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Carbon dioxide (CO 2 ) is considered to be an important factor during incubation of eggs. Effects attributed to higher CO 2 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 CO 2 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 CO 2 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 CO 2 concentration (both P = 0.001). Furthermore, no effect of CO 2 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 CO 2 . 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 CO 2 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., Competing Interests: Financial support statement This research received no specific grant from any funding agency, commercial or not-for-profit section., (Copyright © 2021 The Author(s). Published by Elsevier B.V. All rights reserved.)

Eggshell temperature (EST) during incubation greatly affects embryo development, chick quality at hatch, and subsequently various broiler physiological systems. Until now, a constant EST of 37.8°C seems optimal. Data on effects of EST patterns on immune organ development and subsequent broiler immune response are, however, scarce. A higher EST of 38.9°C in week 2 and/or a lower EST of 36.7°C in week 3 of incubation potentially positively affect embryo immune organ development and broiler immune response post hatch. Broiler eggs (n = 468) were incubated at 4 different EST patterns (n = 117 eggs/treatment) from week 2 of incubation onward. Week 1 (embryonic age (E)0 < E7) EST was 37.8°C for all eggs. Week 2 (E7 < E14) EST was either 37.8°C (Control) or 38.9°C (Higher), and week 3 (E14 - /hatch) EST was either Control or 36.7°C (Lower). At hatch, histology of bursal follicles and jejunum villi and crypts were determined as well as heterophil to lymphocyte ratio (H:L) (n = 49). Posthatch, both sexes were grown in 8 pens/treatment for 6 wk (n = 320). Natural antibodies (NAb) were determined at day 14, 22, and slaughter (day 41 or 42) as an indicator of immunocompetence and response to a Newcastle disease (NCD) vaccination was determined by antibody levels at day 22 and slaughter (n = 128). Results showed no interaction EST week 2 × EST week 3, except for jejunum histology. Higher EST in week 2 resulted in lower cell density within bursal follicles (P = 0.02) and a tendency for lower H:L (P = 0.07) at hatch, and higher NCD titers at slaughter (P = 0.02) than Control EST. Lower EST in week 3 resulted at hatch in higher cell density within bursal follicles, higher H:L (both P < 0.05), and a tendency for a higher posthatch mortality rate than control EST (P = 0.10). In conclusion, higher EST in week 2 during incubation may benefit embryonic immune organ development and posthatch broiler immunocompetence, while lower EST in week 3 showed opposite indications., (Copyright © 2020. Published by Elsevier Inc.)

This study was designed to determine effects of eggshell temperature (EST) pattern in week 2 and week 3 of incubation on tibia development of broiler chickens at slaughter age. A total of 468 Ross 308 eggs were incubated at an EST of 37.8°C from incubation day (E) 0 to E7. Thereafter, a 2 × 2 factorial arrangement with 2 EST (37.8°C and 38.9°C) from E8 to E14 and 2 EST (36.7°C and 37.8°C) from E15 till hatch was applied. After hatching, chickens were reared until slaughter age with the 4 EST treatments and 8 replicates per treatment. At day 41 and 42, one male chicken per replicate per day was selected, and hock burn and food pad dermatitis were scored. Rotated tibia, tibia dyschondroplasia, epiphyseal plate abnormalities, bacterial chondronecrosis with osteomyelitis, and epiphysiolysis were assessed. Tibia weight, length, thickness, head thickness, and robusticity index were determined. X-ray analyses (osseous volume, pore volume, total volume, volume fraction, mineral content, and mineral density) and a 3-point bending test (ultimate strength, yield strength, stiffness, energy to fracture, and elastic modulus) were performed. A high EST (38.9°C) in week 2 of incubation, followed by a normal EST (37.8°C) in week 3 resulted in higher mineral content (P = 0.001), mineral density (P = 0.002), ultimate strength (P = 0.04), yield strength (P = 0.03), and stiffness (P = 0.05) compared with the other 3 EST groups (week 2 × week 3 interaction). A high EST (38.9°C) in week 2 of incubation, regardless of the EST in week 3, resulted in a higher tibia weight (P < 0.001), thickness (P = 0.05), osseous volume (P < 0.001), and total volume (P < 0.001) than a normal EST (37.8°C). It can be concluded that 1.1°C higher EST than normal in week 2 of incubation appears to stimulate tibia morphological, biophysical, and mechanical characteristics of broiler chickens at slaughter age. Additionally, a 1.1°C lower EST in week 3 of incubation appears to have negative effects on tibia characteristics, particularly in interaction with the EST in week 2 of incubation., (Copyright © 2020. Published by Elsevier Inc.)

The objective of this study was to investigate the effect of eggshell temperature (EST) and carbon dioxide (CO2) concentration during only the hatching phase on embryonic development and chick quality. Three batches of eggs were incubated at an EST of 37.8°C until d of incubation (E) 19. From E19, embryos were incubated at low (36.7°C), normal (37.8°C), or high (38.9°C) EST and at low (0.2%) or high (1%) CO2 concentration. Organ growth and embryo and chick quality were measured at E19, internal pipping (IP), hatch, and 12 h after hatch. A few interactions between EST and CO2 were found at IP, hatch, and 12 h after hatch, but all of these interactions were temporary and in most cases weak. High EST resulted in a lower relative heart weight compared with low ( = 0.05) and normal EST ( = 0.06) at IP, compared with low ( = 0.11) and normal EST ( = 0.08) at hatch, and compared with low ( = 0.11) and normal EST ( = 0.08) at 12 h after hatch. At hatch, high EST resulted in a lower YFBM compared with low EST ( = 0.65). At 12 h after hatch, high EST resulted in a lower relative liver weight compared with low EST ( = 0.12). At low EST, greater relative intestinal weight was found compared with normal ( = 0.41) and high EST ( = 0.37). The effect of CO2 solely was found at 12 h after hatch at which a higher relative heart weight ( = 0.05) and a higher relative lung weight ( = 0.0542) was found at high CO2 compared with low CO2. High EST during only the hatching phase negatively affected chick development, mainly expressed by the lower relative heart weight at IP, hatch, and 12 h after hatch and lower YFBM at hatch. The resolving effect of CO2 demonstrates that CO2 only seem to have a temporary effect during the hatching phase.

The objective of this study was to investigate the effect of eggshell temperature (EST) and carbon dioxide concentration during only the hatching phase on physiological characteristics of embryos and chicks. Three groups of eggs were incubated at an EST of 37.8°C until d 19 of incubation (E19). From E19, embryos were incubated at a low (36.7°C), normal (37.8°C), or high (38.9°C) EST and at a low (0.2%) or high (1.0%) CO2 concentration. For E19, internal pipping (IP), hatch, and 12 h after hatch, blood parameters were analyzed and hepatic glycogen was determined. At IP, hatch, and 12 h after hatch, interactions were found between EST and CO2, but all these interactions were temporary and in most cases weak. High EST resulted in a lower hepatic glycogen concentration compared with low ( = 21.1) and normal EST ( = 14.43) at IP, and a lower hepatic glycogen concentration compared with low EST ( = 6.24) at hatch. At hatch, high EST resulted in lower hematocrit value ( = 2.4) and higher potassium ( = 0.5) compared with low EST. At 12 h after hatch, high EST resulted in a higher lactate concentration compared with low ( = 0.77) and normal EST ( = 0.65). And high EST resulted in higher potassium compared with low ( = 0.4) and normal EST ( = 0.3). An effect of CO2 solely was only found at IP, at which high CO2 resulted in a lower pH ( = 0.03) and a lower hepatic glycogen concentration ( = 7.27) compared with low CO2. High EST during only the hatching phase affected embryo and chick physiology, indicated by the lower hepatic glycogen levels at IP and hatch. High CO2 affected pH and hepatic glycogen at IP. Effects of CO2 were only found at low EST, which emphasizes the large effect of EST during the hatching phase.

Previous studies have shown that RH during incubation of chicken eggs influences water loss from the egg and embryonic mortality. In those studies, eggshell temperatures (EST) were not monitored or controlled. Because RH influences the egg's heat loss through evaporation, EST might have been different between RH treatments, influencing embryonic mortality and development. To eliminate the effect of EST, in the current study eggs were incubated at an EST of 37.8°C from embryonic d (E) 0 until E18 and at a high (55 to 60%) or low (30 to 35%) RH from E2 until hatch. Embryonic mortality, hatch curve, and several chick quality characteristics (length, weight, navel quality, organ weights, and DM of the yolk free body mass and yolk) were determined on E18 and at hatch. Low RH increased egg weight loss between E0 and E18 (+3.0%) and third week embryonic mortality (+3.0% of fertile eggs) and decreased hatch of fertile eggs (-2.9% of fertile eggs) compared with high RH. Hatch duration and chick quality characteristics did not differ between RH treatments. To assess the effect of RH during incubation on posthatch performance under suboptimal conditions, hatchlings were brooded at a normal (35.0°C at d 0, decreasing to 27.0°C at d 4) or cold (27.8°C at d 0, decreasing to 25.6°C at d 4) temperature until 4 d posthatch. Incubation RH and brooding temperature significantly interacted with posthatch growth but not development. Both low and high RH × cold brooding temperature resulted in lower (-6.9 and -6.0 g, respectively) BW than high RH × normal brooding temperature at 4 d of age. The cold brooding temperature resulted in lower daily feed intake (-1.3 g/chick) than the normal brooding temperature. In conclusion, incubating eggs at a low RH compared with a high RH and maintaining the EST at 37.8°C decreased hatch of fertile eggs, but had little effect on chick quality or posthatch performance.

This study evaluated the influence of incubation conditions on the developmental and physiological status of birds in the perinatal period, which spans the end of incubation until the early posthatch period. Embryos were incubated at a normal (37.8°C) or high (38.9°C) eggshell temperature (EST) and a low (17%), normal (21%), or high (25%) O(2) concentration from d 7 until 19 of incubation. After d 19 of incubation, EST was maintained, but O(2) concentrations were 21% for all embryos. Body and organ weights, and hepatic glycogen levels were measured at d 18 of incubation and at 12 and 48 h after emergence from the eggshell. In addition, blood metabolites were measured at 12 and 48 h after emergence from the eggshell. Embryos incubated at a high EST and low O(2) concentration had the highest mortality in the last week of incubation, which may be related to their low yolk-free body mass (YFBM) or a reduced nutrient availability for hatching (i.e., hepatic glycogen). High EST, compared with normal EST, decreased YFBM. This may be due to the shorter incubation duration of 8 h, the lower weight of supply organs (i.e., heart and lung), or a lack of glucose precursors. Because of this lack of glucose precursors, embryos incubated at high EST may have used proteins for energy production instead of for body development at the end of incubation. The YFBM at d 18 of incubation increased with an increase in O(2) concentration. However, differences between the normal and high O(2) concentration disappeared at 12 and 48 h after emergence, possibly because the high O(2) concentration had difficulties adapting to lower O(2) concentrations in the perinatal period. Blood metabolites and hepatic glycogen were comparable among O(2) concentrations, indicating that the physiological status at hatch may be related to the environment that the embryo experienced during the hatching process. In conclusion, EST and O(2) concentration differentially influence the developmental and physiological status of broilers during the perinatal period.

High eggshell temperatures (EST; ≥38.9°C) during the second half of incubation are known to decrease the body and organ development of broiler hatchlings. In particular, relative heart weights are decreased by a high EST, and this may increase the incidence of metabolic disorders that are associated with cardiovascular development, such as ascites. The current study investigated the effects of a high EST on chick quality, subsequent performance, and the incidence of ascites later in life. Eggs were incubated at a normal (37.8°C) or high (38.9°C) EST from d 7 of incubation onward. After hatching, the chickens were housed per EST in pens, and a normal or cold temperature schedule was applied during the grow-out period. Hatchability, hatchling quality, BW, feed conversion ratio, total mortality, mortality associated with ascites, slaughter characteristics, and ascites susceptibility at 6 wk of age were evaluated. Except for total ventricle weight, no interaction was found between EST and the grow-out temperature. Hatchability was comparable between the EST treatments, but the percentage of second-grade chickens was 0.7% higher at the high EST. Yolk-free body mass was 3.0 g lower, and heart weights were 26% lower at hatch in the high compared with the normal EST treatment. Body weight continued to be less during the grow-out period after the high EST incubation. However, breast meat yield was 1.0% higher in the high than in the normal EST. Feed conversion ratio did not differ between EST treatments. Total mortality was 4.1% higher and mortality associated with ascites was 3.8% higher in the high compared with the normal EST treatment. The ratio between the right and total ventricle was 1.1% higher in the high compared with the normal EST treatment at slaughter age. In conclusion, a high EST from d 7 of incubation onward decreased hatchling quality and growth performance, but increased breast meat yield. Furthermore, high EST incubation increased the incidence of ascites, which may be related to the reduced heart development at hatch.

To investigate the effect of incubation conditions on layer hatchlings, an experiment was performed in which layer eggs were incubated at a normal (37.8 degrees C) or high (38.9 degrees C) eggshell temperature (EST) and a hole was punctured in the air cell of half of the eggs in both EST treatments from d 14 of incubation onward. Chick development, plasma metabolites, and hepatic glycogen were measured at 12 h after emergence from the eggshell. Embryo mortality was not affected by the EST or hole treatment. At the high EST, yolk-free body mass was 0.7 g lower and residual yolk weight was 0.7 g higher than at the normal EST. This may be related to the shorter incubation duration at the high EST. Relative heart, lung, stomach, liver, spleen, and intestinal weights were lower in the high EST than in the normal EST group. Yolk-free body mass did not differ between eggs with or without a hole, but residual yolk weight was slightly lower in eggs with a hole (0.3 g). Relative lung weights were higher in eggs with than without a hole, whereas no effect on other organs was found. Plasma glucose, lactate, and uric acid concentrations did not differ between the EST or hole treatments. Hepatic glycogen was lower in the high EST (7.3 mg) than in the normal EST group (11.2 mg) at 12 h after emergence from the eggshell, and this effect may be related to the shorter hatching process at the high EST. Hepatic glycogen levels were lower in eggs with a hole (8.6 mg) compared with eggs without a hole (10.0 mg), and this may be related to the longer period between external pipping and hatching in eggs with a hole. In conclusion, the EST and hole treatment did not interact, and neither treatments affected embryonic survival. High EST negatively affected hatchling development and seemed to change the carbohydrate metabolism in layer embryos. The effect of a hole in the air cell was limited.