Interaction between eggshell temperature and carbon dioxide concentration after day 8 of incubation on broiler chicken embryo development

Carbon dioxide (CO₂) is considered to be an important factor during incubation of eggs. Effects attributed to higher CO₂ concentrations during experiment might be due to confounding effects of other environmental conditions, such as incubation temperature. To disentangle effects of eggshell temperature (EST) and CO₂ concentration, an experiment was conducted. A total of 630 Cobb 500 hatching eggs from 37 to 45 wk commercial breeder flocks were collected and incubated according to treatments. The experiment was setup as a complete randomized 2 × 3 factorial design, resulting in 6 treatments. From day 8 of incubation onward, broiler eggs were exposed to one of two EST (37.8 or 38.9 °C) and one of three CO₂ concentrations (0.1, 0.4 or 0.8%). Eggs were incubated in climate-respiration chambers and metabolic heat production was determined continuously. At day 18 of incubation and at 6 h after hatching, embryo and chicken quality were determined by evaluation of organ weights, navel condition, blood metabolites and hepatic glycogen. Hatching time and chicken length at 6 h after hatching showed an interaction between EST and CO₂ concentration (both P = 0.001). Furthermore, no effect of CO₂ concentration was found on embryo development or chicken quality. Metabolic heat production between day 8 and 18 of incubation was not affected by either EST or CO₂. At day 18 of incubation, an EST of 38.9 °C resulted in a higher egg weight loss, longer embryos, higher yolk free body mass (YFBM) and lower heart weight than an EST of 37.8 °C (all P < 0.008). At 6 h after hatching, an EST of 38.9 °C resulted in a higher residual yolk weight and lower YFBM, liver weight and heart weight than an EST of 37.8 °C (all P < 0.003). Lactate, uric acid and hepatic glycogen were not affected by EST at either day 18 of incubation or at hatch. Glucose was not affected by EST at day 18 of incubation, but at hatch, it was higher at an EST of 37.8 °C than at an EST of 38.9 °C (P = 0.02). It can be concluded that effects of CO₂ concentration (at concentrations ≤0.8%) on embryonic development and chicken quality appear to be limited when EST is maintained at a constant level. Moreover, a higher EST from day 8 of incubation onward appears to negatively affect chicken quality at hatch.     

Artificial incubation of trumpeter swan eggs: Selected factors affecting hatchability

Eggs collected from captive trumpeter swans (Cygnus buccinator) in 1993 (n = 33) and 1994 (n = 42) were artificially incubated with careful monitoring to identify factors contributing to the low hatch success reported by the Ontario Trumpeter Swan Restoration Program. Fertility was > 80% in both years, whereas hatch success of fertile eggs was 14.3% (n = 4) of 28 eggs in 1993 and 37.1% (n = 13) of 35 eggs in 1994. Necropsy of non‐viable eggs indicated a high incidence of embryonic mortality during early and late incubation. Early embryonic mortality was associated with egg storage times exceeding 7 days (P < 0.05) and bacterial contamination of eggs (P < 0.01). Late mortality was associated with (P < 0.001) increased weight loss during incubation period and may have resulted from incubator temperature and humidity fluctuations. We established patterns of weight loss for eggs and determined that hatched eggs lost 11–15% of initial mass and that weight loss >15% resulted in embryo mortality. Results from this study indicate that collection and handling of eggs before incubation and precise control of the incubator environment are critical to hatchability of eggs. Zoo Biol 18:403–414, 1999. © 1999 Wiley‐Liss, Inc.     

Embryonic heart rate and oxygen pulse in two procellariiform seabirds,Diomedea immutabilis and Puffinus pacificus

Developmental patterns of embryonic heart rate were measured non-invasively in two procellariiform seabirds, the Laysan albatross (Diomedea immutabilis) and wedge-tailed shearwater (Puffinus pacificus), during prepipping and after pipping. The O₂ pulse, defined as the O₂ consumption per single heart beat, was calculated using the previously reported O₂ consumption for these species. The embryonic heart rate of the albatross was not changed by internal pipping (initial pipping event in this species), remained unchanged during the prolonged internal pipping period and tended to increase, although insignificantly, with the initiation of external pipping (second pipping event). Heart rate in the shearwater remained unchanged during the late prepipping stages, did not change with external pipping (initial pipping event), but increased during the prolonged internal pipping period (second pipping event) and reached a maximum on the last day of incubation. The developmental pattern of heart rate in the shearwater was very similar to that reported previously for the brown noddy, a member of the order Charadriiformes but with the same pipping sequence. Developmental patterns of embryonic O₂ pulse were also different between the two procellariiform seabirds. However, those of the shearwater and the noddy were similar. The sequence of access to atmospheric O₂ during development and hatching may be an important factor determining the developmental patterns of embryonic heart rate and O₂ pulse.Abbreviations A-VO₂ difference, arteriovenous oxygen difference - BCG ballistocardiogram of egg - CV coefficient of variation - EP external pipping - HR heart rate - IP internal pipping - mass mass of freshly laid egg - SD standard deviation     

Effect of hatching time on time to first feed intake,organ development,enzymatic activity and growth in broiler chicks hatched on-farm

The conventional commercial hatcheries used today do not allow the newly hatched chicks to consume feed or water. Combined with natural variation in hatching time, this can lead to early hatched chicks being feed-deprived for up to 72 h before being unloaded at the rearing site. This study investigated the effects of hatching time on time to first feed intake and development of organs, digestive enzymes and productivity in terms of growth and feed conversion ratio in chicks hatched on-farm. Chicks were divided into three hatching groups (early, mid-term and late), and assessed over a full production cycle of 34 days. The results revealed that chicks remain inactive for a considerable amount of time before engaging in eating-related activities. Eating activity of 5% (i.e. when 5% of birds in each hatching group were eating or standing close to the feeder) was recorded at an average biological age (BA) of 25.4 h and a proportion of 50% birds with full crop was reached at an average BA of 30.6 h. Considering that the hatching window was 35 h in this study, the average chick probably did not benefit from access to feed and water immediately post-hatch in this case. At hatch, mid-term hatchlings had a heavier small intestine (30.1 g/kg bw) than both early (26.4 g/kg bw) and late (26.0 g/kg bw) hatchlings. Relative length of the small intestine was shorter in late hatchlings (735 cm/kg bw) than in mid-term (849 cm/kg bw) and early (831 cm/kg bw) hatchlings. However, the relative weight of the bursa fabricii was greater in mid-term (1.30 g/kg bw) than in early hatchlings (1.01 g/kg bw). At hatch, late hatchlings were heavier than early and mid-term hatchlings (P < 0.05), but by 3 days of age early hatchlings were heavier than mid-term and late hatchlings (P < 0.01). The only effect persisting throughout the study was a difference in the relative weight of the small intestine, where late hatchlings had heavier intestines than early hatchlings (P < 0.05). Thus, while there were differences between hatching groups, this study showed that the hatchlings seemed capable of compensating for these as they grew.     

Incubation and hatch management: consequences for bone mineralization in Cobb 500 meat chickens

From ~35 days of age fast growing meat chickens spend extended periods sitting or lying and less time standing. In a fast-feathering parent line lower early incubation temperatures which delayed chick hatch time, improved bone ash and extended their standing time. This incubation study assessed the consequences of incubation temperatures, hatch time and chick management at hatch/take off on femoral bone ash (BA) in Cobb 500 meat chickens. Embryos were incubated under either Control (between 37.8°C and 38.2°C egg shell temperature (EST)) or a Slow start (from 37.2°C at sett (the start of incubation), reaching 37.8°C EST at day 13 incubation), temperatures. Hatched chicks were identified at 492 h (20.5 days of incubation – classified as early (E)) or, between >492 and ⩽516 h (>20.5 and ⩽21.5 days of incubation – classified as late (L)), from setting. The E hatch chicks were allocated across three post-hatch treatments; treatment 1: E hatch chicks that were sampled E at 492 h from setting; treatment 2: E hatch chicks that were fed for a further 24 h in a floorpen before being sampled L at 516 h from setting; treatment 3: E hatch chicks that spent a further 24 h in the incubator before being sampled L at 516 h from setting. All L hatch chicks formed one treatment group which was sampled L at 516 h (i.e. L hatch chicks sampled L). It is not possible to sample L hatching chicks E hence this treatment is absent from the experimental design. Slow start incubation resulted in a higher total hatch percentage with a greater proportion of chicks hatching L, compared with the Control incubation. The L hatching chicks had significantly higher BA than the E hatching chicks. Of the E hatching chicks, those sampled both E and L had significantly lower BA than E hatching chicks fed for 24 h before L sampling. The E hatch, fed and sampled L chicks had the numerically highest BA, which was not significantly different from the BA of the L hatching chicks sampled L These results demonstrate that BA at hatch can be improved, either by extending the incubation period through a Slow start incubation profile, inducing L hatch, or alternatively, via the prompt provision of feed to E hatching chicks.     

The effect of timing of thermal conditioning during incubation on embryo physiological parameters and its relationship to thermotolerance in adult broiler chickens

Previously, we reported that thermal conditioning at 39°C on days 13–17 of incubation of broiler eggs enabled thermotolerance during post-hatch growth (J. Therm. Biol. 28 (2003) 133). Tolerance to a temperature of 30°C was accompanied by changes in thyroid hormones and metabolic parameters. In the current study, we determined the mechanism of epigenetic heat adaptation during embryonic age by measuring blood physiological parameters that may be associated with the ultimate effects of thermal conditioning. Hatching eggs from Ross breeders were subjected to heat treatment of 39°C at days 13, 14, 15, 16 and 17 of incubation for 2 h per day. Control eggs were incubated at 37.6°C. Samples of eggs were withdrawn on each day of thermal conditioning and at internal pipping (IP) to obtain blood samples from embryos. The remaining eggs were weighed at day 18 and transferred to hatchers. The timing of IP, external pipping (EP) and hatching were monitored every 2 h. At hatch, chicks were weighed and hatchability was determined. Blood samples were obtained from samples of day-old chicks. T3, T4, corticosterone, pCO₂, pO₂ levels were determined in the blood. Blood pH was measured and T3/T4 ratios were calculated. Heat conditioning significantly increased corticosterone and pO₂ levels and blood pH but depressed pCO₂ at day 14. These were followed by a significant depression of T4 level on day 15. Remarkably, at day 16, all these parameters were back to normal as in the control embryos. Hatching was delayed by thermal conditioning probably as a result of the depressed corticosterone levels at IP. Hatchability was also lower in the heat-treated group but 1-day old chick weights were comparable to those of the controls. The result suggests that epigenetic thermal conditioning involves changes in these physiological parameters and probably serve as a method for epigenetic temperature adaptation since the same mechanisms are employed for coping with heat during post-embryonic growth. It also suggests that days 14–15 may be the optimal and most sensitive timing for evoking this mechanism during embryonic development. The adverse effects of heat treatment observed in this study may have been due to the continued exposure to heat until day 17. Fine-tuning thermal conditioning to days 14–15 only may improve these production parameters.     

Embryonic Development and Rates of Metabolic Activity in Early and Late Hatching Eggs of the Major Malaria Vector Anopheles gambiae

Anopheles gambiae eggs generally hatch at the completion of embryo development; two-three days post oviposition. However, staggered or delayed hatching has been observed whereby a single batch of eggs shows marked variation in time-to-hatch, with some eggs hatching 18 days post oviposition or later. The mechanism enabling delayed hatch has not been clearly elucidated but is likely mediated by environmental and genetic factors that either induce diapause or slow embryo development. This study aimed to compare metabolic activity and embryonic development between eggs collected from sub-colonies of the baseline Anopheles gambiae GAH colony previously selected for early or late time-to-hatch. Egg batches from early and late hatch sub-colonies as well as from the baseline colony were monitored for hatching. For both time-to-hatch selected sub-colonies and the baseline colony the majority of eggs hatched on day two post oviposition. Nevertheless, eggs produced by the late hatch sub-colony showed a significantly longer mean time to hatch than those produced by the early hatch sub-colony. The overall proportions that hatched were similar for all egg batches. CO₂ output between eggs from early and late hatch sub-colonies showed significant differences only at 3 and 7 days post oviposition where eggs from the early hatch and the late hatch sub-colony were more metabolically active, respectively. No qualitative differences were observed in embryo development between the sub-colonies. It is concluded that all viable embryos develop to maturity at the same rate and that a small proportion then enter a state of diapause enabling them to hatch later. As it has previously been shown that it is possible to at least partially select for late hatch, this characteristic is likely to involve genetic as well as environmental factors. Delayed hatching in An. gambiae is likely an adaptation to maximise reproductive output despite the increased risk of desiccation in an unstable aquatic environment.     

Identification of QTL for live weight and growth rate using DNA markers on chromosome 3 in an F2 population of Japanese quail

The Japanese quail (Coturnix japonica) is an important agricultural species and is an animal model for genetic researches. This study was conducted to identify quantitative trait loci (QTL) affecting live weight and growth rate on chromosome 3 in quail. Two strains of Japanese quail including wild and white were crossed reciprocally and F₁ generation was created. The birds from F₂ generation were measured for growth traits and all of 472 birds (8 pairs from the parental strains, 34 F₁ birds and 422 F₂ birds) were genotyped for microsatellite markers on chromosome 3. The results indicated chromosome wide significant QTL for hatching weight (P < 0.01) and weight at 1, 2, 3 and 4 weeks of age, average daily gain from hatch to 1, 1–2 and 3–4 weeks of age and Kleiber ratio (P < 0.05), an indirect criterion of feed efficiency. The highest QTL additive and imprinting effects (2.72 and 0.79 % of the trait variation in the F₂ population, respectively) were related to hatching weight. The identified QTL for this trait (at 7 cM relative to the centromeric region of the chromosome) had significant interaction with sex and hatch (P < 0.01). The dominance effect of QTL was significant (P < 0.05) for bodyweight at one week of age accounting for 1.69 % of the trait variation in the F₂ population.     

Gas exchange and energy metabolism of the ostrich (Struthio camelus) embryo

We measured oxygen consumption ( ) and carbon dioxide emission ( ) rates, air-cell gas partial pressures of oxygen (P_AO₂) and CO₂ (P_ACO₂), eggshell water vapour conductance and energy content of the ostrich (Struthio camelus) egg, ‘true hatchling’ and residual yolk, and calculated RQ and total oxygen consumption ( ) for ostrich eggs incubated at 36.5°C and 25% relative humidity. The pattern showed a drop of approximately 5% before internal pipping. just prior to internal pipping agrees with allometric calculations. Despite the higher incubation temperature compared to other studies, and the resultant shorter incubation duration (42 days), (91.7 l kg⁻¹) was similar to a previously reported value. RQ values during the second half of incubation (approx. 0.68) were lower than expected for lipid catabolism. Prior to internal pipping, P_AO₂ and P_ACO₂ were 98 and 48.3 torr (13.1 and 6.4 kPa), respectively. The growth pattern of the ostrich embryo is different from the typical precocial pattern, showing a time delay in the rapid growth phase. As a result, the lowered overall energy expenditure for tissue maintenance, as compared to other species, is reflected in the low yolk utilization and high residual yolk fraction of the whole hatchling dry mass. These could also result from the relatively short incubation period of the ostrich egg, thereby evading desiccation by excess water loss.     

Effects of in ovo feeding of creatine pyruvate on the hatchability,growth performance and energy status in embryos and broiler chickens

The effects of in ovo feeding (IOF) of creatine pyruvate (CrPyr) on the hatchability, growth performance and energy status of embryos and broilers (Arbor Acres) were investigated. Five treatments were arranged as non-injected treatment (Control), 0.6 ml physiological saline (0.75%) injected treatment (Saline), and IOF treatments injected with 0.6 ml physiological saline (0.75%) containing 3, 6 or 12 mg CrPyr (CrPyr₃, CrPyr₆ or CrPyr₁₂) into the amnion per fertile egg on day 17.5 of incubation. After hatching, 80 male chicks from each treatment with similar weight close to the average BW of their pooled group were selected and randomly assigned into eight replicates of 10 chicks each. The results showed that the hatchability was not affected among groups, whereas the hatching weight of broilers in CrPyr₁₂ was significantly higher than the control and saline groups (P<0.05). At 21 day post-hatch, the BWs of broilers in CrPyr₆ and CrPyr₁₂ were increased relative to the control and saline groups (P<0.05). Chickens in CrPyr₆ and CrPyr₁₂ exhibited higher BW gain and feed intake than the control and saline groups during 8 to 21 days post-hatch and the entire experiment period (P<0.05). Compared with the control and saline groups, the total and relative weight of pectoral muscle of embryos or chickens were greater in CrPyr₆ and CrPyr₁₂ at 19^th day of incubation (19 E), hatch, 3 and 21 days post-hatch (P<0.05). The concentrations of glucose and glycogen in liver were increased in CrPyr₆ and CrPyr₁₂ at 19 E and hatch (P<0.05). Neither glycogen nor glucose concentration in pectoral muscle was altered among treatments (P>0.05). Irrespective of dosage, the concentrations of creatine and phosphocreatine, and activities of creatine kinase in embryos were enhanced in CrPyr treatments at 19 E when compared with the control and saline groups (P<0.05). The activities of glucose-6-phosphatase in liver in CrPyr₆ and CrPyr₁₂ treatments were higher than the control and saline groups at 19 E (P<0.05). In conclusion, these results indicated that IOF of CrPyr, especially at the level of 12 mg/egg, could improve energy status of embryos and hatchlings, which was useful for enhancing hatching weight, BW and pectoral muscle weight until the end of the experiments at 21 days post-hatch in broilers.     

Physiological status of broiler chicks at pulling time and the relationship to duration of holding period

Newly hatched chicks may be held longer than 48 h and experience long periods of fasting in commercial hatcheries. Limited information is known about the physiological status of chicks in such situations, due to the difficulty of precisely recording time of hatch. This study investigated the effect of the time from hatch to pulling (holding period) on physiological measures/parameters in 109 broiler chicks. Fertile Ross 308 eggs were incubated in a custom built small-scale incubator. The individual hatching time of each focal chick was determined using eggshell temperature monitoring. At ‘pulling’ (512 h of incubation time), the quality of focal chicks was assessed using the chick scoring method and physiological parameters were measured including BW, organ (heart, liver and stomach) weights, blood values and plasma corticosterone level. The time from hatch to pulling varied from 7.58 to 44.97 h. Egg weight at setting was significantly correlated with chick BW and weight of organs at pulling, but had no effect on chick quality, blood values and plasma corticosterone. Relative BW at pulling was negatively associated with the duration of holding period (P=0.002). However, there was a positive correlation between relative stomach weight and the duration of the holding period (P<0.001). As the holding period duration increased, there was a trend that blood partial pressure of oxygen, haematocrit and haemoglobin also increased, and blood partial pressure of carbon dioxide, total carbon dioxide and bicarbonate decreased (P<0.05). A wide range of plasma corticosterone was observed from chicks that had experienced different durations of holding period. We conclude that shortening the hatch window and minimising the number of chicks that experience a long holding period before pulling may improve chick quality and physiological status, which may be due to unfavourable environmental conditions that include feed and water deprivation.     

Effects of in ovo feeding of l-arginine on breast muscle growth and protein deposition in post-hatch broilers

In ovo feeding (IOF) of l-arginine (Arg) can affect growth performance of broilers, but the response of IOF of Arg on breast muscle growth is unclear, and the mechanism involved in protein deposition remains unknown. Hense, this experiment was conducted to evaluate the effects of IOF of Arg on breast muscle growth and protein-deposited signalling in post-hatch broilers. A total of 720 fertile eggs were collected from 34-week-old Arbor Acres breeder hens and distributed to three treatments: (1) non-injected control group; (2) 7.5 g/l (w/v) NaCl diluent-injected control group; (3) 0.6 mg Arg/egg solution-injected group. At 17.5 days of incubation, fertile eggs were injected 0.6 ml solutions into the amnion of the injected groups. Upon hatching, 80 male chicks were randomly assigned to eight replicates of 10 birds each and fed ad libitum for 21 days. The results indicated that IOF of Arg increased relative breast muscle weight compared with those of control groups at hatch, 3-, 7- and 21-day post-hatch (P<0.05). In the Arg-injected group, the plasma total protein and albumen concentrations were higher at 7- and 21-day post-hatch than those of control groups (P<0.05). The alanine aminotransferase activity in Arg group was higher at hatch than that of control groups (P<0.05). The levels of triiodothyronine at four time points and thyroxine hormones at hatch, 7- and 21-day post-hatch in Arg group were higher than those of control groups (P<0.05). In addition, IOF of Arg increased the amino acid concentrations of breast muscle at hatch, 7- and 21-day post-hatch (P<0.05). In ovo feeding of Arg also enhanced mammalian target of rapamycin, ribosomal protein S6 kinase-1 and eIF4E-bindingprotein-1 messenger RNA expression levels at hatch compared with those of control groups (P<0.05). It was concluded that IOF of Arg treatment improved breast muscle growth, which might be associated with the enhancement of protein deposition.     

Seasonal variations in larval biomass and biochemical composition of brown shrimp, Crangon crangon (Decapoda, Caridea), at hatching

The “brown shrimp”, Crangon crangon (Linnaeus 1758), is a benthic key species in the North Sea ecosystem, supporting an intense commercial fishery. Its reproductive pattern is characterized by a continuous spawning season from mid-winter to early autumn. During this extended period, C. crangon shows significant seasonal variations in egg size and embryonic biomass, which may influence larval quality at hatching. In the present study, we quantified seasonal changes in dry weight (W) and chemical composition (CHN, protein and lipid) of newly hatched larvae of C. crangon. Our data revealed significant variations, with maximum biomass values at the beginning of the hatching season (February–March), a decrease throughout spring (April–May) and a minimum in summer (June–September). While all absolute values of biomass and biochemical constituents per larva showed highly significant differences between months (P < 0.001), CHN, protein and lipid concentrations (expressed as percentage values of dry weight) showed only marginally significant differences (P < 0.05). According to generalized additive models (GAM), key variables of embryonic development exerted significant effects on larval condition at hatching: The larval carbon content (C) was positively correlated with embryonic carbon content shortly after egg-laying (r ² = 0.60; P < 0.001) and negatively with the average incubation temperature during the period of embryonic development (r ² = 0.35; P < 0.001). Additionally, water temperature (r ² = 0.57; P < 0.001) and food availability (phytoplankton C; r ² = 0.39; P < 0.001) at the time of hatching were negatively correlated with larval C content at hatching. In conclusion, “winter larvae” hatching from larger “winter eggs” showed higher initial values of biomass compared to “summer larvae” originating from smaller “summer eggs”. This indicates carry-over effects persisting from the embryonic to the larval phase. Since “winter larvae” are more likely exposed to poor nutritional conditions, intraspecific variability in larval biomass at hatching is interpreted as part of an adaptive reproductive strategy compensating for strong seasonality in plankton production and transitory periods of larval food limitation.     

The in ovo administration of l-trans pyrrolidine-2,4-dicarboxylic acid regulates small intestinal growth in chicks

Glutamate, which is one of the most important contributors to oxidative metabolism in the intestinal mucosa, is mainly transported by the excitatory amino acids transporters (EAATs) that are expressed in enterocytes. The objective of this study was to evaluate the effects of in ovo administration of l-trans pyrrolidine-2,4-dicarboxylic acid (l-trans-PDC), a potent competitive inhibitor of glutamate uptake by EAATs, on the growth of the small intestine in chicks. Two series of experiments were conducted with hatching eggs; 100 μl of various l-trans-PDC solutions (0, 0.075 or 0.225 mg/egg for the Control group, low-dose l-trans pyrrolidine 2,4-dicarboxylic acid group (L-PDC) or high-dose l-trans pyrrolidine 2,4-dicarboxylic acid group (H-PDC), respectively) was injected into the albumen sac of these hatching eggs before incubation. Hatchlings were sacrificed by cervical dislocation to determine the embryonic development in Experiment I, whereas the birds in Experiment II were raised or sampled at hatching, days 7 and 14 (D7 and D14) for further study. Gene expression in the small intestines was determined by real-time RT-PCR; and serum concentration of free amino acids was determined by an amino acid analyzer. The results showed that the hatchability was decreased by in ovo administration of l-trans-PDC. The small intestinal weights of the H-PDC group were decreased (P<0.05) at hatching and increased (P<0.05) on D7 and D14 compared with those in the Control group. In addition, the gene expression of EAAT2 in the completed or segmental small intestines was not changed (P>0.05); EAAT3 gene expression in the duodenum (P<0.05), jejunum (P=0.084) and ileum (P=0.060) on D14 was lower in the H-PDC group than in the Control group. Furthermore, the serum concentrations of free proline, threonine and phenylalanine but not glutamate or aspartate were increased (P<0.06) in H-PDC group. In conclusion, this paper is the first to report that in ovo administration of l-trans-PDC induces small intestinal growth retardation during the embryonic period and catch-up growth after hatching.     

Effects of ocean acidification increase embryonic sensitivity to thermal extremes in Atlantic cod,Gadus morhua

Thermal tolerance windows serve as a powerful tool for estimating the vulnerability of marine species and their life stages to increasing temperature means and extremes. However, it remains uncertain to which extent additional drivers, such as ocean acidification, modify organismal responses to temperature. This study investigated the effects of CO₂‐driven ocean acidification on embryonic thermal sensitivity and performance in Atlantic cod, Gadus morhua, from the Kattegat. Fertilized eggs were exposed to factorial combinations of two PCO₂ conditions (400 μatm vs. 1100 μatm) and five temperature treatments (0, 3, 6, 9 and 12 °C), which allow identifying both lower and upper thermal tolerance thresholds. We quantified hatching success, oxygen consumption (MO₂) and mitochondrial functioning of embryos as well as larval morphometrics at hatch and the abundance of acid–base‐relevant ionocytes on the yolk sac epithelium of newly hatched larvae. Hatching success was high under ambient spawning conditions (3–6 °C), but decreased towards both cold and warm temperature extremes. Elevated PCO₂ caused a significant decrease in hatching success, particularly at cold (3 and 0 °C) and warm (12 °C) temperatures. Warming imposed limitations to MO₂ and mitochondrial capacities. Elevated PCO₂ stimulated MO₂ at cold and intermediate temperatures, but exacerbated warming‐induced constraints on MO₂, indicating a synergistic interaction with temperature. Mitochondrial functioning was not affected by PCO₂. Increased MO₂ in response to elevated PCO₂ was paralleled by reduced larval size at hatch. Finally, ionocyte abundance decreased with increasing temperature, but did not differ between PCO₂ treatments. Our results demonstrate increased thermal sensitivity of cod embryos under future PCO₂ conditions and suggest that acclimation to elevated PCO₂ requires reallocation of limited resources at the expense of embryonic growth. We conclude that ocean acidification constrains the thermal performance window of embryos, which has important implication for the susceptibility of cod to projected climate change.     

Supplementation of 17β-estradiol and progesterone in the co-culture medium of bovine oviductal epithelial cells and ovine spermatozoa reduces the sperm kinematics and capacitation

This study investigated the effect that bovine oviductal epithelial cell (BOEC) and ovine spermatozoa co-culture exposed to different hormonal environments had on ram sperm function over the course of a 24-h incubation period. Ram cooled-stored spermatozoa were selected by swim-up and then co-cultured separately for 24?h at 38.5?°C under 5% CO₂ with either: (1) Fert-TALP medium (positive control [POSControl]), (2) Fert-TALP medium supplemented with 17β-estradiol (E2) and progesterone (P4) at concentrations similar to follicular phase (Follicular NEGControl), (3) Fert-TALP medium supplemented with E2 and P4 concentrations similar to luteal phase (Luteal NEGControl), (4) BOEC cultured in the same medium as that of the Follicular NEGControl group (Follicular BOEC group), or (5) BOEC cultured in the same medium as that of the Luteal NEGControl group (Luteal BOEC group). The sperm kinematics, capacitation status, and plasma membrane (PM) integrity were evaluated at different intervals. Sperm PM integrity was not affected (P ? 0.05) by BOEC co-culture, regardless of the phase of the estrous cycle. After 4?h of incubation, the Luteal BOEC group presented lower (P?<? 0.05) progressive motility and total motility than the Luteal NEGControl group while the Follicular BOEC group showed lower (P?<? 0.05) velocimetric parameters and progressive motility than the Follicular NEGControl group. Throughout the incubation period, both BOEC co-culture groups showed a decrease (P?<? 0.05) in their capacitation rate in comparison to the POSControl group. Conversely, the Luteal BOEC group presented a higher (P?<? 0.05) non-capacitated rate than both the POSControl and Luteal NEGControl groups. In conclusion, BOEC co-culture with ovine spermatozoa at either the follicular or luteal phase decreases sperm kinematics and delays sperm capacitation.     

Differences in egg deposition of corticosterone and embryonic expression of corticosterone metabolic enzymes between slow and fast growing broiler chickens

Glucocorticoids (GCs) are vital for embryonic development and their bioactivity is regulated by the intracellular metabolism involving 11β-hydroxysteroid dehydrogenases (11β-HSDs) and 20-hydroxysteroid dehydrogenase (20-HSD). Here we sought to reveal the differences in egg deposition of corticosterone and embryonic expression of corticosterone metabolic enzymes between slow and fast growing broiler chickens (Gallus gallus). Eggs of fast-growing breed contained significantly higher (P < 0.05) corticosterone in the yolk and albumen, compared with that of a slow-growing breed. 11β-HSD1 and 11β-HSD2 were expressed in relatively higher abundance in the liver, kidney and intestine, following similar tissue-specific ontogenic patterns. In the liver, expression of both 11β-HSD1 and 11β-HSD2 was upregulated (P < 0.05) towards hatching, yet 20-HSD displayed distinct pattern showing a significant decrease (P < 0.05) on posthatch day 1 (D1). Hepatic mRNA expression of 11β-HSD1 and 11β-HSD2 was significantly higher in fast-growing chicken embryos at all the embryonic stages investigated and so was the hepatic protein content on embryonic day of 14 (E14) for 11β-HSD1 and on E14 and D1 for 11β-HSD2. 20-HSD mRNA was higher in fast-growing chicken embryos only on E14. Our data provide the first evidence that egg deposition of corticosterone, as well as the hepatic expression of glucocorticoid metabolic enzymes, differs between fast-growing and slow-growing chickens, which may account, to some extent, for the breed disparities in embryonic development.     

Embryonic developmental plasticity of the chick: increased CO(2) during early stages of incubation changes the developmental trajectories during prenatal and postnatal growth

This study investigated the effect of non-ventilation of the incubator during the first 10 days of incubation on carbon dioxide (CO(2)) concentrations in the incubator and its effects on the embryonic and post-hatch development of the chicken (Gallus gallus). Two different incubation conditions were created, one incubator was kept at standard conditions, with adequate ventilation (V) and a second incubator was non-ventilated (NV) during the first ten days of incubation, allowing the CO(2) to rise. After the first 10 days, both incubations were continued under standard conditions. The experiment was repeated twice with different ages of the breeders (45 and 60 wks) which resulted in different CO(2) levels at ED10 (1.5 and 1%). The CO(2) concentration in the V incubators remained below 0.1% in these first 10 days. The eggs of the NV incubation showed higher pCO(2) levels in the air cell from ED10 until ED14 compared to the eggs of the V group. The NV embryos had significantly higher absolute and relative (to egg weight) body weights from ED10 until ED18, pointing to an accelerated embryonic growth. At internal pipping, the NV chick embryos had higher plasma corticosterone and T(3) levels and higher pCO(2) in the air cell. Chicks incubated under NV conditions hatched 10 h earlier in the first and 15 h earlier in the second experiment and the spread of hatch was narrower. During the post-hatch period, the NV chickens had a higher body weight compared to the V chickens. From these results, it is clear that higher levels of CO(2) during the first ten days of incubation have persistent (epigenetic) effects during the incubation and early post-hatch period.     

Effects of rumen-protected betaine supplementation on meat quality and the composition of fatty and amino acids in growing lambs

Rumen-protected betaine (RPB) can enhance betaine absorption in the small intestine of ruminants, while betaine can alter fat distribution and has the potential to affect the meat quality of livestock. Hence, we hypothesized that RPB might also affect the meat quality of lambs. Sixty male Hu sheep of similar weight (30.47 ± 2.04 kg) were selected and randomly subjected to five different treatments. The sheep were fed a control diet (control treatment, CTL); 1.1 g/day unprotected-betaine supplemented diet (UPB); or doses of 1.1 g/day (low RPB treatment; L-PB), 2.2 g/day (middle RPB treatment; M-PB) or 3.3 g/day (high RPB treatment; H-PB) RPB-supplemented diet for 70 days. Slaughter performance, meat quality, fatty acid and amino acid content in the longissimus dorsi (LD) muscle, shoulder muscle (SM) and gluteus muscle (GM) were measured. Compared with CTL, betaine (including UPB and RPB) supplementation increased the average daily weight gain (ADG) (P < 0.05) and average daily feed intake (P < 0.01) of lambs. Rumen-protected betaine increased ADG (P < 0.05) compared with UPB. With increasing RPB doses, the eye muscle area of the lambs linearly increased (P < 0.05). Compared with CTL, betaine supplementation decreased water loss (P < 0.05) in SM and increased pH₂₄ in the SM (P < 0.05) and GM (P < 0.05). Compared with UPB, RPB decreased water loss in the GM (P < 0.01), decreased shear force (P < 0.05) in the LD and SM and increased the pH of the meat 24 h after slaughter (pH₂₄). With increasing RPB doses, the shear force and b* value in the LD linearly decreased (P < 0.05), and the pH₂₄ of the meat quadratically increased (P < 0.05). Compared with CTL, betaine supplementation increased the polyunsaturated fatty acid in the GM (P < 0.05). Compared with UPB, RPB supplementation decreased the saturated fatty acid (SFA) content in the LD (P < 0.05) and increased the unsaturated fatty acids (UFA), mono-unsaturated fatty acids and UFA/SFA ratio in the LD (P < 0.05). Compared with CTL, the content of histidine in the LD increased with betaine supplementation. Compared with UPB, RPB supplementation increased the content of total free amino acids and flavor amino acids in the LD of lambs (P < 0.05). With increasing RPB, the isoleucine and phenylalanine contents in the LD linearly increased (P < 0.05). Overall, the data collected indicated that the meat quality of lambs (especially in the LD) improved as a result of betaine supplementation, and RPB showed better effects than those of UPB.