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O presente trabalho objetivou a produção e caracterização de um anticorpo monoclonal (PST 33) reativo com os timócitos e linfócitos T de surubim híbrido. PST 33 foi produzido por meio de imunização de camundongos com timócitos de surubim e moléculas isoladas das membranas celulares destas células. PST 33 mostrou-se reativo com a maioria dos timócitos e com linfócitos T periféricos. Por citometria de fluxo foi possível quantificar os linfócitos T no sangue periférico (34,1%), baço (36,6%), rim (19,1%) e timo (87,5%). Esse marcador reagiue com uma molécula de 215 kDa expressa na maioria dos linfócitos T. Seu uso em imunohistoquímica permitiu também a localização de linfócitos T no timo e baço, demonstrando assim o seu potencial para estudar a ontogenia do sistema imune dos surubins. Em imuno-electron microscopia PST 33 reagiu com células de aparência linfocítica. Morfologicamente essas células apresentaram-se como pequenos linfócitos de elevada relação núcleo/citoplasmaA monoclonal antibody against hybrid surubim thymocytes and circulating T-cells (PST 33) was produced and characterized. The mAb was obtained by immunizing mice with isolated membrane molecules of hybrid surubim thymocytes, and whole thymocytes. Flow cytometric analysis showed that mAb PST 33 reacted with 87.5% of the thymocytes, 34.1% of circulating, 19.1% of pronephros, and 36.6% of spleen lymphoid cells. Immunohistochemistry demonstrated that PST 33-immunoreactive cells are mostly present in the medullar region of the thymus, and in the vicinity of arterioles spread throughout the parenchyma of the spleen. PST 33 appeared as a suitable marker for immunohistochemistry, and should prove useful in studying the ontogeny of surubim immune system. Immunogold labeling revealed that PST 33+ cells were lymphoid-like cells. Morphologically they appeared as small lymphocytes with a high nucleus/cytoplasm ratio. Western blotting of non-reduced membrane lysates showed that PST 33 reacted with a 215 kDa molecule

Monoclonal antibodies (mAb) against hybrid surubim immunoglobulin (IgM) were produced. Screening, with an indirect enzyme-linked immunosorbent assay (ELISA), revealed three probes: PSIg 10, PSIg 13 and PSIg 28 specifically reacting to surubim IgM. No species cross-reactivity was found. Isotyping revealed these probes as belonging to the IgG2b/κ subclass, and recognising, exclusively, the heavy chain of surubim IgM on immunoblotting. The most interesting mAb, PSIg 13, was employed to detect, for the first time, Ig-bearing cells of surubim in peripheral blood, spleen and head-kidney by FACS analysis and immunocytochemistry. The mAb raised here recognises the antigen in its native form, as denatured heavy-chain subunits and when expressed in living and fixed cells.

This paper describes the results of our initial studies with hybrid surubim catfish leucocytes and suggests that the techniques used may have further application. According to size, shape, staining patterns and ultrastructure, five blood leucocyte types were identified in the peripheral blood of surubim: lymphocytes, thrombocytes, monocytes, neutrophils and eosinophils. A tentative plasma cell was also observed by electron microscopy. Haematocrit and leucocrit were also analysed and reference values for these parameters are proposed. The results obtained in the present study should be useful to support further evaluations, and to develop or refine methods to properly evaluate the health status of this fish.

Foram analisadas amostras sangüíneas de seis exemplares de Pseudoplatystoma corruscans. As amostras foram coletadas por punção caudal e foram determinados os valores e as médias de hematócrito, leucócrito, eritrócitos e leucócitos e das concentrações protéicas plasmáticas. O plasma sangüíneo foi submetido a eletroforese, para determinação das diferentes frações protéicas. A partir de extensões sangüíneas coradas com métodos rotineiros, cinco tipos celulares puderam ser distinguidos sob microscopia ótica. A média das células sangüíneas totais foi de 1.29 x 106 células.ml-1, enquanto que a concentração protéica média do plasma sangüíneo foi 11.82g.l-1, constituindo-se de cinco frações principais. Essas características hematológicas foram estabelecidas no intuito de se obter um primeiro quadro hematológico da espécie, no qual serão baseados estudos mais aprofundados.The ranges and mean values of haematocrit, leucocrit, red cell, white cell and differential cell counts and serum protein concentration of young growing Pseudoplatystoma corruscans (n=6) were measured, and serum electrophoretic patterns on cellulose acetate described. Blood was obtained by puncture of the caudal vessels. Using routine staining methods, five different cell types could be distinguished by light microscopy. Mean total cell count was 1.29 x 106 cells.ml-1, while mean plasma protein content was 11.82g.l-1 consisting of five major protein fractions. These haematological characteristics were established as a “working document” upon which to base more extensive studies.

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2013-08-22T19:01:06Z No. of bitstreams: 1 S0103-90162006000600002.pdf: 58705 bytes, checksum: a2326f44ec1caaa6153bbe1149b25e7e (MD5) Made available in DSpace on 2013-08-22T19:01:06Z (GMT). No. of bitstreams: 1 S0103-90162006000600002.pdf: 58705 bytes, checksum: a2326f44ec1caaa6153bbe1149b25e7e (MD5) Previous issue date: 2006-12-01 Made available in DSpace on 2013-09-30T20:00:42Z (GMT). No. of bitstreams: 2 S0103-90162006000600002.pdf: 58705 bytes, checksum: a2326f44ec1caaa6153bbe1149b25e7e (MD5) S0103-90162006000600002.pdf.txt: 31243 bytes, checksum: a796e8e95d545c5e5c1117d450933794 (MD5) Previous issue date: 2006-12-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:18:11Z No. of bitstreams: 2 S0103-90162006000600002.pdf: 58705 bytes, checksum: a2326f44ec1caaa6153bbe1149b25e7e (MD5) S0103-90162006000600002.pdf.txt: 31243 bytes, checksum: a796e8e95d545c5e5c1117d450933794 (MD5) Made available in DSpace on 2014-05-20T13:18:11Z (GMT). No. of bitstreams: 2 S0103-90162006000600002.pdf: 58705 bytes, checksum: a2326f44ec1caaa6153bbe1149b25e7e (MD5) S0103-90162006000600002.pdf.txt: 31243 bytes, checksum: a796e8e95d545c5e5c1117d450933794 (MD5) Previous issue date: 2006-12-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Apesar da possível influência do tanino sobre o valor nutritivo das forrageiras da Caatinga, poucos são os estudos que avaliam a concentração de taninos nestas plantas. O objetivo do presente estudo foi caracterizar os taninos condensados presentes nas espécies Mimosa hostilis (Jurema Preta), Mimosa caesalpinifolia (Sabiá) e Bauhinia cheilantha (Mororó), em três fases do ciclo fenológico. As concentrações de tanino solúvel (TS), tanino ligado ao resíduo (TL) e tanino total (TT) foram determinadas pelo método butanol-HCl; a adstringência foi avaliada pelo método de difusão radial e a composição de monômeros dos taninos purificados através do sistema de cromatografia líquida de alta resolução, utilizando delfinidina, cianidina e pelargonidina como padrões. A concentração e adstringência dos taninos condensados purificados, assim como sua composição monomérica, variou entre as espécies e, em alguns casos, entre os ciclos fenológicos. Os valores foram superiores aos considerados benéficos a digestão ruminal (5%). Jurema Preta apresentou os maiores valores (30,98% TT e 22% de adstringência na vegetação plena) e Mororó os menores valores observados (10,38 TT e 14% de adstringência na frutificação). A Jurema Preta apresentou uma relação prodelfinidina (PD):procianidina (PC) média de 97:3 que se mostrou pouco variável, indicando uma alta capacidade adstringente dos taninos desta espécie em todas as fases do ciclo fenológico. O Sábia apresentou uma relação de 90:20 nas fases de vegetação plena e floração, diminuindo para 40:50 na fase de frutificação. A relação PD:PC do Mororó foi mais equilibrada, oscilando em torno de 40:50 nas fases de vegetação plena e floração e reduzindo para 35:60 durante a frutificação. A propelargonidina esteve ausente ou em pequena concentração nas espécies estudadas. Despite the possible influence of tannins on the nutritional value of the forages from Caatinga vegetation, there are few studies that evaluated their tannin concentration. This study was conducted to characterize condensed tannins present in the legumes species Mimosa hostilis (Jurema Preta), Mimosa caesalpinifolia (Sabiá) and Bauhinia cheilantha (Mororó), at three stages of their phenological cycle. The concentration of soluble tannin (ST), bound tannin (BT) and total tannin (TT) were determined using the butanol-HCL method; astringency was by the radial diffusion method, and the monomeric composition of purified tannins by a high-performance liquid chromatograph with delphinidin, cyanidin and pelargonidin as standards. Concentration and astringency of purified condensed tannins, as well as their monomeric composition varied between species, and in some cases among phenological cycles. The values observed were always above the limits considered beneficial for ruminal digestion (i.e. 5%). Jurema Preta presented the highest values (30.98% TT and 22% astringency at full growth stage), and Mororó the lowest (10.38% TT and 14% astringency during fructification). Jurema Preta presented a mean relationship prodelfinidin (PD): procyanidin (PC) of 97:3, which did not vary during the phenological cycle, showing the high astringent capacity of these tannins. Sabiá presented a relationship of 90:20 during full growth and flowering stages, decreasing to 40:50 at fructification. In Mororó the PD:PC relationship was more equilibrated, around 40:50 during full growth and flowering stages, decreasing to 35:60 During fructification. Propelargonidin was not detected or was present at low concentration in the three species. UFPB CCA Depto. de Zootecnia UNESP FCAV Depto. de Zootecnia Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA) Caprinos UNESP FCAV Depto. de Zootecnia

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2013-08-22T18:55:47Z No. of bitstreams: 1 S0102-09352006000500029.pdf: 227095 bytes, checksum: ab4d65b44cb0bf41a513439abe7acf75 (MD5) Made available in DSpace on 2013-08-22T18:55:47Z (GMT). No. of bitstreams: 1 S0102-09352006000500029.pdf: 227095 bytes, checksum: ab4d65b44cb0bf41a513439abe7acf75 (MD5) Previous issue date: 2006-10-01 Made available in DSpace on 2013-09-30T19:49:01Z (GMT). No. of bitstreams: 2 S0102-09352006000500029.pdf: 227095 bytes, checksum: ab4d65b44cb0bf41a513439abe7acf75 (MD5) S0102-09352006000500029.pdf.txt: 26268 bytes, checksum: a230b96c9390307868b31945af0c157f (MD5) Previous issue date: 2006-10-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:18:12Z No. of bitstreams: 2 S0102-09352006000500029.pdf: 227095 bytes, checksum: ab4d65b44cb0bf41a513439abe7acf75 (MD5) S0102-09352006000500029.pdf.txt: 26268 bytes, checksum: a230b96c9390307868b31945af0c157f (MD5) Made available in DSpace on 2014-05-20T13:18:12Z (GMT). No. of bitstreams: 2 S0102-09352006000500029.pdf: 227095 bytes, checksum: ab4d65b44cb0bf41a513439abe7acf75 (MD5) S0102-09352006000500029.pdf.txt: 26268 bytes, checksum: a230b96c9390307868b31945af0c157f (MD5) Previous issue date: 2006-10-01 Avaliaram-se o crescimento da população, a atividade in vitro da enzima 1,4-b-endoglucanase e a taxa de digestão de celulose em culturas de Ruminococcus flavefaciens FD1 na presença de 50, 100, 200 e 400µg/ml de taninos purificado das leguminosas Mimosa hostilis (Jurema Preta), Mimosa caesalpinifolia (Sabiá) e Bauhinia cheilantha (Mororó). O crescimento bacteriano, a atividade da endoglucanase e a digestão de celulose foram fortemente inibidos pela presença dos taninos condensados purificados das três espécies, entretanto, a intensidade da inibição foi variável em função da espécie da leguminosa e da concentração de tanino. The effects of tannin levels in three Brazilian browse legumes Mimosa hostilis (Jurema Preta), Mimosa caesalpinifolia (Sabiá) e Bauhinia cheilantha (Mororó) on growth and endoglucanase activity of Ruminococcus flavefaciens FD1 were evaluated. Isolated cultures of Ruminococcus flavefaciens FD1 were used to assess cellular growth, 1, 4-beta-endoglucanase activity, and cellulose digestion in the presence of 50, 100, 200 and 400µg/ml of purified tannins from the three legume species. Growth of FD1, 1, 4-beta-endoglucanase activity and cellulose digestion were highly inhibited by the presence of purified tannins of the three legume species. However, the intensity of inhibition varied in according to the legume species and tannin concentration. UFAL Centro de Ciências Agrárias Universidade Estadual Paulista Mississippi State University Universidade Estadual Paulista