Effect of four eggshell repair materials on weight loss during incubation of white leghorn chicken eggs.

Egg weight loss during incubation is a key indicator used to monitor successful egg development and is closely related to hatchability and chick survival. Artificial incubation is one of the most important captive breeding techniques used in conservation efforts to bolster avian populations. To repair damage to the eggshell and ensure embryonic viability during incubation, a variety of repair coverings can be applied. This study tested the impact of four repair materials (nail polish, synthetic glue, medical dressing, and molten wax film) on egg weight loss during incubation. We found no impact on weight loss for coverings smaller than 35% of the eggshell surface, nor did we find any differences between covering types. The average egg weight loss decreased as the coverage area increased, and the weight loss did not differ when blunt versus sharp-end coverings were compared. Given the relative insensitivity of egg weight loss and survival to the type of patch material used, we concluded that the selection of material for the purpose of weight loss management could be based on practical considerations, such as ease of application and availability.

Winter breeding of black swan Cygnus atratus in a solar greenhouse with a water pond.

Winter propagation of black swans Cygnus atratus is introduced at Kwangpho on the east coast of the Democratic People's Republic of Korea located in the temperate zone of northeast Asia. The natural lake Kwangpho was selected as a rearing site to provide the environment which matched most closely to black swan's natural habitats. Comparison of the climate data of Kwangpho with Australia and New Zealand led to the conclusion that it is necessary to increase winter temperature 10 °C for artificial rearing. In order to overcome the low temperature with renewable energy, a passive solar greenhouse with a water pond was introduced, where black swans were reared simultaneously while observing the microclimate such as air temperature, humidity and water temperature in winter. When the pond depth was 0.4 m and 0.8 m, there was no significant difference in indoor temperature, the depth fixed at 0.4 m to reduce pumping power by more than 50%. During the study, 80 mature black swans wintered safely and 13 pairs of them laid 74 eggs, the artificial hatching rate was 65% and the natural hatching rate was 53%. It was confirmed that the passive solar greenhouse with a pond can raise the winter minimum temperature 8-10 °C higher than outside, and in this microclimate it is possible to propagate the black swan and it is believed that the greenhouse has abundant potential to enable the artificial breeding as well as wintering of black swans, in temperate zone occupying wide area of the earth.

Breeding history and husbandry of the Raggiana Bird-of-paradise (Paradisaea raggiana).

The Raggiana Bird-of-paradise (Paradisaea raggiana) has been held by the San Diego Zoo for more than 50 years, but the population remained in low numbers until the mid-1990s. A previous article published in 1997 documented the beginning of the rise in the population; however, no new information has been published since that time. Over the past 25 years, behavioral observations by animal staff have led to improvements in husbandry techniques, which includes artificial incubation of eggs and hand-rearing of chicks. The ability to simulate a lek with various housing arrangements, in which the males can display with each other, be given access to a female for copulation, and then be separated again has proven paramount for breeding success. Additionally, females are allowed mate choice, further mimicking natural behaviors observed by this species. The refinement of these methods has led to a greater number of fertile eggs as well as higher hatchability and survivability, which in turn has resulted in a significant increase in the captive population.

A new candling procedure for thick and opaque eggs and its application to avian conservation management.

The ability to monitor developing avian embryos and their associated vascular system via candling enables the application of important reproductive management techniques. Egg candling facilitates the confirmation of egg viability throughout the incubation process and identification of a precise position on a vein for the safe extraction of blood. Blood samples may then be analysed to retrieve vital health and genetic information to assist in conservation management. However, the thick or opaque egg shell characteristics of some avian species prevents the observation of egg contents using traditional candling methods, thus limiting management options. This paper tests a novel method of preparing thick-shelled or opaque eggs so that traditional egg candling and blood extraction methods may be applied. Eggs from captive emu (Dromaius novaehollandiae, Latham 1790) and southern cassowary (Casuarius casuarius johnsonii, Linnaeus 1758) were obtained, and partial fenestration was performed on two areas of shell either before incubation or at ⅓ of incubation. Hatchability and weight loss were examined as a measure of effect of the fenestration process on the developing embryo. Clear observation of vascular development was successful in 97% of viable fenestrated eggs, without affecting hatchability or weight loss. Blood samples were taken from developing embryos and DNA was successfully extracted for proof of concept of this new technique. The ability to observe vascular development and monitor the developing embryo in thick and opaque eggs will significantly improve both in situ and ex situ population management options such as in ovo sexing in species of concern.

Very Early Corona Treatment-Mediated Artificial Incubation of Silkworm Eggs and Germline Transformation of Diapause Silkworm Strains.

Diapause is an important biological characteristic for many insect species to adapt to adverse environmental conditions and maintain the continuity of the race. Compared with the traditional hydrochloric acid or/and cold storage treatment methods, the artificial corona incubation technology of silkworm (Bombyx mori) eggs has many advantages including, the absence of pollution, easy operation and safety. However, this technology has not yet been applied in sericulture. In this study, we developed a novel artificial corona instrument to successfully disrupt the diapause of newly laid and refrigerated eggs from various Chinese and Japanese lineage silkworm strains. Subsequently, we invented a very early corona treatment (VECT) strategy to prevent the diapause of newly laid silkworm eggs within 4 h of oviposition. The hatching rates of the larvae were more than 95% in all diapause silkworm strains, which was comparable to the effect of the traditional HCl treatment strategy. In addition, we developed a combination strategy of VECT and pre-blastoderm microinjection and successfully created transgenic silkworms in various diapause strains. The results of the current study can aid in improving the corona artificial incubation technology and promote its application in sericulture.

Monitoring of Unhatched Eggs in Hermann's Tortoise (Testudo hermanni) after Artificial Incubation and Possible Improvements in Hatching.

The causes of embryonic mortality in Hermann's tortoises (Testudo hermanni) during artificial incubation were determined. Total egg failure at the end of the hatching period was investigated. The hatching artefacts represented 19.2% (N = 3557) of all eggs (N = 18,520). The viability rate of incubated eggs was 80.8%. The eggs, i.e., embryos, were sorted according to the cause of unsuccessful hatching and subsequently analyzed. Some of the eggs were divided into two or more groups. Unfertilized eggs were confirmed in 61.0%, infected eggs in 52.5%, and eggs in various stages of desiccation in 19.1%. This group also included mummified embryos. Pseudomonas aeruginosa, Bacillus sp., Purpureocillium lilacinum, and Escherichia coli were frequently confirmed in infected eggs. Embryos were divided into three groups: embryos up to 1.0 cm-group 1 (2.2%), embryos from 1.0 cm to 1.5 cm-group 2 (5.4%) and embryos longer than 1.5 cm-group 3 (7.3%) of all unhatched eggs. Inability of embryos to peck the shell was found in 1.3%. These tortoises died shortly before hatching. Embryos still alive from the group 2 and group 3 were confirmed in 0.7% of cases. Dead and alive deformed embryos and twins were detected in the group 3 in 0.5% and 0.1% of cases, respectively. For successful artificial hatching, it is important to establish fumigation with disinfectants prior to incubation and elimination of eggs with different shapes, eggs with broken shells, and eggs weighted under 10 g. Eggs should be candled before and periodically during artificial incubation, and all unfertilized and dead embryos must be removed. Heartbeat monitor is recommended. Proper temperature and humidity, incubation of "clean" eggs on sterile substrate and control for the presence of mites is essential. Monitoring of the parent tortoises is also necessary.

The exploration of artificial incubation of Cherax quadricarinatus eggs.

The redclaw crayfish, Cherax quadricarinatus, is an economically valuable freshwater crayfish. However, some production obstacles, such as a low egg hatching rate and asynchronous hatching, are hindering its development in the aquaculture industry. Artificial incubation of eggs may solve these problems. This study explored the technology of artificial incubation of redclaw crayfish eggs. The following results were obtained: 1) 75% alcohol as a disinfectant for 60 s had a preferable antibacterial effect and promoted the hatching rate; 2) densities of 300 and 600 eggs/incubator resulted in significantly higher hatching and survival rates than a density of 900 eggs/incubator; 3) at a density of 600 eggs/incubator, the optimum number of net pieces for attachment of freshly hatched juveniles was 20 per incubator; 4) with a density of 600 eggs/incubator and 20 net pieces/incubator, the hatching rate was 82.05% ± 4.09%, the survival rate was 55.12% ± 7.51%, and a total of 129,200 SPF (specific pathogen-free) seedlings were cultivated. This artificial incubation system was maintained at close to an aseptic state, with an absence of white spot virus, iridovirus, Vibrio and ciliates; this was true for all water sources used and for incubation of both eggs and juvenile crayfish. In conclusion, if we implement reasonable methods of disinfection, SPF detection and pathogen isolation and utilize optimal egg densities and incubation systems, large-scale production of SPF seedlings of Cherax quadricarinatus is possible.

Sound exposure and its beneficial effects on embryonic growth and hatching of broiler chicks.

1. The physical factors that involve artificial incubation determine embryonic growth and hatching of broiler chicks, and have important implications for the poultry production chain and chick welfare. This study explored the effects of sound exposure on incubation responses (hatching, embryonic development and chick quality).2. A 2 × 2 factorial experiment with the factors 'sound pressure level' (at 70 and 90 dB) and 'species-specific vocalisations' (at 75dB - 85dB) was performed in four experimental incubators (loaded with 90 eggs each) for eight consecutive cycles of incubation, totalling 2880 Cobb-500® broiler breeder eggs.3. Embryonic growth was not influenced by any experimental factor, but the highest sound pressure level (90 dB) led to earlier hatching, higher hatchability, better chick quality and lower weight at hatching. Additional effects of species-specific vocalisations were found at 70 dB noise, which caused early hatching and better chick quality.4. It was concluded that the loud noise found in commercial hatcheries (at 90 dB) can be beneficial. Furthermore, the presence of species-specific vocalisations could improve the hatching time and chick quality from experimental incubators (at 70 dB). However, this would not provide additional benefits in commercial hatcheries, due to the machinery noise masking them.

Sex-Specific Effects of Incubation Temperature on Embryonic Development of Zebra Finch (Taeniopygia guttata) Embryos.

In oviparous species, the embryonic environment-particularly temperature-can alter phenotype and survival of an individual by affecting its size as well as its metabolic rate. Previous studies have shown that incubation temperatures can affect sex ratio in birds; specifically, low incubation temperatures were shown to produce a male-biased sex ratio in zebra finches (Taeniopygia guttata) possibly because of a higher pre- or postnatal mortality rate in females. We hypothesized that sexes respond differently to suboptimal incubation temperature, leading to a male-biased sex ratio. To test this hypothesis, zebra finch eggs were incubated at 36.1°, 37.5°, or 38.5°C and hatching success, hatchling mass, residual yolk mass, and pectoralis mass were measured. We found that while hatchling mass was similar between the sexes at 37.5°C, female hatchlings were heavier at 36.1°C, and male hatchlings were heavier at 38.5°C. Pectoralis muscle mass was similar between the sexes at 36.1°C; however, at 37.5°C, female pectoralis mass was heavier at hatching than that of males. Females at 37.5°C also had lower residual yolk at hatching compared with males, reflecting a higher use of energy by female embryos compared with male embryos at this temperature. In contrast, residual yolk was similar between the sexes at 36.1° and 38.5°C. Our results suggest that there are sex differences in how incubation temperature alters organ mass and yolk energy reserve; this can lead to a difference in survival at different incubation temperatures between the sexes. Taken together with previous studies showing that females alter incubation behavior with ambient temperature, rising ambient temperatures could impact phenotype and survival of avian offspring in a sex-specific manner.

Captive-rearing of Gunnison sage-grouse from egg collection to adulthood to foster proactive conservation and recovery of a conservation-reliant species.

Gunnison sage-grouse (Centrocercus minimus) are distributed across southwestern Colorado and southeastern Utah, United States. Their distribution has decreased over the past century and the species has been listed as threatened by the U.S. Fish and Wildlife Service. Reduced genetic diversity, small population size, and isolation may affect Gunnison sage-grouse population persistence. Population augmentation can be used to counteract or mitigate these issues, but traditional translocation efforts have yielded mixed, and mostly unsuccessful, results. Captive-rearing is a viable, although much debated, conservation approach to bolster wild conservation-reliant species. Although there have been captive-rearing efforts with greater sage-grouse (C. urophasianus), to date, no information exists about captive-rearing methods for Gunnison sage-grouse. Therefore, we investigated techniques for egg collection, artificial incubation, hatch, and captive-rearing of chicks, juveniles, subadults, and adults for Gunnison sage-grouse. In 2009 we established a captive flock that produced viable eggs. From 2009-2011, we collected and artificially incubated 206 Gunnison sage-grouse eggs from 23 wild and 14 captive females. Our hatchability was 90%. Wild-produced eggs were heavier than captive-produced eggs and lost mass similarly during incubation. We produced 148 chicks in captivity and fed them a variety of food sources (e.g. invertebrates to commercial chow). Bacterial infections were the primary cause of chick mortality, but we successfully reduced the overall mortality rate during the course of our study. Conservationists and managers should consider the utility in developing a captive-rearing program or creating a captive population as part of a proactive conservation effort for the conservation-reliant Gunnison sage-grouse.

Embryological staging of the Zebra Finch, Taeniopygia guttata.

Zebra Finches (Taeniopygia guttata) are the most commonly used laboratory songbird species, yet their embryological development has been poorly characterized. Most studies to date apply Hamburger and Hamilton stages derived from chicken development; however, significant differences in development between precocial and altricial species suggest that they may not be directly comparable. We provide the first detailed description of embryological development in the Zebra Finch under standard artificial incubation. These descriptions confirm that some of the features used to classify chicken embryos into stages are not applicable in an altricial bird such as the Zebra Finch. This staging protocol will help to standardize future studies of embryological development in the Zebra Finch.