Angiogenesis and angioregression gene expression analyses in swine corpus luteum

The corpus luteum (CL) lifespan is characterized by a rapid growth, differentiation and controlled regression of the luteal tissue, accompanied by an intense angiogenesis and angioregression. Indeed, the CL is one of the most highly vascularised tissue in the body with a proliferation rate of the endothelial cells 4- to 20-fold more intense than in some of the most malignant human tumours. This angiogenic process should be rigorously controlled to allow the repeated opportunities of fertilization. After a first period of rapid growth, the tissue becomes stably organized and prepares itself to switch to the phenotype required for its next apoptotic regression. In pregnant swine, the lifespan of the CLs must be extended to support embryonic and foetal development and vascularisation is necessary for the maintenance of luteal function. Among the molecules involved in the angiogenesis, Vascular Endothelial Growth Factor (VEGF) is the main regulator, promoting endothelial cells proliferation, differentiation and survival as well as vascular permeability and vessel lumen formation. During vascular invasion and apoptosis process, the remodelling of the extracellular matrix is essential for the correct evolution of the CL, particularly by the action of specific class of proteolytic enzymes known as matrix metalloproteinases (MMPs). Another important factor that plays a role in the processes of angiogenesis and angioregression during the CL formation and luteolysis is the isopeptide Endothelin-1 (ET-1), which is well-known to be a potent vasoconstrictor and mitogen for endothelial cells. The goal of the present thesis was to study the role and regulation of vascularisation in an adult vascular bed. For this purpose, using a precisely controlled in vivo model of swine CL development and regression, we determined the levels of expression of the members of VEGF system (VEGF total and specific isoforms; VEGF receptor-1, VEGFR-1; VEGF receptor-2, VEGFR-2) and ET- 1 system (ET-1; endothelin converting enzyme-1, ECE-1; endothelin receptor type A, ET-A) as well as the activity of the Ca++/Mg++-dependent endonucleases and gelatinases (MMP-2 and MMP-9). Three experiments were conducted to reach such objectives in CLs isolated from ovaries of cyclic, pregnant or fasted gilts. In the Experiment I, we evaluated the influence of acute fasting on VEGF production and VEGF, VEGFR-2, ET-1, ECE-1 and ET-A mRNA expressions in CLs collected on day 6 after ovulation (midluteal phase). The results indicated a down-regulation of VEGF, VEGFR-2, ET-1 and ECE-1 mRNA expression, although no change was observed for VEGF protein. Furthermore, we observed that fasting stimulated steroidogenesis by luteal cells. On the basis of the main effects of VEGF (stimulation of vessel growth and endothelial permeability) and ET-1 (stimulation of endothelial cell proliferation and vasoconstriction, as well as VEGF stimulation), we concluded that feed restriction possibly inhibited luteal vessel development. This could be, at least in part, compensated by a decrease of vasal tone due to a diminution of ET-1, thus ensuring an adequate blood flow and the production of steroids by the luteal cells. In the Experiment II, we investigated the relationship between VEGF, gelatinases and Ca++/Mg++-dependent endonucleases activities with the functional CL stage throughout the oestrous cycle and at pregnancy. The results demonstrated differential patterns of expression of those molecules in correspondence to the different phases of the oestrous cycle. Immediately after ovulation, VEGF mRNA/protein levels and MMP-9 activity are maximal. On days 5–14 after ovulation, VEGF expression and MMP-2 and -9 activities are at basal levels, while Ca++/Mg++-dependent endonuclease levels increased significantly in relation to day 1. Only at luteolysis (day 17), Ca++/Mg++-dependent endonuclease and MMP-2 spontaneous activity increased significantly. At pregnancy, high levels of MMP-9 and VEGF were observed. These results suggested that during the very early luteal phase, high MMPs activities coupled with high VEGF levels drive the tissue to an angiogenic phenotype, allowing CL growth under LH (Luteinising Hormone) stimulus, while during the late luteal phase, low VEGF and elevate MMPs levels may play a role in the apoptotic tissue and extracellular matrix remodelling during structural luteolysis. In the Experiment III, we described the expression patterns of all distinct VEGF isoforms throughout the oestrous cycle. Furthermore, the mRNA expression and protein levels of both VEGF receptors were also evaluated. Four novel VEGF isoforms (VEGF144, VEGF147, VEGF182, and VEGF164b) were found for the first time in swine and the seven identified isoforms presented four different patterns of expression. All isoforms showed their highest mRNA levels in newly formed CLs (day 1), followed by a decrease during mid-late luteal phase (days 10–17), except for VEGF182, VEGF188 and VEGF144 that showed a differential regulation during late luteal phase (day 14) or at luteolysis (day 17). VEGF protein levels paralleled the most expressed and secreted VEGF120 and VEGF164 isoforms. The VEGF receptors mRNAs showed a different pattern of expression in relation to their ligands, increasing between day 1 and 3 and gradually decreasing during the mid-late luteal phase. The differential regulation of some VEGF isoforms principally during the late luteal phase and luteolysis suggested a specific role of VEGF during tissue remodelling process that occurs either for CL maintenance in case of pregnancy or for noncapillary vessel development essential for tissue removal during structural luteolysis. In summary, our findings allow us to determine relationships among factors involved in the angiogenesis and angioregression mechanisms that take place during the formation and regression of the CL. Thus, CL provides a very interesting model for studying such factors in different fields of the basic research.

Milk fatty acid profile related to energy balance in dairy cows

Milk fatty acid (FA) profile is a dynamic pattern influenced by lactational stage, energy balance and dietary composition. In the first part of this study, effects of the energy balance during the proceeding lactation [weeks 1-21 post partum (pp)] on milk FA profile of 30 dairy cows were evaluated under a constant feeding regimen. In the second part, effects of a negative energy balance (NEB) induced by feed restriction on milk FA profile were studied in 40 multiparous dairy cows (20 feed-restricted and 20 control). Feed restriction (energy balance of -63 MJ NEL/d, restriction of 49 % of energy requirements) lasted 3 weeks starting at around 100 days in milk. Milk FA profile changed markedly from week 1 pp up to week 12 pp and remained unchanged thereafter. The proportion of saturated FA (predominantly 10:0, 12:0, 14:0 and 16:0) increased from week 1 pp up to week 12 pp, whereas monounsaturated FA, predominantly the proportion of 18:1,9c decreased as NEB in early lactation became less severe. During the induced NEB, milk FA profile showed a similarly directed pattern as during the NEB in early lactation, although changes were less marked for most FA. Milk FA composition changed rapidly within one week after initiation of feed restriction and tended to adjust to the initial composition despite maintenance of a high NEB. C18:1,9c was increased significantly during the induced NEB indicating mobilization of a considerable amount of adipose tissue. Besides 18:1,9c, changes in saturated FA, monounsaturated FA, de-novo synthesized and preformed FA (sum of FA >C16) reflected energy status in dairy cows and indicated the NEB in early lactation as well as the induced NEB by feed restriction.

Performance and metabolic profile of dairy cows during a lactational and deliberately induced negative energy balance with subsequent realimentation

Homeorhetic and homeostatic controls in dairy cows are essential for adapting to alterations in physiological and environmental conditions. To study the different mechanisms during adaptation processes, effects of a deliberately induced negative energy balance (NEB) by feed restriction near 100 d in milk (DIM) on performance and metabolic measures were compared with lactation energy deficiency after parturition. Fifty multiparous cows were studied in 3 periods (1=early lactation up to 12 wk postpartum; 2=feed restriction for 3 wk beginning at 98+/-7 DIM with a feed-restricted and control group; and 3=a subsequent realimentation period for the feed-restricted group for 8 wk). In period 1, despite NEB in early lactation [-42 MJ of net energy for lactation (NE(L))/d, wk 1 to 3] up to wk 9, milk yield increased from 27.5+/-0.7 kg to a maximum of 39.5+/-0.8 kg (wk 6). For period 2, the NEB was induced by individual limitation of feed quantity and reduction of dietary energy density. Feed-restricted cows experienced a greater NEB (-63 MJ of NEL/d) than did cows in early lactation. Feed-restricted cows in period 2 showed only a small decline in milk yield of -3.1+/-1.1 kg and milk protein content of -0.2+/-0.1% compared with control cows (30.5+/-1.1 kg and 3.8+/-0.1%, respectively). In feed-restricted cows (period 2), plasma glucose was lower (-0.2+/-0.0 mmol/L) and nonesterified fatty acids higher (+0.1+/-0.1 mmol/L) compared with control cows. Compared with the NEB in period 1, the decreases in body weight due to the deliberately induced NEB (period 2) were greater (56+/-4 vs. 23+/-3 kg), but decreases in body condition score (0.16+/-0.03 vs. 0.34+/-0.04) and muscle diameter (2.0+/-0.4 vs. 3.5+/-0.4 mm) were lesser. The changes in metabolic measures in period 2 were marginal compared with the adjustments directly after parturition in period 1. Despite the greater induced energy deficiency at 100 DIM than the early lactation NEB, the metabolic load experienced by the dairy cows was not as high as that observed in early lactation. The different effects of energy deficiency at the 2 stages in lactation show that metabolic problems in early lactating dairy cows are not due only to the NEB, but mainly to the specific metabolic regulation during this period.

Endocrine changes and liver mRNA abundance of somatotropic axis and insulin system constituents during negative energy balance at different stages of lactation in dairy cows

The liver has an important role in metabolic regulation and control of the somatotropic axis to adapt successfully to physiological and environmental changes in dairy cows. The aim of this study was to investigate the adaptation to negative energy balance (NEB) at parturition and to a deliberately induced NEB by feed restriction at 100 days in milk. The hepatic gene expression and the endocrine system of the somatotropic axis and related parameters were compared between the early and late NEB period. Fifty multiparous cows were subjected to 3 periods (1=early lactation up to 12 wk postpartum, 2=feed restriction for 3 wk beginning at around 100 days in milk with a feed-restricted and a control group, and 3=subsequent realimentation period for the feed-restricted group for 8 wk). In period 1, plasma growth hormone reached a maximum in early lactation, whereas insulin-like growth factor-I (IGF-I), leptin, the thyroid hormones, insulin, and the revised quantitative insulin sensitivity check index increased gradually after a nadir in early lactation. Three days after parturition, hepatic mRNA abundance of growth hormone receptor 1A, IGF-I, IGF-I receptor and IGF-binding protein-3 (IGFBP-3) were decreased, whereas mRNA of IGFBP-1 and -2 and insulin receptor were upregulated as compared with wk 3 antepartum. During period 2, feed-restricted cows showed decreased plasma concentrations of IGF-I and leptin compared with those of control cows. The revised quantitative insulin sensitivity check index was lower for feed-restricted cows (period 2) than for control cows. Compared with the NEB in period 1, the changes due to the deliberately induced NEB (period 2) in hormones were less pronounced. At the end of the 3-wk feed restriction, the mRNA abundance of IGF-I, IGFBP-1, -2, -3, and insulin receptor was increased as compared with the control group. The different effects of energy deficiency at the 2 stages in lactation show that the endocrine regulation changes qualitatively and quantitatively during the course of lactation.

Short communication: Circulating and milk adiponectin change differently during energy deficiency at different stages of lactation in dairy cows

Adiponectin, one of the most abundant adipokines in circulation, is known for its role in regulation of body metabolism. The aim of this study was to evaluate the effects of a negative energy balance (NEB) at 2 stages of lactation (lactational NEB at the onset of lactation and an induced NEB by feed restriction near 100 d of lactation) on circulating adiponectin concentrations. We also investigated the effect of feed restriction on adiponectin concentrations in milk and the relationships of blood and milk adiponectin with selected plasma or milk variables and with measures of body condition. Plasma adiponectin was measured in 50 multiparous Holstein dairy cows throughout 3 experimental periods [i.e., period 1=3 wk antepartum up to 12 wk postpartum, period 2=3 wk of feed restriction starting at around 100 d in milk with a control (n=25) and feed-restricted group (50% of energy requirements; n=25), and period 3=subsequent realimentation period for 8 wk]. Milk adiponectin was investigated among 21 multiparous cows at wk 2 and wk 12 of period 1 and wk 2 of period 2. Adiponectin concentrations in plasma and skim milk were measured using an in-house ELISA specific for bovine adiponectin. Major changes in circulating adiponectin concentrations were observed during the periparturient period, whereas energy deficiency during established lactation at around 100 d in milk and subsequent refeeding did not affect plasma adiponectin. Together with lower adiponectin concentrations in milk (µg/mL), the reduction in milk yield led to decreased adiponectin secretion via milk (mg/d) at the second week of feed restriction. Irrespective of time and treatment, milk adiponectin represented about 0.002% of total milk protein. Mean adiponectin concentrations in milk (0.61 ± 0.03 µg/mL) were about 92% lower than the mean plasma adiponectin concentrations (32.1 ± 1.0 µg/mL). The proportion of the steady-state plasma adiponectin pool secreted daily via milk was 2.7%. In view of the similar extent of NEB in both periods of energy deficiency, decreasing adiponectin concentrations seems important for accomplishing the adaptation to the rapidly increasing metabolic rates in early lactation, whereas the lipolytic reaction toward feed restriction-induced NEB during established lactation seems to occur largely independent of changes in circulating adiponectin.

Response of the cholesterol metabolism to a negative energy balance in dairy cows depends on the lactational stage.

The response of cholesterol metabolism to a negative energy balance (NEB) induced by feed restriction for 3 weeks starting at 100 days in milk (DIM) compared to the physiologically occurring NEB in week 1 postpartum (p.p.) was investigated in 50 dairy cows (25 control (CON) and 25 feed-restricted (RES)). Blood samples, liver biopsies and milk samples were taken in week 1 p.p., and in weeks 0 and 3 of feed restriction. Plasma concentrations of total cholesterol (C), phospholipids (PL), triglycerides (TAG), very low density lipoprotein-cholesterol (VLDL-C) and low density lipoprotein-cholesterol (LDL-C) increased in RES cows from week 0 to 3 during feed restriction and were higher in week 3 compared to CON cows. In contrast, during the physiologically occurring NEB in week 1 p.p., C, PL, TAG and lipoprotein concentrations were at a minimum. Plasma phospholipid transfer protein (PLTP) and lecithin:cholesterol acyltransferase (LCAT) activities did not differ between week 0 and 3 for both groups, whereas during NEB in week 1 p.p. PLTP activity was increased and LCAT activity was decreased. Milk C concentration was not affected by feed restriction in both groups, whereas milk C mass was decreased in week 3 for RES cows. In comparison, C concentration and mass in milk were elevated in week 1 p.p. Hepatic mRNA abundance of sterol regulatory element-binding factor-2 (SREBF-2), 3-hydroxy-3-methylglutaryl-coenzyme A synthase 1 (HMGCS1), 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGCR), and ATP-binding cassette transporter (ABCA1) were similar in CON and RES cows during feed restriction, but were upregulated during NEB in week 1 p.p. compared to the non-lactating stage without a NEB. In conclusion, cholesterol metabolism in dairy cows is affected by nutrient and energy deficiency depending on the stage of lactation.

Efecto del sistema de alimentación sobre los resultados técnicos, calidad de la canal, de la carne y de la grasa de cerdos ibéricos

La presente tesis doctoral pretende ampliar conocimientos de los sistemas de producción y tecnología asociada del cerdo Ibérico. Los trabajos de campo fueron llevados a cabo en el CIA ‘El Dehesón del Encinar’ (Oropesa, Toledo, España) con cerdos Ibéricos de la estirpe Torbiscal. En los dos primeros experimentos, utilizando dos grupos de cerdos, se estudió el efecto de la restricción de la alimentación en premontanera sobre las características de la canal y sobre la composición de la grasa al sacrificio de los cerdos posteriormente cebados en montanera. Los animales restringidos en premontanera crecieron significativamente menos que los no restringidos durante dicho periodo, y manifestaron un crecimiento compensatorio durante la montanera. Este crecimiento compensatorio no fue suficiente, y los animales restringidos llegaron con pesos significativamente inferiores al sacrificio, aunque esto no supuso diferencias para las características de la canal entre ambos grupos de animales. También al sacrificio, los animales restringidos durante la premontanera tuvieron concentraciones significativamente diferentes de ácidos grasos, sobre todo en la capa interna de la grasa subcutánea y en la fracción de lípidos polares de la grasa intramuscular; asimismo, tuvieron mayores concentraciones de tocoferoles en el músculo Longissimus dorsi. En un tercer experimento, utilizando cuatro grupos de animales, se estudió el efecto del tiempo de estancia de los cerdos en montanera exclusiva sobre las características de la canal y de la grasa al sacrificio. La duración de la montanera afectó significativamente al crecimiento, a los pesos y porcentajes de los productos del despiece y al perfil de ácidos grasos de la grasa subcutánea. Los animales cebados con pienso en confinamiento tuvieron mayor crecimiento, mejor calidad de la canal, y peor calidad de grasa que los otros tres grupos de animales que se acabaron en montanera exclusiva durante 46, 83 y 111 días. En el último experimento se evaluó si la impedancia bioeléctrica es un método adecuado para realizar un estudio tisular de los jamones de cerdos Ibéricos, aspecto especialmente relevante para la industria. La impedancia bioeléctrica resultó ser un método no invasivo adecuado, que puede ser aplicado para predecir la composición tisular de los jamones, aunque se necesitan más ensayos antes de llevar este método a la práctica cotidiana de la industria. ABSTRACT This Thesis aims to expand knowledge on production systems and technology associated to Iberian pigs. Fieldwork was conducted in the CIA ‘El Dehesón del Encinar’ (Oropesa, Toledo, Spain) with Iberian pigs Torbiscal line. In first two experiments, using two groups of pigs, the effect of feed restriction during the period previous to free-range fattening on growth and carcass characteristics in Iberian pigs finished under free-range conditions was studied. The pigs fed low feeding levels during the period previous to free-range finishing period, had significantly lower growth than those feed with a high level during that period, and showed a compensatory growth during free-range period. However, this compensatory growth was not enough to reach the same weight than the animals fed with a high level, and the slaughter weights were significantly different, although no differences were observed for the carcass characteristics between both groups of pigs. At slaughter, animals fed low feeding levels during the period previous to free-range finishing period, had significantly different fatty acid proportions, especially in the inner backfat layer and in polar lipids fraction of the intramuscular fat; also, the tocopherol concentration found in Longissimus dorsi muscle from pigs fed low feeding levels was higher than those fed high feeding levels. In a third experiment, using four groups of animals, the effect of duration under freerange exclusive conditions on carcass and fat characteristics at slaughter was studied. The growth, weights and percentages of carcass cuts, and fatty acid profile of the backfat were significantly affected by the duration under free-range exclusively conditions. The animals fed in confinement with concentrate diets during the whole fattening period had high growth, better carcass quality and worse fat quality than the other three groups of animals fed under free-range exclusive conditions during 46, 83 and 111 days. In the last experiment, it assessed whether the bioelectrical impedance is a suitable method for perform a tissue study of Iberian pig hams, particularly important aspect for industry. The bioelectrical impedance has proved to be a non-invasive suitable method, which can be applied to predict the tissues composition of the hams, but more trials are needed before carrying this method to everyday practice in the industry.

Farelo do bagaço de caju na restrição alimentar qualitativa para suínos em terminação

The objective of this study was to evaluate the effects of the use of cashew bagasse bran (CBB) as food ingredient in qualitative feed restriction programs on the carcass traits, meat quality, organs weight and intestinal morphometry of barrows and gilts. Twenty – four crossbred pigs were used (12 barrows and 12 gilts) with an average initial body weight of 57.93 ± 3.67 kg/LW. The experimental designs was in randomized blocks 3x2 factorial arrangement with three level (0%, 15% e 30% CBB), two genders (barrows and gilts) and four repetition. A total of twenty-four instalments. The treatments were composed of basal diet (BD) formulated with corn, soybean meal and commercial base mix for finishing pigs, being containing different levels of CBB. At the end of the trial period the animals were slaughtered for the evaluation of the meat quality, traits carcass, Absolute Weight (AW) and Relative Weight (RW) of the organs and morphometric study of small intestine fragment. The inclusion of (CBB) in the diets did not affect the traits carcass of gilts, but interfered in the traits carcass of the barrow positively, increasing the yield of meat into cold carcass and reducing the thickness of subcutaneous fat, without affecting the fatty acid profile. However, we observed increased weight of organs and partial volume of absortiva mucosa of gilts. In the comparison between sex was observed a greater liver weight (AW) and (RW), and surface density of absortiva mucosa of barrow. The use of CBB was considered as ingredient to be used in programs of qualitative feed restriction for finishing pigs.

Farelo do bagaço de caju na restrição alimentar qualitativa para suínos em terminação

The objective of this study was to evaluate the effects of the use of cashew bagasse bran (CBB) as food ingredient in qualitative feed restriction programs on the carcass traits, meat quality, organs weight and intestinal morphometry of barrows and gilts. Twenty – four crossbred pigs were used (12 barrows and 12 gilts) with an average initial body weight of 57.93 ± 3.67 kg/LW. The experimental designs was in randomized blocks 3x2 factorial arrangement with three level (0%, 15% e 30% CBB), two genders (barrows and gilts) and four repetition. A total of twenty-four instalments. The treatments were composed of basal diet (BD) formulated with corn, soybean meal and commercial base mix for finishing pigs, being containing different levels of CBB. At the end of the trial period the animals were slaughtered for the evaluation of the meat quality, traits carcass, Absolute Weight (AW) and Relative Weight (RW) of the organs and morphometric study of small intestine fragment. The inclusion of (CBB) in the diets did not affect the traits carcass of gilts, but interfered in the traits carcass of the barrow positively, increasing the yield of meat into cold carcass and reducing the thickness of subcutaneous fat, without affecting the fatty acid profile. However, we observed increased weight of organs and partial volume of absortiva mucosa of gilts. In the comparison between sex was observed a greater liver weight (AW) and (RW), and surface density of absortiva mucosa of barrow. The use of CBB was considered as ingredient to be used in programs of qualitative feed restriction for finishing pigs.

Effect of feed quality restriction followed by realimentation on growth, nutrient utilization, biochemical changes and haematological profiles of Indian major carp, rohu (Labeo rohita H.)

To investigate the effect of protein restriction with subsequent re-alimentation on nutrient utilization, hematological and biochemical changes of Indian major carp, Rohu (Labeo rohita H.), 150 acclimatized Rohu fingerlings (average 20.74 ± 0.13 g) divided into five experimental groups (30 fingerlings in each groups with three replications with 10 fingerlings in each) for experimental trial of 90 days using completely randomized design. Control group (T sub(CPR)) was fed with feed having 30% crude protein at 3% of body weight for 90 days trial period. Other experimental groups T sub(1PR) was alternatively 3 days fed with feed having 20% CP and 30% CP at 3% of body weight, T sub(2PR) was alternatively 7 days fed with feed having 20% CP and 30% CP at 3% of body weight, T sub(3PR) was alternatively 15 days fed with feed having 20% CP and 30% CP at 3% of body weight and T sub(4PR) was alternatively 25 days fed with feed having 20% CP and 30% CP at 3% of body weight during 90 days trial period with daily ration in two equal halves at morning and afternoon. It was noticed that retention of different nutrients was almost similar among all treatment groups indicated improvement of digestibility of nutrients might not be the mechanisms for recovery growth in carps. Increased percent feed intake of body weight (hyperphagia) (4.14 ± 0.30 or 4.94 ± 0.46 and 3.33 ± 0.29), improved specific growth rate (1.86 ± 0.09 or 2.26 ± 0.05 and 1.43 ± 0.01), absolute growth rate (1.57 ± 0.08 or 1.84 ± 0.18 and 1.36 ± 0.12), protein efficiency ratio (1.19 ± 0.11 or1.16 ± 0.12 and 1.05 ± 0.09) were the important mechanism showing better performance index (21.60 ± 1.09 or 23.80 ± 0.21 and 19.45 ± 0.37) through which the experimental groups which were protein restricted and re-alimented at 3 or 7 days alternatively during 90 days trial period could able to compensate the growth retardation and to catch up the final body weight of control (128.68 ± 11.53 g/f) but other experimental groups failed to compensate during 90 days trial period. Result of the present study indicated that deprived fish i.e., fish received alternate 3 or 7 days protein restriction and re-alimentation showed recovery growth had still lower values of Hb (10.21 ± 0.02, and 9.88 ± 0.04 g/dl), hematocrit value (30.62 ± 0.05 and 26.64 ± 0.11%), total erythrocytic count (3.40 ± 0.01 and 3.29 ± 0.01 X10super(6) mm³), plasma glucose (126.93 ± 0.20 and 126.67 ± 0.05 mg/dl), total plasma lipid (1.04 ± 0.01 and 1.02 ± 0.01 g/dl) and liver glycogen (290.10 ± 0.80 and 288.99 ± 0.95 mg/kg) in comparison to control (10.56 ± 0.08 g/dl, 31.68 ± 0.24%, 3.52 ± 0.03 X10super(6) mm³, 128.23 ± 0.25 mg/dl, 1.07 ± 0.01g/dl and 292.00 ± 0.23 mg/kg) at the end of 90 days trial but total plasma protein in deprived group was compensated with advancement of trial period. All hematological and biochemical parameters studied were proportionately lowered in the experimental group got higher degree of deprivation. These findings suggested that with the increase of trial length complete compensation of hematological and biochemical profiles of rohu might be achieved. The results indicated that the implementation of alternative 7 days low and high protein diet feeding during aquaculture of carps could make economize the operation through minimizing the feed input cost.

Vaccination with an Insulin-like Growth Factor Binding Protein-1 (IGFBP-1)-bBased Vaccine Improves Growth in Feed-Restricted Brahman Steers

Dry-season weight loss in grazing cattle in northern Australia has been attenuated using a number of strategies (Hunter and Vercoe, 1987, Sillence et al. 1993, Gazzola and Hunter, 1999). Furthermore, the potential to improve efficiency of feed utilisation (and thus, dry-season performance) in ruminants through conventional modulation of the insulin-like growth factor (IGF) axis (Oddy and Owens, 1997, Hill et al., 1999) and through immunomodulation of the IGF axis (Hill et al., 1998a,b) has been demonstrated. The present study investigated the use of a vaccine directed against IGFBP-1 in Brahman steers which underwent a period of nutritional restriction followed by a return to wet-season grazing.

Dietary restriction: Metabolic shifting for cardiac health

Purpose: To determine the effect of dietary restriction on metabolic pathways and the relationship of the metabolic shifting on antioxidant enzymes in cardiac tissue. Design: Randomized, controlled study. Male rats at 60 days old were randomly divided into four groups. Materials and Methods: The rats of control groups C30 and C60 were given free access to the diet over 30 and 60 days. The rats of the DR30 group were fed 60% of the chow consumed by the control groups over 30 days. The animals of the DR60 group ate 60% of the amount consumed by the C60 group over 60 days. Serum was used for total protein, lactate dehydrogenase (LDH), alanine aminotransferase (ALT) and aspartate aminotransferase (AST). Protein, glycogen, total lipids, superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), LDH, AST and ALT were determined in cardiac tissue. Results: Dietary restriction induced diminished serum and cardiac LDH activities. AST activities were lower in the serum and cardiac muscle of the DR60 animals. Dietary restriction induced elevated total lipid concentrations in cardiac muscle. No significant differences were observed in total protein and glycogen content among the groups. Antioxidant enzyme determinations demonstrated increased cardiac GSH-Px activities in the DR60 animals and increased SOD activities in the cardiac tissue of both feed-restricted groups. Conclusions: Dietary restriction was protective against oxidative stress in the heart by improving cardiac endogenous antioxidant defences and shifting the metabolic pathway for energy production.

Short periods of food restriction do not affect growth, survival or muscle development on pacu larvae

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Physiological Response to Dolops carvalhoi (Crustacea: Branchiura) Infection by Pacu, Piaractus mesopotamicus, Subjected to Short Cycles of Food Restriction and Refeeding

Feeding strategies that reduce feed and promote compensatory growth could be an interesting tool to reduce costs in the fish production. However, fish health must be monitored to evaluate if their physiological response to adverse conditions, such as parasite infection, does not become compromised. A 12-wk growth trial was conducted to determine the physiological responses of pacu, Piaractus mesopotamicus, that were subjected to different fasting/refeeding cycles and infected with the Dolops carvalhoi. The schemes were: (i) control group fish (FD), (ii) food-restricted and controlled refeeding group (FR/Rc), and (iii) food-restricted and refeeding to satiation group (FR/Rs). After 84 d, the fish were exposed to D. carvalhoi for 30 h. The fish subjected to food restriction did not exhibit compensatory growth. Cortisol levels decreased in all groups within 30 h after infection. Glucose levels increased 6 h after the D. carvalhoi in the FR/Rs and 30 h after infection in the FD. In all of the fish groups, the hematocrit values were reduced after infection, and it was associated with a reduction in the mean corpuscular volume and erythrocytes. At 30 h after infection, the number of erythroblasts increased. The use of the feeding schemes does not indicate a failure of the pacu physiological responses.

Accurate adjustment of feed formulation for broiler chickens on performance and lipid composition of meat

Most consumers consider the fat of chicken meat undesirable for a healthy diet, due to the high levels of saturated fatty acids and cholesterol. The purpose of this experiment was to investigate the influence of changes in dietary metabolizable energy level, associated with a proportional nutrient density variation, on broiler chickens performance and on the lipid composition of meat. Males and females Cobb 500 broilers were evaluated separately. Performance evaluation followed a completely randomized design with factorial 6x3 arrangement - six energy levels (2,800, 2,900, 3,000, 3,100, 3,200 and 3,300 kcal/kg) and three slaughter ages (42, 49 and 56 days). Response surface methodology was used to establish a mathematical model to explain live weight, feed intake and feed conversion behavior. Total lipids and cholesterol were determined in skinned breast meat and in thigh meat, with and without skin. For lipid composition analysis, a 3x3x2 factorial arrangement in a completely randomized design - three ration’s metabolizable energy levels (2,800, 3,000 and 3,300 kcal/kg), three slaughter ages (42, 49 and 56 days) and two sexes - was used. The reduction in the diet metabolizable energy up to close to 3,000 kcal/kg did not affect live weight but, below this value, the live weight decreased. Feed intake was lower when the dietary energy level was higher. Feed conversion was favored in a direct proportion to the increase of the energy level of the diet. The performance of all birds was within the range considered appropriate for the lineage. Breast meat had less total lipids and cholesterol than thigh meat. Thigh with skin had more than the double of total lipids of skinned thigh, but the cholesterol content did not differ with the removal of the skin, suggesting that cholesterol content is not associated with the subcutaneous fat. Intramuscular fat content was lower in the meat from birds fed diets with lower energy level. These results may help to define the most appropriate nutritional management. Despite the decrease in bird’s productive performance, the restriction of energy in broiler chickens feed may be a viable alternative, if the consumers are willing to pay more for meat with less fat.