PESQUISA VETERINÁRIA BRASILEIRA

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.- 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. 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.- Barbosa R.R., Pacífico da Silva I. & Benito Soto-Blanco B. 2008. Development of conditioned taste aversion to Mascagnia rigida in goats. Pesquisa Veterinária Brasileira 28(12):571-574. Programa de Pós-Graduação em Ciência Animal, Universidade Federal Rural do Semi-Árido, BR 110 Km 47, Mossoró, RN 59625-900, Brazil. E-mail: benito.blanco@pq.cnpq.br The aim of this study was to determine whether goats could be averted from consuming Mascagnia rigida, a toxic plant found in the semiarid region of northeastern Brazil. Fourteen male goats not previously familiarized to M. rigida were randomly allocated to two treatment groups: control (treated with 5.5mL water orally by a drenching gun) and lithium group (treated with 100mg LiCl/kg body weight orally by a drenching gun). For conditioning, goats were allowed to feed on M. rigida leaves for 15 min, followed by LiCl or water administration. The time spent on eating M. rigida leaves was measured. The conditioning was repeated daily until the LiCl-treated goats stopped eating M. rigida. On the 10th, 17th, and 24th day after conditioning, extinction trials of the M. rigida aversion were performed in goats by using single-choice tests. There was no difference between the two treatment groups with respect to the consumption of M. rigida on the first day of aversion conditioning, however, controls ingested increasing amounts of the plant on consecutive conditioning days. On the second day, five out of the seven goats in the lithium group did not eat the leaves, but on the third day, all the goats in the lithium group did not ingest M. rigida. This aversion persisted throughout all evaluated days. This indicates that goats can be easily conditioned by using lithium chloride to avoid eating M. rigida temporarily

• Poisonous plants Mascagnia rigida Malpighiaceae conditioned food aversion taste aversion diet selection plant poisoning goats

Abstract in Portuguese:

ABSTRACT.- Barbosa R.R., Pacífico da Silva I. & Benito Soto-Blanco B. 2008. Development of conditioned taste aversion to Mascagnia rigida in goats. Pesquisa Veterinária Brasileira 28(12):571-574. Programa de Pós-Graduação em Ciência Animal, Universidade Federal Rural do Semi-Árido, BR 110 Km 47, Mossoró, RN 59625-900, Brazil. E-mail: benito.blanco@pq.cnpq.br The aim of this study was to determine whether goats could be averted from consuming Mascagnia rigida, a toxic plant found in the semiarid region of northeastern Brazil. Fourteen male goats not previously familiarized to M. rigida were randomly allocated to two treatment groups: control (treated with 5.5mL water orally by a drenching gun) and lithium group (treated with 100mg LiCl/kg body weight orally by a drenching gun). For conditioning, goats were allowed to feed on M. rigida leaves for 15 min, followed by LiCl or water administration. The time spent on eating M. rigida leaves was measured. The conditioning was repeated daily until the LiCl-treated goats stopped eating M. rigida. On the 10th, 17th, and 24th day after conditioning, extinction trials of the M. rigida aversion were performed in goats by using single-choice tests. There was no difference between the two treatment groups with respect to the consumption of M. rigida on the first day of aversion conditioning, however, controls ingested increasing amounts of the plant on consecutive conditioning days. On the second day, five out of the seven goats in the lithium group did not eat the leaves, but on the third day, all the goats in the lithium group did not ingest M. rigida. This aversion persisted throughout all evaluated days. This indicates that goats can be easily conditioned by using lithium chloride to avoid eating M. rigida temporarily

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.- Costa M.M., Peixoto R.M., Boijink C.L., Castagna L., Meurer F., & Vargas A.C. 2008. [Antimicrobial sensibility of bacterial isolates from jundiá (Rhamdia quelen).] Sensibilidade antimicrobiana de bactérias isoladas de jundiá (Rhamdia quelen). Pesquisa Veterinária Brasileira 28(10):477-480. Universidade Federal do Vale do São Francisco, Rua da Simpatia 179, Petrolina, PE 56304-440, Brazil. E-mail: mateus.costa@univasf.edu.br Aiming the evaluation of sensitivity profiles of pathogen bacteria responsible for fish diseases, 51 bacterial isolates from Jundiá (Rhamdia quelen) belonging to the genus Acinetobacter spp. (8), Aeromonas spp. (15), Edwardsiella spp. (2), Enterobacter spp. (2), Klebsiella spp. (1), Plesiomonas spp. (5), Pseudomonas spp. (1), Staphylococcus spp. (11), and Vibrio spp. (6), were tested against antimicrobial agents used for treatment of bacterial fish diseases. All samples were processed at the Laboratory of Bacteriology, Department of Preventive Veterinary Medicine, UFSM. From 51 bacteria isolated from jundiá fishes (Rhamdia quelen) 51 (100%) were sensitive to gentamycin, 49 (96,08%) to sulphazotrin, 47(92,16%) to chloramphenicol, 43 (84,31%) to tetracylin, 43 (84,31%) to naldixic acid, 31 (60,78%) to nitrofurantoin, 22 (43,14%) to erytromycin, 22 (43,14%) to ampicillin, 15 (29,41%) spiramycin, 13 (25,50%) to cholystin, and 5 (3%) to penicillin G. With exception of an isolate of Staphylococcus spp., the bacteria analyzed in the present study were resistant to one or more antimicrobial agents tested. Knowledge of the sensitivity profile of bacteria involved in infectious processes in fish will allow rational antimicrobial treatment that will contribute to the control of fish diseases in Rhamdia quelen without causing great risks to public health and the environment.

• Pathogen bacterial antimicrobial drugs Rhamdia quelen

Abstract in Portuguese:

ABSTRACT.- Costa M.M., Peixoto R.M., Boijink C.L., Castagna L., Meurer F., & Vargas A.C. 2008. [Antimicrobial sensibility of bacterial isolates from jundiá (Rhamdia quelen).] Sensibilidade antimicrobiana de bactérias isoladas de jundiá (Rhamdia quelen). Pesquisa Veterinária Brasileira 28(10):477-480. Universidade Federal do Vale do São Francisco, Rua da Simpatia 179, Petrolina, PE 56304-440, Brazil. E-mail: mateus.costa@univasf.edu.br Aiming the evaluation of sensitivity profiles of pathogen bacteria responsible for fish diseases, 51 bacterial isolates from Jundiá (Rhamdia quelen) belonging to the genus Acinetobacter spp. (8), Aeromonas spp. (15), Edwardsiella spp. (2), Enterobacter spp. (2), Klebsiella spp. (1), Plesiomonas spp. (5), Pseudomonas spp. (1), Staphylococcus spp. (11), and Vibrio spp. (6), were tested against antimicrobial agents used for treatment of bacterial fish diseases. All samples were processed at the Laboratory of Bacteriology, Department of Preventive Veterinary Medicine, UFSM. From 51 bacteria isolated from jundiá fishes (Rhamdia quelen) 51 (100%) were sensitive to gentamycin, 49 (96,08%) to sulphazotrin, 47(92,16%) to chloramphenicol, 43 (84,31%) to tetracylin, 43 (84,31%) to naldixic acid, 31 (60,78%) to nitrofurantoin, 22 (43,14%) to erytromycin, 22 (43,14%) to ampicillin, 15 (29,41%) spiramycin, 13 (25,50%) to cholystin, and 5 (3%) to penicillin G. With exception of an isolate of Staphylococcus spp., the bacteria analyzed in the present study were resistant to one or more antimicrobial agents tested. Knowledge of the sensitivity profile of bacteria involved in infectious processes in fish will allow rational antimicrobial treatment that will contribute to the control of fish diseases in Rhamdia quelen without causing great risks to public health and the environment.

Abstract in English:

ABSTRACT.- Abreu S.R.O., Mota R.A., Rosinha G.M.S., Forner O., Pinheiro Júnior J.W., Pereira R.R.B., Castro R.S., Elisei C., Soares C.S., Araújo F.R. & Madureira R.C. 2008. [Genotypic comparison between Corynebacterium pseudotuberculosis samples obtained from sheep and goats with caseous lymphadenitis, raised in the semi-arid region of Pernambuco.] Comparação genotípica de isolados de Corynebacterium pseudotuberculosis de caprinos e ovinos do sertão de Pernambuco. Pesquisa Veterinária Brasileira 28(10):481-487. Clínica Escola de Medicina Veterinária, Faculdade de Ciências Biológicas e da Saúde, Centro de Ensino Superior de Maceió, Rodovia Divaldo Suruagy s/n, Quadra 4, Lote 4, Praia do Francês, Marechal Deodoro, AL 57160-000, Brazil. E-mail: silviobiotec@yahoo.com.br The objective was to genotypically compare 35 samples of Corynebacterium pseudotuberculosis obtained from abscesses of sheep and goats diagnosed with caseous lymphadenitis originated from 5 different municipalities in the semi-arid region of Pernambuco, Brazil. The RFLP-PCR technique with Hpy-Ch4 and Msp I and Pst I Msp I restriction enzimes was used to fingerprint the genes rpoB and pld, respectively. The results demonstrate that there was no difference on the fragments banding pattern among samples, independently of the host species or geographic area studied, defining a homogeneous profile of C. pseudotuberculosis responsible for superficial abscesses for the region.

• Genotyping RFLP-PCR Corynebacterium pseudotuberculosis goats sheep

Abstract in Portuguese:

ABSTRACT.- Abreu S.R.O., Mota R.A., Rosinha G.M.S., Forner O., Pinheiro Júnior J.W., Pereira R.R.B., Castro R.S., Elisei C., Soares C.S., Araújo F.R. & Madureira R.C. 2008. [Genotypic comparison between Corynebacterium pseudotuberculosis samples obtained from sheep and goats with caseous lymphadenitis, raised in the semi-arid region of Pernambuco.] Comparação genotípica de isolados de Corynebacterium pseudotuberculosis de caprinos e ovinos do sertão de Pernambuco. Pesquisa Veterinária Brasileira 28(10):481-487. Clínica Escola de Medicina Veterinária, Faculdade de Ciências Biológicas e da Saúde, Centro de Ensino Superior de Maceió, Rodovia Divaldo Suruagy s/n, Quadra 4, Lote 4, Praia do Francês, Marechal Deodoro, AL 57160-000, Brazil. E-mail: silviobiotec@yahoo.com.br The objective was to genotypically compare 35 samples of Corynebacterium pseudotuberculosis obtained from abscesses of sheep and goats diagnosed with caseous lymphadenitis originated from 5 different municipalities in the semi-arid region of Pernambuco, Brazil. The RFLP-PCR technique with Hpy-Ch4 and Msp I and Pst I Msp I restriction enzimes was used to fingerprint the genes rpoB and pld, respectively. The results demonstrate that there was no difference on the fragments banding pattern among samples, independently of the host species or geographic area studied, defining a homogeneous profile of C. pseudotuberculosis responsible for superficial abscesses for the region.

Abstract in English:

ABSTRACT.- Campos T.A., Lago J.C., Nakazato G., Stehling E.G., Brocchi M., Castro A.F.P. & Silveira W.D. 2008. Occurrence of virulence-related sequences and phylogenetic analysis of commensal and pathogenic avian Escherichia coli strains (APEC). Pesquisa Veterinária Brasileira 28(10):533-540. Departamento de Microbiologia e Immunologia, Instituto de Biologia, Unicamp, Cidade Universitrária Zeferino Vaz s/n, Campinas, SP 13081-862, Brazil. E-mail: wds@unicamp.br The presence of iron uptake (irp-2, fyuA, sitA, fepC, iucA), adhesion (iha, lpfAO157/O141, lpfAO157/O154, efa, toxB) and invasion (inv, ial-related DNA sequences and assignment to the four main Escherichia coli phylogenetic groups (A, B1, B2 e D) were determined in 30 commensal E. coli strains isolated from healthy chickens and in 49 APEC strains isolated from chickens presenting clinical signs of septicemia (n=24) swollen head syndrome (n=14) and omphalitis (n=11) by PCR. None of the strains presented DNA sequences related to the inv, ial, efa, and toxB genes. DNA sequences related to lpfAO157/O154, iucA, fepC, and irp-2 genes were significantly found among pathogenic strains, where iucA gene was associated with septicemia and swollen head syndrome and fepC and irp-2 genes were associated with swollen head syndrome strains. Phylogenetic typing showed that commensal and omphalitis strains belonged mainly to phylogenetic Group A and swollen head syndrome to phylogenetic Group D. Septicemic strains were assigned in phylogenetic Groups A and D. These data could suggest that clonal lineage of septicemic APEC strains have a multiple ancestor origin; one from a pathogenic bacteria ancestor and other from a non-pathogenic ancestor that evolved by the acquisition of virulence related sequences through horizontal gene transfer. Swollen head syndrome may constitute a pathogenic clonal group. By the other side, omphalitis strains probably constitute a non-pathogenic clonal group, and could cause omphalitis as an opportunistic infection. The sharing of virulence related sequences by human pathogenic E. coli and APEC strains could indicate that APEC strains could be a source of virulence genes to human strains and could represent a zoonotic risk.

• Avian colibacillosis Escherichia coli APEC virulence related-genes pathogenic clones

Abstract in Portuguese:

ABSTRACT.- Campos T.A., Lago J.C., Nakazato G., Stehling E.G., Brocchi M., Castro A.F.P. & Silveira W.D. 2008. Occurrence of virulence-related sequences and phylogenetic analysis of commensal and pathogenic avian Escherichia coli strains (APEC). Pesquisa Veterinária Brasileira 28(10):533-540. Departamento de Microbiologia e Immunologia, Instituto de Biologia, Unicamp, Cidade Universitrária Zeferino Vaz s/n, Campinas, SP 13081-862, Brazil. E-mail: wds@unicamp.br The presence of iron uptake (irp-2, fyuA, sitA, fepC, iucA), adhesion (iha, lpfAO157/O141, lpfAO157/O154, efa, toxB) and invasion (inv, ial-related DNA sequences and assignment to the four main Escherichia coli phylogenetic groups (A, B1, B2 e D) were determined in 30 commensal E. coli strains isolated from healthy chickens and in 49 APEC strains isolated from chickens presenting clinical signs of septicemia (n=24) swollen head syndrome (n=14) and omphalitis (n=11) by PCR. None of the strains presented DNA sequences related to the inv, ial, efa, and toxB genes. DNA sequences related to lpfAO157/O154, iucA, fepC, and irp-2 genes were significantly found among pathogenic strains, where iucA gene was associated with septicemia and swollen head syndrome and fepC and irp-2 genes were associated with swollen head syndrome strains. Phylogenetic typing showed that commensal and omphalitis strains belonged mainly to phylogenetic Group A and swollen head syndrome to phylogenetic Group D. Septicemic strains were assigned in phylogenetic Groups A and D. These data could suggest that clonal lineage of septicemic APEC strains have a multiple ancestor origin; one from a pathogenic bacteria ancestor and other from a non-pathogenic ancestor that evolved by the acquisition of virulence related sequences through horizontal gene transfer. Swollen head syndrome may constitute a pathogenic clonal group. By the other side, omphalitis strains probably constitute a non-pathogenic clonal group, and could cause omphalitis as an opportunistic infection. The sharing of virulence related sequences by human pathogenic E. coli and APEC strains could indicate that APEC strains could be a source of virulence genes to human strains and could represent a zoonotic risk.