Search

Ambient conditions during chicken embryogenesis, such as insufficient oxygen or changes in temperature, are expected to cause permanent phenotypic changes and affect their posthatch performance. Decades of genetic selection for high growth rate resulted with various physiological and morphological changes that can affect the broiler fitness under environmental stress. To evaluate the selection effect on responses to environmental challenge during embryonic development, and the long-term implications, we have used a unique genetic line, that was not selected for over 30 yr (since 1986), as control for the modern commercial genetic line. At embryonic day 5 (E5), broiler embryos from these 2 genetic lines were divided into 2 treatments: 1) control; 2) 15% O2 concentration for 12 h/day from E5 through E12 the embryonic period of chorioallantoic membrane formation. Embryos and hatched chicks were characterized for physiological and morphological parameters. Significant differences in relative embryo weight and yolk consumption were found between the 2 lines. The modern line was characterized by a higher metabolic rate and rapid growth, supported by higher hemoglobin levels and hematocrit concentrations, whereas the 1986 line had slower metabolism, lower levels of hematocrit and hemoglobin, higher oxygen volume per 1 g of embryonic tissue indicating higher oxygen availability. Both lines exhibited changes in heart rate, and blood parameters corresponding to cardiovascular system adaptation after hypoxic exposure, seemingly implemented to increase oxygen-carrying capacity to the embryo tissues. Our finding stand in agreement that the genetic selection for high growth rate that led to higher metabolism without a fit of the cardiovascular system, increased the imbalance between oxygen supply and demand.

Adequate ambient temperature and oxygenation are necessary to maintain normal embryonic development of broilers, however, hypoxia challenge during incubation can aid in improving regulatory plasticity and lead to different phenotypes later in life. This study aimed to examine the effects of moderate hypoxia (O2 17%) during the plateau phase on the embryonic physiological parameters and on post-hatch performance (growth rate, feed consumption and feed conversion) up to the age of poultry marketing. The study included examined embryos exposed to O2 17% for 12 h per day (h/d) from E16 through E18 (designated as 12H), or O2 17% continuously, from E16 through E17 (designated as 48H) and a standard incubation control group (21% O2). Physiological and morphological parameters of embryos and hatched chicks were measured. Male Chicks from all three treatment groups were raised under recommended temperature regime, and body weight, feed intake and FCR were recorded on a weekly basis. The intermittent hypoxia protocol (12H), allowed embryos to properly adapt to the shortage of oxygen, compensate for the gap in body mass that developed following the first exposure window, and hatch with characteristics similar to those of the control embryos. In contrast, while the 48H embryos were able to adapt to the hypoxic stress, the prolonged exposure prevented them from catching up with both control and 12H embryos. Broilers that were subjected to hypoxia showed hatchling body weights and growth rates similar to those of controls, throughout the entire growth phase. During the fifth week, lower feed consumption was observed in the 12H and 48H groups and became significantly lower than the control chicks in the sixth week of growth. Following hypoxia exposure, chicks managed to reach normal body weight with less feed, with the 12H group demonstrating lower and more efficient FCR during the last two weeks of growth. Broiler embryos reacted to plateau-phase hypoxia challenge with numerous physiological and metabolic modifications. The prudent alterations in metabolism and cardiovascular system during exposure to hypoxia and post-hatch, resulted in more efficient energy utilization in broilers, which may have a long-lasting enhancing effect on post-hatching thermotolerance and sustainability in chicks reared under sub-optimal environmental conditions.

Early life encounters with stress can lead to long-lasting beneficial alterations in the response to various stressors, known as cross-tolerance. Embryonic heat conditioning (EHC) of chicks was previously shown to mediate resilience to heat stress later in life. Here we demonstrate that EHC can induce cross-tolerance with the immune system, attenuating hypothalamic inflammation. Inflammation in EHC chicks was manifested, following lipopolysaccharide (LPS) challenge on day 10 post-hatch, by reduced febrile response and reduced expression of LITAF and NFκB compared to controls, as well as nuclear localization and activation of NFκB in the hypothalamus. Since the cross-tolerance effect was long-lasting, we assumed that epigenetic mechanisms are involved. We focused on the role of ten-eleven translocation (TET) family enzymes, which are the mediators of active CpG demethylation. Here, TET transcription during early life stress was found to be necessary for stress resilience later in life. The expression of the TET family enzymes in the midbrain during conditioning increased in parallel to an elevation in concentration of their cofactor α-ketoglutarate. In-ovo inhibition of TET activity during EHC, by the α-ketoglutarate inhibitor bis-2-(5-phenylacetamido-1,3,4-thiadiazol-2-yl) ethyl sulfide (BPTES), resulted in reduced total and locus specific CpG demethylation in 10-day-old chicks and reversed both thermal and inflammatory resilience. In addition, EHC attenuated the elevation in expression of the stress markers HSP70, CRHR1, and CRHR2, during heat challenge on day 10 post-hatch. This reduction in expression was reversed by BPTES. Similarly, the EHC-dependent reduction of inflammatory gene expression during LPS challenge was eliminated in BPTES-treated chicks. Thus, TET family enzymes and CpG demethylation are essential for the embryonic induction of stress cross-tolerance in the hypothalamus.

In all vertebrates, hypoxia plays an important role in fetal development, driving vasculogenesis, angiogenesis, hematopoiesis, and chondrogenesis. Therefore, the ability to sense and respond to changes in the availability of oxygen (O2) is crucial for normal embryonic development as well as for developmental plasticity. Moderate levels of hypoxia trigger a regulated process which leads to adaptive responses. Regulation of angiogenesis by hypoxia is an important component of homeostatic control mechanisms that link the cardio-pulmonary-vascular O2 supply to metabolic demands in local tissues. Hypoxia leads to the activation of genes that are important for cell and tissue adaptation to low O2 conditions, such as hypoxia-inducible factor 1. Previous studies have shown a dose-response effect to hypoxia in chicken embryos, with lower and/or prolonged O2 levels affecting multiple mechanisms and providing a spectrum of responses that facilitate the ability to maintain O2 demand despite environmental hypoxia. In chicken embryos, mild to extreme hypoxia during embryogenesis improves chorioallantoic membrane and cardiovascular development, resulting in an increase in O2 carrying capacity and leading to developmental plasticity that may affect post-hatch chick performance and improve adaptation to additional environmental stresses at suboptimal environmental conditions.

The prenatal circulatory system is adaptive and capable of plasticity designed for the needs of the growing tissue. When a broiler embryo is faced with hypoxic stress, the process of angiogenesis in tissues begins. Exposure to hypoxic conditions of 17% oxygen during the chorioallantoic membrane (CAM) development (E5 to E12) affected the circulatory system and contributed to an increase in the blood oxygen carrying capacity. The present study aimed to evaluate the effects of hypoxic exposure during CAM development on post-hatch performance of broilers and to examine whether hypoxic exposure improved sustainability of birds exposed to acute heat stress. Two consecutive trials, with male broilers from each of the incubation treatments—optimal conditions and exposure to hypoxia of 15 or 17% oxygen, for 12 h/day, during CAM development—were conducted. In experiment 1, 60 male chicks from each group were raised in individual cages. In experiment 2, 160 male chicks from each group were raised in 40-chick pens until marketing. On d 35, 20 birds from each group were transferred to individual cages kept at a temperature of 23°C for 72 h, and then birds were exposed to 35°C for 5 hours. Body temperatures were measured at 0, 2, and 5 h of the heat exposure. In both experiments BW, feed intake, and FCR were recorded. At marketing, chicks were slaughtered, and relative weights of breast muscle, abdominal fat pad, heart, and liver were calculated. Hypoxia treatment resulted in a FCR advantage. Food intake was similar in all treatments, but groups exposed to hypoxia grew better than controls until the age of 35 days. Hypoxia-treated groups had higher relative breast, heart, and liver weights than controls. Body temperatures of hypoxia-treated chickens remained lower during heat stress exposure, and their mortality rate was lower as well. Intermittent exposure to moderate hypoxia during CAM development confers advantages to broilers in feed utilization efficiency and in coping with heat stress. It may be considered as a mitigating step in incubation to facilitate broilers in achieving their full growth potential.

The chick embryo employs several adaptive responses to hypoxic challenges, affecting both metabolism and oxygen (O2) transport. The present study assessed the effects of hypoxic conditions (17% O2) during the plateau phase on embryonic metabolic rate, cardiovascular parameters, and development up to hatching. The study was divided into 2 experiments: (1) Control; 17% O2 for 6h/d on E16-E18 (6H), and 17% O2 for 12h/d on E16-E18 (12H), and (2) Control; 12H, and 17% O2 continuously for 72h on E16-E18, (72H). Hypoxic embryos exhibited a significant increase in heart rate and an upward trend starting on E17 in hematocrit and hemoglobin levels. We observed a decrease in metabolism in 12H and 72H embryos during the plateau period; their oxygen consumption as well as yolk consumption were lower compared to Control and they hatched with a significantly lower body temperature, indicating lower heat production. There was no evidence of adaptation or long-term effects of exposure to 17% O2 for 6h/d. Exposure to 72h of hypoxic conditions led to significant physiological changes and had a detrimental influence on embryonic development and growth. In contrast, exposure to 12h/d produced moderate hypoxic changes, which helped the embryo to cope with the stress without significant influences on its growth and development. The decrease in metabolism may represent a metabolic adaptation through a decrease in resting metabolic rate and lower heat production. Such alterations may affect post-hatch performance and energy allocation between maintenance and growth, especially under stress when there is increased oxygen demand.

Tibial dyschondroplasia (TD) is a skeletal abnormality that can cause economic losses and animal welfare concerns. Thiram-induced TD is characterized by enlarged, unvascularized growth plates, low levels of the vascular endothelial growth factor receptor Flk-1, abnormal chondrocyte differentiation, and lameness. Recently we reported the involvement of heat-shock protein 90 (Hsp90) in chondrocyte differentiation and growth-plate vascularization. Inhibition of Hsp90 activity in thiram-induced TD resulted in increased Flk-1 levels, re-instated normal growth-plate angiogenesis and morphology, and abrogated lameness. In the present study, we evaluated the efficacy of various concentrations of 17-(dimethylaminoethylamino)-17-demethoxygeldanamycin (17-DMAG), an inhibitor of Hsp90 activity, in preventing growth-plate histopathology and lameness in TD-affected chicks. Low doses of 17-DMAG (2 injections, each of 100 or 300 μg) did not prevent TD development even though Flk-1 levels were restored, which suggests that Flk-1 is not the only rate-limiting factor in growth-plate angiogenesis. High doses of 17-DMAG (2 injections, each of 600 or 900 μg) prevented BW loss, decreased the TD score, reduced lesion width, restored proper chondrocyte differentiation, increased blood vessel invasion, and eliminated lameness. To assess the specificity of Hsp90, we evaluated the efficacy of the flavonoid quercetin, an inhibitor of Hsp70 synthesis, in preventing TD development; it decreased Hsp70 levels but not those of Hsp90 in the control growth plates and prevented upregulation of Hsp70 in the TD-affected growth plates. Dietary quercetin (at 100 or 500 ppm) did not prevent the hypoxia that is characteristic of the TD-affected growth plate or development of thiram-induced TD and lameness. The present results demonstrate the specificity and the major role of Hsp90 in chondrocyte differentiation and growth-plate vascularization. In contrast to the anti-angiogenic effect of 17-DMAG observed in mammals, inhibition of Hsp90 activity in the unvascularized TD-affected growth plates resulted in activation of the angiogenic switch and restored normal growth-plate morphology.

Birds dissipate considerable heat through respiratory-evaporative and cutaneous-evaporative mechanisms and sensible heat loss (SHL) via radiation, convection, and conduction. The significance of SHL in laying hens is still to be confirmed. This study aimed to elucidate the effect of ventilation on egg production and quality during exposure to high ambient temperature. Lohman laying hens were raised outdoors up to age 35 wk, and 300 hens with similar egg production were divided among 5 treatments each comprising 4 replicates of 15 hens. Birds in 4 treatments were kept in computerized controlled-environment rooms acclimated to 35°C and 50% RH, with ventilation flow rates of 0.5, 1.5, 2.0, and 3.0 m/s, respectively, and those in the control were kept outdoors. Hens were acclimated to the controlled environment rooms for 1 wk and to the targeted environmental conditions for another week, and then were subjected to measurements for 2 wk. Egg production, mass, and shell density, and feed and water consumption were monitored. Body temperature, SHL, and plasma thyroid hormone concentrations were measured at the end of the experiment. The high environmental temperature impaired egg production and quality: whereas exposure of hens to ventilation flows of 2.0 and 3.0 m/s elicited significant recovery of these parameters with time, exposure to a rate of 0.5 m/s negatively affected these parameters throughout the experimental period. The highest feed intake and water consumption were observed in hens exposed to 2.0 and 3.0 m/s, respectively, and the highest SHL was observed in those exposed to 3.0 m/s. It can be concluded that ventilation rate significantly affected hens exposed to high ambient temperature: high ventilation (3.0 m/s) improved egg production whereas low ventilation (0.5 m/s) negatively affected production and quality.

The aim of this study was to fine-tune previous acute cold exposure treatments of broiler embryos during late embryogenesis to improve lifelong cold resistance and performance. Six hundred Cobb hatching eggs were incubated under standard conditions and then exposed to 3 treatments: control; cold treatment in which embryos were exposed to 15°C for 30 min on d 18 and 19 of incubation (30 × 2); and cold treatment similar to 30 × 2 but with 60-min exposures (60 × 2). Egg shell temperature (T(egg)) and heart rate (HR) were monitored pre- and posttreatment. Upon hatching, hatchability, body weight, and body temperature were recorded. From 14 to 35 d of age, three quarters of the chickens in each treatment were raised under ascites-inducing conditions (AIC) and the remaining birds were raised under standard brooding conditions (SBC). The T(egg) and HR decreased significantly in response to increased exposure time on d 18 of incubation. On d 19 of incubation, before the second cold exposure, the 30 × 2 group showed greater T(egg) and HR than the controls, and during the second exposure they maintained these parameters better than the 60 × 2 embryos. No treatment effect on hatchability was observed. At 35 d of age ascites incidence among 30 × 2 chickens under AIC was significantly less than that among the controls (P < 0.01), and body weight of these chickens under either SBC or AIC was significantly higher than that of the controls. Under SBC relative breast muscle weight was significantly higher in 60 × 2 chickens, whereas the relative heart weight was higher in both cold-treated groups than in the controls. It can be concluded that repeated short acute cold exposures during late embryogenesis significantly reduced ascites incidence and improved growth rate under either SBC or AIC. These results may be related to a prenatal epigenetic adaptation of the thermoregulatory and cardiovascular systems to low ambient temperature.

Rapid growth rate has presented broiler chickens with serious difficulties when called on to thermoregulate efficiently in hot environmental conditions. Altering the incubation temperature may induce an improvement in the acquisition of thermotolerance (AT). This study aimed to elucidate the effect of thermal manipulations (TM) during the development of the thyroid and adrenal axis of broiler embryos on the potential of broilers to withstand acute thermal stress at marketing age. Cobb broiler embryos were subjected to TM at 39.5 degrees C and 65% RH from embryonic day 7 to 16 (inclusive), either continuously (24 h) or intermittently (12 h). After hatching chicks were raised under standard conditions to 35 d of age and then subjected to thermal challenge (35 degrees C for 5 h). Continuous TM caused a significant decline in hatchability, coupled with significantly lower BW and body temperature at hatching. The intermittent (12-h) chicks showed results similar to the controls but had significantly lower body temperature. Thermal challenge at marketing age demonstrated a significant improvement in AT in both the 12- and 24-h TM-treated broilers, which was characterized by a significantly lower level of stress (as evidenced by the level of plasma corticosterone) and rate of mortality. It was concluded that TM during the portion of embryogenesis when the thyroid and adrenal axis develop and mature had a long-lasting effect and improved the AT of broiler chickens. Whereas intermittent TM had no significant effect on hatchability and performance parameters, continuous TM negatively affected these parameters.

Tibial dyschondroplasia (TD) is one of the most prevalent skeletal abnormalities in avian species; it causes economic losses and is an animal welfare problem. It has been hypothesized that the absence of vasculature in the lesion of the TD growth plates at the ends of the long bones is involved in the etiology of the disease. We evaluated the hypoxia status of normal and thiram-induced TD growth plates by immunostaining the protein adducts after pimonidazole hydrochloride administration. In addition, we evaluated the expression of hypoxia-inducible factor-1alpha (HIF-1alpha), the major regulator of the hypoxic response that is essential for chondrogenesis, and that of heat-shock proteins (Hsp) downstream from HIF-1alpha. We demonstrated that, in contrast to the normal growth plates, those afflicted by TD were hypoxic. A major increase in hypoxia was observed in the proliferative, hypertrophic, and calcified zones. In the normal growth plate, HIF-1alpha was expressed in chondrocytes of the articular cartilage and of the maturation zone, whereas in cases of TD, HIF-1alpha was also expressed in chondrocytes below the lesion. The expression level of HIF-1alpha was related to the severity of the disease, but was independent of its cause; the same pattern of expression was observed in growth plates of chicks selected for a high incidence of TD. No differentiation-dependent expression of HIF-1alpha was observed in response to hypoxia, as demonstrated by the use of primary cultures of growth plate chondrocytes. In the normal growth plates, Hsp90 and Hsp70 were localized to the maturation zone. More cells expressed both Hsp in the TD lesion. In conclusion, we demonstrated that the TD growth plate, in contrast to the normal one, is hypoxic, probably because of the lack of vascularization. Hypoxia leads to an increase in the transcription factor HIF-1alpha, causing increases in the levels of Hsp90 and Hsp70.

Efficient ventilation affects thermoregulation and, thereby, the performance of domestic fowl. This became crucial as genetic selection for growth development significantly coupled with increased metabolic rate. The specific aims of this study were to elucidate a) the effects of different rates of ventilation on young turkey performance during exposure to constant 35, 30, and 25 degrees C and b) their effects on body temperature and surface temperature. In 3 separate experiments, turkeys were raised under regular conditions up to 3 wk of age. Thereafter, they were acclimated for 1 wk to the targeted ambient temperatures (T(a)) and to air velocities of 0.8, 1.5, 2.0, or 2.5 m/s and raised under those conditions up to 6 wk of age. Turkeys exposed to 35 degrees C performed optimally at an air velocity (AV) of 2 m/s; they exhibited significantly higher feed intake and significantly lower body temperature. At 30 degrees C, performance was optimal at AV of 1.5 to 2.5 m/s and significantly lower at 0.8 m/s. Performance of turkeys exposed to 25 degrees C did not vary with AV. Comparison of BW and feed intakes of turkeys exposed to the 3T(a) levels revealed significantly higher feed intake at 25 degrees C but similar BW compared with those exposed to 30 degrees C, meaning that those exposed to 25 degrees C used more energy for maintenance than for growth. In general, surface temperature of the body declined significantly with T(a), whereas that of the face and legs was significantly lower at 25 degrees C. It can be concluded that AV affects the performance of young turkeys. The range of AV within which BW was optimal expanded as T(a) declined. It can be further concluded that the combination of 30 degrees C with AV from 1.5 to 2.5 m/s was optimal for young turkeys.

Recent decades have seen significant progress in the genetic selection of fast-growing meat-type broiler chickens. However, fast growth has coincided with inferior development of the visceral syste...

1. The effect of age on ovarian function was studied in 245-, 350-, 500-, 700- and 800-d-old Lohmann hens. The effect of three different methods for moult induction on ovarian function and corticosterone concentration was studied in 500-d-old hens. 2. No significant reductions in ovarian weight or in number of follicles before the age of 700 d were found. The ability to produce progesterone and oestradiol-17beta was unchanged up to the age of 700 d and the circadian secretion of these two steroids was identical in young (225 d) and old hens (600 d). 3. The effects of induced moulting by feed withdrawal (FW) and a high Zn (HZn) diet on body weight and ovarian function were very similar; those of a moderate Zn with low Ca (MZn/LCa) diet were smaller. 4. The first significant effect of moulting was a decrease in oestradiol-17beta plasma concentration (d 2). Plasma progesterone decreased more gradually than oestradiol-17beta, and reached a nadir on d 6 in FW- and HZn-treated hens and on d 9 in MZn/LCa-treated ones. 5. Hens treated with either FW or the MZn/LCa, but not those with the HZn diet, showed a very sharp rise in corticosterone concentration on d 2 of treatment. Thus the MZn/LCa diet was less efficient than the other treatments in induction of ovarian involution, but had a similar effect on stress induction, as indicated by increases in plasma corticosterone.

The significant developments in the genetic selection of fast-growing broiler chickens cause difficulties for broilers in coping with extreme environmental conditions. This study was conducted to elucidate the effect of repetitive short-term increases in incubation temperature on hatchability and chick's body weight (BW) and thermoregulation immediately after hatch. Thermal manipulation (TM) had no effect on hatchability when the three trials were combined for statistical analysis, and had no effect on chick's BW. It caused a significant reduction in chicks T b , and significant decline in plasma thyroid hormones concentration, but had no effect on plasma corticosterone concentration. It can be concluded that TM did not affect BW but had a positive effect on thermoregulation, most probably in reducing metabolic rate.

Shortening daylight (to 10.5 to 11 h/d) slightly reduced the rest period (the interval between last egg and first clutch), whereas feeding a low-protein diet during the 22-d period following 8 d of feed withdrawal (FW) (recovery period) markedly extended it. Feed withdrawal accompanied by a short-daylight regime and a long recovery period led to the best postmolt production. However, production per hen housed during the whole experimental period was only slightly increased. Thus, a long rest period may mask the positive effects of short daylight and recovery diet. At least 140 to 170 d were needed to enable the molted hens to compensate for the loss of eggs during the rest period. Therefore, under certain economic conditions, rearing of nonmolting hens for 640 to 700 d should be an economic consideration. Ten days of feeding of a diet containing 0.06% nicarbazin (NICR) arrested egg production and caused a 22-d rest period but not a typical molt. Few variables of production or shell quality were improved by NICR but to a lesser extent than by FW or Zn feeding. Five days of feeding on a high-Zn diet (20 g Zn/kg; HZn) improved postmolt performances similarly to FW. Ten days of feeding on a modest-Zn (2.8 g Zn/kg), low-Ca, and low-P diet (Zn-CaP) affected postmolt performances inconsistently. In one out of two trials (trial 2), the effects of Zn-CaP were similar to those of FW or HZn; in the other (trial 3), the effects were less pronounced, more time was required for egg arrest, and more eggs were laid occasionally during the rest period. In trial 2, only the Zn-CaP diet was accompanied by short daylight. In both trials, feed intake during the induction period was only slightly reduced. Zn feeding increased the yolk Zn content slightly in eggs laid during the induction period and at the onset of production. In trial 2, only Zn-CaP markedly increased yolk Zn of eggs laid during the first 5 d of production.

The ascites syndrome in broiler chickens is attributed to metabolic burdening, which results from intensive genetic selection for rapid growth coupled with exposure to extreme environmental conditions, such as low ambient temperature. These conditions impose on the broilers difficulties in fulfilling tissue demands for oxygen, and the birds exhibit a decrease in blood oxygen saturation and high hematocrit values. It is unknown whether the increase in hematocrit results from alteration in erythropoiesis or from fluid exudation out of the blood system to the abdominal cavity. The present study was conducted to examine the association between abnormal stress response and erythropoiesis process in ascitic broilers. Ascitic chickens revealed a uniquely continuous stress response: expressing an increase (P < or = 0.05) in plasma corticosterone concentration 2 to 3 wk before death. At 5 wk of age, ascitic broilers exhibited an increase (P < 0.05) in hematocrit, blood cell count, and packed cells and blood volumes, with no significant change in plasma volume. These results confirm an accelerated erythropoiesis process in ascitic birds. Increased blood cell production in ascitic birds was matched by an increase (P < 0.05) in the proportion of immature red blood cells (23%) in comparison with broilers that remained healthy (7%), and by decreased (P < 0.05) hemoglobin content relative to red blood cells. We conclude that continually increased corticosterone concentrations, as an inducer of erythropoiesis proliferation and differentiation arrest, in ascitic chickens, resulted in increased production of red blood cells (partially immature) with decreased hemoglobin content; this decrease in hemoglobin might have contributed to enhanced development of hypoxemia and to aggravation of the syndrome.

Leptin, the key regulator of mammalian energy balance, has been at the center of a great controversy in avian biology for the last 15 years since initial reports of a putative leptin gene (LEP) in chickens. Here, we characterize a novel LEP in rock dove (Columba livia) with low similarity of the predicted protein sequence (30% identity, 47% similarity) to the human ortholog. Searching the Sequence-Read-Archive database revealed leptin transcripts, in the dove's liver, with 2 noncoding exons preceding 2 coding exons. This unusual 4-exon structure was validated by sequencing of a GC-rich product (76% GC, 721 bp) amplified from liver RNA by RT-PCR. Sequence alignment of the dove leptin with orthologous leptins indicated that it consists of a leader peptide (21 amino acids; aa) followed by the mature protein (160 aa), which has a putative structure typical of 4-helical-bundle cytokines except that it is 12 aa longer than human leptin. Extra residues (10 aa) were located within the loop between 2 5'-helices, interrupting the amino acid motif that is conserved in tetrapods and considered essential for activation of leptin receptor (LEPR) but not for receptor binding per se. Quantitative RT-PCR of 11 tissues showed highest (P.05) expression of LEP in the dove's liver, whereas the dove LEPR peaked (P.01) in the pituitary. Both genes were prominently expressed in the gonads and at lower levels in tissues involved in mammalian leptin signaling (adipose; hypothalamus). A bioassay based on activation of the chicken LEPR in vitro showed leptin activity in the dove's circulation, suggesting that dove LEP encodes an active protein, despite the interrupted loop motif. Providing tools to study energy-balance control at an evolutionary perspective, our original demonstration of leptin signaling in dove predicts a more ancient role of leptin in growth and reproduction in birds, rather than appetite control.

The present study examined the association between thyroid hormones and the development of ascites on one hand and the ability to predict ascites from growth rate and hematocrit on the other hand. Ascites syndrome was induced in broiler chickens in two trials by exposing the chicks to low ambient temperature (Ta) and by supplying a pellet form of diet. Weight gain, hematocrit, hemoglobin, and plasma thyroxin (T4) and triiodothyronine (T3) concentrations were measured weekly for each bird, and comparisons were made between birds that eventually died from ascites and those that did not. Mortality from ascites amounted to 24.3 and 24.2% in Trials 1 and 2, respectively. Weight gain did not differ between ascitic and healthy chickens up to approximately 2 wk before death but was significantly lower in the ascitic broilers 1 to 2 wk before death. Hematocrit was significantly higher in broilers with ascites with the exception of ascitic broilers that died at the age of 7 wk (Trial 1). In ascitic broilers, T4 and T3 concentrations declined significantly during the week of death. The present findings raise the question of whether the association between low levels of thyroid hormones and the development of ascites is one of the physiological responses in the syndrome cascade, or whether the failure to maintain thyroid hormones concentration is one of the triggers of the syndrome initiation. This question requires further investigation. It can be concluded that a high rate of weight gain is not always a good predictor of ascites development. Hematocrit and thyroid hormones can provide a good indication but only during the last week of life, and not in all cases. None of these parameters, however, can predict the development of ascites at an early age.

1. The effects of relative humidity (rh=40% to 70%) at high ambient temperature (Ta) on the performance of laying hens at different ages (8 to 10 months, Trial 1; and 16 to 18 months, Trial 2) was evaluated. Laying hens were exposed to 25 degrees C (control) for 3 weeks and thereafter acclimated for 1 week to 35 degrees C and 4 different rh. 2. Body weight declined significantly in young and older hens exposed to 60% or 70% and 70% rh, respectively: Food intake declined with increasing Ta, except in the case of older hens exposed to 60% rh, for which it remained relatively constant. Water consumption, however, increased with increasing Ta but the increase was significant in young hens exposed to 70% rh only. 3. Egg production was not affected by the changes in Ta. However, a decrease in egg production was observed in older hens exposed to 60% rh. 4. Egg weight (EW), shell weight (SW) and shell thickness (ST) were significantly reduced by exposure to elevated Ta, whereas % breakage significantly increased. In young hens, a response to rh was exhibited in ST which was significantly higher in hens exposed to the low rh (40% to 45%) than in those exposed to the highest rh (70% to 75%). 5. It can be concluded that Ta is the main environmental factor affecting young and older laying hens while the effect of rh is minor.

We studied the balance of Na+, K+, Cl-, and water in six high yielding (> 39 kg/d of milk) cows between wk 2 to 1 prepartum and at 2 and 7 wk postpartum during winter in Israel. Cows were fed complete diets; Na+ and Cl- contents exceeded dietary recommendations, and K+ content was equal to dietary recommendations. Milk yield was related positively and significantly to retention of Cl- and K+, indicating that ions that are the main constituents of sweat can limit the ability of cows to express full genetic potential. The highest ion retention was recorded for cows that had the highest dry matter intake and, hence, the highest ion intake. Retention of Cl- was highest for cows that were most efficient in retaining Cl- in the kidney. In hot climates, increasing the concentrations of ions in the diet of early lactation cows according to the actual dry matter intake could prevent or reduce the severity of ion deficiencies. Water turnover rate of the cows was dependent on dry matter intake, milk yield, and respiratory-cutaneous water loss. The milk-free water balance (water turnover rate minus water secreted in milk) could be very efficiently predicted for lactating and nonlactating cows by the following equation: milk-free water balance (kilograms per day) = digestible energy intake (megacalories per day) x 0.58 + respiratory-cutaneous loss (kilograms per day) x 0.97 (n = 18; R2 = 0.97). This formula provides a tool to assess the evaporative-cutaneous water loss from feed and water intake measurements to evaluate the severity of heat stress.

A procedure to measure poly(ethylene glycol) (PEG) binding to plant samples is described. The amount of PEG bound to a sample was found to be reproducible with a small coefficient of variation (0.5%). The PEG-binding capacity of plant samples was not affected by predrying of the samples at 90 °C, unlike the colorimetric methods for determination of tannins. Both the PEG-binding and the protein precipitation capacity of the plant samples were found to be useful in predicting the negative effects of tannins on ruminal degradation of the plant material. However, PEG binding to plant samples had an advantage over protein precipitation in samples in which formation of strong tannin−protein complexes results in low extractability of tannins. For this reason, the binding method was found to be preferable to the protein precipitation method in predicting the negative effect of tannins on ruminal degradation of forage plants. Keywords: Poly(ethylene glycol) (PEG); tannin; rumen; digestibility; forage plants; herbi...

Embryo development is a dynamic process, determined by both the genetic background of the organism and the environment in which it develops. Environmental alterations during an organism's embryogenesis may induce changes in the development of some physiological regulatory systems, thereby causing permanent phenotypic changes in the embryo. The present study aimed to assess the effect of 17% O2 concentration during chorioallantoic membrane (CAM) development on a) CAM development, b) cardiovascular parameters, and c) embryo development postexposure and up to hatch. Two replicated trials, each with 840 fertile Cobb eggs, were conducted. At embryonic d 5 (E5) eggs were divided into 2 treatments: 1) control, and 2) 17% O2 concentration for 12 h/d from E5 through E12 (12H). The 12H embryos exhibited a clear and significant increase in the vascular area of the CAM, which grew to 6.8% larger than that of the control. Hematocrit and hemoglobin levels, as measured on E13 and E14, increased in response to the hypoxic treatments, but these differences were not maintained subsequently. Heart rate and relative heart weight were not affected by hypoxic exposure, but eggshell temperature in the 12H treatment was higher than that of the control, indicating higher heat production, which is consistent with the elevated plasma concentrations of triiodothyronine and thyroxin and with the enhanced oxygen consumption and residual yolk intake rate that followed exposure to hypoxic conditions. These findings indicate that embryos adapted to hypoxic condition enhance angiogenesis processes, which subsequently increase their blood oxygen-carrying capacity, enabling the increase of oxygen consumption, which positively affects their growth development and maturation compared with the control embryos. Such alterations may affect posthatch performance and the ability of broilers cardiovascular system to meet elevated oxygen demand.

The lack of affordable techniques for gene transfer in birds has inhibited the advancement of molecular studies in avian species. Here we demonstrate a new approach for introducing genes into chicken somatic tissues by administration of a lentiviral vector, derived from the feline immunodeficiency virus (FIV), into the chorioallantoic membrane (CAM) of chick embryos on embryonic day 11. The FIV-derived vectors carried yellow fluorescent protein (YFP) or recombinant alpha-melanocyte-stimulating hormone (α-MSH) genes, driven by the cytomegalovirus (CMV) promoter. Transgene expression, detected in chicks 2 days after hatch by quantitative real-time PCR, was mostly observed in the liver and spleen. Lower expression levels were also detected in the brain, kidney, heart and breast muscle. Immunofluorescence and flow cytometry analyses confirmed transgene expression in chick tissues at the protein level, demonstrating a transduction efficiency of ∼0.46% of liver cells. Integration of the viral vector into the chicken genome was demonstrated using genomic repetitive (CR1)-PCR amplification. Viability and stability of the transduced cells was confirmed using terminal deoxynucleotidyl transferase (dUTP) nick end labeling (TUNEL) assay, immunostaining with anti-proliferating cell nuclear antigen (anti-PCNA), and detection of transgene expression 51 days post transduction. Our approach led to only 9% drop in hatching efficiency compared to non-injected embryos, and all of the hatched chicks expressed the transgenes. We suggest that the transduction efficiency of FIV vectors combined with the accessibility of the CAM vasculature as a delivery route comprise a new powerful and practical approach for gene delivery into somatic tissues of chickens. Most relevant is the efficient transduction of the liver, which specializes in the production and secretion of proteins, thereby providing an optimal target for prolonged study of secreted hormones and peptides.

Extreme thermal conditions may dramatically affect the performance of broilers and other domestic animals, thereby impairing animal welfare and causing economic losses. Although body core temperature is the parameter that best reflects a bird's thermal status, practical and physiological obstacles make it irrelevant as a source of information on the thermal status of commercial flocks. Advances in the technology of infrared thermal imaging have enabled highly accurate, noncontact, and noninvasive measurements of skin surface temperature. Providing that skin surface temperature correlates with body temperature, this technology could enable acquisition of reliable information on the thermal status of animals, thereby improving diagnoses of environmental stress in a flock. This study of broiler chickens found a strong positive correlation between body core temperature and facial surface temperature, as recorded by infrared thermal imaging. The correlation was equally strong at all ages from 8 to 36 d during exposure to acute heat stress with or without proper ventilation and after acclimation to chronic heat exposure. A similar correlation was found by measurements in commercial flocks of broilers. Measurements of blood plasma concentrations of corticosterone, thyroid hormones, and arginine vasotocin confirmed that metabolic activity was low after acclimation to chronic exposure to heat, whereas ventilation was at least as efficient as acclimation in reducing thermal stress but did not impair metabolism. In light of these novel results, commercial benefits of infrared thermal imaging technology are suggested, especially in climate control for commercial poultry flocks. The application of this technique to other domestic animals should be investigated in future experiments.

Ascites syndrome (AS) is manifested in flocks of contemporary broilers that are allowed to fully manifest their genetic potential for rapid growth. After successful selection, a pair of divergent lines was established, AS-susceptible (AS-S) and AS-resistant (AS-R). These lines facilitate comparisons between genetically resistant and susceptible healthy young broilers when reared under standard brooding conditions (SBC). The aim of the present study was to look for predictive indicators for AS susceptibility by comparing relevant physiological parameters in the AS-S and AS-R lines under SBC and after exposure to extreme ascites-inducing conditions (AIC). In this design, a trait differing significantly between the 2 lines under SBC is expected to be a reliable indicator for selection against AS susceptibility in breeding stocks when reared under noninducing conditions. Males from the AS-S and AS-R lines were reared together under SBC to 19 d of age, then under the AIC protocol. Cumulative incidence of AS mortality was 93.2% in the AS-S line and only 9% in the AS-R line, confirming the genetic divergence between the lines. Exposure to AIC enhanced the imbalance between oxygen demands and supply in the AS-S birds and induced differences in blood parameter level between the 2 lines. The AS-S birds exhibited elevated hematocrit and red blood cell counts and a decline in oxygen saturation in the arterial blood. No difference in hemoglobin concentration was found, but calculation of hemoglobin content per 1,000 red blood cells revealed a significant reduction in hemoglobin content in the AS-S birds. Under SBC, there were no significant differences between the lines for hematocrit, red blood cell count, hemoglobin concentration, hemoglobin count per 1,000 red cells, and blood oxygen saturation. However, heart rate during the first week of life was significantly higher in the AS-S birds than in the AS-R birds on d 1 and 7, suggesting that high heart rate may potentially serve as an early criterion for selection against AS susceptibility.

The time just before hatch is critical, because the embryo shifts toward internal and external pipping. This study aimed to determine the beneficial effect of repeated acute reductions of the incubation temperature during the last phase of broiler embryogenesis on posthatch cold tolerance and on the development of ascites syndrome. Fertile eggs were incubated at 37.8 degrees C and 56% RH. At 18 and 19 d of incubation, 3 treatments were conducted, comprising 2 or 3 exposures to 15 degrees C for 30 or 60 min each. During these cold exposures, egg temperature was measured by infrared thermography to determine sensible heat loss from the eggs. At hatch, BW and body temperature were measured. At 3 and 14 d of age, chicks were challenged by cold exposure to 10 degrees C for 3 h. From 14 d of age onward, three-quarters of the chicks were raised under ascites-inducing conditions (AIC) and the others were raised under regular conditions. The sensible heat loss from the eggs was 512 +/- 66 cal and 718 +/- 126 cal for 30 and 60 min of cold exposure, respectively. No effect of treatment on hatchability was observed, but body temperature and BW were greater to significantly greater in the treated chicks. Cold challenges at 3 and 14 d of age revealed a relative thermoregulatory advantage of embryos exposed to cold for 60 min. Under AIC, fewer treated chickens than controls developed ascites. At 38 d of age, BW and relative breast muscle weight were numerically to significantly greater in the treated chicks than in the control chicks when both were raised under regular conditions, whereas no differences were observed among the chicks raised under AIC. Repeated brief acute cold exposures during the last phase of embryogenesis appeared to improve the ability of growing broilers to withstand low ambient temperatures during their life span. Moreover, chickens treated during embryogenesis improved their performance under regular growth conditions.

The potential to induce improved thermotolerance in broiler chickens is of great importance. Thermal conditioning is one of the management tools used to improve thermotolerance, enabling broilers to cope with extreme environmental conditions. This study investigated the effects of exposing chicks to low ambient temperature (T(a)) on-chick body (T(b)), surface (T(s)) temperatures and total sensible heat loss (SHL) by convection and radiation from the body and from 2 main radiative organs, the face and the legs. At 3, 4, or at both 3 and 4 d of age, chicks were exposed to 5 degrees C for 1.5 h a day (to avoid mortality) or to 10 or 15 degrees C for 3 h a day. In general, in all treatments, the results during exposure to cold differed significantly from the control. A second cold exposure (on d 4 after a first exposure on d 3) clearly enhanced the chicks' ability to maintain on-chick body surface temperatures during exposure to 15 degrees C and to recover much faster from cold exposure. A dramatic decline in average surface temperature was observed during the first 15 min of chicks' exposure to the various low ambient temperatures in all ages, reaching the lowest values in the 5 degrees C treated chicks. The face responded immediately to cold exposure by significantly increasing its SHL to a level that then remained relatively steady (15 degrees C) or declined moderately with time (10 and 5 degrees C). In the legs, however, a significant and continuous decline in SHL was exhibited in all ages. The dynamics of SHL from the legs differed from that from the face, suggesting that the legs are a major organ for vasomotor responses, whereas the face is a more conservative vasoregulatory organ. It is concluded that repetitive exposure to cold may enhance thermotolerance, and that this is partially related to the vasomotor responses. This is the first report quantifying the differentiation between the legs as a responsive vasomotor organ and the face as a conservative vasomotor one.

Agonists of membranal melanocortin 3 and 4 receptors (MC3/4Rs) are known to take part in the complex control mechanism of energy balance. In this study, we compared the physiological response to an exogenous MC3/4R agonist and the hypothalamic expression of proopic melanocortin (POMC) gene, encoding few MC3/4R ligands, between broiler and layer chicken strains. These strains, representing the two most prominent commercial strains of chickens grown for meat (broilers) and egg production (layers), differ in their food intake, fat accumulation, and reproductive performance and, therefore, form a good model of obese and lean phenotypes, respectively. A single i.v. injection of the synthetic peptide melanotan-II (MT-II; 1 mg/kg body weight) into the wing vein of feed-restricted birds led to attenuation of food intake upon exposure to feeding ad libitum in both broiler and layer chickens. A study of the POMC mRNA encoding the two prominent natural MC3/4R agonists, α-MSH and ACTH, also revealed a general similarity between the strains. Under feeding conditions ad libitum, POMC mRNA levels were highly similar in chicks of both strains and this level was significantly reduced upon feed restriction. However, POMC mRNA down-regulation upon feed restriction was more pronounced in layers than in broilers. These results suggest: (i) a role for MC3/4R agonists in the control of appetite; (ii) that the physiological differences between broilers and layers are not related to unresponsiveness of broiler chickens to the satiety signal of MC3/4R ligands. Therefore, these findings suggest that artificial activation of this circuit in broiler chicks could help to accommodate with their agricultural shortcomings of overeating, fattening, and impaired reproduction.

This study aimed at elucidating the effects of thermal manipulation (TM) during different periods of embryogenesis on hatchability, body weight, and thermoregulation of Ross chicks at hatch, and on the chick's ability to cope with thermal challenge (TC) at 3 d of age. Control embryos were incubated at 37.8 degrees C and 56% RH, whereas the TM embryos were treated for 3 h at 39.5 (1) or 41.0 degrees C (2) and 65% RH, during early embryogenesis (EE), E8 to E10, and late embryogenesis (LE) at E16 to E18. At hatch, after feather drying, BW and body temperature (Tb) were measured, blood was drawn from the jugular vein to measure plasma concentrations of thyroxine (T4), triiodothyronine (T3), and corticosterone. These parameters were also measured in 3-d-old chicks during exposure to TC at 41 degrees C for 6 h. Hatchability was significantly higher in the LE1-treated group than in the other treatments. EE1- and LE1-treated embryos exhibited significantly lower Tb at hatch than the LE2 chicks. Significant hyperthermia was noted in the control chicks during 6 h of TC; it coincided with a higher plasma T3 concentration (P = 0.074). During TC, the LE1-treated chicks exhibited the lowest T3 to T4 ratio and a significantly reduced plasma corticosterone concentration. It can be concluded that TM at 39.5 degrees C for 3 h during E16 to E18 of incubation improved chick's thermotolerance acquisition and reduced the corticosterone level of chicks exposed to TC at the age of 3 d.

Air velocity (AV) is one of the main environmental factors involved in thermoregulation, especially at high ambient temperatures. To elucidate the effect of AV on performance and thermoregulation of 4- to 7-wk-old broiler chickens, an experiment was conducted using 4 different AV (0.8, 1.5, 2.0, and 3.0 m/s) at constant ambient temperatature (35 +/- 1.0 degrees C) and RH (60 +/- 2.5%). BW, feed intake, and fecal and urinary excretions were monitored in individuals and were used to calculate the amount of energy expended for maintenance. Infrared thermal imaging radiometry was used to measure surface temperatures for the calculation of heat loss by radiation and convection. Brachial vein blood was collected for plasma osmolality and arginine vasotocin analysis. Broilers performed optimally at an AV of 2.0 m/s. Energy expenditure for maintenance was significantly higher under these conditions, suggesting the ability to direct a sufficient amount of energy to control body temperature, while maintaining relatively high growth rates. Convective heat loss increased significantly with increasing AV, whereas radiative heat loss was not affected. Sensible heat loss, expressed as a percentage of energy expenditure for maintenance, was significantly higher at 2.0 m/s compared with 0.8 m/s but significantly lower than that of 3.0 m/s. The high level of heat loss observed at 3.0 m/s probably affected body water balance, as supported by significantly higher plasma osmolality, arginine vasotocin concentration, and the hyperthermic status of these birds. It can be concluded that AV of 2.0 m/s enables broilers to maintain proper performance together with efficient thermoregulation and water balance under harsh environmental conditions.

Three series of experiments were conducted with fast-growing chickens in order: to evaluate the effects of dietary Ca and P on cholecalciferol metabolism and expression; to determine dietary Ca requirements; to determine dietary P requirements. The results of the first series confirmed previous results on the effects of dietary Ca and P on some variables of vitamin D metabolism and expression, Ca homeostasis and P metabolism in the young chicken (1- to 21-d-old), and extended them to older birds (22- to 43-d-old). The bone formation rate and the duodenal calbindin content were maintained at high levels until the age of 43 d. Dietary Ca or P restriction increased duodenal calbindin and decreased bone ash in both 22- and 43-d-old chickens, but the effect on bone ash was less pronounced in the 43-d-old birds than in the younger ones. These results suggest that: (a) the capabilities for adaptation to dietary Ca and P restriction remain high during the whole growing period; (b) the growing broilers express a high adaptive capability even when the diet contains the recommended Ca and P contents. The results of the second and third series of experiments suggest that: (c) unlike the Ca requirements of the 1- to 22-d-old chick, P requirements for growth and bone ash are similar, and are as high in the older chicks as in the younger ones (7·4–8·3 g P/kg or 4·8–5·7 g non-phytate P/kg diet); (d) although growth and bone ash in the 29- to 43-d-old chickens appear to be less sensitive to dietary Ca content, within a range close to the calculated P requirement, 10 g Ca/kg diet appears to be required for best tibia mineralization, and to a lesser extent for better growth at this age.

Summary A chicken gene orthologous to human leptin receptor (LEPR) has been characterized and found to be active in leptin signaling in vitro in response to a variety of recombinant leptins and leptin-containing blood samples. However, the endogenous ligand of chicken LEPR (cLEPR) - the putative chicken leptin - has been reported by us and others to be undetectable at the DNA, mRNA, protein and activity levels. These reports have raised questions as to cLEPR's role. Here we analyzed the effects of a pegylated superactive mouse leptin antagonist (PEG-SMLA) in chicken. We showed that the leptin antagonist efficiently and specifically blocks leptin signaling through the cLEPR in vitro. The effect of the leptin antagonist was then studied in vivo by daily administration of 10 mg/kg for 10 consecutive days to White Leghorn female chickens (G. gallus), at the age of two weeks. Despite the efficient attenuation of the cLEPR in vitro, no effect was observed on body weight, feed intake, feed efficiency or fat accumulation in the treated birds. Since similar treatment in rodents leads to a highly pronounced increase in appetite and body weight that are observed from the first day of treatment, it is concluded that the cLEPR is not implicated in the control of appetite or adipose homeostasis in chickens.

The effect of air velocity (AV) on weight gain, feed intake, and feed efficiency was studied in male broiler chickens exposed to a constant 35 C, 60% RH, and continuous illumination from 4 through 7 wk of age. In two separate trials, AV of 0.5, 1.5, 2.0, and 3.0 m/s (Trial 1) and 1.0, 1.5, 2.0, and 2.5 m/s (Trial 2) were used. The fifth week was used for acclimation to the targeted environmental conditions. Weight gain was significantly higher in broilers exposed to AV of 1.5 and 2.0 m/s in both trials, which coincided with significantly higher feed intake and feed efficiency. Exposure to the low AV (0.5 and 1.0 m/s) resulted in weight gain, feed intake, and feed efficiency that were less than those of broilers exposed to high AV (2.5 and 3.0 m/s). In all treatments, broilers developed hyperthermia, but there were no significant differences in body temperature (Tb) among the birds. It can be concluded that AV has a significant effect on broiler performance; the optimal AV was found to be 1.5 to 2.0 m/s.

Stress and stress related hormones such as glucocorticoids inhibit lactation in cows. In the present study we propose a novel mechanism connecting stress with plasminogen-plasmin system (PPS) (an enzymatic mechanism in milk, which leads to the breakdown of the major milk protein casein). We show that stress activates the PPS leading to an increase in plasmin activity, and that a distinct plasmin-induced beta-casein breakdown product (fraction 1-28) is a potent blocker of potassium channels in mammary epithelia apical membranes. The reduction in milk production due to dehydration stress or glucocorticoid (dexamethsone) was correlated with the activities of plasmin and channel blocking activity in the milk of the tested cows. The notion that the axis Stress-PPS-beta-casein fraction 1-28 is responsible for the reduction in milk yield is supported by the results of experiments showing that injecting solution composed of casein digest enriched with beta-casein fraction 1-28 to the udder lumen leads to a transient reduction in milk production. Furthermore, injecting a pure beta-casein fraction 1-28 to the udder lumen of goat's lead also to a transient reduction in milk production with kinetics that was similar to the kinetics observed in cows.