Objetivou-se avaliar a influência da monensina sódica sobre o acúmulo hepático de cobre em ovinos. Haja vista, que ionóforos podem alterar a absorção, disponibilidade e distribuição tecidual de minerais em muitos tecidos do organismo. Foram utilizados 24 ovinos mestiços das raças Santa Inês x Dorper com média de três meses de idade e peso medio de 30 kg ao início do experimento, que receberam dieta basal calculada em 2,75% do peso corpóreo, sendo 50% de concentrado comercial (7,13 ppm de Cu) e 50% de feno de capim coast-cross (3,72 ppm de Cu) e 10 gramas de sal mineral (5,9 ppm de Cu). Os cordeiros foram distribuídos em quatro grupos distintos, sendo: Controle apenas dieta basal; Monensina (Mon) - dieta basal e 30 ppm de monensina sódica; Cobre (Cu) - dieta basal acrescida de 10 mg/kg P.V./dia de cobre, na forma de sulfato de cobre pentahidratado; e grupo Monensina+Cobre (MonCu) - dieta basal acrescida de 10 mg/kg P.V./dia de cobre e 30 ppm de monensina sódica. O período experimental teve duração de 14 semanas. Foram coletadas amostras de figado e bile no início por biópsia, e ao final do experimento por necropsia, para determinação de minerais, assim como amostras semanais de sangue para avaliação bioquímica, hematológica e mineral. Foi realizada análise estatística por meio de fatorial 2x2 considerando como fatores a inclusão ou não de cobre e monensina na dieta. Somente um animal apresentou quadro clínico de intoxicação cúprica acumulativa. As concentrações de cobre no fígado no início do experimento variaram entre 243,8 e 345,4 ppm não havendo diferença entre os grupos experimentais. As concentrações médias e desvios-padrão em ppm do cobre hepático ao final do experimento foram 3.363 ± 576 para o grupo MonCu; 2560 ± 524 para o grupo Cu; 562 ± 168 para o grupo Mon e 253 ± 131 para o grupo Controle. Ao final do estudo a concentração de cobre hepático foi influenciada pela suplementação com cobre (P = 0,0001) e monensina (P = 0,0003). Os grupos suplementados com cobre tiveram redução dos teores de ferro hepático (P = 0,0287) e aumento das concentrações de cobre na bile. A avaliação bioquimica demonstrou elevação da atividade sérica da GGT e AST (P < 0,05) nos grupos Cu e MonCu a partir da décima primeira semana do experimento em relação aos grupos Controle e Mon. O aumento da atividade destas enzimas foram influenciadas pela suplementação de cobre (P = 0,0340), enquanto os grupos suplementados com monensina apresentaram menores valores de creatinina quinase. Os resultados obtidos permitem concluir que a monensina interfere positivamente com o acúmulo hepático de cobre sendo que a sumplentação deste aditivo pode predispor a quadros de intoxicações cúprica em ovinos. / The aim of this study was to verify if monensin can change the accumulation of cooper in the liver of sheep, in view of ionophores can alter the absorption, availability, tiissue distribution of many minerals in the body. Twenty four crossbred Dorper X Santa Inês lambs wiht mean age of 3 months and wheitgh 30 kg at the begining of trial recived a basal diete calculated by 2,75% of body wheigth of 50% concentrade (7,13 ppm of Cu) and 50% cost-cross hay (3,72 ppm of Cu) and 10 g of mineral salt (5,9 ppm of Cu). Lambs were randomly distributed in four groups: Control: basal diet, Mon Group reciving basal diet plus 30 mg of monensin; Cu group: basal diet plus 10 mg Cu B.W of CuSO4.5H2O solution; Group CuMon: basal diet plus 30 mg of monensin and 10 mg of Cu B.W. The exeperimental period lasted for fourteen weeks. Liver samples were obtined by liver biopsy initially and at the end by necropsy when lambs were slaughterd. Blood samples were obtained weekly for determination of biochemical, hematological and mineral. Statistical analysis using a 2x2 factorial was conducted considering the factors copper and monensin in the diet and the interaction between them to evaluate the hepatic copper at the beginning and at the end of the study. Only one of animal of group MonCu exhibit clinical sings of copper toxicosis. Copper concentrations in the liver at the beginning of the experiment ranged between 243,8 and 345,4 ppm with no difference between experimental groups. Mean concentrations and standard deviations in ppm of hepatic copper at the end of the experiment were 3,363 ± 576 for MonCu group; 2560 ± 524 for Cu group; 562 ± 168 for the Mon group and 2538 ± 131 for the control group. At the end of the study liver concentrations and total hepatic copper accumulation were influenced both by the amount of dietary copper (P = 0.0001) and of monensin (P = 0.0003). The groups supplemented with copper had reduced levels of hepatic iron (P = 0.0287) and increased concentrations of copper in the bile. The biochemical evaluation demonstrated elevated serum AST and GGT activity (P <0.05) with Cu and MonCu from the eleventh week of the experiment compared to the control groups and Mon. The increased activity of these enzymes were influenced by copper supplementation (P = 0.0340), while group monensin had lower values of creatinine kinase. The results indicate that monensin interferes positively with hepatic copper accumulation and supplementation of this additive can predispose cupric poisoning in sheep.

Submitted by Sandra Pereira (srpereira@ufrrj.br) on 2017-01-26T10:49:56Z No. of bitstreams: 1 2015 - Rafaela Scalise Xavier de Freitas.pdf: 1344462 bytes, checksum: 1e3669fc0b4da746d7a41a31525f3492 (MD5) / Made available in DSpace on 2017-01-26T10:49:56Z (GMT). No. of bitstreams: 1 2015 - Rafaela Scalise Xavier de Freitas.pdf: 1344462 bytes, checksum: 1e3669fc0b4da746d7a41a31525f3492 (MD5) Previous issue date: 2015-06-26 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / This survey was divided into two chapters, the first one producing and characterizing Urochloa humidicola methanol extract in order to present secondary metabolites classes and bromathological and chemical composition. These metabolites have several functions within the plant and are associated to defense system, protecting the environment where they have been living. These compounds have been used on animal feed in the reason presenting antimicrobial properties that could be employed for inducing ruminal fermentation. Phytochemical screening tests and chemical composition of U. humidicola methanol extract and in natura plant were carried out on this present survey. The following secondary compounds classes: saponins, tannins, flavonoids, non-protein amino acids, cardiotive glycosides, steroids, tripernoids, catechins and saccharides were indentified U. humidicola methanol extract in regarding to in natura plant showed 10,20% and 5,17% crude protein concentrations, 35% and 1,51% lipids and 9,59% aand 39,92% non-fibrous carbohydrates, respectively. These results might be explained by percolation with methanol extraction methods extracting only soluble constituents transporting silted protein, lipids and ash to the extract. The second chapter of this survey aimed evaluating U. humidicola extract addition effect containing saponin associated to U. brizantha assessing gases production, (methane and carbon dioxide), ruminal kinetics, dry matter degradation and short chain fatty acids production (SCFA: acetate, propionate and butyrate), as well. Plant extracts have been an alternative inducing ruminal fermentation by secondary metabolites in the reason they are from natural sources and with no residue hazards in products like meat and milk. Ruminal fermentation induction could reduce methane production, as well as, increase acetate: propionate ratio and improve food degradation. Four U. humidicola methanol extract concentrations (0, 75, 150 and 250 g/L) on U. brizantha degradability by in vitro gases production were tested. At 150 g/L extract concentration gas production from fibrous carbohydrates was 118,21 mL. However, the highest fiber concentration rate occurred at 150 g/L. Increasing extract concentrations (75, 150 and 250 g/L) soluble fraction values were: 10,27; 7,46 and 14,07%, respectively. Effective ruminal degradability at 75, 150 and 250 g/L concentrations for passage rates for an animal in maintenance were 38,53%, 27,71% and 20,30%, respectively. Extract concentrations increase exerted a linear effect (P<0.05) on ruminal pH values being more evident at high extract concentration (250 g/L) as 5,73 and 5,43 at 12 and 24 hs, respectively. CO2 averages in regarding to incubated and degraded dry matter did not differ with extract concentrations increase at 12 hs. Methane averages in regarding to incubated and degraded dry matter were no significative by regression analysis. Treatment at 250 g/L concentration presented the lowest value for methane at 12 hs. At 75 g/L concentration, total SCFA (acetic, propionic and butyric acid) increase at 12 and 24 hs was reported. U. humidicola methanol extract different concentration addition improved U. brizantha fermentation kinects parameters at 150 g/L and 250 g/L concentrations. However, negative effect on U. brizantha dry matter degradation and ruminal pH values according to extract concentrations increase was reported. Strong correlation between pH values and dry matter degradation (p=0,61, P<0,05) was presented. Carbon dioxide concentration increased, as well as, methane production decreased U. humidicola crude methanol extract presented potential for use as ruminal fermentation promoter. New studies about U. humidicola extract employing animals for justifying its efficiency as food additive should, furthermore, be developed. / Este trabalho foi dividido em dois cap?tulos, o primeiro teve por objetivo produzir e caracterizar o extrato metan?lico de Urochloa humidicola, com o intuito de conhecer as classes de metab?litos secund?rios presentes e a composi??o qu?mico-bromatol?gica. Estes metab?litos possuem diversas fun??es dentro dos vegetais e est?o associados ao sistema de defesa, os protegendo no ambiente que vivem. Estes compostos est?o sendo utilizados na alimenta??o animal por apresentarem propriedades antimicrobianas que podem ser empregadas para induzir a fermenta??o ruminal. Para este estudo foram realizados os testes de prospec??o fitoqu?mica e as an?lises de composi??o bromatol?gica do extrato metan?lico de U. humidicola e da U. humidicola in natura. Foram identificadas as seguintes classes de compostos secund?rios: saponinas, taninos, flavonoides, amino?cidos n?o proteicos, glicos?deos cardioativos, ester?ides e tripern?ides, catequinas e sacar?deos. O extrato metan?lico de U. humidicola em rela??o ? planta in natura, apresentaram concentra??es de prote?na bruta de 10,20% e 5,17%, e mat?ria mineral de 16,14% e 8,14%, extrato et?reo de 35% e 1,51%, carboidrato n?o fibroso, 9,59% e 39,92%, respectivamente. Esse resultado pode ser explicado pelo m?todo de extra??o que foi por percola??o com metanol, extraindo somente os constituintes sol?veis carreando somente prote?na, lip?deos e cinzas para o extrato. No segundo cap?tulo deste trabalho teve como objetivo avaliar o efeito da adi??o de extrato de U. humidicola contendo saponina, associada ? Urochloa brizantha, avaliando a produ??o de gases (metano e de di?xido de carbono), a cin?tica ruminal, a degrada??o da mat?ria seca e produ??o de ?cidos graxos de cadeia curta (AGCC; acetato, propionato e butirato). Os extratos vegetais de plantas s?o uma alternativa para induzir da fermenta??o ruminal por possu?rem metab?litos secund?rios, por serem de fontes naturais e sem riscos de res?duos nos produtos como carne e leite. A indu??o da fermenta??o ruminal pode reduzir a produ??o de metano, aumentar a rela??o de acetato: propionato e melhorar a degrada??o do alimento. Foram testados quatro concentra??es de extrato metan?lico de U. humidicola (0, 75, 150 e 250 g/L) sobre a degradabilidade da U. brizantha pela produ??o de gases in vitro. Na concentra??o de 150 g/L do extrato, a produ??o de g?s proveniente dos carboidratos fibrosos, foi de 118,21 mL. No entanto, a maior taxa de degrada??o dos carboidratos fibrosos ocorreu na concentra??o 150 g/L. Com o aumento das concentra??es de extrato (75, 150 e 250 g/L) os valores da fra??o sol?vel foram de 10,27; 7,46 e 14,07% respectivamente. A degradabilidade ruminal efetiva nas concentra??es de (75, 150 e 250 g/L) para as taxas de passagem para um animal em manten?a foram de 38,53%, 27,71% e 20,30%, respectivamente. O aumento das concentra??es de extrato exerceu um efeito linear (P<0,05) sobre os valores de pH ruminal, sendo mais evidente na alta concentra??o de extrato (250 g/L) que foi de 5,73 e 5,43 nos tempos de 12 e 24 horas, respectivamente. As m?dias de CO2 com rela??o ? mat?ria seca incubada e degradada n?o diferiram entre si com o aumento das concentra??es de extrato nos tempos de 12 horas. As m?dias de metano com base na mat?ria seca incubada e degradada n?o apresentaram signific?ncia para an?lise de regress?o. O tratamento com a concentra??o de 250 g/L de extrato apresentou menor valor para metano no tempo de 12 horas. A concentra??o de extrato (75 g/L) proporcionou um aumento do total de AGCC, ?cido ac?tico, ?cido propi?nico e ?cido but?rico tanto no tempo de 12 e 24 horas. A adi??o das diferentes concentra??es de extrato metan?lico de U. humidicola melhorou os par?metros da cin?tica da fermenta??o da U. brizantha nas concentra??es de 150 e 250 g/L. Mas causou um efeito negativo sobre a degrada??o da mat?ria seca da U. brizantha e no pH ruminal com o aumento das concentra??es de extrato. Existe uma forte correla??o entre os valores de pH e degrada??o da mat?ria seca (?=0,61, P<0,05). Aumentou as concentra??es de g?s carb?nico e reduziu a produ??o de metano. O extrato metan?lico bruto de U. humidicola tem potencial para uso como indutor da fermenta??o ruminal. ? necess?rio ? purifica??o e o isolamento da saponina do extrato para comprovar o efeito ben?fico sobre a fermenta??o ruminal. S?o imprescind?veis novos estudos com o extrato de U. humidicola, utilizando animais para se comprovar a efici?ncia na utiliza??o como aditivo alimentar

Ingestion and dispersal of seeds of desirable species by domestic livestock is potentially important as a range improvement practice, but the passage of seed by livestock has only been studied in a fragmented way, particularly for species adapted to rangelands of western North America. The objectives of this research were to examine the effects of different periods of exposure to in vitro and in vivo digestion processes in cattle on the germinability of several grass species, and determine if the in vitro incubation technique is a good predictor of seed fate following passage through the ruminant digestive tract. Seeds of 13 grass species adapted to the Intermountain West were exposed to in vitro incubation for 24, 48, and 72 hours, and then tested for germination at an optimal temperature regime (10°C night/20°C day) in a controlled environment chamber. Germination responses varied considerably among grass species with changes in length of exposure to in vitro incubation, but germination decreased for incubated seed compared to untreated seed for all species. Five species with the highest germination in in vitro incubation trials were fed to Holstein steers in in vivo digestion trials. Approximately 20% of the ingested seeds were recovered for all species 6 days after feeding, and the highest recovery occurred 2 and 3 days after feeding. Germination of undamaged, recovered seeds decreased as passage time through the digestive tract increased. Of the species tested, seeds of Psathyrostachys juncea, Thinopyrum ponticum, Agropyron cristatum X A. desertorum, and Elytrigia repens X Pseudoroegneria spicata have the greatest potential to survive passage through the digestive tract and germinate in appreciable numbers. The in vitro incubation technique may be used as a crude indicator of seed fate following passage through the digestive tract.

Three experiments were conducted to evaluate low quality forages treatment by anhydrous ammonia (NH3) and(or) hydrogen peroxide (H2O2). In experiment I, a control and three treatments of barley straw were compared: NH3, NH3 after rehydration of the straw with water (NH3+H2O), and NH3 after rehydration with a H2O2 solution (NH3+H2O2). Forages were fed, with a supplement, at two levels of intake (ad libitum and 75% of ad lib.). Treatments were administered in a split-plot in a Latin square design to 8 ewes. Total collections and rumen digesta measurements were made. Ammoniation, rehydration, and H2O2 increased primarily dry matter intake (DMI) but also digestibility of different nutrients. A similar trend was observed in rumen fermentation characteristics. Dry matter (DM) digestibility was slightly raised by limiting DMI. Digestible DMI (DDMI) was correlated with both the increase of forage CP content and the decrease of forage neutral detergent fiber. Water intake and output were highly correlated with fiber. Intake and digestibility better than acid detergent lignin insoluble ash (ADLIA). Both markers were adequate in determining hemicellulose digestibility. In experiment II, ten solutions were prepared to rehydrate wheat straw. Six solutions were adjusted (with NaOH) to a pH of 9, 11, or 13 and contained 2% H2O2. Four solutions had a pH of 7 or 11 and contained no H2O2. Half of the straws were treated with NH3. In situ DM disappearance (DMD) of the different straws was measured at different times in 10 Holstein cows and three periods (Incomplete block design). The evolution of DMD was slow and almost linear. The positive effect of ammoniation on DMD was consistently apparent at all pH levels though depressed at pH 11. The effect of H2O2 was minor, but was complementary with ammoniation. In experiment III, mature baltic rush (Juncus balticus Willd.) was either treated or not with NH3. Eight wethers had ad libitum access to the nonsupplemented forages in a cross-over design. total collections and rumen digesta measurements were made. DMI and DDMI were not affected by treatment, but DM digestibility was decreased by ammoniation.

Despite existing mad cow disease surveillance efforts in the United States, in place since the 1980s, a cow that tested positive for mad cow disease was granted entrance into the U.S. in December, 2003. The cow that tested positive, according to witnesses, displayed no symptoms that are synonymous with advanced bovine spongiform encephalopathy, BSE. This occurrence had detrimental effects on the U.S. beef export market, as many countries banned American beef. Estimates of the damage inflicted reach into the billions of dollars. BSE in the U.S. has the potential of causing damages in other aspects as well. Aside from the fact that BSE is a public health issue, it has caused political rifts between nations, particularly between Canada and the U.S. It can undermine confidence in the USDA and confidence in the governments ability to handle emergencies. BSE can imperil American good that contain beef or beef products. Finally, it can undermine trust in scientists to provide useful guidance. The subtle changes in U.S. BSE surveillance efforts in the 1980’s were greatly surpassed by the changes that were made when a BSE-positive cow was discovered in Washington State in 2003. However, there remains room for much needed improvement in U.S. BSE surveillance efforts. These changes include: increased testing to include all cows slaughtered in the U.S. and all imported beef products, a nationwide animal tracking program, increased proficiency in training of inspectors, and the implementation of strict rules governing the ingredients of animal feed. The implementation of regulations based on economics instead of public health concerns has the potential to leave loopholes in regulations that the BSE agent might exploit. By enacting the recommendations made in this thesis, the U.S. will greatly increased its' odds of stopping the entrance and proliferation of BSE within its’ borders.

Grazing cattle on alfalfa (Medicago sativa L.) would be economically beneficial, but its rapid initial rate of protein degradation results in pasture bloat, low efficiency of protein utilization and excessive N pollution into the environment. Introducing a gene that stimulates the accumulation of mono/polymeric anthocyanidins might reduce the ruminal protein degradation rate and reduce bloat related foam stability. The overall objective of this thesis was to evaluate newly developed anthocyanidin-accumulating Lc-alfalfa progeny for nutritional properties (composition, site of degradation and molecular structure), environmental emissions and bloat characteristics. The objective of the first study was to determine survival and phytochemical and chemical profiles of Lc-alfalfa progeny (BeavLc1, RambLc3 and RangLc4) and their non-transgenic (NT) parental cultivars (Beaver, Rambler and Rangelander). Lc-alfalfa forage accumulated enhanced amounts of anthocyanidin, with an average concentration of 197.4 µg/g DM, while proanthocyanidin (i.e. condensed tannins) were not detected. Both of these metabolites were absent in the NT-parental varieties. Lc-alfalfa progeny had ~3 % less crude protein (CP) and ~3 % more carbohydrates (CHO), which resulted in their 11 g/kg lower N:CHO ratio compared with NT-alfalfa. Total rumen-degradable N:CHO ratio based on chemical analysis was 12.9 g/kg lower in Lc-alfalfa compared with NT-alfalfa. The objective of the second study was to evaluate in vitro degradation, fermentation and microbial-N partitioning of three forage color phenotypes [green, light purple-green (LPG) and purple-green (PG)] within Lc-progeny and their parental green NT-alfalfa varieties. Purple-green-Lc alfalfa accumulated more anthocyanidin than Green-Lc with LPG-Lc intermediate. Gas, methane and ammonia accumulation rates were slower for the two purple-Lc phenotypes compared with NT-alfalfa with Green-Lc intermediate. Effective degradable DM and N were lower in the three Lc-phenotypes compared with NT-alfalfa. Anthocyanidin concentration correlated negatively with gas and methane production rates and effective degradability of DM and N. The objectives of the third study were to evaluate in situ ruminal degradation characteristics and synchronization ratios, and to model protein availability to dairy cattle and net energy for lactation of three Lc-alfalfa progenies, BeavLc1, RambLc3 and RangLc4 and the cultivar AC Grazeland (selected for a low initial rate of ruminal degradation). Anthocyanidin accumulation was on average 163.4 ìg/g DM in the three Lc-progeny while AC Grazeland did not accumulate anthocyanidin. The basic chemical composition of the original samples, soluble and potentially degradable fractions and degradation characteristics of crude protein and carbohydrates were similar in Lc-alfalfa and AC Grazeland. The undegradable in situ crude protein and neutral detergent fiber fraction were, respectively, 1.3 %CP and 4.8 %CHO lower in the three Lc-progeny compared with AC Grazeland. Lc-alfalfa had a 0.34 MJ/kg DM higher net energy for lactation and tended to have a 11.9, 6.9 and 8.4 g/kg DM higher rumen degradable protein, rumen degraded protein balance and intestinal available protein, respectively, compared with AC Grazeland,. The hourly rumen degraded protein balance included an initial and substantial peak (over-supply) of protein relative to energy which was highest in RangLc4 and lowest in RambLc3. The hourly rumen degraded protein balance between 4 and 24 h was similar and more balanced for all four alfalfa populations. The objective of the fourth study was to determine foam formation and stability in vitro from aqueous leaf extracts of three Lc-alfalfa progeny (BeavLc1, RambLc3, RangLc4), parental NT-alfalfa and AC Grazeland (bloat reduced cultivar) harvested in the field at 07:00 or 18:00 h. Anthocyanidin accumulation averaged 247.5 ìg/g DM in the leaves of the three Lc-progeny. There was an interaction between population and harvest time for the foam parameters. Initial foam volume (0 min) and final foam volume (150 min) at 07:00 h were lower for AC Grazeland compared with all other treatments and lower for RangLc4 compared with the other two Lc-progeny at 0 min and NT-alfalfa at 150 min; while from the 18:00 h harvest, initial foam volume was larger for NT-alfalfa and final foam volume was larger for RambLc3 compared with AC Grazeland, BeavLc1 and RangLc4. Foam formation correlated positively (R = 0.30 to 0.44) with leaf DM content, leaf extract protein and ethanol-film content, spectroscopic vibration intensity due to all carbohydrates (CHOVI) and amide I:amide II ratio and negatively (R = -0.33 and -0.34; P<0.05) with á-helix:â-sheet ratio and amide I:CHOVI. Final foam volume correlated negatively (R = -0.53 to -0.25; P<0.05) with leaf extract pH, spectroscopic vibration intensity due to all protein structures, structural carbohydrates (SCVI) and lipids (CH2 and CH3 asymmetric stretching) and amide I:CHOVI ratio and corelated positively (R = 0.39 to 0.44; P<0.05) with CHOVI, amideI:SCVI ratio and CHOVI:SCVI ratio. In conclusion, all Lc-alfalfa progeny and phenotypes accumulated anthocyanidin in their forage. Lc-alfalfa progeny had lower protein and higher carbohydrate content which improved the nitrogen to carbohydrate balance compared to their parental NT-alfalfa cultivars. Rate of fermentation and effective degradability in vitro reduced for both purple anthocyanidin-accumulating Lc-alfalfa phenotypes compared with NT-alfalfa. Intestinal protein availability tended to be higher and net energy for lactation was higher from Lc-alfalfa progeny for dairy cattle compared with AC Grazeland. Foaming properties were reduced in Lc-alfalfa progeny compared with parental non-transgenic alfalfa but not compared with AC Grazeland. However, differences between the Lc-alfalfa progeny and other cultivars were small. Therefore, further increases in mono/polymeric anthocyanidin accumulation in alfalfa are required in order to develop an alfalfa cultivar with superior nutritional and bloat preventing characteristics compared to currently available alfalfa cultivars.

Grazing cattle on alfalfa (Medicago sativa L.) would be economically beneficial, but its rapid initial rate of protein degradation results in pasture bloat, low efficiency of protein utilization and excessive N pollution into the environment. Introducing a gene that stimulates the accumulation of mono/polymeric anthocyanidins might reduce the ruminal protein degradation rate and reduce bloat related foam stability. The overall objective of this thesis was to evaluate newly developed anthocyanidin-accumulating Lc-alfalfa progeny for nutritional properties (composition, site of degradation and molecular structure), environmental emissions and bloat characteristics. The objective of the first study was to determine survival and phytochemical and chemical profiles of Lc-alfalfa progeny (BeavLc1, RambLc3 and RangLc4) and their non-transgenic (NT) parental cultivars (Beaver, Rambler and Rangelander). Lc-alfalfa forage accumulated enhanced amounts of anthocyanidin, with an average concentration of 197.4 µg/g DM, while proanthocyanidin (i.e. condensed tannins) were not detected. Both of these metabolites were absent in the NT-parental varieties. Lc-alfalfa progeny had ~3 % less crude protein (CP) and ~3 % more carbohydrates (CHO), which resulted in their 11 g/kg lower N:CHO ratio compared with NT-alfalfa. Total rumen-degradable N:CHO ratio based on chemical analysis was 12.9 g/kg lower in Lc-alfalfa compared with NT-alfalfa. The objective of the second study was to evaluate in vitro degradation, fermentation and microbial-N partitioning of three forage color phenotypes [green, light purple-green (LPG) and purple-green (PG)] within Lc-progeny and their parental green NT-alfalfa varieties. Purple-green-Lc alfalfa accumulated more anthocyanidin than Green-Lc with LPG-Lc intermediate. Gas, methane and ammonia accumulation rates were slower for the two purple-Lc phenotypes compared with NT-alfalfa with Green-Lc intermediate. Effective degradable DM and N were lower in the three Lc-phenotypes compared with NT-alfalfa. Anthocyanidin concentration correlated negatively with gas and methane production rates and effective degradability of DM and N. The objectives of the third study were to evaluate in situ ruminal degradation characteristics and synchronization ratios, and to model protein availability to dairy cattle and net energy for lactation of three Lc-alfalfa progenies, BeavLc1, RambLc3 and RangLc4 and the cultivar AC Grazeland (selected for a low initial rate of ruminal degradation). Anthocyanidin accumulation was on average 163.4 ìg/g DM in the three Lc-progeny while AC Grazeland did not accumulate anthocyanidin. The basic chemical composition of the original samples, soluble and potentially degradable fractions and degradation characteristics of crude protein and carbohydrates were similar in Lc-alfalfa and AC Grazeland. The undegradable in situ crude protein and neutral detergent fiber fraction were, respectively, 1.3 %CP and 4.8 %CHO lower in the three Lc-progeny compared with AC Grazeland. Lc-alfalfa had a 0.34 MJ/kg DM higher net energy for lactation and tended to have a 11.9, 6.9 and 8.4 g/kg DM higher rumen degradable protein, rumen degraded protein balance and intestinal available protein, respectively, compared with AC Grazeland,. The hourly rumen degraded protein balance included an initial and substantial peak (over-supply) of protein relative to energy which was highest in RangLc4 and lowest in RambLc3. The hourly rumen degraded protein balance between 4 and 24 h was similar and more balanced for all four alfalfa populations. The objective of the fourth study was to determine foam formation and stability in vitro from aqueous leaf extracts of three Lc-alfalfa progeny (BeavLc1, RambLc3, RangLc4), parental NT-alfalfa and AC Grazeland (bloat reduced cultivar) harvested in the field at 07:00 or 18:00 h. Anthocyanidin accumulation averaged 247.5 ìg/g DM in the leaves of the three Lc-progeny. There was an interaction between population and harvest time for the foam parameters. Initial foam volume (0 min) and final foam volume (150 min) at 07:00 h were lower for AC Grazeland compared with all other treatments and lower for RangLc4 compared with the other two Lc-progeny at 0 min and NT-alfalfa at 150 min; while from the 18:00 h harvest, initial foam volume was larger for NT-alfalfa and final foam volume was larger for RambLc3 compared with AC Grazeland, BeavLc1 and RangLc4. Foam formation correlated positively (R = 0.30 to 0.44) with leaf DM content, leaf extract protein and ethanol-film content, spectroscopic vibration intensity due to all carbohydrates (CHOVI) and amide I:amide II ratio and negatively (R = -0.33 and -0.34; P<0.05) with á-helix:â-sheet ratio and amide I:CHOVI. Final foam volume correlated negatively (R = -0.53 to -0.25; P<0.05) with leaf extract pH, spectroscopic vibration intensity due to all protein structures, structural carbohydrates (SCVI) and lipids (CH2 and CH3 asymmetric stretching) and amide I:CHOVI ratio and corelated positively (R = 0.39 to 0.44; P<0.05) with CHOVI, amideI:SCVI ratio and CHOVI:SCVI ratio. In conclusion, all Lc-alfalfa progeny and phenotypes accumulated anthocyanidin in their forage. Lc-alfalfa progeny had lower protein and higher carbohydrate content which improved the nitrogen to carbohydrate balance compared to their parental NT-alfalfa cultivars. Rate of fermentation and effective degradability in vitro reduced for both purple anthocyanidin-accumulating Lc-alfalfa phenotypes compared with NT-alfalfa. Intestinal protein availability tended to be higher and net energy for lactation was higher from Lc-alfalfa progeny for dairy cattle compared with AC Grazeland. Foaming properties were reduced in Lc-alfalfa progeny compared with parental non-transgenic alfalfa but not compared with AC Grazeland. However, differences between the Lc-alfalfa progeny and other cultivars were small. Therefore, further increases in mono/polymeric anthocyanidin accumulation in alfalfa are required in order to develop an alfalfa cultivar with superior nutritional and bloat preventing characteristics compared to currently available alfalfa cultivars.

Continuous culture fermentation (CCF) was used to test the hypotheses that: marine microalgae (MA) and macroalgae (seaweeds) alter rumen microbial metabolism; MA types differ in abilities to provide rumen escape n-3 polyunsaturated fatty acids (PUFA); and algae have the potential to reduce enteric methane emission. The CCF system of Teather and Sauer (1988) was modified to reduce clogging, refrigerate effluent, and allow for determination of gas production. The CCF systems were inoculated with pooled rumen fluid from 4 cows. Total mixed ration was fed at the rate of 30 g DM d-1. Temperature was maintained at 39 oC, and buffered with artificial saliva to maintain pH 6.2. Response variables were measured from effluent digesta (fatty acids, NH4+-N, digestibility), fermentor contents (CCF density, volatile fatty acids), and the gas phase (CO2, CH4). The experimental design for MA testing was a 3**3 factorial. Treatments consisted of heterotrophic and photoautotrophic MA as well as a 1:1 blend with protection levels of zero, 33 and 50 % of encapsulation (w/w), and fluid turnover rates of 5, 7.5, and 10 % h-1. The seaweed treatments consisted of a PEI shoreweed mix containing Laminaria longicruris and Fucus vesiculosus tested as a component of the mix, and Chondrus crispus and Furcellaria fastigiata tested individually. The design for seaweed testing was an unbalanced 5*5 Latin square. The heterotrophic MA destabilized the digesta mat while the autotroph improved stability. Biohydrogenation was extensive for C18 FA in the basal ration (> 90 %) and less for C22:6n3 (75 %) from the heterotroph and C20:5n3 (60 %) from the photoautotroph. The recovery of PUFA was improved by encapsulation, however PUFA in the MA were not greatly affected and digestibility was improved by turnover rate. Seaweeds had no effect on CCF stability, however they reduced CH4 production without reduction in OM digestibility. The heterotroph reduced overall fermentation resulting in diminished density and volatile fatty acids and NH4+-N concentrations. Seaweed supplementation decreased NH4+-N, CO2 and CH4 production, and increased density.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The effects of aflatoxin B1 (AFB1) (1μg/mL) were evaluated and sorbents β-glucans derived from Saccharomyces cerevisae with 65% of active ingredient (β-glu) (1mg/mL) and montmirillonite (MMT) (5mg/mL) under ruminal fermentation. Two in vitro assays were conducted. On the first assay the objectives were to determine the production of short chain fatty acids, the production of ammonia during 24-h in vitro. While on the second assay the production of methane (CH4) and kinetic parameters based on gas production date were determined (GPmax= maxim gas production in t time; Lag= lag phase before gas production commenced; S=gas production rate (h-1)) during 72-h in vitro incubation. In each assay six repetitions were made for the following treatments: CONT: control (without AFB1 or sorbents); AF: AFB1 (1μg/mL); β-glu (1mg/mL); β-glu + AF: (1mg/mL of β-glu + 1μg/mL of AFB1); MMT: (5mg/mL); MMT + AF: (5mg/mL of MMT + 1μg/mL of AFB1). The amount of AGCC produced by the β-glu (67,7 mM) treatment was significantly higher about CONT (57,72 mM) and MMT (53,3 mM). treatments. On the other hand, MMT clay reduced the production of NH3 (9,6 mM) about CONT (11,4 mM), AF (12,6 mM) and β-glu (12,2 mM). The amount of GPmax by the β-glu treatment was 103,4 mL, significantly higher about produced CONT (89,0 mL) e MMT (91,6 mL). There was also higher gas production rate by the β-glu treatment. The montmorilonite raised the lag phase and reduced the CH4 production. The results of this study suggest that AFB1 (1μg/mL) has no toxic effect on ruminal fermentation. Whereas the β-glu impacts the ruminal fermentation by increase the AGCC produced. The montmorilonite can delay the bacterial colonization but does t effect the quantity of AGCC produced. / Foram avaliados os efeitos da aflatoxina B1 (AFB1) (1μg/mL) e os adsorventes β-glucanas devivadas de Saccharomyces cerevisae com 65% de princípio ativo (β-glu) (1mg/mL) e montmorilonita (MMT) (5mg/mL) sobre a fermentação ruminal. Foram conduzidos dois ensaios in vitro, no primeiro ensaio o propósito foi determinar a produção de ácidos graxos de cadeia curta, a produção de amônia em 24h de incubação. Enquanto no segundo ensaio determinou-se a produção de metano (CH4) e os parâmetros da cinética da produção de gases (Vf = volume final de gás (ml) no tempo t; L = tempo de colonização; S = taxa de degradação (h-1)) no período de 72h de incubação. Em cada ensaio seis repetições foram realizadas para os seguintes tratamentos: CONT: controle (sem AFB1 ou adsorventes); AF: AFB1 (1μg/mL); β-glu (1mg/mL); β-glu + AF: (1mg/mL de β-glu + 1μg/mL de AFB1); MMT: (5mg/mL); MMT + AF: (5mg/mL de MMT + 1μg/mL de AFB1). A quantidade produzida de AGCC foi significativamente maior no tratamento β-glu (67,7 mM) em relação aos tratamentos CONT (57,72 mM) e MMT (53,3 mM). A MMT reduziu significativamente a produção de NH3 (9,6 mM) em relação aos tratamentos CONT (11,4 mM), AF (12,6 mM) e β-glu (12,2 mM). O tratamento β-glu produziu maior volume de gás (103, 4 mL) em relação aos tratamentos CONT (89,0 mL) e MMT (91,6 mL). Também o tratamento β-glu teve maior taxa de degradação em relação aos demais tratamentos. A montmorilonita aumentou o tempo de colonização e reduziu a produção de CH4. Os resultados deste estudo sugerem que a AFB1 (1μg/mL) não é tóxica a fermentação ruminal. Enquanto, o uso do β-glu impacta a fermentação ruminal, aumentando a produção de AGCC. O uso de montmorilonita pode retardar a colonização bacteriana no alimento porém, não interfere significativamente na quantidade total de AGCC produzidos.

Orientadora: Silvana Martinez Baraldi Artoni / Banca: Carlos Alberto Vicentini / Banca: Marcos Lania de Araújo / Banca: Maria Rita Pacheco / Banca: Vanessa Sobue Franzo / Resumo: Neste trabalho foram utilizados 14 ovinos machos adultos, seis castrados e oito não castrados da raça Santa Inês com o peso médio de 32 kg. Foi realizado estudo macroscópico (topográfico e biométrico) e microscópico (histológico) das glândulas vesiculares e bulbouretrais com o objetivo de averiguar o comportamento dessas glândulas nos animais castrados e não castrados. Para o estudo macroscópico realizou-se a descrição destas duas glândulas, e avaliou-se a biometria após a dissecação e mensuração do peso, comprimento, altura e a largura das respectivas glândulas. Para a histologia foram obtidos fragmentos das respectivas glândulas os quais foram recortados, obteu-se cortes histológicos de cinco micrômetros sendo diafanizados em benzol e processados para inclusão do material em paraplast e corados com Hematoxilina-eosina e fotodocumentados em fotomicroscópio Olympus BX 50. Morfologicamente pode-se documentar que toda a sua topografia apresentou semelhança à espécie bovina, e na biometria evidenciouse medidas macroscópicas menores (p<0,05) das glândulas de ovinos castrados em relação aos não castrados, exceto para o peso vivo dos animais e peso relativo das glândulas vesiculares (p>0,05). Histologicamente verificou-se que a glândula vesicular é um tubo enrolado sobre si mesmo seccionado em diferentes posições. Separando-o há uma delgada camada de tecido conjuntivo frouxo. A mucosa é pregueada e o epitélio é prismático simples com células altas ricas. A lâmina própria possui fibras elásticas e está envolta por uma camada muscular lisa, constituída por duas lâminas: uma interna, de fibras circulares e outra externa, de fibras longitudinais. A camada adventícia é constituída por tecido conjuntivo frouxo. Os ductos excretores principais estão revestidos por epitélio prismático biestratificado... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: In this project, 14 adult male sheep were used. six of them were castrated and eight non-castrated, and they were all Santa Ines breed, weighing 32kg in average. A macroscopic (topographic and biometric) and microscopic (histologic) study of the vesicular and bulbourethral glands was carried out with the aim of investigating the behavior of these glands in the castrated and non-castrated animals. For the macroscopic study, the description of these two glands was carried out, and the biometry after the dissecation, and weight, length, height and width measurement of the respective glands was evaluated. For the histology, fragments of these glands were obtained and cut. The obtained histological cuts of five micrometers were diaphanized in benzol and processed for the inclusion of the material in paraplast and stained with Hematoxylin and eosin and photo documented in photomicroscope Olympus BX 50. Morphologically, all its topography can be documented presenting similarity to the bovine species, and in the biometry it has evidenced smaller macroscopic measurements (p<0,05) comparing the glands of the castrated ovines and the non-castrated ones, except for the live weight of the animals and relative weight of the vesicular glands (p>0,05). Histologically, it has been verified that the vesicular gland is a tube coiled round itself sectioned at different positions. There is a thin layer of loose connective tissue separating it. The mucosa is wrinkled and the simple prismatic epithelium is composed of high cells that are rich. The blade itself has elastic fibers and is wrapped by a layer of smooth muscle, composed by two blades: an internal one, composed of circular fibers and an external one, composed of longitudinal fibers. The tunica adventitia is composed by loose connective tissue. The main excretory ducts are covered by bistratified prismatic epithelium... (Complete abstract click electronic access below) / Doutor