O objetivo deste trabalho foi avaliar o efeito da ractopamina, cromo-metionina e suas combinações nas dietas de suínos em crescimento e terminação sobre o desempenho, as características de carcaça e a qualidade da carne. Sessenta e quatro suínos da linhagem Topigs, com peso médio inicial de 26,56 ± 3,11 kg, foram distribuídos em um delineamento experimental de blocos casualizados em esquema fatorial 2 x 2, sendo dois níveis de ractopamina (0 e 5 ppm) e dois níveis de cromo-metionina (0 e 400 ppb), perfazendo quatro tratamentos. Os tratamentos foram: C controle: dieta basal constituída por milho e farelo de soja; Rac ractopamina: dieta basal com a inclusão de 5 ppm de ractopamina dos 70,32 aos 114,80 kg de PV; Cr-Met cromometionina: dieta basal com a inclusão de 400 ppb de cromo-metionina dos 26,56 aos 114,80 kg de PV; Rac + Cr-Met ractopamina + cromo-metionina: dieta basal com a inclusão de 400 ppb de cromo-metionina dos 26,56 aos 114,80 kg de PV e 5 ppm de ractopamina dos 70,32 aos 114,80 kg de PV. Foram utilizados oito repetições (blocos) por tratamento e dois animais por unidade experimental (um macho castrado e uma fêmea). O período experimental foi dividido em três fases, de acordo com o peso dos animais: crescimento I: 26,56 aos 50,51 kg de PV; crescimento II: 50,51 aos 70,32 kg de PV e a terminação: 70,32 aos 114,80 kg de PV. Ao atingirem o peso vivo médio de 114,80 ± 4,98 kg, os animais foram submetidos a jejum sólido de 24 horas e, posteriormente, encaminhados ao abate. As meias-carcaças foram resfriadas por cerca de 24 horas e avaliadas para o rendimento de carcaça fria, comprimento de carcaça, espessura de toucinho, área de olho-de-lombo e relação gordura:carne. Amostras do músculo Longissimus dorsi de cada animal, foram coletadas 24 horas post mortem para as mensurações da cor, pH e perda de peso por gotejamento da carne. A suplementação de ractopamina melhorou a conversão alimentar (P=0,05) e algumas características de carcaça [rendimento de carcaça fria (P=0,05), área de olho-de-lombo (P<0,004) e relação gordura:carne (P<0,04)], sem, contudo, alterar a espessura de toucinho (P>0,05). Além disso, a adição de ractopamina na dieta dos suínos acarretou diminuição na perda de peso por gotejamento (P<0,05), bem como nos valores de a* (P<0,004) e b* (P<0,05) da carne. O cromo-metionina, embora tenha ocasionado redução no consumo diário de ração (P<0,03), não alterou as demais variáveis de desempenho (P>0,05), nem as características de carcaça (P>0,05) e a qualidade da carne (P>0,05) dos suínos. A suplementação associada de ractopamina com cromo-metionina não afetou o desempenho (P>0,05), as características de carcaça (P>0,05) e, tampouco, e qualidade da carne dos animais (P>0,05). Portanto, o presente estudo mostrou que apenas a inclusão de ractopamina na dieta contribuiu para a melhoria do desempenho, características de carcaça e até mesmo da qualidade da carne dos suínos. / The purpose of this study was to evaluate the effect of ractopamine, chromium-methionine and their combinations in growing and finishing diets on performance, carcass traits and meat quality of pigs. Sixty-four pigs of Topigs genetic with 26.56 ± 3.11 kg initial live weight were used in a randomized complete block design experiment, in a 2 x 2 factorial arrangement [two ractopamine levels (0 and 5 ppm) and two chromium-methionine levels (0 and 400 ppb)], resulting four treatments. The treatments were: C control: basal diet with corn and soybean meal; Rac ractopamine: basal diet with the inclusion of 5 ppm de ractopamine dos 70.32 to 114.80 kg; Cr-Met chromium-methionine: basal diet with the inclusion of 400 ppb of chromium-methionine dos 26,56 to 114,80 kg; Rac + Cr-Met ractopamine + chromiummethionine: basal diet with the inclusion of 400 ppb of chromium-methionine dos 26,56 to 114,80 kg and 5 ppm of ractopamine dos 70.32 to 114.80 kg. Eight replications (blocks) per treatment and two animals per experimental unit (one barrow and one gilt) were used. The experimental period was divided in three phases according to average live weight of animals: growing I: 26.56 to 50.51 kg BW, growing II: 50.51 to 70.32 kg BW and finishing: 70.32 to 114.80 kg BW. When pigs reached 114.80 ± 4.98 kg BW, a 24-hour-fasting period was given to animals before slaughter. Pig carcasses were stored for 24 hours in cold camera and then data of cold carcass yield, carcass length, backfat thickness, loin eye area and fat area/loin eye area ratio were registered. Samples of Longissimus dorsi of each animal were taken 24 hours postmortem to determine colour, pH and drip loss of meat. Ractopamine improved feed conversion (P=0.05) and some carcass traits [cold carcass yield (P=0.05), loin eye area (P<0.004), fat area/loin eye area ratio (P<0.04), but not affect the backfat thickness (P>0.05)]. Ractopamine reduced water drip loss (P<0.05), a* (P<0.004) and b* (P<0.05) values of meat. Chromium-methionine decreased daily feed intake (P<0.03), but did not affect the carcass traits (P>0.05) and meat quality (P>0.05) of pigs. Combination of ractopamine and chromium-methionine did not affect performance (P>0.05), carcass traits (P>0.05) and meat quality (P>0.05) of animal. Therefore, this study showed that only ractopamine can enhance performance, carcass traits and some meat quality traits of pigs.

O objetivo do trabalho foi estudar os efeitos de probióticos, prebióticos e extratos vegetais como alternativas ao uso de agentes antimicrobianos como promotores do crescimento de leitões recém-desmamados, por meio da morfologia de órgãos, frequência de diarréia, microbiologia intestinal, histologia de epitélio intestinal, digestibilidade dos nutrientes e desempenho. Foram realizados dois experimentos em blocos casualizados, nos quais foram testados cinco tratamentos: controle - ração basal; antimicrobiano - basal com 50 ppm de bacitracina de zinco + 50 ppm de olaquindox; probiótico - basal com 1300 ppm de probiótico à base de Bacillus subtilis e Bacillus licheniformis; prebiótico - basal com 3000 ppm de mananoligossacarídeo; extrato vegetal – basal com 500 ppm de extrato vegetal (alho, cravo, canela, pimenta, tomilho, cinamaldeído e eugenol). No Experimento I utilizaram-se 40 leitões desmamados aos 21 dias de idade, distribuídos a 20 gaiolas suspensas com dois leitões (um macho castrado e uma fêmea) em cada gaiola (unidade experimental) e quatro repetições por tratamento. Durante a última semana do período experimental foi realizado o ensaio de digestibilidade dos nutrientes, por meio da metodologia da coleta parcial de fezes, tendo o óxido crômico (Cr2O3) como marcador. Ao 35º dia, um animal de cada unidade experimental foi abatido para a coleta de dados referentes à morfometria de órgãos, de amostras do epitélio intestinal para análise histológica e de amostras do conteúdo do jejuno/íleo para análise microbiológica. No Experimento II, foram utilizados 120 leitões desmamados aos 21 dias de idade, alocados em 60 baias com dois animais (um macho castrado e uma fêmea) por baia (unidade experimental), perfazendo 12 repetições por tratamento. As variáveis de consumo diário de ração, ganho diário de peso e conversão alimentar foram coletadas e analisadas nos períodos de 1-14, 15-35 e 1-35 dias de experimentação. Também foi avaliada a frequência de diarréia. Apesar da diferença não ser significativa (P>0,05), em valores numéricos, os agentes antimicrobianos melhoraram o ganho diário de peso nos períodos de 15-35 (+22%) e 1-35 (+21,4%) dias de experimentação. Também melhoraram (P<0,05) a digestibilidade aparente de nutrientes (P<0,05) em relação ao tratamento controle. O prebiótico mananoligossacarídeo foi a melhor alternativa aos antimicrobianos, proporcionando um ganho diário de peso, numericamente (P>0,05), 25,7% superior ao tratamento controle e equivalente ao tratamento antimicrobiano, nos primeiros 14 dias de experimentação. Entretanto, o prebiótico mananoligossacarídeo não melhorou a conversão alimentar (P>0,10). Nos períodos de 15-35 dias e no período total, o desempenho dos animais do tratamento prebiótico foi intermediário entre o tratamento controle e antimicrobiano. O probiótico não melhorou o desempenho (P>0,10) e não proporcionou nenhuma alteração nas outras variáveis estudadas. Os extratos vegetais aumentaram o peso relativo do pâncreas (P<0,05) e melhoraram a digestibilidade aparente da matéria seca da dieta (P<0,05). No entanto, os extratos vegetais não melhoraram (P>0,10) o desempenho dos leitões. / The purpose of this work was to evaluate the probiotics, prebiotics and herbal extracts as alternatives to antimicrobial agents as growth promoters of weanling pigs, based on the morphometry of organs, fecal score, intestinal microbiology, intestinal histology, nutrient digestibility and performance. Two randomized complete block design experiments were carried out to compare five treatments: control - basal diet; antimicrobial - basal diet plus Zn bacitracin and olaquindox (50 ppm of each); probiotic - basal diet plus 1300 ppm of probiotic (Bacillus subtilis and Bacillus licheniformis); prebiotic - basal diet plus 3000 ppm of mannanoligosaccharide; herbal extract - basal diet plus 500 ppm of herbal extract (garlic, clove, cinnamon, pepper, thyme, cinnamaldehyde and eugenol). In Experiment I, forty 21-d-weaned pigs were alloted to 20 suspended pens, with two pigs (a barrow and a gilt) per pen (experimental unit) and four replications per treatment. In the last week, the digestibility assay was carried out, using the method of the partial faeces collection, with the chromium oxide (Cr2O3) as fecal marker. On 35th day, an animal of each experimental unit was slaughtered to collect the morphometry data, the intestinal epithelium samples for histological analysis, and the content of the jejunum/ileum for microbiological analysis. In Experiment II, one hundred and twenty 21-d-weaned pigs were used, with 12 replications per treatment and two animals (a barrow and a gilt) per experimental unit. The daily feed intake, the average daily gain and the feed conversion were analyzed for 1-14, 15-35 and 1-35 days of experimental periods. The fecal score was also evaluated. Although not significant (P>.05), the antimicrobial agents improved the average daily gain of weanling pigs, during 1-35 (+21.4%) and 15-35 (+22%) days of experimental periods, compared to pigs fed control diet. The nutrient apparent digestibility of pigs fed antimicrobial were higher (P<.05) than of those fed control. The prebiotic mannanoligosaccharide showed to be the most efficient alternative to antimicrobials. The average daily gain of pigs fed mannanoligosaccharide was, in numerical values (P<.05), 25.7% higher than of those fed control (P>.05) and similar (P>.05) to the animals of the antimicrobial treatment, during 1-14 days of experimental period. However, the feed conversion was not improved (P>.10). During 15-35 and 1-35 days of experimental periods, the performance of animals fed mannanoligosaccharide was intermediate between of those fed control and antimicrobial diet. The probiotic did not improve the growth performance of pigs (P>.10) and did not cause any effect. The herbal extracts increased the relative weight of pancreas (P<.10) and improved the dry matter apparent digestibility of the diet (P<.05), compared to control treatment. However, the herbal extracts did not improve pig performance (P>.10) in any period.

O presente trabalho teve como objetivo avaliar o efeito do emurchecimento da forragem associado ou não ao uso do aditivo bacteriano-enzimático na ensilagem do capim Tifton 85 (Cynodon spp.). Foi realizada avaliação de perdas nos painéis das silagens experimentais, assim como da composição bromatológica e do tamanho de partículas das mesmas. Ocorreram observações periódicas de comportamento ingestivo, consistência ruminal, consumo voluntário, parâmetros de fermentação ruminal, digestibilidade dos nutrientes, taxa de passagem de sólidos e líquidos ruminais em novilhos que receberam rações contendo 80% desses volumosos na base seca. Foram utilizados sete novilhos da raça Nelore, distribuídos ao acaso em um experimento do tipo quadrado latino 7 x 7. Os tratamentos avaliados foram resultantes da combinação de 3 concentrações de matéria seca (MS) na silagem do capim Tifton 85 (25, 45 e 65%) associadas (CA) ou não (SA) ao aditivo bacteriano-enzimático. Um tratamento adicional contendo 55% MS sem aditivo também foi avaliado. O aditivo utilizado foi o produto comercial SIL–ALL®, contendo cepas de Streptococcus faecium, Lactobacillus plantarum, Pediococcus acidilactici e as enzimas hemicelulase, celulase e amilase, aplicadas na dosagem de 250 g de SIL–ALL® diluídas em 50 litros de água por tonelada de forragem, imediatamente antes da ensilagem. A técnica de emurchecimento possibilitou elevar o teor de matéria seca mas, da mesma maneira que a adição do inoculante bacteriano-enzimático, provocou alterações pouco pronunciadas na composição química da silagem. Nas silagens SA o aumento da concentração de MS foi acompanhado de elevação no teor de FDN resultando em redução do teor de carboidratos não fibrosos (CNF). A fração protéica B3 nas silagens apresentou tendência de aumento com a elevação na concentração de MS, em função da progressiva elevação da fração N insolúvel no FDN, simultaneamente à redução N insolúvel no FDA. A avaliação dos silos após abertura, indicou aumento de perdas com a elevação da concentração de MS que, entretanto, podem ter sido decorrentes de uma menor taxa de utilização dos fardos contendo silagens com menor umidade. Tanto a consistência ruminal, avaliada como o tempo de ascensão do pêndulo no interior do rúmen (1194 seg.), como os parâmetros de cinética de passagem de sólidos (3,09% hora -1 ) e de líquidos (4,37% hora -1 ) foram similares para os tratamentos avaliados. O comportamento ingestivo dos animais não foi alterado com a elevação da concentração de MS da silagem, sendo observados tempos (minutos dia -1 ) e taxas (min kg MS –1 ) de ingestão de MS (324; 47), ruminação (518; 75) e mastigação (841; 123), respectivamente. O consumo diário de MS das rações, pelos animais, foi semelhante para os tratamentos avaliados, resultando em média de 6,95 kg equivalente a 1,88% do peso corpóreo. As concentrações molares (mM) individuais e totais (140,29) dos ácidos graxos voláteis, a relação acetato:propionato (4,2:1), concentração de nitrogênio amoniacal (6,31 mg dL -1 ) e o pH (6,76), do fluido ruminal, não foram alterados pelos tratamentos estudados. As digestibilidades da matéria orgânica (71,5%), proteína bruta (69,1%), FDN (71,3%), FDA (64,9%) e hemicelulose (83,3%) não foram, em geral, fortemente influenciadas pela concentração de MS da silagem do capim Tifton 85, aditivadas ou não. No entanto, a digestibilidade da MS (71,0%) apresentou um comportamento quadrático (P<0,05), com ligeira elevação ao redor de 45% de concentração de MS, nas silagens. Diante destas observações, pode-se concluir que o emurchecimento da forragem e o uso de aditivo bacteriano-enzimático não resultaram em alterações expressivas nos parâmetros ruminais, digestivos e no comportamento ingestivo dos animais. / The present trial aimed to study the effects of forage wilting associated or not with a bacterial-enzymatic inoculant on ensiling Tifton 85 grass. Front panel of experimental silos (325 kg square wrapped bales) were daily scored for fungi development and weekly sampled for chemical composition analysis and particle chop length measurements. In a randomized 7 x 7 Latin square design, seven ruminal cannulated growing beef steers were assigned to diets (treatments) containing 80% silage. Periodically chewing behavior, feed intake, ruminal mat consistency, ruminal fermentation parameters, nutrient digestibility and ruminal rate of passage of solids and liquids phases were evaluated. Treatments combined 3 planned dry matter (DM) concentrations (25, 45 and 65%) of Tifton 85 grass silage associated (CA) or not (SA) with the bacterial-enzymatic (BE) inoculant source. An additional treatment containing wilted silage (55% DM) without (BE) was also studied. SIL-ALL®, the BE source used, was sprayed onto chopped forage just before silage packing at a rate of 250g diluted in 50L of deionized water per ton of wet forage. The commercial BE showed the following composition: Streptococus faecium, Lactobacillus plantarum, Pediococcus acidilactici plus hemicellulase, cellulase and starch degrading enzymes. The wilting of forage allowed to increase the DM concentration of silages, however, as well as BE addition, it was not effective to promote major changes on chemical composition of silages. On SA silages, higher NDF levels were observed as the DM level of the wilted silage increased, which resulted in lower non-fiber carbohydrate (NFC) fraction. The B3 protein fraction (NDIN minus ADIN) showed an increase as the DM concentration of wilted silage was raised, probably as result of an opposite trend between neutral detergent insoluble nitrogen (NDIN) and acid detergent insoluble nitrogen (ADIN) across DM levels. Silage losses, measured as both, fungi scored at bale front panel and percentage of spoiled silage, indicated higher levels associated with increased DM concentration. It might be explained by the lower bale unloading rate observed during the feeding trial, mainly in higher DM wilted silages. Both, ruminal mat consistency, measured as time required to weight ascension into the rumen (1194 sec.) as well as ruminal kinetics, evaluated as passage rate of solid (3.09% hour -1 ) and liquid (4.37% hour -1 ) phases were similar for all treatments. Animal ingestive behavior measured as total time (minutes day -1 ) and rate (minutes DM kg -1 ) was not changed across silages DM concentration, averaging DM eating (324; 47), DM ruminating (518; 75) and DM chewing (841; 123), respectively. Residual daily time was spent with drinking and idling activities. The DM intakes observed among experimental diets, containing wilted silages, were not statistically different and averaged 6.95 kg day -1 or 1.88% as body weight basis. Individual and total (140.29) molar concentration (mM) of volatile fatty acids (VFA), acetate:propionate ratio (4,2:1), ammonia-N (6.31 mg dL -1 ) and average daily pH (6.76) of ruminal fluid were similar across silages diets. Nutrient digestibilities were, also, not significantly affected by the increased DM concentration in wilted silages or by the BE inoculation, as follows: organic matter (71.5%), crude protein (69.1%), NDF (71.3%), ADF (64.9%) and hemicellulose (83.3%). DM digestibilities, however, performed a quadratic pattern (P<0.05), with a slight increase at 45% DM concentration in silages. According to the results it may be concluded that neither the increase of DM concentration of grass silages nor the addition of bacterial-enzymatic inoculant changed animal ingestive behavior or improved ruminal and digestive parameters analyzed.

<p>Thyroid hormone is essential for regulating adult metabolism and proper fetal development. Under normal conditions, maternal and fetal thyroid hormones are subject to metabolism at the placenta and within fetal tissues through deiodination and sulfation to regulate fetal exposure to the bioactive hormone. Disruptions of the thyroid hormone system can result in non-thyroidal illness syndrome (NTIS), which is classified as the dysregulation of thyroid hormone homeostasis. The exact cause of the alterations in circulating thyroid hormone levels during NTIS is not well- known. In comparison, hypothyroidism results from the absence of thyroid hormone production and presents as low thyroid hormone levels.</p> <p>Porcine reproductive and respiratory syndrome virus (PRRSV) crosses the late gestation placenta and causes suppression of circulating maternal and fetal thyroid hormone. Chapter 2 investigates the potential role of thyroid hormone metabolism in this disruption. Pregnant gilts were challenged with PRRSV2 (n=22) or sham inoculated (n=5) at gestation day 85. Samples were collected on day 106, and viral load was assessed in fetal serum and thymus. From the entire fetal population, three distinct subsets of fetuses representing biological extremes were identified, including uninfected with no detectable viral load (UNIF), high viral load viable (HV-VIA), or high viral load with severe meconium staining (HV-MEC). In addition, control fetuses from sham inoculated gilts (CON) were used as a reference group. Samples of fetal liver, kidney, and the corresponding fetal placenta and maternal endometrium for n=10 fetuses per group were then used to evaluate gene expression. A total of 11 genes associated with thyroid hormone metabolism including deiodinases (DIO1,2,3), sulfotransferases (SULT1A3,1B1,1C2,1E1,2A1), sulfatase (STS), and solute carriers (SLC16A2,16A10) were quantified using absolute quantification qPCR. Evidence of fetal decompensation was observed within the high viral fetuses in the form of decreased DIO1 expression within the fetal liver and increased DIO3 expression in both components of the placenta. Circulating levels of T4 and inactive thyroid hormone metabolites, reverse-triiodothyronine (rT3) and two diiodothyronines (3,5-T2 and 3,3’-T2), were measured in fetal serum. While T4 was depressed, no change was observed in circulating rT3 levels, and neither T2 metabolite reached the lower detection limit. This may suggest that alterations in thyroid hormone metabolism generate a localized effect on hormone metabolites in the respective tissues.</p> <p>Alternatively, the low levels of available T3 and T4 limit the production of downstream metabolites to be found in serum.</p> <p>The cause-and-effect relationship between PRRSV infection, fetal thyroid disruption, and the effects on fetal thyroid hormone metabolism are unclear. Therefore, Chapter 3 developed a non-pathogenic model using methimazole (MMI) to induce hypothyroidism in the late gestation fetus and evaluate the impact on fetal development and thyroid hormone metabolism. Pregnant gilts were either treated with oral methimazole or equivalent sham from gestation day 85-106 (n=4/group), followed by classification of all fetuses as live, live but meconium stained, or dead. Fetuses exposed to MMI in-utero were notably hypothyroid with significantly suppressed serum T3 and T4 and histological evidence of goiter. Surprisingly, fetuses from MMI-treated dams were substantially larger but appeared to exhibit non-allometric growth with an increase in girth but not length. The liver, kidney, and the corresponding fetal placenta and maternal endometrium were collected from a subset of 16 fetuses per group to evaluate the relative expression of five genes associated with thyroid hormone metabolism, including three deiodinases and two solute carriers known to transport thyroid hormone. Compensatory transcription of DIO3 was observed in all tissues evaluated, suggesting increased vertical transfer of maternal thyroid hormone at the placenta and decreased breakdown of thyroid hormone within fetal organs.</p> <p>The evaluation of thyroid hormone metabolism within the fetus and within the placenta has allowed us to differentiate suppressed thyroid hormone levels of the pig fetus under pathogenic and non-pathogenic conditions. In the context of PRRSV infection, the observed decompensation of thyroid hormone metabolism would further exacerbate the hypothyroid state and is therefore consistent with NTIS. In contrast, fetuses with induced thyroid hormone suppression following maternal exposure to MMI showed compensatory thyroid hormone metabolism in the same tissues. This indicates true hypothyroidism and clearly demonstrates a fetal capacity to respond to such endocrine disruption.</p>

Two consecutive experiments were conducted to test a hypothesis that mixed grazing outcome is influenced by the type of stocking system applied. The objective of both experiments was to investigate the influence of co-grazing with sheep on cattle liveweight gain (LWG) under continuous (C) and rotational (R) stocking, where sheep weekly liveweight change under the two stocking systems was kept similar. In experiment I nine yearling heifers (266 ± 4.5 kg liveweight) and 27 ewe hoggets (54±0.9 kg liveweight) were continuously stocked for 19 weeks on an irrigated perennial ryegrass-white clover pasture (2.95 ha) maintained at a sward surface height (SSH) of 5cm by adding or removing additional animals in a fixed ratio (1: 1 W⁰.⁷⁵ cattle:sheep). An equal area of pasture was rotationally stocked by a similar group of animals where they received a new area of pasture daily and also had access to the grazed area over the previous 2 days. The size of the new area provided daily was such that the weekly liveweight change of rotationally co-grazed sheep was equal to that of those continuously co-grazed with cattle. Similar groups of animals were used in the second experiment with additional group of 9 heifers grazed alone on C and R pastures. Liveweight of animals was recorded weekly and final fasted weight was determined after 24-hour total feed restriction. SSH on both treatment swards was recorded daily. There were three intake measurement periods spread over the trial period. Organic matter intake (OMI) was predicted from the ratio of N-alkanes in faeces and herbage. Diet composition was determined by dissecting oesophageal extrusa samples. Grazing behaviour (bite rates and grazing time) were also recorded. The mean SSH for C pasture was 5.1±0.09 cm. Overall pre- and post-grazing SSH for R pasture was 15.9 ±0.12 and 5.6 ±0.07 cm, respectively. As determined by the protocol average daily LWG of sheep was similar between C and R (147 (±5.8) vs 138 (±6.7) g day⁻¹; (P>0.05). In contrast, cattle continuously stocked with sheep grew 200 g day⁻¹ slower than those rotationally stocked with sheep (800 (±41.6) vs 1040 (±47.7) g day⁻¹, P<0.0l). R heifers achieved 30 kg higher final fasted liveweight than C heifers (350 vs 381 kg; P<0.01). Overall LWG per ha was also 6 % higher under R than C stocking (674 vs 634 kg ha⁻¹). The OMD of both sheep (73.5 vs 75.8 %) and cattle (75.8 vs 78.0 %) diets was similar under continuous and rotational stocking. There was no significant difference OMI data also concurred with the L WG data (Cattle: 7.94 vs 6.31 (±0.32) kg day⁻¹ (P<0.05); sheep: 1.40 vs 1.44 (±0.04) kg day⁻¹ for Rand C treatments, respectively). There was no difference in clover content of cattle diet under C and R treatments. C heifers had higher number of bites per minute than R heifers (62 vs 56; P<0.05). Proportion of heifers seen grazing (every 15-minute) during four 24-hour observations was greater on C than R pasture (0.44 vs 0.31 (±0.03); P<0.05). The similarity coefficient between sheep and cattle diet was 0.61 and 0.76 under C and R stocking, respectively. The lower daily LWG of C heifers was attributed to (a) the lower SSH under C than R stocking and/or (b) the inability of cattle to compete well with sheep where there is small, continual renewal of resources (C) in contrast to a large periodic renewal under R stocking. This experiment showed that the outcome of mixed gruing can be influenced by the stocking system chosen. But it was not possible to apportion the difference in LWG of cattle between mixed grazing per se and the difference in mean grazed sward height (5.1 for C vs 10.8 cm for R). A second experiment was conducted to determine the relative performance of cattle co-grazed with sheep (CS) and grazed alone (CA) under each stocking system. Hence, there were four treatments. CA- continuous stocking (CA-C), CS- continuous stocking (CS-C), CA- rotational stocking (CAR) and CS- rotational stocking (CS-R). A total area of 4.42 ha was allocated to each stocking system. Under C stocking, 2.95 ha (2/3) was assigned to CS-C and 1.47 ha (1/3) to CA-C, and SSH on both treatments was kept at 4 cm by adding or removing extra animals. Under R stocking, CA-R and CS-R grazed side by side separated by an electric fence. They were given a fresh area daily, the size of which was varied such that the weekly LW change of R sheep was equal to that of the C sheep. CA-R received one-third of the new area though the size was adjusted regularly to achieve the same post-grazing SSH with CS-R. Measurements included: weekly liveweight change, OMI (two periods) and diet composition (using N-alkanes). The mean SSH of CA-C and CS-C swards was 4.27 and 4.26 (±0.02) cm, respectively. CA-R and CS-R swards had mean pre-grazing SSH of 14.9 and 15.2 (±0.08) cm and post-grazing heights of 4.87 and 4.82 cm (±0.03), respectively. The proportion of areas infrequently grazed was higher for CA-C than CS-C swards (0.22 vs 0.17, respectively). C and R sheep daily LWG: 155 (±0.6) and 147 (±0.7) g, and OMI: 1.96 and 2.04 (±0.ll) kg, respectively, were not significantly different. They also had similar diet composition. In comparison, CS-C heifers grew only at 69 % of the daily LWG achieved by CS-R heifers (706 vs 1028 (±72) g; P<0.05). LWG of CA-C and CA-R was 916 and 1022 (±72) g day⁻¹, respectively. The difference in LWG between CS-R and CS-C (D₁) heifers was due to difference in mean sward height, stocking system and mixed grazing, while D₂ (difference in LWG between CA-R and CAC) was due to difference in mean sward height and stocking system. D₁-D₂ (the effect of stocking system on mixed grazing) was 216 g and made up 67 % of the total difference between CS-R and CS-C. There was a significant stocking system-species mixture interaction in the final fasted LW achieved by heifers. Final fasted LW was significantly lower for CS-C than CA-C heifers (283 vs 323 (±9.7) kg), but did not differ between CS-R and CA-R (332 vs 330 (±9.7) kg, respectively). The digestibility of diet OM was similar for both continuously and rotationally stocked sheep (84.4 vs 83.2 %, respectively). Cattle diet OMO was 76.5, 74.7, 79.4 and 77.8 for CA-C, CS-C, CA-R and CS-R respectively (P>0.05). Differences in OMI followed a similar pattern to daily LWG. Mean daily OMI was 8.98, 6.24, 8.80 and 9.45 (±0.40) kg for CA-C, CS-C, CA-R and CS-R, respectively. Clover content of the diet of CA-C heifers was three times higher than that of CS-C heifers (30.7 vs 10.4 % OM; P<0.05); there was no difference in clover content of diets of CS-R and CA-R heifers (21.5 vs 23.9 % OM, respectively). In both stocking systems LWG per ha was higher on CA than CS treatments. These results suggested that the disadvantage of selective clover grazing by sheep outweighed the advantages of sheep grazing around cattle dung patches under continuous stocking. Under rotational stocking, rapid diurnal changes in sward conditions probably limited selective grazing by both sheep and cattle such that there was no disadvantage to CS cattle. The results do not provide a basis for recommending grazing cattle with sheep rather than cattle alone, but do provide some basis for recommending co-grazing of sheep and cattle using rotational rather than continuous stocking.

This thesis describes two approaches for studying of lipoprotein metabolism in sheep. The first approach involves the assay of lipoprotein lipase (LPL) activity to determine the role of lipoprotein-triacylglycerol fatty acids in fat deposition in sheep. This enzyme is the rate limiting enzyme in the hydrolysis of fatty acids from lipoprotein-triacylglycerol. The second approach was to characterize and quantify in vivo lipoprotein metabolism using iodinated very low density lipoprotein (¹²⁵I-VLDL) and low density lipoprotein (¹³¹I-LDL). Cross-bred lambs were divided into two treatment groups and either weaned early at 5 weeks of age or remained suckling. Lambs were slaughtered at 12 or 23 weeks at which time the body composition and adipose tissue LPL activity were determined. The differences in rearing led to differences in body composition. The suckled lambs were larger and fatter than weaned lambs. The increased fatness in the suckled lambs was associated with increased LPL activity (U/mg protein) in subcutaneous adipose tissue and was reflected in higher LPL activity in post-heparin plasma (PHP) taken 2 days prior to slaughter. The role of insulin in the regulation of LPL activity was investigated by either infusing a subset of the weaned and suckled lambs with insulin for 7 or 18 weeks or using the euglycemic clamp technique to study the effect of short insulin infusions. The long term infusion of insulin had no significant effect on PHP LPL or on adipose tissue LPL (U/g tissue). However, after infusing insulin for 6h at 6.3 mU.kg⁻·⁷⁵.h⁻¹ during the euglycemic clamps, a two fold increase in LPL activity in biopsied subcutaneous adipose tissue was observed. In the second approach, in vivo lipoprotein metabolism was investigated in 4 lambs using apolipoprotein B as a marker. Following the simultaneous injection of ¹²⁵I VLDL and ¹³¹I VLDL, the specific activities of apoB in VLDL, IDL and LDL fractions were determined. ApoB specific activity curves demonstrated that VLDL is metabolised to IDL and subsequently to LDL. The turnover of VLDL-B (3.45mg.d⁻¹.kg⁻¹) and LDL-B (4.8mg.d⁻¹.kg⁻¹) was calculated by fitting the VLDL-¹²⁵I-B and LDL-¹³¹I-B specific activity data to a mono-exponential equation. The metabolism of lipoproteins, inferred from the study of apoB, was shown to be similar in sheep to that reported in other animals although the amount of lipoprotein synthesised was low. A model to describe the kinetics of apoB metabolism in sheep was developed using SAAM. The proposed model features a three pool delipidation chain for VLDL, and subsystems containing two pools for IDL and LDL. IDL may be catabolised to LDL or cleared directly from the plasma. The developed model can now be used to compare the metabolism of lipoproteins in different physiological states and to design new experiments to study lipoprotein metabolism further.

Submitted by Claudinei Pracidelli (cpracide@ipen.br) on 2016-06-22T11:39:54Z No. of bitstreams: 0 / Made available in DSpace on 2016-06-22T11:39:54Z (GMT). No. of bitstreams: 0 / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Tese (Doutorado em Tecnologia Nuclear) / IPEN/T / Instituto de Pesquisas Energeticas e Nucleares - IPEN-CNEN/SP / FAPESP:11/21708-6

Submitted by Claudinei Pracidelli (cpracide@ipen.br) on 2016-06-22T11:39:54Z No. of bitstreams: 0 / Made available in DSpace on 2016-06-22T11:39:54Z (GMT). No. of bitstreams: 0 / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / A deficiência de hormônio de crescimento (DGH) é a deficiência mais comum entre os hormônios pituitários. A terapia utilizada atualmente consiste de injeções diárias de hormônio de crescimento humano recombinante (r-hGH), entretanto esta terapia apresenta alguns inconvenientes, como a necessidade de frequentes injeções de r-hGH durante um longo período de vida, dependendo da severidade da deficiência, e o alto custo do hormônio, em razão dos dispendiosos processos de purificação. Uma alternativa ao tratamento padrão seria aquele no qual fossem evitados estes tipos de inconvenientes e o processo de liberação da proteína fosse sustentável, por um longo período e promovesse níveis normais e sustentáveis do fator de crescimento semelhante à insulina I (IGF-I), o principal mediador dos efeitos do GH. Uma alternativa é a terapia gênica in vivo, baseada na administração de DNA plasmidial em diversos órgãos/tecidos, seguida de eletroporação. É considerada uma metodologia bastante promissora e que tem sido alvo de vários estudos para diversos tipos de deficiências sistêmicas. Neste trabalho foram realizadas diversas administrações de um plasmídeo contendo o gene do hormônio de crescimento humano, nos músculos quadríceps exposto ou tibial anterior sem exposição, seguidas de eletroporação, em camundongos anões e imunodeficientes (lit/scid) com 40-80 dias de idade, na tentativa de obter uma correção fenotípica do nanismo, mediante a avaliação de parâmetros de crescimento. A administração deste plasmídeo no músculo tibial anterior, em camundongos com a idade inicial de 40 dias, foi capaz de proporcionar uma normalização dos níveis de mIGF-I, quando comparados aos dos camundongos não-deficientes de GH. Além disso, foram obtidos valores de catch-up dos parâmetros de crescimento longitudinal de 36-77%. Visando uma maior eficiência na expressão de GH, foram construídos plasmídeos parentais, e a partir destes, foram produzidos minicírculos de DNA com os promotores do CMV e Ubiquitina C e com os cDNAs de hGH e mGH. Estes minicírculos de DNA foram transfectados em células HEK 293 e foram até 2 vezes mais eficientes em relação aos plasmídeos convencionais com o promotor do CMV. Estes dados são bastantes promissores e abrem caminho para ensaios mais eficientes, utilizando este tipo de protocolo de terapia gênica para a DGH, visando uma normalização de todos os parâmetros de crescimento. / Tese (Doutorado em Tecnologia Nuclear) / IPEN/T / Instituto de Pesquisas Energeticas e Nucleares - IPEN-CNEN/SP / FAPESP:11/21708-6

<p dir="ltr">Newborn animals undergo a lot of early-life stress that heavily impact on their long-term growth, performance, and welfare. Typically, the stress would indirectly interfere with the capacity of these neonates to utilize dietary nutrients and consequently impact tissue growth and development. In piglets, weaning is a stressful situation characterized by disruption of intestinal epithelial cell development which causes poor digestion of solid feed and a negative impact on absorption of nutrients especially in the post-gastric region. In addition, weaning in piglets could cause an increase in cellular assault by reactive oxygen species thereby potentially causing gut leakiness and paracellular loss of nutrients along the intestinal tract. In broiler chickens, access to feed may take up to 72 h following hatching which may affect their gut development as well as their gut microbiota. After the first feed ingestion, there is a sharp increase in the gut microbiota which triggers an increase in the development of the immune system as well as the gut. There is continuous attention on the strategies and nutritional interventions to mitigate or ameliorate the adverse effects of early life stressors in these food animals, especially in broiler chickens and piglets. In the studies described in this dissertation, myo-inositol (purely supplemented or phytase-induced) and luteolin were tested as nutritional strategies to mitigate the effects of early-life stressors on growth and the potential mechanisms by which myo-inositol and luteolin regulate growth were investigated.</p><p dir="ltr">In study I, the effect of myo-inositol on growth in 128 postweaning piglets fed protein-deficient corn-soy diets was tested. There were 4 dietary treatments in a randomized complete block design with body weight as the blocking factor. The treatments consisted of 1 positive control (PC) diet formulated to meet all the nutrient requirements of the piglets with a 20% crude protein (CP); the remaining 3 diets were the negative control (NC) diets with a 3% reduction in CP, a 2 g/kg myo-inositol supplemented negative control diet (NC+INO), and phytase (3,000 FTU/kg) supplemented negative control (NC+PHY) diet. The results showed that phytase enhanced the apparent total tract digestibility (ATTD) of P in the weanling pigs. Myo-inositol supplementation in a protein-deficient diet improved (P < 0.05) porcine plasma myo-inositol concentration while an in vitro myo-inositol incubation with intestinal epithelial cells increased the expression of genes that encode for Claudin-1, Claudin-3, Claudin-4, ZO-1, NaPiIIb, GLUT2, and SLC7A2. The in vitro analysis of tight junction integrity in the IPEC-J2 cells indicated by the transepithelial electrical resistance and FITC-Dextran permeability showed an enhancement in response to myo-inositol treatment. Although the in vivo study found that myo-inositol did not improve growth performance or ATTD, the in vitro myo-inositol enhanced markers of gut health and function.</p><p dir="ltr">In study II, the effect of myo-inositol on the growth of broiler chickens was tested. In this study, there were 6 experimental treatments based on two dietary protein levels (PC and NC) and three supplement types (BASAL, INO, and PHY) resulting in a 2 x 3 factorial arrangement in a completely randomized design. A total of 384 broiler chickens comprising 6 treatments with eight replicates per treatment and 8 birds per replicate were used. The birds were fed a common starter diet for the initial 7 days after they arrived at the poultry unit followed by a 14-day trial. The protein-deficient diet decreased the feed efficiency of the birds. Phytase addition increased (P < 0.05) the apparent ileal digestibility (AID) and ATTD of P and Ca in both PC and NC groups. The jejunal gut morphology was enhanced by supplemental phytase as indicated by an increase in villus height and the ratio of the villus height-to-crypt depth, coupled with an increase in serum myo-inositol concentration caused by both myo-inositol and phytase. In conclusion, myo-inositol showed a differential influence on growth performance, nutrient digestibility, and gut morphology.</p><p dir="ltr">In study III, the effects of luteolin on weanling pigs and IPEC-J2 cells were examined. A total of 48 piglets were randomly allotted to two dietary treatments consisting of a control group and a luteolin (LUT)-supplemented dietary group for a 4-week trial. A weekly assessment of the growth performance and expression of specific proteins in the jejunal mucosa was performed. In each dietary group, 8 piglets were slaughtered at weeks 1, 2, and 4 postweaning to collect blood, jejunal and ileal mucosa, and tissues. Luteolin supplementation numerically improved the ADG and G:F of the pigs. Luteolin feeding altered the jejunal and ileal gut morphology with increased villi height (P < 0.05) and villus height-to-crypt depth ratio (VCR, P < 0.05) in the jejunum and decreased crypt depth in the ileum. The effect of luteolin on IPEC-J2 global proteome and phosphor-proteome showed that luteolin could potentially improve intestinal barrier integrity by enhancing the abundance of proteins important in cell growth and survival. </p><p dir="ltr">In summary, dietary supplementation with myo-inositol and luteolin could regulate growth and nutrient digestibility in broiler chickens and weanling pigs by enhancing the integrity of intestinal cells and facilitating the expression of nutrient transporters that are significant in the uptake of nutrients across the lining of the gastrointestinal tract. Phytase supplementation improves the P release from phytate in the diets thereby alleviating its loss.</p>

Prolificacy in sheep, under most types of flock management, may exert an overwhelming influence on profitability. Three major classes of sheep farming are found in New Zealand, namely Extensive farming, on high country and droughty areas where wool is the chief product, Store sheep farming, on harder hill country, where income is derived from sales of both wool and surplus stock, and Fat lamb farming, in the easier and improved areas, where sales of fat stock almost exclusively dictate the size of the income. In all three types, ewe fertility is of paramount importance. This investigation was undertaken as a pilot attempt to demonstrate, with more accuracy, the source of, or reason for, the additional lambs which result from flushing ewes, in as far as this practice may increase both ovulation rate and subsequent mortality or merely reduce mortality in developing ova at some as yet underdefined stage of early pregnancy. The nature of the experiment was such that a study of the time-relationships of ovum loss and embryo mortality at various stages in early pregnancy could be made. Thus the matings of 225 ewes in two separate mobs were observed and slaughter dates were measured for individual ewes from mating times. By this means a considerable collection of both field and laboratory data was made available for a study of comparative individual and group reactions to the flushing treatment applied.