PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

ABSTRACT.- Ståhl K., Benito A., Felmer R., Zuñiga J., Reinhardt G., Rivera H., Baule C. & Moreno-López J. 2009. Genetic diversity of bovine viral diarrhoea virus (BVDV) from Peru and Chile. Pesquisa Veterinária Brasileira 29(1):41-44. Joint Virology Research and Development Division, National Veterinary Institute and Swedish University of Agricultural Sciences Uppsala, Sweden. E-mail: Karl.Stahl@bvf.slu.se Twenty-five BVDV strains, detected in serum from persistently infected cattle from Peru (n=15) and Chile (n=10) were genetically characterized. The phylogenetic analysis based on the 5’ UTR showed that all 25 strains belonged to genotype 1. Twenty-three of the strains could further be subdivided into subtype 1b, and two out of ten Chilean strains into subtype 1a. In conclusion, in total 23 out of 25 strains analyzed were of genotype 1, subtype 1b. This is the predominant BVDV subtype in many countries all over the world, including USA. The close homology with previously described strains reflects the influence of livestock trade on the diversity of BVDV circulating within and between countries and continents. Peru and Chile have imported large numbers of cattle from USA and Europe, mostly with insufficient or lacking health documentation.

• Bovine viral diarrhoea virus BVDV diversity pestivirus phylogenetic analysis

Abstract in Portuguese:

ABSTRACT.- Ståhl K., Benito A., Felmer R., Zuñiga J., Reinhardt G., Rivera H., Baule C. & Moreno-López J. 2009. Genetic diversity of bovine viral diarrhoea virus (BVDV) from Peru and Chile. Pesquisa Veterinária Brasileira 29(1):41-44. Joint Virology Research and Development Division, National Veterinary Institute and Swedish University of Agricultural Sciences Uppsala, Sweden. E-mail: Karl.Stahl@bvf.slu.se Twenty-five BVDV strains, detected in serum from persistently infected cattle from Peru (n=15) and Chile (n=10) were genetically characterized. The phylogenetic analysis based on the 5’ UTR showed that all 25 strains belonged to genotype 1. Twenty-three of the strains could further be subdivided into subtype 1b, and two out of ten Chilean strains into subtype 1a. In conclusion, in total 23 out of 25 strains analyzed were of genotype 1, subtype 1b. This is the predominant BVDV subtype in many countries all over the world, including USA. The close homology with previously described strains reflects the influence of livestock trade on the diversity of BVDV circulating within and between countries and continents. Peru and Chile have imported large numbers of cattle from USA and Europe, mostly with insufficient or lacking health documentation.

Abstract in English:

ABSTRACT.- Hussni C.A., Dornbusch P.T., Yoshida W.B., Alves A.L.G., Nicolett, J.L.M., Mamprim M.J. & Vulcano L.C. 2009. [Thrombectomy with Fogarty’s catheter as a treatment of induced jugular thrombophlebitis in horses.] Trombectomia com cateter de Fogarty no tratamento da tromboflebite jugular experimental em eqüinos. Pesquisa Veterinária Brasileira 29(1):45-51. Departamento de Cirurgia e Anestesiologia Veterinária, Faculdade de Medicina Veterinária e Zootecnia, Universidade Estadual Paulista, Botucatu, SP 18618-000, Brazil. E-mail: cahussni@fmvz.unesp.br Thrombosis of jugular vein is a common problem in the equine medicine, implying frequently in fatal outcomes. The diagnosis is relatively simple, based on the clinical findings, angiographics images and ultrasonographycs. The therapeutic employed to a large extent of the cases is unsatisfactory. The purpose of this study was to evaluate the applicability of the thrombectomy with Fogarty’s catheter in horses. This technique is routinely used in medicine, in the reestablishment of the vascular perviousness. Ten horses were allocated in two groups (five animals each) and induced to an unilateral thrombosis of right jugular vein, through the surgical access and an application of stenotic suture and glucose 50% injection. In the control group evolution of the thrombophlebitis without any therapeutical intervention was evaluated. The animals of the treatment group were submitted to the thrombectomy with Fogarty’s catheter. General clinical parameters were analyzed at the moment of the preinduction (MPRE), induction of thrombosis (MTI), and at the 10th day of thrombosis evolution (M10). The procedure induced thrombophlebitis that completely obstructed a segment of the jugular vein in all animals. In the animals of the control group, the thrombus totally obstructed the vascular lumen until the end of the period of evaluation, and parotid edema and vascular dilated, cranial to the thrombophlebitis of jugular vein were observed. The treatment group presented all veins pervious in the end of the experiment, with total remission of the clinical signs, confirmed by angiographic and ultrasonographic examinations. So far, it was concluded that the technique of thrombectomy with Fogarty’s catheter was effective in removal of the thrombosis obstruction experimentally induced in the jugular vein.

• Horses jugular vein thrombophlebitis thrombectomy Fogarty’s catheter

Abstract in Portuguese:

ABSTRACT.- Hussni C.A., Dornbusch P.T., Yoshida W.B., Alves A.L.G., Nicolett, J.L.M., Mamprim M.J. & Vulcano L.C. 2009. [Thrombectomy with Fogarty’s catheter as a treatment of induced jugular thrombophlebitis in horses.] Trombectomia com cateter de Fogarty no tratamento da tromboflebite jugular experimental em eqüinos. Pesquisa Veterinária Brasileira 29(1):45-51. Departamento de Cirurgia e Anestesiologia Veterinária, Faculdade de Medicina Veterinária e Zootecnia, Universidade Estadual Paulista, Botucatu, SP 18618-000, Brazil. E-mail: cahussni@fmvz.unesp.br Thrombosis of jugular vein is a common problem in the equine medicine, implying frequently in fatal outcomes. The diagnosis is relatively simple, based on the clinical findings, angiographics images and ultrasonographycs. The therapeutic employed to a large extent of the cases is unsatisfactory. The purpose of this study was to evaluate the applicability of the thrombectomy with Fogarty’s catheter in horses. This technique is routinely used in medicine, in the reestablishment of the vascular perviousness. Ten horses were allocated in two groups (five animals each) and induced to an unilateral thrombosis of right jugular vein, through the surgical access and an application of stenotic suture and glucose 50% injection. In the control group evolution of the thrombophlebitis without any therapeutical intervention was evaluated. The animals of the treatment group were submitted to the thrombectomy with Fogarty’s catheter. General clinical parameters were analyzed at the moment of the preinduction (MPRE), induction of thrombosis (MTI), and at the 10th day of thrombosis evolution (M10). The procedure induced thrombophlebitis that completely obstructed a segment of the jugular vein in all animals. In the animals of the control group, the thrombus totally obstructed the vascular lumen until the end of the period of evaluation, and parotid edema and vascular dilated, cranial to the thrombophlebitis of jugular vein were observed. The treatment group presented all veins pervious in the end of the experiment, with total remission of the clinical signs, confirmed by angiographic and ultrasonographic examinations. So far, it was concluded that the technique of thrombectomy with Fogarty’s catheter was effective in removal of the thrombosis obstruction experimentally induced in the jugular vein.

Abstract in English:

ABSTRACT.- Ribeiro M.G., Geraldo J.S., Langoni H., Lara G.H.B., Siqueira A.K., Salerno T. & Fernandes M.C. 2008. [Pathogenic microorganisms, somatic cell count and drug residues evaluation in organic bovine milk.] Microrganismos patogênicos, celularidade e resíduos de antimicrobianos no leite produzido no sistema orgânico. Pesquisa Veterinária Brasileira 29(1):52-58. Disciplina de Enfermidades Infecciosas dos Animais, Departamento de Higiene Veterinária e Saúde Pública, Faculdade de Medicina Veterinária e Zootecnia, Universidade Estadual Paulista, Botucatu, SP 18618-000, Brazil. E-mail: mgribeiro@fmvz.unesp.br In last years increase the importance of milk quality and conditions of bovine milking. Simultaneously, increase the interest about organic milk and derivates. The aim of present study was investigate the milk pathogens, sensitivity and multiple drug resistance of isolates, somatic cell count and residues of drugs in milk, from cattle with and without mastitis, come from four little organic dairy farms in State of São Paulo, Brazil. Were used 148 cattle on the middle period of lactation. From these, two showed clinical mastitis, 72 subclinical mastitis and 74 without signs of mammary inflammation (control). Staphylococcus aureus (25.7%), Streptococcus spp. (21.4%), Corynebacterium bovis (12.9%), Streptococcus agalactiae (4.3%) and Staphylococcus spp (4,3%) were the more-frequent microorganisms isolated from animals with mastitis. Aspergillus spp. was isolated from one animal. Ceftiofur (95.2%), oxacillin (84.2%), gentamicin (76.3%) and cefoperazone (70.3%) were the more effective drugs. High resistance of isolates were found to penicillin (53.5%), ampicillin (41.6%) and neomycin (38.6%). Multiple drug resistance to three or more drug was observed in 40 (39.6%) isolates. Media of somatic cell count encountered in animals with mastitis and controls were 175,742.67cs/mL and 58,227.6 cs/mL, respectively. Antimicrobials residues in milk were detected in four (2.7%) animals. The present findings showed the low somatic cell count of animals, indicative of good quality of milk. However, pointed the need of control measures for contagious pathogens of bovine mastitis and more attention for prohibition of antimicrobial use in organic dairy farms.

• Organic milk mastitis bovine somatic cell count antimicrobials multiresistance

Abstract in Portuguese:

ABSTRACT.- Ribeiro M.G., Geraldo J.S., Langoni H., Lara G.H.B., Siqueira A.K., Salerno T. & Fernandes M.C. 2008. [Pathogenic microorganisms, somatic cell count and drug residues evaluation in organic bovine milk.] Microrganismos patogênicos, celularidade e resíduos de antimicrobianos no leite produzido no sistema orgânico. Pesquisa Veterinária Brasileira 29(1):52-58. Disciplina de Enfermidades Infecciosas dos Animais, Departamento de Higiene Veterinária e Saúde Pública, Faculdade de Medicina Veterinária e Zootecnia, Universidade Estadual Paulista, Botucatu, SP 18618-000, Brazil. E-mail: mgribeiro@fmvz.unesp.br In last years increase the importance of milk quality and conditions of bovine milking. Simultaneously, increase the interest about organic milk and derivates. The aim of present study was investigate the milk pathogens, sensitivity and multiple drug resistance of isolates, somatic cell count and residues of drugs in milk, from cattle with and without mastitis, come from four little organic dairy farms in State of São Paulo, Brazil. Were used 148 cattle on the middle period of lactation. From these, two showed clinical mastitis, 72 subclinical mastitis and 74 without signs of mammary inflammation (control). Staphylococcus aureus (25.7%), Streptococcus spp. (21.4%), Corynebacterium bovis (12.9%), Streptococcus agalactiae (4.3%) and Staphylococcus spp (4,3%) were the more-frequent microorganisms isolated from animals with mastitis. Aspergillus spp. was isolated from one animal. Ceftiofur (95.2%), oxacillin (84.2%), gentamicin (76.3%) and cefoperazone (70.3%) were the more effective drugs. High resistance of isolates were found to penicillin (53.5%), ampicillin (41.6%) and neomycin (38.6%). Multiple drug resistance to three or more drug was observed in 40 (39.6%) isolates. Media of somatic cell count encountered in animals with mastitis and controls were 175,742.67cs/mL and 58,227.6 cs/mL, respectively. Antimicrobials residues in milk were detected in four (2.7%) animals. The present findings showed the low somatic cell count of animals, indicative of good quality of milk. However, pointed the need of control measures for contagious pathogens of bovine mastitis and more attention for prohibition of antimicrobial use in organic dairy farms.

Abstract in English:

ABSTRACT.- Scarpelli L., Lopes W.D.Z., Migani M., Bresciani K.D.S. & Costa A.J. 2009. Toxoplasma gondii in experimentally infected Bos taurus and Bos indicus semen and tissues. Pesquisa Veterinária Brasileira 29(1):59-64. Departamento de Medicina Veterinária Preventiva, Faculdade de Ciências Agrárias e Veterinárias, Universidade Estadual Paulista, Via de Acesso Prof. Paulo Donatto Castellani s/n, Jaboticabal, SP 14884-900, Brazil. E-mail: wdzlopes@hotmail.com Eighteen young steers were inoculated with Toxoplasma gondii and randomly distributed into three groups of six animals each: GI, 2.5x105 “P” strain oocysts, GII, 5.0x106 “RH” strain tachyzoites, and GIII (Control). Clinical, serological and parasitemia exams were realized. Parasite investigation by bioassay and PCR was realized on semen and fragments of skeletal musculature, lymph nodes, brain, retina, spleen, liver, lung, testicle, epididymis and seminal vesicle. Blood and semen samples were collected on days -2, -1, 1, 3, 5, 7, 14 and weekly thereafter, up to postinfection day (PID) 84. The inoculated steers (GI and GII) presented hyperthermia from PID 3 to 16. Antibodies against T. gondii were detected through the indirect fluorescence antibody test (IFAT) on PID 5 (1:16) in both inoculated groups (oocysts and tachyzoites), reaching peaks of 1:4096 on PID 7. Parasitemia outbursts occurred in all infected bovines, principally from PID 7 to 28, independent of the strain and inoculate used. Bioassays revealed the presence of parasites in semen samples of animals infected with oocysts (GI) and tachyzoites (GII) on several experimental days between PID 7 and 84. Tissue parasitism by T. gondii was diagnosed by bioassay and the PCR technique in several organ and tissue fragments. These findings suggest the possibility of sexual transmission of T. gondii in the bovine species.

• Cattle PCR semen Toxoplasma gondii Apicomplexa

Abstract in Portuguese:

ABSTRACT.- Scarpelli L., Lopes W.D.Z., Migani M., Bresciani K.D.S. & Costa A.J. 2009. Toxoplasma gondii in experimentally infected Bos taurus and Bos indicus semen and tissues. Pesquisa Veterinária Brasileira 29(1):59-64. Departamento de Medicina Veterinária Preventiva, Faculdade de Ciências Agrárias e Veterinárias, Universidade Estadual Paulista, Via de Acesso Prof. Paulo Donatto Castellani s/n, Jaboticabal, SP 14884-900, Brazil. E-mail: wdzlopes@hotmail.com Eighteen young steers were inoculated with Toxoplasma gondii and randomly distributed into three groups of six animals each: GI, 2.5x105 “P” strain oocysts, GII, 5.0x106 “RH” strain tachyzoites, and GIII (Control). Clinical, serological and parasitemia exams were realized. Parasite investigation by bioassay and PCR was realized on semen and fragments of skeletal musculature, lymph nodes, brain, retina, spleen, liver, lung, testicle, epididymis and seminal vesicle. Blood and semen samples were collected on days -2, -1, 1, 3, 5, 7, 14 and weekly thereafter, up to postinfection day (PID) 84. The inoculated steers (GI and GII) presented hyperthermia from PID 3 to 16. Antibodies against T. gondii were detected through the indirect fluorescence antibody test (IFAT) on PID 5 (1:16) in both inoculated groups (oocysts and tachyzoites), reaching peaks of 1:4096 on PID 7. Parasitemia outbursts occurred in all infected bovines, principally from PID 7 to 28, independent of the strain and inoculate used. Bioassays revealed the presence of parasites in semen samples of animals infected with oocysts (GI) and tachyzoites (GII) on several experimental days between PID 7 and 84. Tissue parasitism by T. gondii was diagnosed by bioassay and the PCR technique in several organ and tissue fragments. These findings suggest the possibility of sexual transmission of T. gondii in the bovine species.

Abstract in English:

ABSTRACT.- Santos T.R.B., Farias N.A.R., Cunha Filho N.A., Pappen F.G. & Vaz Junior I.S. 2009. [Studies of the management of the tick Rhipicephalus (Boophilus) microplus in southern Rio Grande do Sul, Brazil.] Abordagem sobre o controle do carrapato Rhipicephalus (Boophilus) microplus no sul do Rio Grande do Sul. Pesquisa Veterinária Brasileira 29(1):65-70. Faculdade de Veterinária, Universidade Federal de Pelotas, Campus Universitário s/n, Cx. Postal 354, Pelotas, RS 96010-900, Brazil. E-mail: tsantos@ufpel.edu.br In the southern region of Rio Grande do Sul, cattle become infested with Rhipicephalus (B.) microplus mainly between October and April due to the climatic conditions. In addition to knowing its life cycle, knowledge of parasite’s epidemiology is essential to establish management strategies. Epidemiological studies on resistance to acaricides in Rio Grande do Sul as well as in the rest of Brazil are scarce. Moreover, the large geographical area and the structural deficiency with respect to the use and access to databases make reliable data difficult to obtain. The present study surveyed the perception by cattle breeders in the southern region of Rio Grande do Sul in regards of identification of R. (B.) microplus populations that are difficult to manage using acaricides, as well as the risk factors for the selection of resistant tick populations. Tick management data on beef cattle in 85 properties of seven municipalities were collected. The results revealed that the difficulty in tick management correlated positively with levels of education of the farm owners (up to elementary school, OR = 3.67 and p = 0.01) and with the yearly number of acaricide applications (over four, with OR = 4.05 and p= 0.006). These results also suggest that properties with more than 100 beef cattle under extensive farming conditions in the southern region of Rio Grande do Sul show characteristics that may contribute to longer acaricide lifetimes as compared to other regions in the Country.

• Rhipicephalus (Boophilus) microplus risk factors resistance acaricides South Brazil

Abstract in Portuguese:

ABSTRACT.- Santos T.R.B., Farias N.A.R., Cunha Filho N.A., Pappen F.G. & Vaz Junior I.S. 2009. [Studies of the management of the tick Rhipicephalus (Boophilus) microplus in southern Rio Grande do Sul, Brazil.] Abordagem sobre o controle do carrapato Rhipicephalus (Boophilus) microplus no sul do Rio Grande do Sul. Pesquisa Veterinária Brasileira 29(1):65-70. Faculdade de Veterinária, Universidade Federal de Pelotas, Campus Universitário s/n, Cx. Postal 354, Pelotas, RS 96010-900, Brazil. E-mail: tsantos@ufpel.edu.br In the southern region of Rio Grande do Sul, cattle become infested with Rhipicephalus (B.) microplus mainly between October and April due to the climatic conditions. In addition to knowing its life cycle, knowledge of parasite’s epidemiology is essential to establish management strategies. Epidemiological studies on resistance to acaricides in Rio Grande do Sul as well as in the rest of Brazil are scarce. Moreover, the large geographical area and the structural deficiency with respect to the use and access to databases make reliable data difficult to obtain. The present study surveyed the perception by cattle breeders in the southern region of Rio Grande do Sul in regards of identification of R. (B.) microplus populations that are difficult to manage using acaricides, as well as the risk factors for the selection of resistant tick populations. Tick management data on beef cattle in 85 properties of seven municipalities were collected. The results revealed that the difficulty in tick management correlated positively with levels of education of the farm owners (up to elementary school, OR = 3.67 and p = 0.01) and with the yearly number of acaricide applications (over four, with OR = 4.05 and p= 0.006). These results also suggest that properties with more than 100 beef cattle under extensive farming conditions in the southern region of Rio Grande do Sul show characteristics that may contribute to longer acaricide lifetimes as compared to other regions in the Country.

Abstract in English:

ABSTRACT.- Medeiros E.S., Santos M.V., Pinheiro Jr J.W., Faria E.B., Wanderley G.G., Teles J.A.A. & Mota R.A. 2009. [In vitro evaluation of the efficacy of commercial disinfectants used in pre and post-dipping against Staphylococcus spp. isolated from bovine mastitis.] Avaliação in vitro da eficácia de desinfetantes comerciais utilizados no pré e pós-dipping frente amostras de Staphylococcus spp. isoladas de mastite bovina. Pesquisa Veterinária Brasileira 29(1):71-75. Departamento de Medicina Veterinária, Universidade Federal Rural de Pernambuco, Rua Dom Manoel de Medeiros s/n, Dois Irmãos, Recife, PE 52171-900, Brazil. E-mail: rinaldo.mota@hotmail.com The objective of this investigation was to evaluate the in vitro sensibility of Staphylococcus spp. to several commercially available disinfectants used for pre and post dipping. A total of 60 isolates of Staphylococcus spp., identified as Staphylococcus aureus (50) and Positive coagulase Staphylococcus (10) were obtained from the mammary glands of dairy cows with subclinical mastitis in the regions of Metropolitan Recife, the Agreste and the Zona da Mata of the State of Pernambuco, Brazil. As active ingredients were used a chlorine base (25%), iodine (0.6%), chlorhexidine (2.0%), quaternary ammonium (4.0%), and lactic acid (2.0%) at four specific intervals (15", 30", 60", and 300"). One hundred percent of S. aureus was found to be sensitive to iodine, 93.3% to chlorhexidine, 80% to ammonia, 35.6% to lactic acid, and 97.8% were resistant to chlorine at a 60-minute interval. With respect to the Positive coagulase Staphylococcus (SCP), 100.0% of the isolates were sensitive to iodine, 81.8% to quaternary ammonium, 99.9% to lactic acid, 72.7% to chlorhexidine, and 100% was resistant to chlorine at an interval of 60 minutes. It can be concluded that the highest disinfectant activity in vitro was found to be with iodine and chlorhexidine for S. aureus, and with iodine and lactic acid for SCP. A further conclusion was that it is important to undertake a periodic evaluation of the disinfectants used on the dairy properties in the regions studied, given the variety of sensibilities and resistance to disinfectants used, which may prejudice the control of bovine mastitis caused by Staphylococcus spp.

• Disinfectants mastitis Staphylococcus spp. dairy cows

Abstract in Portuguese:

ABSTRACT.- Medeiros E.S., Santos M.V., Pinheiro Jr J.W., Faria E.B., Wanderley G.G., Teles J.A.A. & Mota R.A. 2009. [In vitro evaluation of the efficacy of commercial disinfectants used in pre and post-dipping against Staphylococcus spp. isolated from bovine mastitis.] Avaliação in vitro da eficácia de desinfetantes comerciais utilizados no pré e pós-dipping frente amostras de Staphylococcus spp. isoladas de mastite bovina. Pesquisa Veterinária Brasileira 29(1):71-75. Departamento de Medicina Veterinária, Universidade Federal Rural de Pernambuco, Rua Dom Manoel de Medeiros s/n, Dois Irmãos, Recife, PE 52171-900, Brazil. E-mail: rinaldo.mota@hotmail.com The objective of this investigation was to evaluate the in vitro sensibility of Staphylococcus spp. to several commercially available disinfectants used for pre and post dipping. A total of 60 isolates of Staphylococcus spp., identified as Staphylococcus aureus (50) and Positive coagulase Staphylococcus (10) were obtained from the mammary glands of dairy cows with subclinical mastitis in the regions of Metropolitan Recife, the Agreste and the Zona da Mata of the State of Pernambuco, Brazil. As active ingredients were used a chlorine base (25%), iodine (0.6%), chlorhexidine (2.0%), quaternary ammonium (4.0%), and lactic acid (2.0%) at four specific intervals (15", 30", 60", and 300"). One hundred percent of S. aureus was found to be sensitive to iodine, 93.3% to chlorhexidine, 80% to ammonia, 35.6% to lactic acid, and 97.8% were resistant to chlorine at a 60-minute interval. With respect to the Positive coagulase Staphylococcus (SCP), 100.0% of the isolates were sensitive to iodine, 81.8% to quaternary ammonium, 99.9% to lactic acid, 72.7% to chlorhexidine, and 100% was resistant to chlorine at an interval of 60 minutes. It can be concluded that the highest disinfectant activity in vitro was found to be with iodine and chlorhexidine for S. aureus, and with iodine and lactic acid for SCP. A further conclusion was that it is important to undertake a periodic evaluation of the disinfectants used on the dairy properties in the regions studied, given the variety of sensibilities and resistance to disinfectants used, which may prejudice the control of bovine mastitis caused by Staphylococcus spp.

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.- Coutinho A.S., Oliveira Filho J.P., Silva D.P.G.S., Oliveira A.P., Marcondes J.S., Chiacchio S.B., Paes A.C., Amorim R.M. & Gonçalves R.C. 2009. [Experimental pneumonic mannheimiosis in calves: Nasal and nasopharingeal swabs for diagnostic.] Mannheimiose pulmonar experimental em bezerros: swab nasal e nasofaringeano como auxílio diagnóstico. Pesquisa Veterinária Brasileira 29(1):83-88. Departamento de Clínica Veterinária, Faculdade de Medicina Veterinária e Zootecnia, Universidade Estadual Paulista, Campus de Botucatu, Distrito de Rubião Júnior s/n, Botucatu, SP 18618000, Brazil. E-mail: zep.filho@hotmail.com An experimental model of bovine pneumonic mannheimiosis (BPM) was used to evaluate the nasal and nasopharynx bacterial species of calves during the course of the disease and for checking the diagnostic efficiency of nasal swab (NS) and nasopharingeal swab (NPS) microbiological exams. A total of 28 calves were randomized into four experimental groups (G1-G4). NS and NPS were obtained 7 days before and 12 (G1), 24 (G2), 48 (G3) e 72 (G4) hours after intrabronchial inoculation of Mannheimia haemolytica. After the induction of BPM, M. haemolytica biotype A was the predominant isolated bacterium in NS and NPS in all evaluated sampling times, except for one NS (harvested 24 hours). There were no significant statistical differences for the rates of Pasteurella multocida isolation in NS and NPS, harvested before and after the induction of BPM. However, this bacterium was isolated more frequently after the induction of BPM, mainly in NPS. Therefore, the microbiological NS and NPS exams were important auxiliary tests for diagnosing BPM.

• Bacterial flora upper airway respiratory diseases bovine Mannheimia haemolytica

Abstract in Portuguese:

ABSTRACT.- Coutinho A.S., Oliveira Filho J.P., Silva D.P.G.S., Oliveira A.P., Marcondes J.S., Chiacchio S.B., Paes A.C., Amorim R.M. & Gonçalves R.C. 2009. [Experimental pneumonic mannheimiosis in calves: Nasal and nasopharingeal swabs for diagnostic.] Mannheimiose pulmonar experimental em bezerros: swab nasal e nasofaringeano como auxílio diagnóstico. Pesquisa Veterinária Brasileira 29(1):83-88. Departamento de Clínica Veterinária, Faculdade de Medicina Veterinária e Zootecnia, Universidade Estadual Paulista, Campus de Botucatu, Distrito de Rubião Júnior s/n, Botucatu, SP 18618000, Brazil. E-mail: zep.filho@hotmail.com An experimental model of bovine pneumonic mannheimiosis (BPM) was used to evaluate the nasal and nasopharynx bacterial species of calves during the course of the disease and for checking the diagnostic efficiency of nasal swab (NS) and nasopharingeal swab (NPS) microbiological exams. A total of 28 calves were randomized into four experimental groups (G1-G4). NS and NPS were obtained 7 days before and 12 (G1), 24 (G2), 48 (G3) e 72 (G4) hours after intrabronchial inoculation of Mannheimia haemolytica. After the induction of BPM, M. haemolytica biotype A was the predominant isolated bacterium in NS and NPS in all evaluated sampling times, except for one NS (harvested 24 hours). There were no significant statistical differences for the rates of Pasteurella multocida isolation in NS and NPS, harvested before and after the induction of BPM. However, this bacterium was isolated more frequently after the induction of BPM, mainly in NPS. Therefore, the microbiological NS and NPS exams were important auxiliary tests for diagnosing BPM.

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.