Early Post-hatch Period Research Articles

In ovo injection of testosterone to yolk sac modulates early posthatching development and physiology of male chick in broilers

The aim of this study was to investigate the effects of in ovo testosterone injection into the yolk sac of embryos on physiology and development of broiler chicks during the early posthatching period. A total of 1,010 hatching eggs were obtained from the Ross genotype. Trial design was conducted with a noninjected group (control) and injection groups in which 100 µL sesame oil, or 100 µL sesame oil + 0.50 µmol testosterone were injected into the yolk sac of the embryo on d 6 or d 12 of incubation. Testosterone hormone level was measured in the egg yolk and albumen at onset of incubation, in the yolk sac on d 19 of incubation and in the residual yolk sac at hatching. Weights of chick, yolk sac and organ, morphological traits (body length, lengths of bilateral traits and beak length), asymmetrical development of bilateral morphological traits and body mass index were measured at hatching and on d 7 after hatching. Testosterone, corticosterone and growth hormone levels were determined in blood plasma obtained from male chicks at hatching and on d 7 of chick age. Chick weight was not affected, plasma testosterone level and brain weight decreased, while body mass index, plasma corticosterone and growth hormone levels increased by administering 0.50 µmol testosterone on d 12 of embryonic age. However, plasma testosterone and growth hormone levels did not change, chick weight increased, while plasma corticosterone level and the chick body length decreased by administering 0.50 µmol testosterone on d 6 of embryonic age. A significant interaction between chick age and in ovo testosterone administration resulted in an increase in lung weight of chicks. In conclusion, this study found that in ovo testosterone administered at different embryonic ages due to age-specific effects of testosterone in the yolk sac of embryo modulates development related to physiological parameters of male broiler chicks during early posthatching period.

The effects of exposure to cold during incubation on developmental stability, fear, growth, and carcass traits in Japanese quails.

The aim of this study was to determine the effects of 6h/day cold (35.0°C) acclimatization between the 9th and 15th days of incubation of Japanese quail embryos on hatchability, livability, chick quality, developmental stability, fear response, live weight, and slaughter-carcass characteristics. Two homologous incubators and a total of 500 hatching eggs were used in the study. Randomly selected half of the eggs were exposed to cold according to the eggshell temperature. The cold acclimation of Japanese quail embryos had no adverse effects on all mentioned traits, except for chick quality. Chicks in the control group had higher Tona scores (99.46) than those exposed to cold (99.00) (P < 0.05). In addition, there were differences among the treatment groups in terms of the parameters of mature weight (β0), instantaneous growth rate (β2), and inflection point coordinates of the Gompertz growth model (P < 0.05 for all). It was found that exposing embryos to cold during the incubation changed the shape of the growth curve. As the development of embryos exposed to cold slows down, a compensatory growth occurs in the early posthatch period. Thus, the growth rate increased in the period before the inflection point of the growth curve.

Influence of In Ovo Leptin Injection into Yolk Sac on Embryo Development, Blood Biochemical Parameters and Lipid Metabolism of Broiler Chicks during Early Post-Hatching Period

The study aimed to investigate the effect of in ovo (IO) leptin injection on yolk sac leptin concentration, embryo development, blood lipid metabolism, and biochemical parameters of broiler chicks during the early post-hatching period. Hatching eggs were weighed and placed in an incubator with 100 eggs/4 replications/group. The groups were: non-injected group (C); and 3 injected groups with 100 μl of phosphate buffer solution (PBS); 0.5 μl leptin (L0.5), and 1 μl leptin (L1) in 100 μl of PBS. Pure leptin hormone dissolved in PBS was injected into the yolk sac on day 7 of incubation. Yolk/yolk sac leptin concentrations at the onset of incubation, on days 7 and 15 of incubation, and at hatching were determined. Embryo/chick development was determined at all ages. Serum leptin, tri-iodothyronine (T3), thyroxine (T4), creatine kinase, total protein, uric acid, and lipid profiles were measured at hatching and day 7 of chick age. The leptin level of the yolk was similar to the leptin level of the yolk sac at hatching. Leptin level of a yolk sac was higher on 7th day than at other ages. IO injection* age was significant for serum leptin level, which was similar to one-day chicks in the L0.5 at day 7 of chick age. T3, T4, creatine kinase, uric acid, lipid profiles, and embryo/chick development did not change with IO leptin. Serum HDL level was higher in the leptin groups than C and PBS groups at hatching. IO leptin application affects blood lipid metabolism depending on dose level for male broiler chicks at the early post-hatch period, without affecting embryo/chick development.Leptin level of yolk was similar to leptin level of yolk sac at hatching. Leptin level of yolk sac was higher on 7th day than other ages. IO injection* age was significant for serum leptin level, which was similar to one-day chicks in the L0.5 at day 7 of chick age. T3, T4, creatine kinase, uric acid, lipid profiles, and embryo/chick development did not change with IO leptin. Serum HDL level was higher in the leptin groups than C and PBS groups at hatching. IO leptin application affects blood lipid metabolism depending on dose level for male broiler chicks at early post-hatch period, without affecting embryo/chick development.

Intestinal Immune Development Is Accompanied by Temporal Deviation in Microbiota Composition of Newly Hatched Pigeon Squabs.

ABSTRACTIdentifying the interaction between intestinal mucosal immune system development and commensal microbiota colonization in neonates is of paramount importance for understanding how early life events affect resistance to disease later in life. However, knowledge about this interaction during the early posthatch development period in altrices is limited. To fill this gap, samples of intestinal content and tissue were collected from newly hatched pigeon squabs at four time points (days 0, 7, 14, and 21) for microbial community analysis and genome-wide transcriptome profiling, respectively. We show that the first week after hatching seems to be the critical window for ileal microbiota colonization and that a potentially stable microbiota has not yet been well established at 21 days of age. Regional transcriptome differences revealed that the jejunum rather than the ileum plays a crucial role in immunity at both the innate and adaptive levels. In the ileum, temporal deviation in innate immune-related genes mainly occurs in the first week of life and is accompanied by a temporal change in microbiota composition, indicating that the ileal innate mucosal immune system development regulated by microbial colonization occurs mainly in this period. Furthermore, we provide evidence that colonization by Escherichia and Lactobacillus within the first week of life is likely one of the causative factors for the induction of proinflammatory cytokine expression in the ileum. We also demonstrate that cellular adaptive immune responses mediated by Th17 cells following commensal-induced proinflammatory cytokine production in the ileum begin as early as the first week posthatch, but this cellular immunity seems to be less effective in terms of maintaining the inflammatory response balance. Because the induction of high levels of mucosal secretory IgA (SIgA) seems to take approximately 3 weeks, we favor the idea that humoral adaptive immunity might be less active, at least, during the first 2 weeks of life. Our data may help to explain the phenomenon of the occurrence of intestinal infections mainly in the ileum of pigeon squabs during the early posthatch period.IMPORTANCE The pigeon (Columba livia), an altricial bird, is one of the most economically important farmed poultry for table purposes. Identifying the interaction between intestinal mucosal immune system development and commensal microbiota colonization in neonates is of paramount importance for understanding how early life events affect resistance to disease and potential productivity later in life. However, knowledge about this interaction during the early posthatch development period in altricial birds is limited. The study described herein is the first to try to provide insights into this interaction. Our data provide evidence on the mutual relationship between intestinal mucosal immune system development and commensal microbiota colonization in pigeon squabs and may help to explain the phenomenon of the occurrence of intestinal infections mainly in the ileum of pigeon squabs during the early posthatch period.

부화 초기 육계의 열 스트레스와 근육발달

Global warming and scorching summer seasons affect the growth ability of broilers and animal welfare. In modern broilers, vital organs, such as the heart and lungs, grow disproportionally under intensive selection, making it difficult to adapt to warmer climates. Changes in environmental temperature can affect muscle formation during embryonic development and the early posthatching period. Satellite cells are highly sensitive to heat stress. Heat stress affects the proliferation and differentiation activity of satellite cells and muscle growth and structure. Therefore, thermal manipulation during broiler chick embryogenesis and environmental temperature management at the beginning of hatching are critical for the development and growth of broiler muscles. This review focuses on the thermoregulation mechanism of birds, the muscle development process of broilers, and the function of satellite cells, the relationship between heat stress and muscle development of chicks shortly after hatching, and studies on heat resistance and muscle growth of broilers.

Supplementation of vitamin E and omega-3 fatty acids during the early posthatch period on intestinal morphology and gene expression differentiation in broilers

Early posthatch nutrition is important for gut health. Vitamin E (VE) and omega-3 (n-3) fatty acids can improve gut health through antioxidative and anti-inflammatory effects. The objective of this study was to identify the effects of VE, n-3 fatty acids, and combination of both during the starter phase (0–10 d) or grower phase (11–24 d) on intestinal morphology and expression of genes associated with gut health. A total of 210 Ross 708 broilers were randomly assigned into 7 treatments with 10 replicates of 3 birds each. The control group was fed a corn–soybean meal–basal diet during the entire study (0–58 d). Supplementation of VE (200 IU/kg), n-3 fatty acids (n-6/n-3 ratio of 3.2:1), and combination of both were fed during the starter phase (0–10 d) or grower phase (11–24 d). All of the broilers were harvested at 58 d of age. Villus height, crypt depth, villus width, distance between villi, and number of intraepithelial lymphocytes were obtained. Expression of 21 genes was measured using NanoString analysis. Expression of solute carrier family 15 member 1 (P = 0.01) associated with peptide transport and mucin 2 (P = 0.03) related with intestinal mucus barrier was increased in the broilers supplemented with n-3 fatty acids in the grower diet compared with the control. Expression of solute carrier family 7 member 1 associated with amino acid transport was decreased in the group supplemented with n-3 fatty acids during the starter phase compared with the group supplemented with n-3 fatty acids (P = 0.01) or VE and n-3 fatty acids during the grower phase (P = 0.03). These data suggest that VE and n-3 fatty acids supplemented during the grower phase have a positive effect on improving nutrient transport with n-3 fatty acids supplementation in the grower diet showing the most beneficial effect. These findings can be used in the development of nutritional management strategies to improve broiler growth performance and meat quality.

Embryonic development, hatching time and newborn juveniles of Octopus tehuelchus under two culture temperatures

The development of cephalopods early life stages is strongly influenced by environmental variables, especially temperature. Octopus tehuelchus (d'Orbigny, 1834) is an Atlantic Patagonian fishery resource currently being studied as a new species for cultivation; however it is not known how temperature modulates its early life stages. In this work, egg masses were artificially incubated at 13 and 16 °C under controlled aquarium conditions. In each thermal treatment, the stages of embryonic development, embryo morphometry and survival throughout embryogenesis, as well as embryogenesis duration were recorded. After hatching, the morphological description of the juveniles was achieved and survival time in starvation was calculated for both temperatures. At 16 °C the mean embryonic duration was 85 days shorter than at 13 °C. For both thermal treatments, the highest mortalities occurred up to the beginning of organogenesis, and no significant differences in hatching success were observed. The temperature also showed the potential to increase or decrease the juvenile performance at the early post hatching period. This resulted in a significant reduction in size and weight of new born juveniles at 13 °C but also in an average increment of 7 days in their survival in starvation when compared to octopus reared at 16 °C. The chromatophore pattern was similar for both thermal treatments and was characteristic of juveniles of this species. The observed differences seem to show adaptive mechanisms that optimize embryos and juveniles viability under the different environmental temperatures that can be found in the northern Atlantic Patagonian coast. From a practical point of view, our findings are important to define the biological parameters and associated procedures for the cultivation of the early life stages of O. tehuelchus.

Timing Is Everything-The High Sensitivity of Avian Satellite Cells to Thermal Conditions During Embryonic and Posthatch Periods.

Myofiber formation is essentially complete at hatch, but myofiber hypertrophy increases posthatch through the assimilation of satellite cell nuclei into myofibers. Satellite cell proliferation and differentiation occur during the early growth phase, which in meat-type poultry terminates at around 8 days posthatch. Thus, any factor that affects the accumulation of satellite cells during late-term embryogenesis or early posthatch will dictate long-term muscle growth. This review will focus on the intimate relationship between thermal conditions during chick embryogenesis and the early posthatch period, and satellite cell myogenesis and pectoralis growth and development. Satellite cells are highly sensitive to temperature changes, particularly when those changes occur during crucial periods of their myogenic activity. Therefore, timing, temperature, and duration of thermal treatments have a great impact on satellite cell activity and fate, affecting muscle development and growth in the long run. Short and mild thermal manipulations during embryogenesis or thermal conditioning in the early posthatch period promote myogenic cell proliferation and differentiation, and have long-term promotive effects on muscle growth. However, chronic heat stress during the first 2 weeks of life has adverse effects on these parameters and may lead to muscle myopathies.

Short communication: Short-term fasting and refeeding induced changes in subcutaneous adipose tissue physiology in 7-day old Japanese quail

Adipose tissue development is influenced by a variety of factors, including nutrition and genetic background. Among avian species, the most is known in chickens and it is unclear if other less-artificially-selected birds are similar during the first week post-hatch. The aim of this study was thus to determine effects of fasting and refeeding on adipose tissue physiology in Japanese quail (Coturnix japonica). On day 7 post-hatch, quail were randomly assigned to fed (control), 6 h of fasting (fasted), or 6 h of fasting followed by 1 h of refeeding (refed) groups. Blood samples were collected for plasma non-esterified fatty acid (NEFA) determination and subcutaneous adipose tissues were harvested for gene expression analyses. Plasma NEFAs were elevated in the fasted state and restored to baseline within 1 h of refeeding, whereas the expression of monoglyceride lipase in subcutaneous adipose tissue was not affected by feeding status. CCAAT/enhancer binding protein α mRNA was decreased by fasting and this change persisted through refeeding, whereas neuropeptide Y receptor 5 mRNA was decreased in refed compared to fasted birds. Our results suggest that fasting promotes lipolysis and gene expression changes in young quail with some of these changes restored to original levels within only 1 h of refeeding. Thus, in quail, adipose tissue physiology is dynamic and influenced by short-term changes in nutritional status during the early post-hatch period.

Selectived and Reshaped Early Dominant Microbial Community in the Cecum With Similar Proportions and Better Homogenization and Species Diversity Due to Organic Acids as AGP Alternatives Mediate Their Effects on Broilers Growth.

Understanding the differences in microbial communities shaped by different food selective forces, especially during early post-hatch period, is critical to gain insight into how to select, evaluate, and improve antibiotic growth promoters (AGPs) alternatives in food animals. As a model system, commercial diet-administered OAs (DOAs) and water-administered OAs (WOAs) were used separately or in combination as Virginiamycin alternatives for broiler feeding during two growth phases: 1–21 days and 22–42 days. Among these three OA-treated groups, the DOA group was most similar to the AGP group in the composition and the proportion of these dominant bacterial communities at the level of phylum, family, and genus in cecal chyme of broilers. Sub-therapeutic Virginiamycin decreased the richness, homogenization, and species diversity of gut microbiota, especially in the early growth stage from days 1 to 21. Among these three OA supplementation schemes, it was clear that DOA supplementation was more likely to increase or maintain the richness, homogenization, species diversity, and predicted gene functions of cecal microbiota in treated broilers than either no supplementation or AGP supplementation during two experimental stages. The interference of DOA treatment with early colonization of probiotics and pathogens in broiler cecum was the most similar to AGP treatment, and OAs did not cause the occurrence of Virginiamycin-resistant strains of Enterococcus at the end of this trial. In terms of the predicted gene functions of the microbiota, AGP and DOA treatments provided a similar selective force for microbial metabolism functions in the cecum of broiler chickens, especially in the early growth stage. Noticeably, the relative abundance of some microbiome that was modified by Virginiamycin or DOA supplementation was significantly correlated with body weight gain and KEGG pathway analysis-annotated gene functions such as replication and repair, translation, nucleotide metabolism, and so on. With the comprehensive analysis of these results and practical application, shortened DOA supplementation, after optimization of the amount of addition, would be a suitable alternative to sub-therapeutic Virginiamycin. It was suggested that the programed intestinal microecology under such early selection forces and the effective addition time may be the key elements to focus on the designed alternate strategies of AGPs in food animals.

Age-related intestinal monosaccharides transporters expression and villus surface area increase in broiler and layer chickens.

In the chicken small intestine, glucose is mainly transported by the apically located sodium/glucose cotransporter 1 (SGLT1) and the basolaterally located glucose transporter 2 (GLUT2). Fructose is transported by the apically located glucose transporter 5 (GLUT5) and similarly by GLUT2. During the early post-hatching period, the intestinal villus surface area (VSA) should be considered as an important factor related to the monosaccharide absorption capacity. Our objective here was to study intestinal monosaccharide absorption by analyzing the effects of age, diet, and breed on monosaccharide transporters and the VSA. The mRNA expression patterns of SGLT1, GLUT2 and GLUT5 genes in broiler and layer chickens were measured from the day of hatching to day 28 using the absolute quantitative real-time PCR. Both the intestinal mRNA expression levels of these genes and the VSA were affected by age. The mRNA expression levels of SGLT1 and GLUT2 were significantly increased from day 1 to day 3 and then decreased from day 3 to day 28. The expression levels of GLUT5 decreased from day 1 to day 7. The broiler chickens VSAs were significantly larger than those of the layer chickens from days 7 to 28. The effect of diet on the gene expression patterns of these monosaccharide transporters and the VSA were not significant. Our results suggest that the expression levels of these monosaccharide transporters are increased rapidly at the beginning of intestinal growth to meet the demands for monosaccharides to support the fast growth of the chick before day 7. As intestinal maturation and VSA increased, the expression levels of these monosaccharide genes decreased to a certain expression level to maintain the intestinal transport capacity and the absorption balance of all other nutrients.

Early-life injury produces lifelong neural hyperexcitability, cognitive deficit and altered defensive behaviour in the squid Euprymna scolopes.

Injury occurring in the neonatal period in mammals is known to induce plasticity in pain pathways that may lead to pain dysfunction in later life. Whether these effects are unique to the mammalian nervous system is not well understood. Here, we investigate whether similar effects of early-life injury are found in a large-brained comparative model, the cephalopod Euprymna scolopes. We show that the peripheral nervous system of E. scolopes undergoes profound and permanent plasticity after injury of peripheral tissue in the early post-hatching period, but not after the same injury given in the later juvenile period. Additionally, both innate defensive behaviour and learning are impaired by injury in early life. We suggest that these similar patterns of nervous system and behavioural remodelling that occur in squid and in mammals indicate an adaptive value for long-lasting plasticity arising from early-life injury, and suggest that injuries inflicted in very early life may signal to the nervous system that the environment is highly dangerous. Thus, neonatal pain plasticity may be a conserved pattern whose purpose is to set the developing nervous system's baseline responsiveness to threat. This article is part of the Theo Murphy meeting issue 'Evolution of mechanisms and behaviour important for pain'.

Early posthatch thermal stress causes long-term adverse effects on pectoralis muscle development in broilers

Broiler chicks in the immediate posthatch handling period are exposed to thermal stress, with potentially harmful consequences for muscle growth and structure (e.g., less protein and more fat deposition). We addressed the effects of broiler chicks' exposure to various ambient temperatures during the first 13 D posthatch on their performance, as well as on muscle development and structure, up to day 35. Body weight and pectoralis muscle growth were lower throughout the entire period in the high-heat-exposed chicks (39°C, Hot) and to a lesser extent in the mild-heat-exposed chicks (35°C, Hot Mild) than in the Control chicks that were raised under a commercial protocol. In the cold-exposed chicks (29oC, Cold), BW and pectoralis muscle absolute growth were similar to the Control group throughout the entire period. The lower body and muscle growth in the Hot and Hot Mild groups were reflected in a lower number of myonuclei expressing proliferating cell nuclear cell in pectoralis major muscle cross sections sampled on day 8, in the distribution of myofibers as the experiment progressed, and in mean myofiber diameter on day 35, whereas in the Cold group, these numbers exceeded that of the Control group. However, TUNEL assay revealed similar cell survival in all groups. Hematoxylin-eosin and Oil red O staining revealed the highest fat deposition in the pectoralis muscle derived from the Hot group, whereas lower fat deposition was observed in the Control Cold group. These results were corroborated by immunostaining for CCAAT/enhancer binding protein β in the pectoralis muscle, the levels of which were significantly higher in the Hot and Hot Mild groups on day 35 than in the Control group. Similar results were observed with Sirius red staining for collagen content in the pectoralis muscle. Together, the results imply long-term effects of chronic heat stress vs. cold stress in the early posthatch period on the broiler's body and muscle growth in general and myodegeneration of the pectoralis muscle in particular.

Effects of in ovo injection of L-histidine on hatch performance and post-hatch development in domestic pigeons (Columba livia)

The objective of this study was to evaluate the effects of in ovo injection of histidine on hatch performance and post-hatch development by determining hatchability, hatching time, BW gain, carcass traits, and intestinal morphology in domestic pigeons (Columba livia). A completely randomized design (n = 3) with a L-histidine injection treatment (His group, 0.55mg histidine dissolved in 200 μL 0.75% saline as 1% Conc compared to total histidine in the egg), a 0.75% saline injection treatment (SC group), and non-injection treatment (NC group) was used. Six squabs from each treatment were randomly sampled at day of hatch (DOH), day 7 (D7), and day 14 (D14) post hatch, respectively. Results showed that in ovo injection of histidine solution increased (P < 0.01) the hatchability in comparison with that of other groups. The hatching time of His group was earlier (P = 0.05) than that of the NC group. In ovo injection of histidine had no influence (P > 0.05) on BW gain and carcass traits, but had a significant effect on the organ index of pigeons. The index of pancreas on DOH, the indices of leg, gizzard, proventriculus, small intestine, and pancreas on D7, and the index of head on D14 were significantly increased (P < 0.05) by injection of histidine. Moreover, in ovo injection of histidine had significant effects on the weight indices of different intestinal segments and jejunal morphology in squabs. Ileum weight index on D7 from His group was significantly higher (P < 0.05) than that of other groups. Compared with the NC group, the jejunal crypt depth on DOH and D7 of His group decreased (P < 0.05), while jejunal villus area and villus crypt ratio on D7 of His group increased (P < 0.05). The results of this study indicate that in ovo injection of histidine may have beneficial effects on squabs' hatchability, intestinal development but have negligible effects on their growth performance during early post-hatch period.

Based serum metabolomics analysis reveals simultaneous interconnecting changes during chicken embryonic development.

Metabolic disorder is a major health problem and is associated with a number of metabolic diseases. Due to native hyperglycaemia and resistance to exogenous insulin, chickens as a model had used in the studies of adipose tissue biology, metabolism and obesity. But no detailed information is available about the comprehensive changes of serum metabolites at different stages of chicken embryonic development. This study employed LC/MS-QTOF to determine the changes of major functional metabolites at incubation day 14 (E14d), 19 (E19d) and hatching day 1 (H1d), and the associated pathways of differential metabolites during chicken embryonic development were analysed using Metabolite Set Enrichment Analysis method. Results showed that 39 metabolites were significantly changed from E14d to E19d and 68 metabolites were significantly altered from E19d to H1d in chicken embryos. Protein synthesis was promoted by increasing the concentrations of L-glutamine and threonine, and gonadal development was promoted through increasing oestrone content from E14d to E19d in chicken embryos, which indicated that serum glutamine, threonine and oestrone contents may be considered as the candidate indicators for assessment of early embryonic development. 2-oxoglutaric acid mainly contributed to enhancing the citric cycle, and it plays an important role in improving the growth of chicken embryos at the late development; the decreasing of L-glutamine, L-isoleucine and L-leucine contents from E19d to H1d in chicken embryonic development implied their possible functions as the feed additive during early posthatch period of broiler chickens to satisfy the growth. These results provided insights into understand the roles of serum metabolites at different developmental stages of chicken embryos, it also provides available information for chicken as a model to study metabolic disease or human obesity.

Comparison of two methods for determination of intestinal villus to crypt ratios and documentation of early age-associated ratio changes in broiler chickens , ,

The determination of intestinal villus to crypt ratios (VCR) is a common method utilized to evaluate effects of various diet regimens on gut microanatomy and for the histologic quantification of intestinal responses to disease processes. Two methods for the determination of small intestinal VCR were compared in early age chickens. A standard method for VCR determination based on 10 villus and crypt length measurements in the jejunal region of the small intestine was employed for the calculation of villus to crypt length ratio (VCLR). That method was compared to a new approach based on a single histomorphometric determination of the crypt and total mucosal areas using image analysis software. Subtraction of the crypt area from the total area provided the villus area and allowed for the subsequent calculation of villus to crypt area ratio (VCAR). At 4 and 18 h posthatch, VCLR was higher than that of VCAR, but there was no significant difference between VCLR and VCAR at 0 h (hatch) and at 168 h (d 7) posthatch. Nevertheless, the pattern of age-associated changes for VCLR and VCAR were comparable throughout the early posthatch period. Furthermore, the new method used in determining VCAR is subject to less human error, allows for an appreciable reduction in the number of measurements required, and facilitates a larger intestinal segment evaluation. Standard linear measurements require the selection of variable numbers of villi and crypts, whereas the area method only requires selection of a single region that incorporates numerous villi and crypts of variable sizes in providing a less subjective approach. This is particularly advantageous in studies on intestinal disease conditions resulting in marked multifocal variation in villus stature. This study further documented age-associated changes occurring in the VCR of the small intestine during the early posthatch period. Across the 2 methods used for VCR determination, a major and highly significant reduction in the VCR was observed to occur between 18 h and 168 h posthatch.

Standard metabolic rates of early life stages of the diamondback terrapin (Malaclemys terrapin), an estuarine turtle, suggest correlates between life history changes and the metabolic economy of hatchlings

I estimated standard metabolic rates (SMR) using measurements of oxygen consumption rates of embryos and unfed, resting hatchlings of the diamondback terrapin (Malaclemys terrapin) three times during embryonic development and twice during the early post-hatching period. The highest observed SMRs occurred during mid to late embryonic development and the early post-hatching period when hatchlings were still reliant on yolk reserves provided by the mother. Hatchlings that were reliant on yolk displayed per capita SMR 135 % higher than when measured 25 calendar days later after they became reliant on exogenous resources. The magnitude of the difference in hatchling SMR between yolk-reliant and exogenously feeding stages was much greater than that attributed to costs of digestion (specific dynamic action) observed in another emydid turtle, suggesting that processing of the yolk was not solely responsible for the observed difference. The pre-feeding period of yolk reliance of hatchlings corresponds with the period of dispersal from the nesting site, suggesting that elevated SMR during this period could facilitate dispersal activities. Thus, I hypothesize that the reduction in SMR after the development of feeding behaviors may reflect an energy optimization strategy in which a high metabolic expenditure in support of development and growth of the embryo and dispersal of the hatchling is followed by a substantial reduction in metabolic expenditure coincident with the individual becoming reliant on exogenous resources following yolk depletion.

High temperature and oxygen supplementation can mitigate the effects of hypoxia on developmental stability of bilateral traits during incubation of broiler breeder eggs

Hypoxia strongly affects embryonic development during the pre-hatch period. This study was conducted to investigate the effects of oxygen supplementation (O) and a 38.5°C high temperature (HT) at high altitude (HA, 1720 m) on morphological traits during a pre-hatch period and on relative fluctuating asymmetry (relative FA) and allometric growth during an early post-hatch period in broilers. A total of 720 eggs were obtained from a 45-week-old Ross 308 broiler breeder flock raised at sea level (2 m). The eggs were divided into six incubation condition (IC) groups and were incubated at HA. O groups were exposed to 23.5% O2 for 1 h daily from either days 0 to 11 (O0–11), days 12 to 21 (O12–21) or days 18 to 21 (O18–21) of incubation. HT groups were exposed to 38.5°C daily from either days 12 to 21 (HT12–21) or days 18 to 21 (HT18–21) of incubation. A control was maintained at 37.8°C and 21% O2. The hatched chicks were raised for 6 days at HA. Embryo/chick and beak lengths and head diameter were measured during pre- and post-hatch periods. The face, middle toe and shank lengths were measured for each chick. The relative asymmetry (RA), mean RA (MRA) and allometric growth of the lengths were computed and the existence of FA was demonstrated. The IC significantly affected the embryo length, with embryos of the O0–11 group shorter than embryos of the other O groups. Chicks were longer in the O and HT groups than those in the control, except for the O0–11. We found significant interactions between the IC and each development period for beak length. During the post-hatch period, the head diameter of the O0-11 was significantly smaller than that of the other groups, but not in O12–21. The interactions among IC, age and sex were significant for the RA of the face and middle toe lengths and for MRA. All the examined bilateral traits were evaluated as allometric growth. The FA for bilateral traits was determined in both sexes. The right (R) – left (L) and IR-Ll were the lowest in females for face length and in males for shank length from the O18–21 and in males for middle toe length from the O0–11 and HT18–21 groups. Therefore, the effects of factors such as HT and O2 could mitigate the adverse effects of HA-induced hypoxia on optimal developmental stability of bilateral traits of broiler.

Early posthatch thermal stress affects breast muscle development and satellite cell growth and characteristics in broilers

Heat or cold stress, can disrupt well-being and physiological responses in birds. This study aimed to elucidate the effects of continuous heat exposure in the first 2 wk of age on muscle development in broilers, with an emphasis on the pectoralis muscle satellite cell population. Chicks were reared for 13 d under either commercial conditions or a temperature regime that was 5°C higher. Body and muscle weights, as well as absolute muscle growth were lower in heat-exposed chicks from d 6 onward. The number of satellite cells derived from the experimental chicks was higher in the heat-treated group on d 3 but lower on d 8 and 13 compared to controls. This was reflected in a lower number of myonuclei expressing proliferating nuclear cell antigen in cross sections of pectoralis major muscle sampled on d 8. However, a TUNEL assay revealed similar cell survival in both groups. Mean myofiber diameter and distribution were lower in muscle sections sampled on d 8 and 13 in heat-treated versus control group, suggesting that the lower muscle growth is due to changes in muscle hypertrophy. Oil-Red O staining showed a higher number of satellite cells with lipids in the heat-treated compared to the control group on these days. Moreover, lipid deposition was observed in pectoralis muscle cross sections derived from the heat-treated chicks on d 13, whereas the controls barely exhibited any lipid staining. The gene and protein expression levels of CCAAT/enhancer binding protein β in pectoralis muscle from the heat-treated group were significantly higher on d 13 than in controls, while myogenin levels were similar. The results suggest high sensitivity of muscle progenitor cells in the early posthatch period at a time when they are highly active, to chronic heat exposure, leading to impaired myogenicity of the satellite cells and increased fat deposition.

Characterization of primary structure and post-hatching increase in chicken cytosolic acetoacetyl-coA thiolase in the liver

Acetoacetyl-CoA thiolase (EC 2.3.1.9) catalyzes the cleavage of acetoacetyl-CoA into acetyl-CoA and its reverse reaction, the synthesis of acetoacetyl-CoA. Cytosolic acetoacetyl-CoA thiolase (CT) is a key enzyme in the initial step of the cholesterol synthesis pathway. In the present study, we characterized the amino acid sequence of chicken CT and the tissue distribution of its mRNA and protein, together with their developmental changes in the liver. The amino acid sequence encoded by the nucleotide sequence of chicken CT cDNA showed a higher overall identity with those of human (74.3%) and rat (74.6%) CTs. Amino acid residues known to participate in enzymatic activity in human CT are conserved in chicken CT. Real-time PCR analysis revealed the expression of CT mRNA in the liver, kidney, adrenal gland, jejunum and ovary of adult hens, with higher levels in the liver, kidney, adrenal gland and ovary. Western blot analysis detected an immunoreactive protein of 41 kDa from cytoplasmic fraction but not particulate fractions of adult chicken liver. The immunoreactive protein was detected in all the tissues. The mRNA levels in the liver rapidly increased after hatching, with a maximum on d 5 post-hatching, after which they gradually decreased to adult levels. A similar change was observed in the protein levels. The increase in transcription and protein synthesis of CT suggests that the synthetic pathway of cholesterol from acetyl-CoA produced by CT replaces the hydrolysis of accumulated cholesteryl ester in the liver, in response to a change in the nutrient source from the lipid-rich yolk to a lower-lipid diet during the early post-hatching period.