Resultado da pesquisa (11)

Termo utilizado na pesquisa DNA

#1 - DNA damage and primordial follicle activation after in vitro culture of sheep ovarian cortex in Morus nigra leaf extract

Abstract in English:

This study evaluated the effect of Morus nigra leaf extract, with or without supplementation, on morphology, activation and DNA damage of preantral follicles cultured within sheep ovarian tissue. Ovaries were collected and divided into fragments, being one fixed for histological and Terminal deoxynucleotidyl transferase (TdT) mediated dUTP nick-end labeling (TUNEL) analysis (fresh control). The remaining fragments were cultured for 7 days in alpha minimum essential media (α-MEM) supplemented with bovine serum albumin (BSA), insulin, transferrin, selenium, glutamine, hypoxanthine and ascorbic acid (α-MEM+; control medium) or into medium composed of M. nigra extract without supplements (0.1; 0.2 or 0.4mg/mL) or supplemented with the same substances described above for α-MEM+ (MN 0.1+; 0.2+ or 0.4+mg/mL). Then, tissues were destined to histological and TUNEL analysis. The α-MEM+ treatment had more morphologically normal follicles than all M. nigra extract treatments. However, α-MEM+ treatment also showed signs of atresia because the percentage of TUNEL positive cells was similar in α-MEM+ and in 0.1mg/mL M. nigra without and with supplements. Moreover, a reduction in the primordial follicles and an increase in the growing ones were observed in all treatments, except 0.2mg/mL M. nigra. In conclusion, the follicles cultured at 0.1mg/mL M. nigra extract were in good condition and able to continue their development, as demonstrated by the same rates of DNA damage and follicular activation as the control medium.

Abstract in Portuguese:

Este estudo avaliou o efeito do extrato das folhas de Morus nigra, com ou sem suplementos, sobre a morfologia, a ativação e o dano ao DNA de folículos pré-antrais cultivados inclusos em tecido ovariano. Os ovários foram coletados e divididos em fragmentos, sendo um fixado para análise histológica e ensaio de marcação de terminações dUTP mediada por desoxinucleotidil transferase terminal (TUNEL) (controle fresco). Os fragmentos restantes foram cultivados durante 7 dias em meio essencial mínimo alfa (α-MEM) suplementado com albumina sérica bovina (BSA), insulina, transferrina, selênio, glutamina, hipoxantina e ácido ascorbico (α-MEM+; meio controle) ou em meio composto de extrato de M. nigra sem suplementos (0,1; 0,2 or 0,4mg/mL) ou suplementado com as mesmas substâncias descritas para α-MEM+ (MN 0,1+; 0,2+ or 0,4+mg/mL). Então, os tecidos foram destinados à análise histológica e TUNEL. O tratamento do α-MEM+ apresentou mais folículos morfologicamente normais que todos os tratamentos do extrato de M. nigra. No entanto, o tratamento com α-MEM+ também mostrou sinais de atresia, pois a porcentagem de células TUNEL positivas foi semelhante em α-MEM+ e em 0,1mg/mL M. nigra sem e com suplementos. Além disso, observou-se uma redução nos folículos primordiais e um aumento nos folículos em crescimento em todos os tratamentos, exceto 0,2mg/mL M. nigra. Em conclusão, os folículos cultivados com 0,1mg/mL de extrato de M. nigra estavam em boas condições e aptos a continuar seu desenvolvimento, como demonstrado pelas taxas de dano ao DNA e de ativação folicular semelhantes ao meio controle.


#2 - Molecular characterization of bovine Deltapapillomavirus (BPV1, 2, and 13) DNA in equine sarcoids, 35(5):431-436

Abstract in English:

ABSTRACT.- De Alcântara B.K., Alfieri A.A., Headley S.A., Rodrigues W.B., Otonel R.A.A., Lunardi M. & Alfieri A.F. 2015. Molecular characterization of bovine Deltapapillomavirus (BPV1, 2, and 13) DNA in equine sarcoids. Pesquisa Veterinária Brasileira 35(5):431-436. Laboratory of Animal Virology, Department of Veterinary Preventive Medicine, Universidade Estadual de Londrina, Rodovia Celso Garcia Cid, Campus Universitário, Cx. Postal 10011, Londrina, PR 86057-970, Brazil. E-mail: alfieri@uel.br Sarcoids are fibroblastic lesions, which are considered as the most common skin tumors of horses; spontaneous regression rarely occurs. The bovine papillomavirus (BPV) types 1 and 2 may be involved in the pathogenesis of sarcoids, and probably the recently described BPV type (BPV13) might be associated with the pathogenesis of this lesion. This study characterized the DNA of BPVs in sarcoids from 15 horses from Brazil by analyzing 20 cutaneous lesions (12 recently collected; 8 from formalin-fixed paraffin-embedded (FFPE) tissues). Histopathology confirmed the proliferative lesions as sarcoids. Three PCRs were performed to amplify papillomavirus (PV) DNA. For screening, the primers IFNR2/IDNT2 were used to amplify a fragment of the PV L1 ORF. The second primer set was complementary to a common sequence of the E5L2 genomic region of BPV1, 2, and 13. The third primer pair (FAP59/FAP64) targeted a fragment of the PVs L1 ORF. The screening and E5L2 PCRs yielded amplicons in all samples evaluated. The FAP amplicons identified BPV1, 2, and 13 only from fresh tissue samples. The phylogenetic analyses of E5L2 resulted in the identification of BPV1, 2, and 13 in 14 (70%), 2 (10%), and 4 (20%) sarcoids, respectively. Two horses demonstrated multiple lesions: the sarcoids of one of these contained only BPV1 DNA and those of the other contained three types of bovine Deltapapillomavirus (BPV1, 2, and 13). This study confirmed the presence of BPV1, 2, and 13 DNA in equine sarcoids. Moreover, these findings represent the first description of three types of BPV diagnosed in the same horse, as well as the first confirmation of BPV1 and 2 in horses from Brazil.

Abstract in Portuguese:

RESUMO.- De Alcântara B.K., Alfieri A.A., Headley S.A., Rodrigues W.B., Otonel R.A.A., Lunardi M. & Alfieri A.F. 2015. Molecular characterization of bovine Deltapapillomavirus (BPV1, 2, and 13) DNA in equine sarcoids. [Caracterização molecular de DNA de Deltapapillomavirus bovino (BPV1, 2 e 13) em sarcoides equinos.] Pesquisa Veterinária Brasileira 35(5):431-436. Laboratory of Animal Virology, Department of Veterinary Preventive Medicine, Universidade Estadual de Londrina, Rodovia Celso Garcia Cid, Campus Universitário, Cx. Postal 10011, Londrina, PR 86057-970, Brazil. E-mail: alfieri@uel.br Sarcoides são tumores fibroblásticos, considerados os tumores de pele mais comuns em pele de equinos e que raramente apresentam regressão espontânea. Papilomavírus bovino (BPV) tipos 1 e 2 são relacionados com a patogenia do sarcoide e, provavelmente, o BPV tipo 13 (BPV13), recentemente descrito, também pode estar associado com a formação dessa lesão. Neste estudo, 20 amostras de lesões cutâneas, sendo 12 constituídas por tecidos frescos e 8 amostras de tecido fixado em formalina e embebido em parafina, provenientes de 15 cavalos foram utilizadas para a identificação do DNA de BPV. A análise histopatológica (HE) confirmou todas as lesões como sarcoide. Para a amplificação do DNA de papilomavírus (PV) foram realizadas três reações de PCR. Como triagem, os primers IFNR2/IDNT2 foram utilizados para amplificar um fragmento da ORF L1 do PV. O segundo par de primers utilizado é complementar a sequência dos genes E5 e L2 de BPVs 1, 2 e 13. O terceiro par de primers (FAP59/FAP64) utilizado tem o gene L1 como alvo. A primeira e a segunda PCRs permitiram amplificar produtos em todas as amostras avaliadas. Entretanto, na terceira reação, na qual foram utilizados os primers FAP, foi possível amplificar produtos com tamanho molecular esperado somente nas amostras constituídas por tecidos frescos. O sequenciamento de nucleotídeos e as análises filogenéticas realizadas nos fragmentos E5L2 resultaram na identificação de BPV1, 2 e 13 em 14 (70%), 2 (10%) e em 4 (20%) amostras de sarcoides, respectivamente. As amostras de sarcoides de um dos animais continha somente o DNA de BPV1. Entretanto, nas amostras provenientes do segundo cavalo foi possível identificar o DNA de três tipos de Deltapapillomavirus bovino (BPV1, 2 e 13) em lesões distintas. Este estudo ratifica a presença do DNA de BPV1, 2 e 13 em lesões de sarcoides em equinos, além de identificar três tipos de BPVs em um mesmo animal e descrever pela primeira vez no Brasil a presença de BPV1 e 2 nesse tipo de lesão.


#3 - Identification of canine papillomavirus type 1 (CPV1) DNA in dogs with cutaneous papillomatosis, 34(12):1223-1226

Abstract in English:

ABSTRACT.- De Alcântara B.K., Alfieri A.A., Rodrigues W.B., Otonel R.A.A., Lunardi M., Headley S.A. & Alfieri A.F. 2014. Identification of canine papillomavirus type 1 (CPV1) DNA in dogs with cutaneous papillomatosis. Pesquisa Veterinária Brasileira 34(12):1223-1226. Laboratório de Virologia Animal, Departamento de Medicina Veterinária Preventiva, Universidade Estadual de Londrina, Rodovia Celso Garcia Cid, Campus Universitário, Cx. Postal 10011, Londrina. PR 86057-970, Brazil. E-mail: alfieri@uel.br Canine oral papillomavirus (COPV), also known as Canine Papillomavirus type 1 (CPV1), induces papillomas at the mucous membranes of the oral cavity and at the haired skin of dogs. The classification of Papillomavirus (PV) types is based on the L1 capsid protein and nucleotide sequence; so far, 14 CPV types have been described in several countries, but the molecular characterization of CPV in Brazil is lacking. This study investigated the presence of the PV in seven papillomas from four mixed breed dogs from Londrina/PR, Southern Brazil, by partial sequencing of the L1 gene. Seven exophytic cutaneous lesions were surgically removed and processed for histopathological and molecular characterization. Histopathology confirmed the lesions as viral papillomas due to typical histological features. Polymerase Chain Reaction (PCR) assay using the FAP59 and FAP64 primers targeted the L1 gene followed by sequence analysis of the amplicons identified CPV1 in all evaluated papilloma samples. This study represents the first description of CPV1 DNA associated with canine papillomatosis in Brazil.

Abstract in Portuguese:

RESUMO.- De Alcântara B.K., Alfieri A.A., Rodrigues W.B., Otonel R.A.A., Lunardi M., Headley S.A. & Alfieri A.F. 2014. Identification of canine papillomavirus type 1 (CPV1) DNA in dogs with cutaneous papillomatosis. [Identificação do DNA de Papillomavirus canino tipo 1 em cães com papilomas cutâneos no Brasil.] Pesquisa Veterinária Brasileira 34(12):1223-1226. Laboratório de Virologia Animal, Departamento de Medicina Veterinária Preventiva, Universidade Estadual de Londrina, Rodovia Celso Garcia Cid, Campus Universitário, Cx. Postal 10011, Londrina. PR 86057-970, Brazil. E-mail: alfieri@uel.br O papilomavírus oral canino (COPV), também denominado Papillomavirus canino tipo 1 (CPV1), tem a capacidade de induzir papilomas na mucosa da cavidade oral e também em pele de cães. A classificação dos tipos de papilomavírus (PV) é baseada na proteína L1 do capsídeo e na sequência de nucleotídeos que a codifica. Atualmente são descritos 14 tipos de CPV, no entanto, ainda faltam estudos moleculares relacionados à identificação dos tipos de CPV no Brasil. O objetivo deste estudo foi investigar a presença de PV em fragmentos de papilomas obtidos de quatro cães sem raça definida, provenientes de Londrina/PR, região sul do Brasil, e definir o tipo viral por meio da análise da sequência parcial de nucleotídeos do gene L1. Sete lesões cutâneas foram cirurgicamente removidas e processadas ​​para a caracterização histopatológica e molecular. O exame histopatológico confirmou as lesões como papilomas. Foi realizada reação em cadeia de polimerase (PCR), utilizando os primers FAP59 FAP64 para a amplificação parcial do gene L1, seguida por análise das sequências dos produtos amplificados, que confirmou a presença do CPV1 em todas as amostras avaliadas. Este estudo representa a primeira identificação do DNA de CPV1 associado com papilomatose canina no Brasil.


#4 - Occurrence of Toxoplasma gondii DNA in sheep naturally infected and slaughtered in abattoirs in Pernambuco, Brazil, 34(4):329-331

Abstract in English:

ABSTRACT.- Bezerra M.J.G., Cruz J.A.L.O., Kung E.S., Silva J.G., Santos A.S., Moraes E.P.B.X., Pinheiro Junior J.W. & Mota R.A. 2014. Occurrence of Toxoplasma gondii DNA in sheep naturally infected and slaughtered in abattoirs in Pernambuco, Brazil. Pesquisa Veterinária Brasileira 34(4):329-331. Laboratory of Infectious Contagious Diseases in Domestic Animals, Universidade Federal Rural de Pernambuco, Rua Dom Manoel de Medeiros s/n, Recife, PE 52171-900, Brazil. E-mail: rinaldo.mota@hotmail.com The aim of the present study was to assess the occurrence of antibodies to Toxoplasma gondii and to detect genomic DNA of the parasite in the reproductive organs, fetuses and fetal membranes of sheep in slaughterhouses in the state of Pernambuco, Brazil. The Indirect Immunofluorescence technique (IFA) was used for screening. The Polymerase Chain Reaction (PCR) was used to detect DNA of T. gondii in the animals that were positive in the serology. In the serology, 13/50 samples were positive and genomic DNA of T. gondii was detected in one uterus, tube, ovary, placenta and fetus (heart, brain and umbilical cord) sample from a sheep that was positive in the serology. The present study provides evidence of the occurrence of T. gondii DNA in the organs of the reproductive system, placenta and fetus of a naturally infected sheep.

Abstract in Portuguese:

RESUMO.- Bezerra M.J.G., Cruz J.A.L.O., Kung E.S., Silva J.G., Santos A.S., Moraes E.P.B.X., Pinheiro Junior J.W. & Mota R.A. 2014. Occurrence of Toxoplasma gondii DNA in sheep naturally infected and slaughtered in abattoirs in Pernambuco, Brazil. [Ocorrência de DNA de Toxoplasma gondii em ovelhas naturalmente infectadas e abatidas em matadouros de Pernambuco.] Pesquisa Veterinária Brasileira 34(4):329-331. Laboratory of Infectious Contagious Diseases in Domestic Animals, Universidade Federal Rural de Pernambuco, Rua Dom Manoel de Medeiros s/n, Recife, PE 52171-900, Brazil. E-mail: rinaldo.mota@hotmail.com Objetivou-se estudar a ocorrência de anticorpos contra Toxoplasma gondii e detectar o DNA genômico do parasito em órgãos reprodutivos, fetos e anexos fetais de ovelhas em matadouros no estado de Pernambuco, Brasil. Foram coletadas amostras de soro sanguíneo, útero, trompas e ovários, além de fetos e placentas. Para a triagem utilizou-se a técnica de Imunofluorescência Indireta (RIFI) e para a detecção do DNA de T. gondii empregou-se a Reação em Cadeia da Polimerase (PCR) nos animais positivos na sorologia e em todos os fetos e anexos fetais. Na sorologia, 13/50 amostras foram positivas e o DNA genômico de T. gondii foi detectado em uma amostra de útero, trompa, ovário, placenta e feto (coração, cérebro e cordão umbilical) de uma ovelha positiva na sorologia. A identidade molecular dos produtos amplificados foi confirmada por sequenciamento. Neste estudo comprova-se a ocorrência do DNA de T. gondii em órgãos do sistema reprodutivo, placenta e feto de ovelha naturalmente infectada.


#5 - Reactivation and distribution of bovine herpesvirus 5 DNA in the brain of latently infected sheep, 31(12):1090-1096

Abstract in English:

ABSTRACT.- Cadore G.C., Anziliero D., Weiblen R. & Flores E.F. 2011. [Reactivation and distribution of bovine herpesvirus 5 DNA in the brain of latently infected sheep.] Reativação e distribuição do DNA do herpesvírus bovino tipo 5 no encéfalo de ovinos latentemente infectados. Pesquisa Veterinária Brasileira 31(12):1090-1096. Departamento de Medicina Veterinária Preventiva, Universidade Federal de Santa Maria, Camobi, Santa Maria, RS 97105-900, Brazil. E-mail: eduardofurtadoflores@gmail.com The biology of latent infection by bovine herpesvirus type 5 (BoHV-5) has been studied in cattle and rabbits, yet many aspects remain poorly understood. We herein investigated the suitability of lambs to investigate aspects of BoHV-5 latency. Thirteen six-month-old lambs inoculated intranasally (IN) with BoHV-5 strain SV-507/99 (titer of 106.8 TCID50/mL) shed the virus in nasal secretions in titers up 105.5 TCID50/mL, during up to 11 days, developing virus neutralizing (VN) titers of 16 to 128 at day 30 post-inoculation (pi). The inoculated animals developed only a mild serous nasal secretion and transient hyperthermia. Examination of brain sections of five lambs euthanized at day 30 pi by PCR revealed the presence of latent DNA in the trigeminal ganglia (TG, 5 out of five), olfactory bulbs (OB, 5/5), pons (2/5), cerebellum (2/5) and cerebral cortex (1/5). Administration of dexamethasone (Dx, n=4) or flumethasone (FluM, n=4) to eight latently infected lambs at day 65 pi resulted in virus reactivation and shedding by 3 out of 4 individuals in each group. Virus shedding in nasal secretions started at day 3 post-treatment and lasted up to five days (1-5) in Dx treated lambs (titers up to 102.8TCID50/mL), was delayed and lasted up to three days (1-3) in FluM-treated lambs (titers up to 102.1 TCID50/mL). PCR examination of the brains of animals submitted to reactivation, at day 30 post-treatment, showed a pattern of distribution of latent viral DNA fairly similar to that found in those not submitted to reactivation. In summary, the ability of BoHV-5 to establish latent infection, the consistent colonization of TGs and OBs by latent viral DNA and virus reactivation induced by corticosteroid treatment are promising findings towards the use of lambs to study selected aspects of BoHV-5 latency.

Abstract in Portuguese:

RESUMO.- Cadore G.C., Anziliero D., Weiblen R. & Flores E.F. 2011. [Reactivation and distribution of bovine herpesvirus 5 DNA in the brain of latently infected sheep.] Reativação e distribuição do DNA do herpesvírus bovino tipo 5 no encéfalo de ovinos latentemente infectados. Pesquisa Veterinária Brasileira 31(12):1090-1096. Departamento de Medicina Veterinária Preventiva, Universidade Federal de Santa Maria, Camobi, Santa Maria, RS 97105-900, Brazil. E-mail: eduardofurtadoflores@gmail.com A biologia da infecção latente pelo herpesvírus bovino tipo 5 (BoHV-5) tem sido estudada em bovinos e coelhos, mas vários aspectos permanecem desconhecidos. Este artigo relata uma avaliação de ovinos jovens como modelo para o estudo da infecção latente pelo BoHV-5. Treze cordeiros com idade entre seis e sete meses, inoculados pela via intranasal (IN) com a cepa SV-507/99 do BoHV-5 (título de 106,8 DICC50/mL) excretaram o vírus em secreções nasais em títulos de até 105,5 DICC50/mL, com duração de até 11 dias, desenvolvendo anticorpos neutralizantes em títulos de 16 a 128 no dia 30 pós-inoculação (pi). Os ovinos inoculados apresentaram apenas secreção nasal serosa leve e hipertermia transitória. O PCR de secções do encéfalo de cinco animais inoculados no dia 30 pi revelou a presença de DNA viral latente nos gânglios trigêmeos (TG, 5 de 5 animais), bulbo olfatório (BO, 5/5), ponte (2/5), cerebelo (2/5), córtex cerebral (1/5). Administração de dexametasona (Dx, n=4) ou flumetasona (FluM, n=4) a oito ovinos no dia 65 pi resultou em reativação e excreção viral por 3 de 4 animais de cada grupo. A excreção viral nas secreções nasais iniciou no dia 3 pós-tratamento e durou entre 1 e 5 dias nos ovinos tratados com Dx (títulos até 102,8TCID50/mL) e foi mais tardia, durando entre 1 e 3 dias nos animais tratados com FluM (títulos de 102,1 TCID50/mL). Uma análise por PCR do encéfalo dos animais submetidos à reativação, no dia 65 pós-infecção, revelou uma distribuição do DNA latente semelhante àquela observada nos animais não submetidos à reativação. Em resumo, a capacidade do BoHV-5 estabelecer infecção latente, a colonização dos TGs a BOs com DNA viral latente e a reativação induzida por corticoides são achados promissores para o uso de cordeiros como modelo para a infecção latente pelo BoHV-5.


#6 - Identification of Staphylococcus strains isolated from bovine mastitis by PCR and 16S rDNA sequencing, 31(1):36-40

Abstract in English:

ABSTRACT.- Lange C.C., Brito M.AV.P., Brito J.R.F., Arcuri E.F., Souza G.N., Machado M.A., Domingues R. & Salimena A.P.S. 2011. [Identification of Staphylococcus strains isolated from bovine mastitis by PCR and 16S rDNA sequencing.] Uso de PCR e sequenciamento do rDNA 16S para identificação de bactérias do gênero Staphylococcus isoladas de mastite bovina. Pesquisa Veterinária Brasileira 31(1):36-40. Embrapa Gado de Leite, Rua Eugênio do Nascimento 610, Juiz de Fora, MG 36030-330, Brazil. E-mail: clange@cnpgl.embrapa.br The objective of this study was to identify the species of 100 isolates of Staphylococcus from mastitis in dairy cows from herds located in the state of Minas Gerais, Brazil. PCR reactions were carried out using specific primers described previously for S. aureus (femA gene), S. intermedius (16S rDNA) and S. hyicus (16S-23S rDNA spacer region). In addition, products of amplification of variable regions of the 16S rDNA gene of the strains were sequenced. According to the results of the PCR, 83 strains were identified as S. aureus, 13 as S. intermedius, two as S. hyicus and two isolates were not identified. The sequencing of 16S rDNA was applied to 23 strains identified by PCR amplifications: six S. aureus and the strains identified as S. intermedius (n=13), S. hyicus (n=2) or not identified (n=2). The sequencing of 16S rDNA confirmed the six strains as S. aureus. The others 17 strains were identified as S. chromogenes (13 isolates) and S. hyicus (four isolates). Each sample was related to a specie according to the smallest E-value and highest similarity (³ 99%). The identification of S. hyicus and S. chromogenes was accomplished only by 16S rDNA sequencing.

Abstract in Portuguese:

RESUMO.- Lange C.C., Brito M.AV.P., Brito J.R.F., Arcuri E.F., Souza G.N., Machado M.A., Domingues R. & Salimena A.P.S. 2011. [Identification of Staphylococcus strains isolated from bovine mastitis by PCR and 16S rDNA sequencing.] Uso de PCR e sequenciamento do rDNA 16S para identificação de bactérias do gênero Staphylococcus isoladas de mastite bovina. Pesquisa Veterinária Brasileira 31(1):36-40. Embrapa Gado de Leite, Rua Eugênio do Nascimento 610, Juiz de Fora, MG 36030-330, Brazil. E-mail: clange@cnpgl.embrapa.br O objetivo deste trabalho foi identificar espécies de Staphylococcus (n=100) isoladas de mastite em rebanhos bovinos do Estado de Minas Gerais. Para esta finalidade foram utilizadas reações de PCR empregando oligonucleotídeos iniciadores descritos anteriormente para amplificar genes específicos de S. aureus (femA), S. intermedius (rDNA 16S) e S. hyicus (rDNA 16S-23S) e o sequenciamento do rDNA 16S. De acordo com as reações de PCR, 83 isolados foram identificados como S. aureus, 13 isolados como S. intermedius, dois como S. hyicus e dois isolados não foram identificados. Foram submetidos ao sequenciamento do rDNA 16S seis isolados identificados como S. aureus e os 17 restantes. Os seis isolados identificados como S. aureus confirmaram essa identificação. Dos outros 17 isolados, 13 foram identificados como S. chromogenes e quatro como S. hyicus, com similaridade igual ou superior a 99%. Baseando-se nos resultados da reação de PCR do gene femA e do sequenciamento do rDNA 16S, foram identificados 83 S. aureus, 13 S. chromogenes e quatro S. hyicus. Neste estudo os oligonucleotídeos iniciadores empregados na reação de PCR para S. intermedius não foram específicos, pois amplificaram também S. chromogenes; e os empregados na reação de PCR para S. hyicus não foram sensíveis, pois falharam na identificação de dois isolados de S. hyicus. A identificação definitiva das duas últimas espécies somente foi possível pelo sequenciamento do rDNA 16S.


#7 - Nuclear and mitochondrial DNA markers in traceability of retail beef samples, 30(9):782-786

Abstract in English:

ABSTRACT.- Cesar A.S.M., Biase F.H., Ripamonte P., Luchiari Filho A., Merighe G.K. & Meirelles F.V. 2010. Nuclear and mitochondrial DNA markers in traceability of retail beef samples. Pesquisa Veterinária Brasileira 30(9):783-786. Laboratório de Morfofisiologia Molecular e Desenvolvimento, Faculdade de Zootecnia e Engenharia de Alimentos, Universidade de São Paulo, Av. Duque de Caxias Norte 225, Pirassununga, SP 13635-900, Brazil. E-mail: meirellf@usp.br Several characteristics are important in a traceability system of animal products, such as age at slaughter, breed composition, besides information of the productive chain. In general, the certification agent records information about the animals and the system which it came from, although cannot guarantee that the slaughtering, meat processing and distribution are error proof. Besides, there is a differential price, at least at the international market, based on sex and breed composition of the animals. Genetic markers allow identification of characteristics controlled in the beef cattle traceability program, as sex and breed composition, in order to correctly identify and appraise the final product for the consumer. The hypothesis of this study was that the majority beef samples retailed in the local market originate from female with a great participation of zebu breeds. Therefore, the objective of this work was to characterize retail beef samples with DNA markers that identify cattle sex and breed composition. Within 10 beef shops localized in Pirassununga, SP, Brazil, 61 samples were collected, all were genotyped as harboring Bos taurus mitochondrial DNA and 18 were positive for the Y chromosome amplification (male). For the marker sat1711b-Msp I the frequency of the allele A was 0.278 and for the marker Lhr-Hha I the frequency of the allele T was 0.417. The results of sat1711b-Msp I and Lhr-Hha I allelic frequencies are suggestive that the proportion of indicus genome compared with the taurine genome in the market meat is smaller than the observed in the Nellore breed. The procedure described in this study identified sex and subspecies characteristics of beef meat samples, with potential application in meat products certification in special as an auxiliary tool in beef cattle traceability programs.

Abstract in Portuguese:

RESUMO.- Cesar A.S.M., Biase F.H., Ripamonte P., Luchiari Filho A., Merighe G.K. & Meirelles F.V. 2010. Nuclear and mitochondrial DNA markers in traceability of retail beef samples. [Marcadores de DNA nuclear e mitocondrial para rastreabilidade da carne bovina comercializada.] Pesquisa Veterinária Brasileira 30(9):783-786. Laboratório de Morfofisiologia Molecular e Desenvolvimento, Faculdade de Zootecnia e Engenharia de Alimentos, Universidade de São Paulo, Av. Duque de Caxias Norte 225, Pirassununga, SP 13635-900, Brazil. E-mail: meirellf@usp.br Várias características são importantes no sistema de rastreabilidade, como o sexo, a idade, a raça e/ou a composição racial dos animais, além de dados da cadeia produtiva. Em geral, a empresa certificadora dispõe das informações do animal que está sendo abatido, porém não tem condições de garantir se houve erro entre abate, desossa, processamento e a distribuição dos produtos. Existe diferenciação no custo e na qualidade dos produtos cárneos, especialmente no mercado internacional, em virtude do sexo e composição racial dos animais. Os marcadores genéticos permitem identificar as características que são controladas num programa de rastreabilidade bovina tais como sexo e composição racial, permitindo identificar e avaliar corretamente para o consumidor, o produto final. A hipótese deste estudo foi que a maioria das amostras de carne bovina vendida no mercado local seria proveniente de fêmeas e com grande participação de raças Zebu. O objetivo deste trabalho foi caracterizar amostras de carne bovina com marcadores de DNA para identificar o sexo e a composição racial. Em dez pontos comerciais da cidade de Pirasssununga, SP, Brasil, foram coletadas 61 amostras e todas foram genotipadas como possuindo DNA mitocondrial Bos taurus e 18 foram positivos para amplificação do cromossomo Y (macho). Para o marcador sat1711b-Msp I a frequência alélica do A foi 0.278 e para o marcador Lhr-Hha I a frequência alélica do T foi 0.417. Os resultados das frequências alélicas do sat1711b-Msp I e Lhr-Hha I apresentaram menor proporção do genoma Bos indicus em relação ao Bos taurus quando comparado ao rebanho Nelore. Com a metodologia descrita neste trabalho foi possível avaliar o sexo e as características de subespécie das amostras de carne bovina, tendo uma importante aplicação para a certificação de produtos cárneos especialmente, em programas de rastreabilidade animal.


#8 - Development and evaluation of a recombinant DNA vaccine candidate expressing porcine circovirus 2 structural protein, p.76-82

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.

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.


#9 - Clonal study of avian Escherichia coli strains by fliC conserved-DNA-sequence regions analysis, p.508-514

Abstract in English:

ABSTRACT.- Campos T.A., Nakazato G., Stehling E.G., Brocchi M. & Silveira W.D. 2008. Clonal study of avian Escherichia coli strains by fliC conserved-DNA-sequence regions analysis. Pesquisa Veterinária Brasileira 28(10):508-514. Departamento de Microbiologia e Imunologia, Instituto de Biologia, Cx. Postal 6109, Universidade Estadual de Campinas, Cidade Universitária Zeferino Vaz s/n, Barão Geraldo, Campinas, SP 3081-862, Brazil. *Corresponding author: wds@unicamp.br The clonal relationship among avian Escherichia coli strains and their genetic proximity with human pathogenic E. coli, Salmonela enterica, Yersinia enterocolitica and Proteus mirabilis, was determined by the DNA sequencing of the conserved 5’ and 3’regions fliC gene (flagellin encoded gene). Among 30 commensal avian E. coli strains and 49 pathogenic avian E. coli strains (APEC), 24 commensal and 39 APEC strains harbored fliC gene with fragments size varying from 670bp to 1,900bp. The comparative analysis of these regions allowed the construction of a dendrogram of similarity possessing two main clusters: one compounded mainly by APEC strains and by H-antigens from human E. coli, and another one compounded by commensal avian E. coli strains, S. enterica, and by other H-antigens from human E. coli. Overall, this work demonstrated that fliC conserved regions may be associated with pathogenic clones of APEC strains, and also shows a great similarity among APEC and H-antigens of E. coli strains isolated from humans. These data, can add evidence that APEC strains can exhibit a zoonotic risk.

Abstract in Portuguese:

ABSTRACT.- Campos T.A., Nakazato G., Stehling E.G., Brocchi M. & Silveira W.D. 2008. Clonal study of avian Escherichia coli strains by fliC conserved-DNA-sequence regions analysis. Pesquisa Veterinária Brasileira 28(10):508-514. Departamento de Microbiologia e Imunologia, Instituto de Biologia, Cx. Postal 6109, Universidade Estadual de Campinas, Cidade Universitária Zeferino Vaz s/n, Barão Geraldo, Campinas, SP 3081-862, Brazil. *Corresponding author: wds@unicamp.br The clonal relationship among avian Escherichia coli strains and their genetic proximity with human pathogenic E. coli, Salmonela enterica, Yersinia enterocolitica and Proteus mirabilis, was determined by the DNA sequencing of the conserved 5’ and 3’regions fliC gene (flagellin encoded gene). Among 30 commensal avian E. coli strains and 49 pathogenic avian E. coli strains (APEC), 24 commensal and 39 APEC strains harbored fliC gene with fragments size varying from 670bp to 1,900bp. The comparative analysis of these regions allowed the construction of a dendrogram of similarity possessing two main clusters: one compounded mainly by APEC strains and by H-antigens from human E. coli, and another one compounded by commensal avian E. coli strains, S. enterica, and by other H-antigens from human E. coli. Overall, this work demonstrated that fliC conserved regions may be associated with pathogenic clones of APEC strains, and also shows a great similarity among APEC and H-antigens of E. coli strains isolated from humans. These data, can add evidence that APEC strains can exhibit a zoonotic risk.


#10 - Genotype characterization of the horn fly Haematobia irritans from different Brazilian geographic regions based on randomly amplified polymorphic DNA (RAPD) analysis, p.1-5

Abstract in English:

ABSTRACT.- Brito L.G., Regitano L.C.A., Huacca M.E.F., Carrilho E. & Moya Borja G.E. 2007. Genotype characterization of the horn fly Haematobia irritans from different Brazilian geographic regions based on randomly amplified polymorphic DNA (RAPD) analysis. Pesquisa Veterinária Brasileira 27(1):1-5. Laboratório de Sanidade Animal, Embrapa Rondônia, BR 364 Km 5,5, Porto Velho, RO 78900-970, Brazil. E-mail: luciana@cpafro.embrapa.br Blood-sucking diptera are important parasites in bovine production systems, especially regarding confinement conditions. Haematobia irritans, the horn fly, is one of the most troublesome species within bovine production systems, due to the intense stress imposed to the animals. An important aspect while studying the variability within a species is the study of the geographic structure of its populations and, attempting to find out the genetic flow of Brazilian populations of horn fly, the RAPD technique, which is suited for this purpose, has been used. The use of molecular markers generated from RAPD made it possible to identify the geographic origin of samples from different Brazilian geographic regions, as well as to estimate the genotypic flow among the different Brazilian populations of the horn fly.

Abstract in Portuguese:

ABSTRACT.- Brito L.G., Regitano L.C.A., Huacca M.E.F., Carrilho E. & Moya Borja G.E. 2007. Genotype characterization of the horn fly Haematobia irritans from different Brazilian geographic regions based on randomly amplified polymorphic DNA (RAPD) analysis. Pesquisa Veterinária Brasileira 27(1):1-5. Laboratório de Sanidade Animal, Embrapa Rondônia, BR 364 Km 5,5, Porto Velho, RO 78900-970, Brazil. E-mail: luciana@cpafro.embrapa.br Blood-sucking diptera are important parasites in bovine production systems, especially regarding confinement conditions. Haematobia irritans, the horn fly, is one of the most troublesome species within bovine production systems, due to the intense stress imposed to the animals. An important aspect while studying the variability within a species is the study of the geographic structure of its populations and, attempting to find out the genetic flow of Brazilian populations of horn fly, the RAPD technique, which is suited for this purpose, has been used. The use of molecular markers generated from RAPD made it possible to identify the geographic origin of samples from different Brazilian geographic regions, as well as to estimate the genotypic flow among the different Brazilian populations of the horn fly.


Colégio Brasileiro de Patologia Animal SciELO Brasil CAPES CNPQ UNB UFRRJ CFMV