Seleção dos animais e cuidados pré, dcurante e pós-cirúrgicos na fistulação ruminal em caprinos e ovinos.

Estudos experimentais em nutrição animal freqüentemente demandam uso de animais canulados para ter acesso ao conteúdo gastrointestinal sendo assim, são submetidos a procedimentos cirúrgicos para criação de uma fístula. Esta técnica experimental trouxe consideráveis avanços para nutrição de ruminantes, pois, foi possível, por exemplo, definir ou mensurar a fração do alimento que sofre degradação ruminal. A determinação desta fração dos alimentos é importante uma vez que atende com mais eficiência os requerimentos dos microrganismos ruminais melhorando a fermentação e conseqüentemente repercutindo positivamente no desempenho animal. O uso de animais em ensaios experimentais para gerar tais informações requer cuidados antes, durantes e após o procedimento cirúrgico e observações constantes a campo. Os resultados obtidos e a longevidade desses animais dependem de tais cuidados. Neste contexto o presente artigo aponta parâmetros adequados para seleção de animais que serão destinados a fistulação do rúmen além de cuidados pré, durante e pós-cirúrgicos.bitstream/CNPC-2010/23058/1/doc92.pd

Ensaio alternativo para triagem de extratos protéicos com atividade celulolítica.

Este comunicado apresenta uma metodologia de difusão radial em gel de agarose (DRGA) desenvolvida por Teather e Wood (1982), modificada por Rehman et al. (2009), e adaptada no Laboratório de Bioquímica da Embrapa Caprinos e Ovinos para avaliar a atividade celulolítica em extratos proteicos de microrganismos ruminais. A técnica baseia-se na difusão radial, em gel de agarose, das proteínas da amostra analisada. O gel contém o substrato específico para celulase, a carboximetilcelulose, que pode ser corada com o corante vermelho congo.bitstream/item/31686/1/UMT-Cot112.pd

Biodegradação do bagaço de cana-de-açúcar por microrganismos ruminais de caprinos e ovinos.

Resumo: No presente estudo objetivou-se avaliar a degradação do bagaço de cana-de-açúcar (BCA) integral (BIN) ou hidrolisado (BH) pela microbiota ruminal de caprinos e ovinos de raças naturalizadas do Nordeste brasileiro e o potencial desses animais como fontes de microrganismos e/ou enzimas celulolíticas para degradação da fibra do BCA. Para hidrólise do BCA foi utilizada uma solução de NaOH a 50%, 30% na matéria seca (MS). Foram determinadas as concentrações de MS, proteína bruta (PB), cinzas (CZ), fibra detergente neutro (FDN), fibra detergente ácido (FDA), celulose (Cel), hemicelulose (Hcel) e lignina (Lig). A degradação in situ da FDN foi determinada pela incubação ruminal em sacos de náilon do BH nos tempos: 0, 6, 24 e 96 horas. A técnica de duas etapas preconizada por Tilley e Terry foi utilizada para determinar a digestibilidade in vitro da MS (DIVMS). Foi coletado conteúdo ruminal dos animais quatro horas após a infusão de 100g de BIN via fístula ruminal para a separação de microrganismos associados às fases líquida e sólida, utilizando tampão fosfato de sódio 50 mM, pH 6,9. A fração sólida foi submetida ao cultivo in vitro com o substrato BIN, por 96 horas, para determinação da taxa de degradação da MS, FDN e atividade celulolítica. O pH foi determinado nos tempos de cultivo de 24, 48 e 96 horas. O pré-tratamento com NaOH aumentou a DIVMS (P0,05). A inclusão de NaOH aumentou a degradação in situ da FDN, apresentando os ovinos menor tempo de colonização (TC). Houve solubilização da Hcel, da Cel e da Lign no BCA pré-tratado com NaOH. A atividade celulolítica se concentrou na fração sólida independente da espécie doadora do inóculo, sendo observado crescimento microbiano no cultivo in vitro do BIN à partir dessa fração. O pH aumentou com o tempo de cultivo in vitro. Microrganismos ruminais de Caprinos e ovinos naturalizados do Nordeste brasileiro colonizaram e degradaram o BIN e BH. O NaOH pode ser utilizado no pré-tratamento alcalino do BCA. [Biodegradation of sugarcane bagasse by ruminal microorganisms from sheep and goats]. Abstract: The purpose of this study was to evaluate the biodegradation of sugar cane bagasse (SCB), whole (WB) or hydrolyzed (HB) by ruminal microorganisms from Brazilian naturalized goat and sheep breeds, and the animals? potential as a source of microorganisms and/or cellulolytic enzymes for the biodegradation of SCB fiber. SCB hydrolysis was performed using 50% sodium hydroxide solution on 30% of dry matter (DM). The concentrations of DM, crude protein (CP), ether extract (EE), ash, neutral detergent fiber (NDF), acid detergent fiber (ADF), cellulose (Cel), hemicellulose (Hcel) and lignin (Lig) were determined in the WB and HB. For the in situ biodegradation of NDF, the HB was placed in nylon bags and incubated in rumen for 0, 6, 24 and 96 hours. The two-step technique recommended by Tilley and Terry was used to determine the in vitro digestibility of DM (IVDDM). Four hours after infusion of WB (100 g) through the ruminal fistula, the rumen content was collected from the animals and the microorganisms associated with the liquid and solid phases were separated, the former by filtration and the latter by extraction with 50 mM sodium phosphate buffer, pH 6.9. The solid fraction was subjected to in vitro culture with WB substrate for 96 hours to determine the biodegradation rate of DM, NDF and cellulolytic activity. The pH was measured after culture times of 24, 48 and 96 hours. Pretreatment with NaOH increased the IVDDM (P 0.05). The addition of NaOH increased the in situ biodegradation rate of NDF, with sheep showing a lower lag time (LT). There was a solubilization of Hcel, Cel and Lig in SCB pretreated with NaOH. Cellulolytic activity was higher in the solid phase, irrespective of the inoculum source, and microbial growth was observed in in vitro cultures using microorganisms from the solid phase and WB as substrate. The pH of the in vitro culture increased over time. Ruminal microorganisms from Brazilian naturalized goat and sheep breeds successfully colonized and biodegraded both WB and HB. Sodium hydroxide proved useful as an alkaline pretreatment for SCB

Degradabilidade in situ do bagaço de cana-de-açúcar para pequenos ruminantes de raças naturalizadas do Nordeste brasileiro.

Resumo: Com o crescimento da indústria sucroalcooleira, buscam-se alternativas de uso dos resíduos gerados por ela. Os pequenos ruminantes de raças naturalizadas do Nordeste brasileiro mostram-se como grupos potenciais para que seja utilizado o bagaço de cana-de-açúcar (BCA) nas dietas. Nesse contexto, objetivou-se determinar a degradabilidade ruminal in situ da matéria seca (MS) e da fibra em detergente neutro (FDN) do BCA e os parâmetros ruminais em caprinos Moxotó e ovinos Morada Nova, fistulados no rúmen. Contidos em sacos de náilon, 3g de BCA foram incubados no rúmen nos tempos seis, 24 e 96 horas, determinando-se o conteúdo de MS e FDN nos resíduos obtidos. Nos tempos zero, seis e 12 horas após a primeira refeição, mediram-se no líquido ruminal pH e nitrogênio amoniacal (N-NH3) ruminal. O CMS não diferiu entre caprinos e ovinos. O potencial de máxima degradação da MS foi semelhante entre espécies, e da FDN foi superior em caprinos. Ovinos apresentaram maiores tempo de colonização, taxa de degradação e degradabilidade efetiva da MS e FDN. O pH não diferiu entre as espécies. Observou-se maior concentração de N-NH3 ruminal em caprinos, no tempo zero. Diante da maior velocidade de degradação da MS do BCA pelos ovinos, essa espécie se mostra detentora de uma microbiota ruminal com crescimento mais eficiente sobre o BCA. [In situ degradability of sugarcane bagasse for naturalized small ruminant breeds from the northeastern Brazil]. Abstract: With the increase of the sugar and alcohol industries, alternative uses for residue are sought, and the small ruminants from the naturalized breeds of northeastern Brazil are potential groups to make use of the sugarcane bagasse (SCB) in their diets. In this context, the objective was to determine the in situ degradability of dry matter (DM) and neutral detergent fiber (NDF) of SCB and ruminal parameters in Moxotó goats and Morada Nova sheep rumen fistulated. Three grams of SCB were placed in naylon bags and incubated in the rumen at 6, 24 and 96 hours, and the residues analyzed for DM and NDF. Rumen fluid was collected at zero, 6 and 12 hours after the first meal and pH and ammonia nitrogen (N-NH3) were determined. DMI did not differ between goats and sheep. The maximum degradation potential of DM was similar between species and the A of NDF was higher in goats. Sheep had higher colonization time, rate of degradation and effective degradability of DM and NDF. The pH did not differ between species. The highest concentration of ruminal N-NH3 in goats was at time zero. Given the higher rate of degradation of bagasse dry matter by sheep, this specie shows a more efficient rumen microbial growth on SCB

Primary refractory follicular lymphoma: a poor outcome entity with high risk of transformation to aggressive B cell lymphoma

Background: Primary refractory (PREF) follicular lymphoma (FL) has a completely different clinical course from that of FL that responds to front-line treatments. In addition to having poor responses to salvage therapies, it seems that patients with PREF are at increased risk of histological transformation (HT). The Aristotle consortium presented the opportunity of investigating the risk of HT in a very large series of cases. Thus, we investigated the risk of HT in patients with PREF FL compared with that of responding patients or in stable disease and ultimately their outcome. Methods: Six thousand three hundred thirty-nine patients from the Aristotle database were included in the analysis. These patients had a histologically confirmed grade 1, 2 or 3a FL diagnosed between 1997 and 2013. The primary end-points were the cumulative incidence (CI) of HT at the first progression or relapse and the survival after transformation. Findings.: The 5-year CI of HT among patients with PREF was 34% (95% confidence interval (CI): 27–43), whilst it was 7.1% (95% CI: 6.0–8.5) in the group of patients with partial response (PR) or stable disease (SD) (PR + SD) and 3.5% (95% CI: 3.0–4.2) in the group of patients achieving complete response (CR). The 5-year survival after relapse (SAR) was 33% (95% CI: 28–39) for the PREF group, 57% (95% CI 54–61) in patients with PR, 51% (95% CI 43–58) in the SD group after first-line therapy and 63% (95% CI: 66–72) in patients with CR after initial treatment (p-value <0.001). The 5-year SAR for those patients with PREF who developed HT was 21% (95% CI: 12–31), clearly diminished when compared with those patients with PREF who did not experience HT (38% [95% CI: 31–44]) (p-value = 0.001). Interpretation.: Patients with PREF FL have a dismal outcome and an associated very high rate of HT that further worsens their poor prognosis

Parâmetros ruminais de caprinos e ovinos nativos do Nordeste brasileiro alimentados com dietas à base de bagaço de cana-de-açúcar in natura.

Resumo: Neste trabalho objetivou-se avaliar o pH e N-NH3 (nitrogênio amoniacal) ruminal de caprinos e ovinos nativos do Brasil alimentados com dietas à base de bagaço de cana in natura. Foram utilizados três caprinos da raça Moxotó e três ovinos da raça Morada Nova submetidos à fistulação ruminal. A relação volumoso:concentrado de 54:46, sendo o volumoso constituindo de feno de tifton-85 e bagaço de cana e o concentrado à base de milho e farelo de soja. A coleta de liquido ruminal foi realizada via cânula ruminal nos tempos 0 (antes da alimentação), 6 e 12 horas após alimentação. Não houve diferença (P>0,05) de pH entre as espécies nos diferentes tempos de coleta. O efeito dentro de espécie foi significativo (P 0.05) in pH between species in different sample times. The effect within species was significant (P <0.05) for goats with higher pH value before the supply of food (6.88) compared with times 6 and 12 (6.33 and 6.05, respectively) not differ. There were differences (P <0.05) for the concentration of N-NH3 of rumen fluid between species and within species. The highest concentration of N-NH3 was observed in animals at time 0 (15.04 mg / dL). The inclusion of sugar cane bagasse in the diet did not affect ruminal fermentation. For evaluation of N-NH3 is recommended to include more time between 0-6 and 6-12 hour

CD8+ T-Cells Expressing Interferon Gamma or Perforin Play Antagonistic Roles in Heart Injury in Experimental Trypanosoma Cruzi-Elicited Cardiomyopathy

In Chagas disease, CD8+ T-cells are critical for the control of Trypanosoma cruzi during acute infection. Conversely, CD8+ T-cell accumulation in the myocardium during chronic infection may cause tissue injury leading to chronic chagasic cardiomyopathy (CCC). Here we explored the role of CD8+ T-cells in T. cruzi-elicited heart injury in C57BL/6 mice infected with the Colombian strain. Cardiomyocyte lesion evaluated by creatine kinase-MB isoenzyme activity levels in the serum and electrical abnormalities revealed by electrocardiogram were not associated with the intensity of heart parasitism and myocarditis in the chronic infection. Further, there was no association between heart injury and systemic anti-T. cruzi CD8+ T-cell capacity to produce interferon-gamma (IFNγ) and to perform specific cytotoxicity. Heart injury, however, paralleled accumulation of anti-T. cruzi cells in the cardiac tissue. In T. cruzi infection, most of the CD8+ T-cells segregated into IFNγ+ perforin (Pfn)neg or IFNγnegPfn+ cell populations. Colonization of the cardiac tissue by anti-T. cruzi CD8+Pfn+ cells paralleled the worsening of CCC. The adoptive cell transfer to T. cruzi-infected cd8−/− recipients showed that the CD8+ cells from infected ifnγ−/−pfn+/+ donors migrate towards the cardiac tissue to a greater extent and caused a more severe cardiomyocyte lesion than CD8+ cells from ifnγ+/+pfn−/− donors. Moreover, the reconstitution of naïve cd8−/− mice with CD8+ cells from naïve ifnγ+/+pfn−/− donors ameliorated T. cruzi-elicited heart injury paralleled IFNγ+ cells accumulation, whereas reconstitution with CD8+ cells from naïve ifnγ−/−pfn+/+ donors led to an aggravation of the cardiomyocyte lesion, which was associated with the accumulation of Pfn+ cells in the cardiac tissue. Our data support a possible antagonist effect of CD8+Pfn+ and CD8+IFNγ+ cells during CCC. CD8+IFNγ+ cells may exert a beneficial role, whereas CD8+Pfn+ may play a detrimental role in T. cruzi-elicited heart injury