., Garcia H.A., Alves H.M., Oliveira I.R.S., Silva A.E., Teixeira M.M.G. & Batista J.S. 2008. [Trypanosoma vivax in testicular and epidydimal tissues of experimentally infected sheep.] Trypanosoma vivax nos tecidos testicular e epididimário de ovinos experimentalmente infectados. Pesquisa Veterinária Brasileira 28(12):575-582. Laboratório de Patologia Veterinária, Departamento de Ciências Animais, Universidade Federal Rural do Semi-árido, BR 110 Km 47, Cx. Postal 147, Mossoró, RN 59625-900, Brazil. E-mail: jaelsbatista@hotmail.com Four adult sheep (number 1, 2, 3 and 4), all males, were inoculated intravenously with 1ml of blood containing 1.25x105 trypomastigotes of Trypanosoma vivax, and Sheep 5, 6, 7 and 8 were used as control. After infection, clinical exams considering rectal temperature, respiratory and cardiac frequencies, and parasitaemia were recorded daily for a 30-day experiment period. Blood samples were obtained for 5-day intervals to hematocrit analysis. At the end of the experimental period, the sheep were orquiectomized. Testes and epididymides from these animals were studied anatomopathologically. Samples from these tissues of Sheep 1, 4 and 5 were taken to polymerase chain reaction (PCR). Clinical parameters remained for the infected group above the values observed in the control group during the experimental period. Parasitaemia was observed on day 3 post-infection, and the highest values occurred between day 6 and 10, and day 15 and 18 post-infection. Sheep 1 and 4 showed severe anemia on day 25 post-infection. All sheep of the infected group showed flabby and palid testes. Histologically, moderate to severe testicular degeneration, multifocal epididymitis and hyperplasia of epididymal epithelium were observed. The result of T. vivax PCR analysis in the testes and epididymal tissues was positive in 100% of the samples of the experimentally infected sheep. Epididymal and testicular lesions associated with the presence of the parasite in these tissues, shown by PCR, suggest the participation of T. vivax in the pathophysiological mechanism of reproductive damage.

• rypanosomosis small ruminant PCR anatomopathology testicular degene-ration epididymitis extravascular foci

Abstract in Portuguese:

ABSTRACT.- Bezerra F.S.B., Garcia H.A., Alves H.M., Oliveira I.R.S., Silva A.E., Teixeira M.M.G. & Batista J.S. 2008. [Trypanosoma vivax in testicular and epidydimal tissues of experimentally infected sheep.] Trypanosoma vivax nos tecidos testicular e epididimário de ovinos experimentalmente infectados. Pesquisa Veterinária Brasileira 28(12):575-582. Laboratório de Patologia Veterinária, Departamento de Ciências Animais, Universidade Federal Rural do Semi-árido, BR 110 Km 47, Cx. Postal 147, Mossoró, RN 59625-900, Brazil. E-mail: jaelsbatista@hotmail.com Four adult sheep (number 1, 2, 3 and 4), all males, were inoculated intravenously with 1ml of blood containing 1.25x105 trypomastigotes of Trypanosoma vivax, and Sheep 5, 6, 7 and 8 were used as control. After infection, clinical exams considering rectal temperature, respiratory and cardiac frequencies, and parasitaemia were recorded daily for a 30-day experiment period. Blood samples were obtained for 5-day intervals to hematocrit analysis. At the end of the experimental period, the sheep were orquiectomized. Testes and epididymides from these animals were studied anatomopathologically. Samples from these tissues of Sheep 1, 4 and 5 were taken to polymerase chain reaction (PCR). Clinical parameters remained for the infected group above the values observed in the control group during the experimental period. Parasitaemia was observed on day 3 post-infection, and the highest values occurred between day 6 and 10, and day 15 and 18 post-infection. Sheep 1 and 4 showed severe anemia on day 25 post-infection. All sheep of the infected group showed flabby and palid testes. Histologically, moderate to severe testicular degeneration, multifocal epididymitis and hyperplasia of epididymal epithelium were observed. The result of T. vivax PCR analysis in the testes and epididymal tissues was positive in 100% of the samples of the experimentally infected sheep. Epididymal and testicular lesions associated with the presence of the parasite in these tissues, shown by PCR, suggest the participation of T. vivax in the pathophysiological mechanism of reproductive damage.

Abstract in English:

ABSTRACT.- Diéguez F.J., Yus E., Sanjuán M.L., Vilar M.J. & Arnaiz I. 2008. Monitoring bovine viral diarrhea virus (BVDV) infection status in dairy herds. Pesquisa Veterinária Brasileira 28(12):588-592. Unidad de Epidemiología y Sanidad Animal, Instituto de Investigación y Análisis Alimentarios, Facultad de Veterinaria s/n, Lugo 27002, Spain. E-mail: franciscojavier.dieguez@usc.es This study was designed to assess the relationship between antibodies against bovine viral diarrhea virus (BVDV) determined in the bulk tank milk (BTM) and the within-herd seroprevalence. We also assessed the efficiency of measuring antibody levels in BTM samples to monitor BVDV infection status in a herd. In the 81 farms included in the study, BTM samples were obtained and blood samples withdrawn from all cattle older than one year. The infection status was then determined in serum and milk using a commercial blocking ELISA based on the detection of anti-p80 antibodies. Apart from these baseline serum and milk samples, another BTM sample was collected from each herd 9 months later, and a third BTM sample obtained 9 months after this. In these second and third milk samples, anti-BVDV antibodies were determined using the same ELISA kit. Statistical tests revealed good agreement between herd seroprevalences (% seropositive animals in the herd) and the antibody levels detected in the BTM samples. During the 18 months of follow-up, the farms with persistently infected cattle at the study outset (14.8% of the herds) showed a significant decrease in BTM antibody titers after virus clearance. Conversely, a significant increase in BTM antibody levels was observed in the herds infected with BVDV during the follow-up period. Our findings indicate that monitoring antibody levels in the BTM is a useful method of identifying changes in the BVDV infection status of a herd.

• Bovine viral diarrhea vírus BVDV antibodies diagnostic milk

Abstract in Portuguese:

ABSTRACT.- Diéguez F.J., Yus E., Sanjuán M.L., Vilar M.J. & Arnaiz I. 2008. Monitoring bovine viral diarrhea virus (BVDV) infection status in dairy herds. Pesquisa Veterinária Brasileira 28(12):588-592. Unidad de Epidemiología y Sanidad Animal, Instituto de Investigación y Análisis Alimentarios, Facultad de Veterinaria s/n, Lugo 27002, Spain. E-mail: franciscojavier.dieguez@usc.es This study was designed to assess the relationship between antibodies against bovine viral diarrhea virus (BVDV) determined in the bulk tank milk (BTM) and the within-herd seroprevalence. We also assessed the efficiency of measuring antibody levels in BTM samples to monitor BVDV infection status in a herd. In the 81 farms included in the study, BTM samples were obtained and blood samples withdrawn from all cattle older than one year. The infection status was then determined in serum and milk using a commercial blocking ELISA based on the detection of anti-p80 antibodies. Apart from these baseline serum and milk samples, another BTM sample was collected from each herd 9 months later, and a third BTM sample obtained 9 months after this. In these second and third milk samples, anti-BVDV antibodies were determined using the same ELISA kit. Statistical tests revealed good agreement between herd seroprevalences (% seropositive animals in the herd) and the antibody levels detected in the BTM samples. During the 18 months of follow-up, the farms with persistently infected cattle at the study outset (14.8% of the herds) showed a significant decrease in BTM antibody titers after virus clearance. Conversely, a significant increase in BTM antibody levels was observed in the herds infected with BVDV during the follow-up period. Our findings indicate that monitoring antibody levels in the BTM is a useful method of identifying changes in the BVDV infection status of a herd.

Abstract in English:

ABSTRACT.- Bonel-Raposo J., Riet-Correa F., Guim T.N., Schuch I.D., Grecco F.G. & Fernandes C.G. 2007. [Acute poisoning and abortions in guinea pigs by the pods of Enterolobium contortisiliquum (Leg. Mimosoideae).] Intoxicação aguda e abortos em cobaias pelas favas de Enterolobium contortisiliquum (Leg. Mimosoideae). Pesquisa Veterinária Brasileira 28(12):593-596. Departamento de Patologia Animal, Faculdade de Veterinária, Universidade Federal de Pelotas, Campus Universitário s/n, Pelotas, RS 96010-900, Brazil. E-mail: bonel-raposo@brturbo.com.br The objective was to study the acute toxicity and the abortive properties of Enterolobium contortisiliquum pods in guinea pigs. Pods of E. contortisiliquum were administered orally to 4 groups of 3 guinea pigs each. Another group of 3 guinea-pigs was used as control. Group 1 and 2 were fed with one dose of 5 and 10g of pods for kg body weight, respectively. The guinea pigs of Group 3 and 4 received 10 and 15g/kg, respectively, divided into daily doses of 5g/kg. One guinea pig from Group 2 and one from Group 4 died 12 and 18 hours after the end of the administration. Gross lesions were hemorrhages of the stomach and of the large and small gut, enlarged liver, and dilated gall bladder. Histologically, the liver had severe vacuolation and necrosis of periportal hepatocytes. In another experiment a ration containing 4% of pods of E. contortisiliquum was fed to 2 groups of 4 guinea-pigs, 35 days after mating. Four of the 8 guinea pigs aborted 6-15 days after the beginning of ingestion. The other 4 guinea pigs were not pregnant. All guinea pigs were euthanized after abortion or at the end of the experiment. Histologically all animals had mild to severe periportal hemorrhagic necrosis. All fetuses had variable degree of autolysis. In 4 fetuses studied no significant histologic lesions were observed. The acute lesions observed in guinea-pigs are similar than those observed in the spontaneous poisoning by Enterolobium spp. in cattle. Similar lesions are observed in guinea-pigs poisoned experimentally with saponins from E. gummiferum. The results of the experiments in pregnant guinea pigs suggest that E. contortisiliquum can be used to study the abortive effect of its pods or its toxic compounds.

• Poisonous plants hepatotoxic plants abortion Enterolobium contortisiliquum Legumiosae Mimosoideae guinea pigs

Abstract in Portuguese:

ABSTRACT.- Bonel-Raposo J., Riet-Correa F., Guim T.N., Schuch I.D., Grecco F.G. & Fernandes C.G. 2007. [Acute poisoning and abortions in guinea pigs by the pods of Enterolobium contortisiliquum (Leg. Mimosoideae).] Intoxicação aguda e abortos em cobaias pelas favas de Enterolobium contortisiliquum (Leg. Mimosoideae). Pesquisa Veterinária Brasileira 28(12):593-596. Departamento de Patologia Animal, Faculdade de Veterinária, Universidade Federal de Pelotas, Campus Universitário s/n, Pelotas, RS 96010-900, Brazil. E-mail: bonel-raposo@brturbo.com.br The objective was to study the acute toxicity and the abortive properties of Enterolobium contortisiliquum pods in guinea pigs. Pods of E. contortisiliquum were administered orally to 4 groups of 3 guinea pigs each. Another group of 3 guinea-pigs was used as control. Group 1 and 2 were fed with one dose of 5 and 10g of pods for kg body weight, respectively. The guinea pigs of Group 3 and 4 received 10 and 15g/kg, respectively, divided into daily doses of 5g/kg. One guinea pig from Group 2 and one from Group 4 died 12 and 18 hours after the end of the administration. Gross lesions were hemorrhages of the stomach and of the large and small gut, enlarged liver, and dilated gall bladder. Histologically, the liver had severe vacuolation and necrosis of periportal hepatocytes. In another experiment a ration containing 4% of pods of E. contortisiliquum was fed to 2 groups of 4 guinea-pigs, 35 days after mating. Four of the 8 guinea pigs aborted 6-15 days after the beginning of ingestion. The other 4 guinea pigs were not pregnant. All guinea pigs were euthanized after abortion or at the end of the experiment. Histologically all animals had mild to severe periportal hemorrhagic necrosis. All fetuses had variable degree of autolysis. In 4 fetuses studied no significant histologic lesions were observed. The acute lesions observed in guinea-pigs are similar than those observed in the spontaneous poisoning by Enterolobium spp. in cattle. Similar lesions are observed in guinea-pigs poisoned experimentally with saponins from E. gummiferum. The results of the experiments in pregnant guinea pigs suggest that E. contortisiliquum can be used to study the abortive effect of its pods or its toxic compounds.

Abstract in English:

ABSTRACT.- Corrêa A.M.R., Zlotowski P., Barcellos D.E.S.N., Cruz C.E.F. & Driemeier D. 2008. Gastric ulcers in pigs affected with postweaning multisystemic wasting syndrome. Pesquisa Veterinária Brasileira 28(12):601-605. Departamento de Patologia Clínica Veterinária, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves 9090, Porto Alegre, RS 91540-000, Brazil. E-mail: davetpat@ufrgs.br Samples of gastric lymph nodes and the stomachs from 24 pigs selected from herds affected by postweaning multisystemic wasting syndrome (PMWS) and sudden death associated with gastric ulcers were studied. Pigs were selected on the basis of unthriftiness, decreased feed intake, and wasting. The stomachs were opened, inverted, and classified into 0-3 score according the severity of the gross lesions present in pars oesophagica (non-glandulargastric mucosa). Selected samples were processed for paraffin embedding and stained with hematoxylin and eosin. Immunohistochemistry using anti-PCV2 (porcine circovírus type 2) antibody, anti-Helicobacter pylori antibody and a wide-spectrum anti-cytokeratin antibody was performed. Gross changes in pars oesophagea were classified according to the severity of lesions as score 3, 2, and 1 in 8, 6, 5 stomachs respectivelly. Microscopically, hyperplastic lymphoid follicles, lymphohistiocytic inflammatory infiltrates and focci of necrosis in the gastric mucosa were common findings. Large amounts of PCV2 antigen were observed in the cytoplasm and nuclei from intralesional cells and debris from the gastric glandular mucosal zone; however, in the fundus, anti-PCV2 immunostaining was restricted to the surface mucosal cells and foveolar compartment. All gastric lymph nodes were positive for PCV2 antigen. Anti-H. pylori immunostaining was seen in eleven cases, mainly in the antrum, on the mucosal surface and foveolar compartment. The association of the anti-PCV2 immunostaining with the glandular mucus-producing cells suggests a role for PCV2 as an additional factor for the swine ulcer development.

• Diseases of swine gastric ulcer PMWS PCV2 Helicobacter pylori

Abstract in Portuguese:

ABSTRACT.- Corrêa A.M.R., Zlotowski P., Barcellos D.E.S.N., Cruz C.E.F. & Driemeier D. 2008. Gastric ulcers in pigs affected with postweaning multisystemic wasting syndrome. Pesquisa Veterinária Brasileira 28(12):601-605. Departamento de Patologia Clínica Veterinária, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves 9090, Porto Alegre, RS 91540-000, Brazil. E-mail: davetpat@ufrgs.br Samples of gastric lymph nodes and the stomachs from 24 pigs selected from herds affected by postweaning multisystemic wasting syndrome (PMWS) and sudden death associated with gastric ulcers were studied. Pigs were selected on the basis of unthriftiness, decreased feed intake, and wasting. The stomachs were opened, inverted, and classified into 0-3 score according the severity of the gross lesions present in pars oesophagica (non-glandulargastric mucosa). Selected samples were processed for paraffin embedding and stained with hematoxylin and eosin. Immunohistochemistry using anti-PCV2 (porcine circovírus type 2) antibody, anti-Helicobacter pylori antibody and a wide-spectrum anti-cytokeratin antibody was performed. Gross changes in pars oesophagea were classified according to the severity of lesions as score 3, 2, and 1 in 8, 6, 5 stomachs respectivelly. Microscopically, hyperplastic lymphoid follicles, lymphohistiocytic inflammatory infiltrates and focci of necrosis in the gastric mucosa were common findings. Large amounts of PCV2 antigen were observed in the cytoplasm and nuclei from intralesional cells and debris from the gastric glandular mucosal zone; however, in the fundus, anti-PCV2 immunostaining was restricted to the surface mucosal cells and foveolar compartment. All gastric lymph nodes were positive for PCV2 antigen. Anti-H. pylori immunostaining was seen in eleven cases, mainly in the antrum, on the mucosal surface and foveolar compartment. The association of the anti-PCV2 immunostaining with the glandular mucus-producing cells suggests a role for PCV2 as an additional factor for the swine ulcer development.

Abstract in English:

ABSTRACT.- Freitas M.F.L., Luz I.S., Silveira-Filho V.M., Júnior J.W.P., Stamford T.L.M., Mota R.A., Sena M.J., Almeida A.M.P., Balbino V.Q. & Leal-Balbino T.C. 2008. Staphylococcal toxin genes in milk samples from cows diagnosed with subclinical mastitis. Pesquisa Veterinária Brasileira 28(12):617-621. Centro de Pesquisas Aggeu Magalhães, Fiocruz, Av. Prof. Moraes Rego s/n, Campus da Cidade Universitária, Recife, PE 50670-420, Brazil. E-mail: cristina@cpqam.fiocruz.br The present study was carried out in 11 dairy herds in four municipal districts of the rural area of the State of Pernambuco, Brazil. Out of 984 quarter milk (246 cows), 10 (1.0%) were positive for clinical mastitis, 562 (57.1%) for subclinical mastitis and 412 (41.9%) were negative. A total of 81 Staphylococcus spp. isolates were obtained from milk samples from the cows diagnosed with subclinical mastitis. From these, 53 (65.0%) were S. aureus, 16 (20.0%) coagulase-positive staphylococci (CPS) and 12 (15.0%) coagulase-negative staphylococci (CNS). The isolates were further investigated for the presence of toxin genes by multiplex and uniplex PCR. The main gene observed was seg followed by seh, sei and sej. The distribution of these observed genes among the isolates obtained from different areas showed a regional pattern for the SEs. The presence of toxin genes in the strains isolated from bovine milk demonstrates a potential problem for public health.

• Staphylococcal toxins toxin genes milk dairy cows

Abstract in Portuguese:

ABSTRACT.- Freitas M.F.L., Luz I.S., Silveira-Filho V.M., Júnior J.W.P., Stamford T.L.M., Mota R.A., Sena M.J., Almeida A.M.P., Balbino V.Q. & Leal-Balbino T.C. 2008. Staphylococcal toxin genes in milk samples from cows diagnosed with subclinical mastitis. Pesquisa Veterinária Brasileira 28(12):617-621. Centro de Pesquisas Aggeu Magalhães, Fiocruz, Av. Prof. Moraes Rego s/n, Campus da Cidade Universitária, Recife, PE 50670-420, Brazil. E-mail: cristina@cpqam.fiocruz.br The present study was carried out in 11 dairy herds in four municipal districts of the rural area of the State of Pernambuco, Brazil. Out of 984 quarter milk (246 cows), 10 (1.0%) were positive for clinical mastitis, 562 (57.1%) for subclinical mastitis and 412 (41.9%) were negative. A total of 81 Staphylococcus spp. isolates were obtained from milk samples from the cows diagnosed with subclinical mastitis. From these, 53 (65.0%) were S. aureus, 16 (20.0%) coagulase-positive staphylococci (CPS) and 12 (15.0%) coagulase-negative staphylococci (CNS). The isolates were further investigated for the presence of toxin genes by multiplex and uniplex PCR. The main gene observed was seg followed by seh, sei and sej. The distribution of these observed genes among the isolates obtained from different areas showed a regional pattern for the SEs. The presence of toxin genes in the strains isolated from bovine milk demonstrates a potential problem for public health.

Abstract in English:

ABSTRACT.- Tortelli P.F., Barbosa J.D., Oliveira C.M.C., Dutra M.D., Cerqueira V.D., Oliveira C.A., Riet-Correa F. & Riet-Correa G. 2008. [Poisoning by Ipomoea asarifolia in cattle and sheep in the Marajó Island, Northern Brazil.] Intoxicação por Ipomoea asarifolia em bovinos e ovinos na Ilha de Marajó. Pesquisa Veterinária Brasileira 28(12):622-626. Central de Diagnóstico Veterinário, Faculdade de Medicina Veterinária, Campus de Castanhal, Universidade Federal do Pará, Rua Maximino Porpino da Silva 1000, Pirapora, Castanhal, PA 68743-080, Brazil. E-mail: griet@ufpa.br To determine plant poisonings occurring on Marajo Island, state of Pará, northern Brazil, 7 farms were visited. All farms had native pastures with low forage availability severely infected by Ipomoea asarifolia. In 5 farms poisoning in sheep by the plant was reported. The disease occurs mainly during the dry period and lambs are more frequently affected than adult sheep. Sheep with clinical signs were observed in 2 farms. In 4 establishments farmers reported the occurrence of I. asarifolia poisoning in cattle. In one farm affected cattle were observed during two visits, one during the dry season and another at the start of the raining season. Deaths are observed mainly in calves. In both species the poisoning occurs every year with variable morbidity and low mortality. One bovine and one sheep with severe clinical signs were euthanized and post-mortem examination was performed. No gross or histologic significant lesions were observed. In all farms buffaloes were raised together with cattle and sheep, and in 3 farms goats were also raised. None of the farmers reported the intoxication in these species.

• Cattle Ipomoea asarifolia sheep toxic plants tremogenic toxins

Abstract in Portuguese:

ABSTRACT.- Tortelli P.F., Barbosa J.D., Oliveira C.M.C., Dutra M.D., Cerqueira V.D., Oliveira C.A., Riet-Correa F. & Riet-Correa G. 2008. [Poisoning by Ipomoea asarifolia in cattle and sheep in the Marajó Island, Northern Brazil.] Intoxicação por Ipomoea asarifolia em bovinos e ovinos na Ilha de Marajó. Pesquisa Veterinária Brasileira 28(12):622-626. Central de Diagnóstico Veterinário, Faculdade de Medicina Veterinária, Campus de Castanhal, Universidade Federal do Pará, Rua Maximino Porpino da Silva 1000, Pirapora, Castanhal, PA 68743-080, Brazil. E-mail: griet@ufpa.br To determine plant poisonings occurring on Marajo Island, state of Pará, northern Brazil, 7 farms were visited. All farms had native pastures with low forage availability severely infected by Ipomoea asarifolia. In 5 farms poisoning in sheep by the plant was reported. The disease occurs mainly during the dry period and lambs are more frequently affected than adult sheep. Sheep with clinical signs were observed in 2 farms. In 4 establishments farmers reported the occurrence of I. asarifolia poisoning in cattle. In one farm affected cattle were observed during two visits, one during the dry season and another at the start of the raining season. Deaths are observed mainly in calves. In both species the poisoning occurs every year with variable morbidity and low mortality. One bovine and one sheep with severe clinical signs were euthanized and post-mortem examination was performed. No gross or histologic significant lesions were observed. In all farms buffaloes were raised together with cattle and sheep, and in 3 farms goats were also raised. None of the farmers reported the intoxication in these species.

Abstract in English:

ABSTRACT.- Pedroso P.M.O., Pescador C.A., Bandarra P.M., Raymundo D.L., Borba M.R., Wouters F., Bezerra Jr P.S. & Driemeier D. 2008. [Standardization of immunohistochemistry technique for detection of rabies virus in formalin-fixed and paraffin-embedded tissue samples from central nervous system of cattle.] Padronização da técnica de imuno-histoquímica para raiva em amostras de tecido do sistema nervoso central de bovinos fixadas em formol e emblocadas em parafina. Pesquisa Veterinária Brasileira 28(12):627-632. Setor de Patologia Veterinária, Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves 9090, Porto Alegre, RS 91540-000, Brazil. E-mail: davetpat@ufrgs.br For standardization of the rabies immunohistochemistry technique, five samples of central nervous system (CNS) of cattle naturally infected with rabies virus were examined. One polyclonal antibody and two monoclonal antibodies were used. The following reagents were evaluated for antigen retrieval: XIV protease, proteinase K and citrate buffer (pH 6.0) boiling at 100°C during 15 minutes in bain-marie. Detection of rabic antigen was possible with the three antibodies tested. The polyclonal antibody was superior to the monoclonal antibodies, demonstrating good results with the three antigen retrieval protocols. The highest intensity staining was obtained with the citrate buffer and heat. The immunohistochemistry technique demonstrated the presence of viral antigens in the cytoplasm of neurons, in form of aggregates or with round or oval shape. The antigens were found as single or multiples inclusion bodies in the neurons. Immunohistochemistry is a fast method that can be used in routine procedures in cases where rabies is suspected, especially when the brain is submitted to the laboratory as formalin-fixed fragments or when samples could not be immediately shipped. The technique is also useful for retrospective studies.

• Diseases of cattle infectious diseases diseases of the central nervous system rabies immunohistochemistry standardization

Abstract in Portuguese:

ABSTRACT.- Pedroso P.M.O., Pescador C.A., Bandarra P.M., Raymundo D.L., Borba M.R., Wouters F., Bezerra Jr P.S. & Driemeier D. 2008. [Standardization of immunohistochemistry technique for detection of rabies virus in formalin-fixed and paraffin-embedded tissue samples from central nervous system of cattle.] Padronização da técnica de imuno-histoquímica para raiva em amostras de tecido do sistema nervoso central de bovinos fixadas em formol e emblocadas em parafina. Pesquisa Veterinária Brasileira 28(12):627-632. Setor de Patologia Veterinária, Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves 9090, Porto Alegre, RS 91540-000, Brazil. E-mail: davetpat@ufrgs.br For standardization of the rabies immunohistochemistry technique, five samples of central nervous system (CNS) of cattle naturally infected with rabies virus were examined. One polyclonal antibody and two monoclonal antibodies were used. The following reagents were evaluated for antigen retrieval: XIV protease, proteinase K and citrate buffer (pH 6.0) boiling at 100°C during 15 minutes in bain-marie. Detection of rabic antigen was possible with the three antibodies tested. The polyclonal antibody was superior to the monoclonal antibodies, demonstrating good results with the three antigen retrieval protocols. The highest intensity staining was obtained with the citrate buffer and heat. The immunohistochemistry technique demonstrated the presence of viral antigens in the cytoplasm of neurons, in form of aggregates or with round or oval shape. The antigens were found as single or multiples inclusion bodies in the neurons. Immunohistochemistry is a fast method that can be used in routine procedures in cases where rabies is suspected, especially when the brain is submitted to the laboratory as formalin-fixed fragments or when samples could not be immediately shipped. The technique is also useful for retrospective studies.