Diagnóstico sorológico da brucelose bovina em animais adultos vacinados com dose reduzida da cepa 19 de Brucella abortus

Com o presente trabalho avaliou-se o uso de dose reduzida da vacina produzida com a amostra 19 de Brucella abortus, em rebanho adulto negativo para a enfermidade, por meio de técnicas de diagnóstico sorológico preconizadas pelo Programa Nacional de Controle e Erradicação da Brucelose e Tuberculose Animal e por um ensaio indireto de imunoadsorção enzimática (ELISA ID). A prova de fixação de complemento detectou 46,77% de positivos, o antígeno acidificado tamponado 67,74%, o 2-mercaptoetanol com soroaglutinação lenta 87,09% e o ELISA ID 100%. A dose reduzida interferiu no diagnóstico sorológico. Nenhuma das técnicas apresentou especificidade adequada para uso em rebanho nestas condições, até 3 meses após a vacinação.

Distribuição da hipovitaminose A no Brasil nas últimas quatro décadas: ingestão alimentar, sinais clínicos e dados bioquímicos

O presente estudo teve como objetivo o registro e a apresentação de trabalhos realizados no Brasil nos últimos 40 anos, relacionados com a investigação sobre a deficiência de vitamina A. Esta deficiência tem sido diagnosticada por um ou mais dos seguintes critérios: ingestão deficiente de alimentos fontes de vitamina A, exame clínico, níveis séricos de retinol abaixo dos aceitos como normais, concentração hepática de retinol, teste de adaptação ao escuro e corante de Rosa Bengala. A deficiência foi diagnosticada em grupos populacionais de vários Estados e capitais brasileiras em cidades grandes e pequenas e em zonas rurais. A maioria dos trabalhos foi desenvolvida em grupos populacionais de baixa renda. Quanto às conseqüências clínicas, relataram-se achados de sinais oculares leves, como cegueira noturna, manchas de Bitot e xerose conjuntival, encontrados principalmente na Região Nordeste. Alguns autores observaram, em menor número de casos, lesões graves, como lesões corneanas e ceratomalácia. Trabalhos da última década indicaram associação entre a hipovitaminose A e o aumento da morbidade e mortalidade, principalmente em crianças pré-escolares.

Avaliação do teste do anel em leite na vigilância epidemiológica da brucelose bovina em rebanhos e em laticínios

Analisaram-se 464 amostras individuais de leite e 54 amostras de leite de latões, oriundos de leite dos mesmos animais, por meio do teste do anel do leite (TAL) visando à sua aplicação no diagnóstico individual e de rebanhos da brucelose bovina. Foram também avaliadas 464 amostras de soro sangüíneo por meio de provas do antígeno tamponado acidificado (ATA), soroaglutinação lenta em tubos (SAL) e 2-mercaptoetanol (2-ME), todas para brucelose. Cento e vinte e três amostras (26,5%) testadas pelo TAL apresentaram resultados positivos. Dessas, 30 resultaram positivas ao ATA, 28 ao ATA, à SAL e ao 2-ME e 18 à SAL. Das amostras positivas ao TAL, 95 pertenciam a animais sorologicamente negativos ao 2-ME, caracterizando 77,2% (95/123) das reações falso-positivas; dos resultados negativos ao TAL, 4 pertenciam a animais sorologicamente positivos, caracterizando 1,2% (4/341) de reações falso-negativas no TAL individual. Das 54 amostras de leite de latões analisadas pelo TAL, 17 foram consideradas positivas, das quais uma foi caracterizada como falso-positivo, pois todos os animais que a compunham foram negativos ao 2-ME. de 37 latões considerados negativos ao TAL, três continham leite de animais positivos ao 2-ME, caracterizando 8,1% de falso-negativos. O TAL individual demonstrou elevado percentual de resultados falso-positivos, enquanto o TAL em amostras de leite obtidas em latões detectou 84,2% de latões contaminados e 75% de rebanhos infectados.

Comparison of infection by Brucella spp. in free-ranging and captive wild animals from São Paulo State, Brazil

The aim of the current study was to evaluate the infection rate by Brucella spp. in wild and in captive animals. Serum samples from 121 animals (94 free-ranging and 27 captive) of different mammal species were evaluated. Sera were submitted to rose Bengal test (RBT) for screening and serum agglutination tests (SAT) and 2-mercaptoethanol test (2-ME) for confirmatory results. Nine animals (five free-ranging and four captive) tested positive in RBT, but negative in the confirmatory tests. Several domestic animal diseases that have control programs are not focused on wild reservoirs, such as brucellosis in Brazil. The study of new reservoirs in wildlife is essential to prevent emerging diseases.

Evaluation of fertilizing potential of frozen-thawed dog spermatozoa diluted in ACP-106 (R) using an in vitro sperm-oocyte interaction assay

The aim of present study was to evaluate frozen canine semen with ACP-106 (R) (Powder Coconut Water) using an in vitro sperm-oocyte interaction assay (SOIA). Ten ejaculates from five stud dogs were diluted in ACP-106 (R) containing 20% egg yolk, submitted to cooling in a thermal box for 40 min and in a refrigerator for 30 min. After this period, a second dilution was performed using ACP-106 (R) containing 20% egg yolk and 12% glycerol. Samples were thawed at 38 degrees C for 1 min. Post-thaw motility was evaluated by light microscopy and by using a computer aided semen analysis (CASA). Plasma membrane integrity and sperm morphology/acrosomal status were evaluated by fluorescent probes (C-FDA/PI) and Bengal Rose respectively. Moreover, frozen-thawed semen was analysed by a SOIA. Subjective post-thaw motility was 52.0 +/- 14.8% and it was significant higher than the total motility estimated by CASA (23.0 +/- 14.8%) because this system considered the egg yolk debris as immotile spermatozoa. Although normal sperm rate and acrosomal integrity evaluated by Bengal Rose stain was 89.6 +/- 3.1 % and 94.3 +/- 3.1 %, respectively, post-thaw percentage of intact plasma membrane was only 35.1 +/- 14.3%. Regarding SOIA, the percentage of interacted oocytes (bound, penetrated and bound and/or penetrated) was 75.3%. Using regression analysis, it was found significant relations between some CASA patterns and data for SOIA. In conclusion, the freezing-thawing procedure using ACP-106 (R) was efficient for maintain the in vitro fertility potential of dog spermatozoa.

Prognostic value of canine frozen-thawed semen parameters on in vitro sperm-oocyte interactions

Combining the data from conventional semen analysis with oocyte penetration assays should improve the assessment of the fertilizing ability of a semen sample. Thus, the objective of the present study was to evaluate the prognostic value of various semen parameters on the in vitro interactions between frozen-thawed canine sperm and homologous oocytes. Ten ejaculates from five stud dogs (two ejaculates/dog) were collected by digital manipulation. Semen samples were evaluated, extended in Tris-egg yolk-glycerol, frozen and stored in liquid nitrogen, and thawed several weeks later. Samples were evaluated for motility and sperm populations by computer-aided semen analysis (CASA), plasma membrane integrity (carboxy-fluorescein diacetate and propidium iodide), and sperm morphology (Bengal Rose). Thawed spermatozoa were also incubated with homologous oocytes for 18 h in an atmosphere of 5% CO2 and 95% air at 38 degrees C and sperm-oocyte interactions were evaluated. Simple linear regression models were calculated, with sperm parameters as independent variables and sperm-oocyte interactions as the dependent variable. There were significant associations between: percentage of oocytes bound to spermatozoa and beat cross frequency (BCF; R-2 = 63%); percentage of oocytes that interacted with spermatozoa and BCF (R-2 = 73%); and number of penetrated spermatozoa and velocity average pathway (VAP; R-2 = 64%) and velocity straight line (VSL; R-2 = 64%). Although plasma membrane integrity and sperm morphology had little prognostic value for in vitro interactions between canine frozen-thawed sperm and homologous oocytes, some motility patterns (evaluated by CASA) were predictive of in vitro sperm-oocyte interactions. (c) 2005 Elsevier B.V. All rights reserved.

Ácaros predadores associados ao ácaro-da-erinose da lichia

O objetivo deste trabalho foi identificar ácaros predadores em plantas de lichia e correlacionar o desenvolvimento populacional dessas espécies com o do ácaro-da-erinose da lichia, Aceria litchii. A pesquisa foi desenvolvida no Município de Casa Branca, SP, com árvores adultas, de 12 anos de idade, da variedade Bengal. Mensalmente, de agosto de 2008 a setembro de 2009, foram coletadas folhas para avaliação dos níveis populacionais de A. litchii e de ácaros predadores. Foram registrados 6.557 indivíduos da família Phytoseiidae. A espécie mais abundante foi Amblyseius compositus (42,6%), seguida por Phytoseius intermedius (31,2%), Euseius concordis (14,1%), Amblyseius herbicolus (8,8%) e Iphiseiodes zuluagai (3,3%). O desenvolvimento populacional de A. compositus, E. concordis e I. zuluagai correlacionou-se positivamente com o de Aceria litchii, o que indica relação de predação.

Imunodifusão em gel de ágar com polissacarídeos de membrana de Brucella abortus 1119-3 no diagnóstico da brucelose bovina

Comparou-se a prova de imunodifusão em gel de ágar (IDGA), utilizando extrato polissacarídico (POLI O), obtido da amostra de B. abortus 1119-3, com os testes de soroaglutinação rápida em placa, de soroaglutinação lenta em tubos, de antígeno acidificado e de 2-mercaptoetanol para o diagnóstico da brucelose bovina. O IDGA mostrou alta especificidade, porém sensibilidade inferior aos métodos convencionais.

Aglutininas anti-Brucella abortus no soro e em secreção de bursite cervical em eqüinos

Fistulous wither secretions from three horses were tested by the plate agglutination (PAT), tube agglutination (SAT), buffered plate-Rose Bengal (RBPT) and 2-mercaptoethanol (2-ME) tests, comparatively with standard agglutination tests. In the modified tests, titers were increased in the PAT, SAT and 2-ME tests and positive reaction was observed in RBPT. Brucella abortus was isolated from the secretion of fistulous withers collected from one animal. These results suggest that the modified tests may be used as alternative tests to diagnose brucellosis in horses with fistulous withers.

Ocorrência de anticorpos anti-Brucella abortus e anti-Brucella canis em cães rurais e urbanos do Município de Monte Negro, Rondônia, Brasil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Perfil sorológico anti-Brucella abortus em bezerras vacinadas com amostras B19

The serological profiles of 33 female bovines, vaccinated at three to eight months of age with the B19 standard strain of Brucella abortus, were studied over a period of 728 days, using the following agglutination procedures: plate agglutination, tube agglutination, rose bengal plate and mercaptoethanol test. Maximum levels of antibodies detected reached by the plate agglutination and tube agglutination tests were found between the 14(th) and 42(nd) day, and with mercaptoethanol test, between the 28(th) and 42(nd) day. Anti-Brucella antibodies decreased thereafter. At 182 days after vaccination, five suspected animals and one positive were detected by the plate agglutination test, while by the tube agglutination test, only one animal was suspected and another one was positive. During the same period, positive reactions were found in six animals by means of the mercaptoethanol test, and five positives by the rose bengal test. By means of tube agglutination and plate agglutination tests, the animals became serologically negative at 245 and 273 days, respectively, after the vaccination, based on the rules adopted for the vaccinated animals. Using the mercaptoethanol and rose bengal plate tests, all the animals were found to be negative at 308 days after vaccination.

Incorporação de mucuna-preta e de restos culturais de milho antes da cultura do feijão (Phaseolus vugaris L.) de inverno

The experiments were conducted during two years (1990/91 and 1992/93), to study the effects of the incorporation of bengal velvetbean and of residues on the development and yield of the winter bean crops. The trial was developed in experimental field at the Faculdade de Engenharia de Ilha Solteira, UNESP, located in Selvíria, MS, Brazil, in dark red latosol covered by savanna vegetation. It was concluded that: the incorporation of residues of corn or corn + bengal velvetbean or only bengal velvetbean had no effects on soil chemical characteristics, evaluated 80 days after the residues incorporation; in yield and bean crop development, no statistical differences were observed among treatments.

Susceptibility of planktonic cultures of Streptococcus mutans to photodynamic therapy with a light-emitting diode

The objective of this study was to evaluate the effect of photodynamic therapy with erythrosine and rose bengal using a light-emitting diode (LED) on planktonic cultures of S. mutans. Ten S. mutans strains, including nine clinical strains and one reference strain (ATCC 35688), were used. Suspensions containing 10 6 cells/mL were prepared for each strain and were tested under different experimental conditions: a) LED irradiation in the presence of rose bengal as a photosensitizer (RB+L+); b) LED irradiation in the presence of erythrosine as a photosensitizer (E+L+); c) LED irradiation only (P-L+); d) treatment with rose bengal only (RB+L-); e) treatment with erythrosine only (E+L-); and f) no LED irradiation or photosensitizer treatment, which served as a control group (P-L-). After treatment, the strains were seeded onto BHI agar for determination of the number of colony-forming units (CFU/mL). The results were submitted to analysis of variance and the Tukey test (p ≤ 0.05). The number of CFU/mL was significantly lower in the groups submitted to photodynamic therapy (RB+L+ and E+L+) compared to control (P-L-), with a reduction of 6.86 log 10 in the RB+L+ group and of 5.16 log 10 in the E+L+ group. Photodynamic therapy with rose bengal and erythrosine exerted an antimicrobial effect on all S. mutans strains studied.

Brazil Anticorpos anti-Brucella canis e anti-Brucella abortus em caes de Araguaina, Tocantins

The aims of the present study were to determine the seroprevalence of infection by Brucella canis and Brucella abortus and to evaluate possible risk factors for infection in dogs from Araguaina, Tocantins, Brazil. Sera from 374 dogs, of the urban zones of the municipality, from both sexes, were submitted to the agar-gel immunodiffusion for Brucella canisantibodies and to rose Bengal test (AAT) and fluorescence polarization assay (FPA) for Brucella abortus-antibodies. From the 374 tested dogs, 21 reacted in the AAT, but no one was positive in the FPA. The seroprevalence of B. canis infection found in Araguaina, Tocantins, Brazil, was 44.53% (95% IC; 39.43 to 49.72). No association was found among seropositivity for B. canis and the risk factors studied. Thus, data from the present study showed that there was no infection by B. abortus among dogs in the sample and that infection by B. canis is widespread and at high prevalence in Araguaina, Tocantins, Brazil.

Lychee propagation (Litchi chinensis Sonn.): A comparative study of rootstocks

The lychee (Litchi chinensis Sonnerat), of the family Sapindaceae, is a highly valued fruit throughout the world. It is native to southern China, but it is cultivated in many parts of the world. In Brazil, the cultivation of the lychee was initiated in the 1970s in the State of São Paulo and has been increasing in production even though the crop size still averages only 1.4 ha per producer. The low crop size is attributed to the high cost of seedlings and the amount of time required for the plant to bear fruit. These factors discourage planting in large areas. The purpose of this research is to study some aspects of what is vitally important for the production of lychee seedlings on a large scale, with low cost and good genetic quality. Tests were performed using rootstocks from seeds of lychee and longan (Euphoria longan Lam.). Scions for grafting were taken from plants of lychee 'Bengal' in the collection of fruit species from FCAV-UNESP, Campus of Jaboticabal, Brazil. The experiments were started in September 2007 and the grafting process was performed in each month of the year finishing in August 2008. During this period the influence of these criteria were evaluated: 1) time of the year; 2) species of rootstock; 3) percentage of grafting success; 4) stem height at grafting point; 5) stem diameter at grafting point. Therefore, through statistical analysis we obtained significant differences in relation to the rootstock used, the months of the year in which the grafting was performed and the interaction between rootstock and the month in which the grafting was performed.