Increased CO2 Levels during the First Half of Incubation at High Altitude Modifies Embryonic Development of Fertile Leghorn Breeder Eggs

The exchange of oxygen (O2) and carbon dioxide (CO2) within an incubator has a significant impact on embryonic development (ED) and hatching processes. This study examines the influence of non-ventilation (NV) conditions during the first ten days of incubation at high altitudes on Leghorn hens hatching eggs. Five hundred four hatching eggs were equally divided into three treatment groups and placed in twelve incubators (R = 4). The first group was subjected to standard ventilated conditions (V) during the setting phase. The ventilation inlet holes of the remaining incubators in the NV treatments were closed with either micropore (M) or polypropylene (P) tape, referred to as NVM and NVP groups, respectively. These two different airtight settings were intended to allow for a gradual rise in CO2 naturally generated by the embryos. Results indicate that carbon dioxide concentration gradually increased during the first half of incubation, reaching 1.42% in the NVM group and 1.20% in the NVP group, while the V condition group remained at 0.15%. From 10 days of incubation onwards, normal V conditions were restored in all incubators. The highest hatchability of fertile eggs (HFE) was shown by the NVP group (55.7%), followed by the V (52.6%) and NVM (38.6%) groups. The NVP group showed a greater yolk-free body mass (YFBM) from 10 days of incubation until the hatch basket transfer. NV conditions during the first 10 days of incubation at high altitude produced higher YFBM with gradually decreasing yolk sac mass. In comparison to the NVM and V conditions, the particular NVP condition showed a beneficial impact on the quality of hatched chicks. Sustaining NVP condition (1.2% of CO2) throughout the first half of incubation at high altitude generated the optimal environment in the incubator ensuring the best hatchability results. This study highlights how important it is for hatchery managers to recognize the influence of low O2 and high levels of CO2 on the development trajectories of Leghorn embryos during early incubation at high altitudes.

Recognition and rejection of foreign eggs of different colors in Barn Swallows

Brood parasitic birds lay eggs in the nests of other birds,and the parasitized hosts can reduce the cost of raising unrelated offspring through the recognition of parasitic eggs.Hosts can adopt vision-based cognitive mechanisms to recognize foreign eggs by comparing the colors of foreign and host eggs.However,there is currently no uniform conclusion as to whether this comparison involves the single or multiple threshold decision rules.In this study,we tested both hypotheses by adding model eggs of different colors to the nests of Barn Swallows(Hirundo rustica)of two geographical populations breeding in Hainan and Heilongjiang Provinces in China.Results showed that Barn Swallows rejected more white model eggs(moderate mimetic to their own eggs)and blue model eggs(highly non-mimetic eggs with shorter reflectance spectrum)than red model eggs(highly nonmimetic eggs with longer reflectance spectrum).There was no difference in the rejection rate of model eggs between the two populations of Barn Swallows,and clutch size was not a factor affecting egg recognition.Our results are consistent with the single rejection threshold model.This study provides strong experimental evidence that the color of model eggs can has an important effect on egg recognition in Barn Swallows,opening up new avenues to uncover the evolution of cuckoo egg mimicry and explore the cognitive mechanisms underlying the visual recognition of foreign eggs by hosts.

Eggs mimicry of Common Cuckoo (Cuculus canorus) utilizing Ashy-throated Parrotbill (Paradoxornis alphonsianus) host

Polymorphism in egg coloration is prominent in the Common Cuckoo (Cuculus canorus) and a common host, the Ashy-throated Parrotbill (Paradoxornis alphonsianus). Egg polymorphism has probably evolved as a consequence of frequency-dependent selection in both host and parasite, and has, according to human vision, resulted in discrete immaculate white, pale blue and blue egg phenotypes within a single population. However, egg mimicry assessment is not always straightforward, and previous studies have shown that human based comparisons applied to the coloration of bird eggs may be inadequate. Here, we objectively quantify egg color of both parasite and host by spectrophotometry and assess egg mimicry of the Common Cuckoo to the eggs of its Ashy-throated Parrotbill host. Our results revealed that egg reflectance spectra agree well with the assessment based on human vision that cuckoo eggs mimic those of the parrotbill host, in both visible (VIS) and ultraviolet (UV) ranges. However, the white cuckoo egg shows slightly poorer mimicry than the blue cuckoo egg in corresponding host clutches. We suggest that the white parrotbill egg morph (and subsequently the whitish cuckoo egg color) may have evolved after the evolution of the blue egg morph due to strong selection from parasites in the cuckoo-parrotbill system.

Study on Effects of Different Factors to Eggs Hatching Rate of Cryptotympana atrata by Orthogonal Design

[Objective] The paper aimed to explore influence of different factors on eggs hatching of Cryptotympana atrata,and optimize technical parameters in practice.[Method] By using hatching rate as index,L16（44）orthogonal design was employed to determine the effects of the four factors including temperature,salinity,photoperiod and pH.[Result] Photoperiod had obviously effects on hatchability of Cryptotympana atrata eggs.The optimal conditions for hatching were temperature 30 ℃,salinity 25%,photoperiod L12∶D12,and pH 6.[Conclusion]The research provided some scientific basis for indoor artificial incubation.

Effects of Varying Temperature and Constant Temperature on Hatching Result of Fertilized Peacock Eggs

In order to investigate the effect of temperature on hatching result of fertilized peacock eggs, they were hatched under conditions of varying temperature and constant temperature respectively. The result shows that the hatching rate in the varying temperature treatment group （Group Ⅰ） is higher than that in the constant temperature treatment group （Group Ⅱ）. It suggests that the varying temperature treatment （high temperature in the early period, medium temperature in the middle period, slightly lower temperature in the late period, and slightly higher temperature in the hulling period） is in line with the optimum temperature variation in the hatching of fertilized peacock eggs, contributing the hatching of fertilized peacock eggs.

Screening of Saprolegniasis Preventive Drugs for Golden Trout (Oncorhynchus mykiss) Fertilized Eggs During Hatching

In order to screen the effective preventive drugs for saprolegniasis occurred during hatching of the fertilized eggs of golden trout, effects of two kinds of self-made Chinese herbal compounds（AB） and two kinds of chemical bactericides（peroxyacetic acid andhydrogen peroxide, CD） with three disinfection concentrations were compared in this study. The fertilized eggs of golden trout were randomly divided into 13 groups, each group with two parallels of 1 000 eggs. Drugs at high,medium and low concentration were respectively used to immerse the fertilized eggs35 min for disinfection once a day. The numbers of live and dead eggs were recorded each day before disinfection and the numbers of survival eyed eggs were counted in each experimental group after finishing the whole test. The results indicated that all of four kinds of drugs used here had a certain effect on prevention and control of saprolegniasis during hatching of fertilized eggs of golden trout, and also showed a correlation between the concentrations and effects. After the selection of several drugs, we found that group B-medium, C-medium and D-high exhibited the most remarkable effects on control of saprolegniasis. However, careful consideration should be given to chemical drugs residues and their potential hazard to the eyed eggs. Therefore, Chinese herbal compound B was suggested to be used for prevention and control of saprolegniasis to achieve the healthy aquaculture. And the concentration and operation time of Chinese herbal compound B should be strictly controlled when used in disinfection of fertilized eggs.

A potential source of recruitment of Acartia pacifica nauplii: viable benthic resting eggs

Many estuarine and coastal planktonic copepods depend on the hatching of benthic resting eggs for recruitment of nauplii to the water column population. The distribution and abundance of viable resting eggs of Acartia pacifica in the Xiamen Bay were determined by the presence of nauplii in the laboratory. The number of viable eggs var/ed temporally and spatially. The maximum number (9.2×10^4m^-2) of viable eggs was in summer. In spring the average abundance of viable eggs was 5.6×10^4 m^-2. The abundances of viable eggs in fall and winter were similar, respectively 2.7×10^4 and 3.3×10^4m^-2, which were the lowest in the year. The numbers of viable eggs inside the stations of the Xiamen Bay were higher than those outside the stations. The viable eggs were found at all depths (0-10 cm),although not in every station. The maximum number did not necessarily occur in the uppermost centimeter of the sediments but often occurred several centimeters below the water-sediment interface. The accumulation of viable eggs in the seabed of a subtropical bay constitutes a potential source of recruitment of nauplii into the pelagic population.

Reduction of recruitment of Acartia pacifica nauplii from benthic resting eggs due to organochlorine pesticides

Many estuarine and coastal planktonic copepods depend on the hatching of benthic resting eggs for recruitment ofnauplii to the water column population. The potential effects of two organochlorine pesticides, hexchloriobinzene （HCH） and dichlorodiphenyltrichloroethane （DDT）, on the recruitment of A ccu＇tla pacifica nauplii from benthic resting eggs in the seabed of Xiamen Bay were experimentally investigated. The abundance of A. pacifica nauplii hatched from the sediment significantly decreased with the increase of pesticide concentration. Trimmed Spearman-Karber analysis gave sediment 96-h LC50 values were 84.81 ng/g for HCH, and 157.94 ng/g for DDT. The median AI （Al50） was -0.77, which suggested that the combined effect of riCH and DDT showed a weak effect than individual effects. There was a positive relationship between mortality and exposure time in DDT treatment, while the relationship was not significant in HCH treatment. The results suggest that organochlorine pesticides can reduce recruitment of A. pacifica nauplii from benthic resting eggs to planktonic population.

Heavy metal exposure reduces hatching success of Acartia pacifica resting eggs in the sediment

The potential effects of three heavy metals （Cu, Pb, and Cd） on hatching success ofAcartia pacifica resting eggs in the sediment of Xiamen Bay were experimentally investigated. The number ofA. pacifica nauplii hatched from the sediment sharply decreased with the increase of metal concentration and exposure time from 3 to 30 d. An increase of the Cu concentration from 34.8 to 348 mg/kg, reduced the number of hatched nauplii by 46.6%-100%. An increase of the Pb concentration from 75.2 to 752 mg/kg, reduced the number of hatched nauplii by 21.4%-78.9%. An increase of the Cd concentration from 0.68 to 6.8 mg/kg, reduced the number of hatched nauplii by 31.6%-94.7%. The number of nauplii also significantly decreased with the increase of mixed-metal concentration and exposure time in the mixed-metal test. Trimmed Spearman-Karber analysis gave sediment metal 72-h LC50 values of 1.25 mmol Cu/kg, 1.73 mmol Pb/kg, and 0.054 mmol Cd/kg, which suggested that Cd was the most toxic to A. pacifica resting eggs in the three tested metals. The results indicate that heavy metals with higher concentrations can reduce recruitment of A. pacifica nauplii from benthic resting eggs to planktonic population.

Spatiotemporal distribution of fish eggs and larvae in the Huanghe(Yellow) River estuary, China in 2005–2016

Decreasing fish resources in estuaries is a subject of anthropogenic activities. Studies of the spatiotemporal distribution of fish eggs and larvae can help identify the status and processes underlying recruitment in a fishery. As the fifth largest river estuary in the world, the Huanghe (Yellow) River estuary (HRE) is a typical estuary that has been seriously aff ected by human activities. Annual surveys on ichthyoplankton and environmental factors were conducted in the months of May of 2005 and 2009-2016 in the HRE to investigate the spatiotemporal distribution of fish eggs and larvae and the associated infl uencing factors. A total of 23 and 20 species of eggs and larvae, respectively, were collected. The dominant orders were Perciformes (51.2%) and Clupeiformes (25.6%). The average number of fish species eggs and larvae were 6.0 and 4.1 in average abundance of 0.91 and 0.13 ind./m^3 , respectively. The dominant species were mainly low-commercial-value small-sized fishes, such as Clupanodon punctatus , Harengula zunasi, and Acanthogobius, whereas certain traditional commercial fishes, such as Trichiurus lepturus , and Clupea pallasii, were not seen. Analysis of the fish egg and larval community revealed four temporal assemblages and two spatial assemblages. Salinity was the main factor on the spatial distribution of ichthyoplankton communities, the species number and Shannon-Weiner diversity index (H′) of the fish egg and larval community near the river mouth with lower salinity were signifi cantly lower than the community far away from the river mouth with higher salinity. In addition, increases of water temperature promoted the number and abundance of fish species eggs, and the areas of abundant prey tended to have a more diversified and abundant of ichthyoplankton species. In overall, overfishing, dam construction, and other human activities were the main drivers that led to the substantial decline in fishery resources in the HRE.

Killing Effect of Mixed Fumigation of Phosphine and Carbon Dioxide on Eggs of Lasioderma serricorne F.

[Objective] This study aimed to investigate the killing effect of mixed fumi- gation of phosphine and carbon dioxide on eggs of Lasioderma serricorne F. [Method] The outside-storage phosphine generator was placed in a tabernacled smoke box, and the mortality rates of L. serricome F. eggs in the smoke box under conditions of different aluminium phosphide usage amount and different fumigation time were studied. In addition, the times needed by tabernacle and smoke box cen- ter to reach the phosphine concentration peak were recorded. [Result] The optimum conditions for killing the eggs of L. serricorne were as follows： temperature of （27± 2） ℃, relatively humidity of （45±5）%, aluminium phosphide usage amount of 1.5 g/m3 and effective exposure time of 96 h. The killing effect of mixed fumigation of phos- phine and carbon dioxide was increased with the extension of fumigation time. The increased usage amount of aluminium phosphide showed no significant effect on killing effect. The tabernacle space and smoke box center all required relatively short time to reach the phosphine concentration peak. If the tabernacle had a good airtightness, the overall fumigation time could be shortened. [Conclusion] The fumi- gation method is reliable, and it can be used for the control of L. serricorne F. in tobacco storage.

Developmental Threshold Temperature and Effective Accumulative Tem- perature of Pupae and Eggs of Holcocerus hippophaecolus

In order to understand the occurrence and the developmental regularity of seabuckthorn carpenterworm (Holcocerus hippophaecolus) and predict its population density, the developmental threshold temperature (C) and effective accumulative tem- perature (K) of the carpenterworm pupae and eggs were analyzed under the conditions of constant and variable temperatures. The results show that the values of C and K of the carpenterworm pupae are (12.1 ± 0.2) °C and (295.2 ± 4.1) day-degrees at constant temperatures, and (15.5 ± 0.4) °C and (202.4 ± 13.1) day-degrees at variable temperatures. However, the values of C and K of the eggs at variable temperatures are (16.7 ± 0.8) °C and (101.5 ± 12.6) day-degrees. The differences of developmental threshold and effective accumulative temperature under the conditions of constant and variable temperatures of the carpenterworm pupae accord with the developmental regularity of most insects in nature. By comparing five different constant temperatures, the conclusion is that the optimum developmental temperature of the pupae is 21 °C when both the pupation of the mature larvae and the eclosion of the pupae are very accordant. Moreover, the percentage of eclosion is over 90%. The average developmental durations of the carpenter- worm pupae and eggs are 31 and 16 d at variable temperatures.

Effects of Predation by Invasive Western Mosquitofish(Gambusia affinis) on Survival of Eggs,Embryos and Tadpoles of Pelophylax nigromaculatus and Duttaphrynus melanostictus in South China

Alien species are one of the most serious threats to the decline and extinction of native amphibian populations. In this study, we examined the predation of invasive Western Mosquitofish Gambusia affinis on the eggs, embryos, and tadpoles of Duttaphrynus melanostictus and Pelophylax nigromaculatus in south China. Our results suggested that the survival of eggs and embryos remaining in the egg capsules of P. nigromaculatus and D. melanostictus was significantly higher than those removed from the egg capsule at 12-h intervals within 72 h in the presence of G. affinis. The survival of P. nigromaculatus eggs and embryos without egg capsules was significantly lower than those of D. melanostictus without egg capsules. The survival of P. nigromaculatus eggs and embryos with egg capsules was significantly higher than those of D. melanostictus with egg capsules from 24 h to 72 h except for 12 h. The survival of D. melanostictus tadpoles was significantly higher than that of P. nigromaculatus tadpoles in the presence of G. affinis. The survival of Gosner stage 26 tadpoles of P. nigromaculatus was significantly higher than that of Gosner stage 30 tadpoles from 12 h to 60 h, but there were no significant differences at 72 h. In contrast, the survival of Gosner stage 26 tadpoles of D. melanostictus was significantly lower than that of Gosner stage 30 tadpoles within 72 h, recording every 12 h. The increasing temperature caused a significant increase in predation by G. affinis on P. nigromaculatus eggs and embryos. The outer jelly capsule surrounding anurans eggs might serve as a mechanical defense against predation by G. affinis due to its large diameter, relatively stationary state and unpalatability. The differences in the vulnerability of P. nigromaculatus and D. melanostictus embryos and tadpoles to G. affinis probably due to differences in the unpalatability, black skin and activity. Based on the magnitude of predation by G. affinis on the eggs, embryos and tadpoles of these two species and the combined impact of temperature, we might speculate that invasive G. affinis and global warming would have more detrimental impacts on population viability of P. nigromaculatus than D. melanostictus in China.

Effect of in ovo zinc injection on the embryonic development, tissue zinc contents, antioxidation, and related gene expressions of broiler breeder eggs

Two experiments were conducted to investigate the effect of in ovo zinc （Zn） injection on the embryonic development, tissue Zn contents, antioxidation and related gene expressions of fertilized eggs of Arbor Acres broiler breeders. Experiment 1 was conducted to determine an optimal embryonic age for early in ovo injection. A total of 720 fertilized eggs with similar weights were randomly allotted to 4 treatments with 6 replicates per treatment and 30 eggs per replicate in a completely randomized design. The eggs were injected with 0.1 mL sterilized water at 3, 6 and 9 embryonic days of incubation （E3, E6 and E9） or non-injection （the control）, respectively. The results from experiment 1 showed that E3 and E6 injections increased （P〈0.05） the embryonic mortalities, and decreased （P〈0.05） hatchabilities compared to the non-injected control, but no differences （P〉0.05） between E9 injection and the non-injected control were observed in either embryonic mortality or hatchability. The findings suggest that the E9 is the optimal embryonic age for early in ovo injection. In experiment 2, a total of 672 fertilized eggs with similar weights were randomly allocated to 7 treatments with 6 replicates per treatment and 16 eggs per replicate in a completely randomized design. The eggs were injected with 0 （the negative control）, 50, 100, 150, 200, or 250 μg Zn/egg as reagent grade ZnSO4.7H20 in a 0.1-mL solution, or non-injection （the positive control）, respectively at E9-10. The results from the experiment 2 demonstrated that no differences （P〉0.05） among 50 and 100 μg Zn/egg groups and the negative control were observed in the embryonic mortality and hatchability, however, the injection of 200 μg Zn/egg increased （P〈0.05） the embryonic mortality, and injections of 150 and 200 μg Zn/egg decreased （P〈0.05） hatchabilities compared with the controls. The embryonic tibia Zn contents at E20 were increased （P〈0.05） by injections of 150, 200 and 250μg Zn/egg. Zinc injection did not affect （P〉0.05） malonaldehyde （MDA） contents, copper- and Zn- containing superoxide dismutase （CuZnSOD） activities and mRNA expression levels in the liver and heart of chick embryos at E15 and E20. Compared with the negative control, injections of 50, 150 and 200 μg Zn/egg up-regulated （P〈0.05） themetallothionein （MT） mRNA expression levels in the embryonic liver at E20. These results indicated that in ovo Zn injections increased Zn contents in the embryonic tibia and MT mRNA expression levels in the embryonic liver at E20, however, injections of 150-200 μg Zn/egg were harmful to the embryonic development.

Pathological Microstructure of a Miocene Ostrich Eggshell from Asia

Struthio linxiaensis is the oldest fossil ostrich from Asia till today. This paper analyzes the microstructure of the eggshell of Struthio linxiaensis. The SEM and light microscopic results show that a pathological structure occurred associated with the formation of pore canal. The feature of this structure is that crystals of the columnar layer are around the lower part of the pore canal, and formed a broad structure. This structure unit contracts upwards more severely than the canal and looks like a ＂garlic＂, which was observed in aumiotes eggshells for the first time. Many evidences indicate that this structure was deposited with the columns simultaneously. The radial section view suggests that the ＂garlic structure＂ is not a result of antiperistaltic waves in the avian oviduct. The most possible process of the ＂garlic structure＂ formation is the column units around the pore canals with imperfect acidifying when calcified, or have some correlations with the changes in the protein profiles of the organic matrix in the calcareous eggshell.

Prediction Model of Weekly Retail Price for Eggs Based on Chaotic Neural Network

This paper establishes a short-term prediction model of weekly retail prices for eggs based on chaotic neural network with the weekly retail prices of eggs from January 2008 to December 2012 in China.In the process of determining the structure of the chaotic neural network,the number of input layer nodes of the network is calculated by reconstructing phase space and computing its saturated embedding dimension,and then the number of hidden layer nodes is estimated by trial and error.Finally,this model is applied to predict the retail prices of eggs and compared with ARIMA.The result shows that the chaotic neural network has better nonlinear fitting ability and higher precision in the prediction of weekly retail price of eggs.The empirical result also shows that the chaotic neural network can be widely used in the field of short-term prediction of agricultural prices.

Statistical Approach for Classification of Dinosaur Eggs from the Heyuan Basin at the Northeast of Guangdong Province

The Zhutian and Dafeng formations （Upper Cretaceous） of the Heyuan Basin in northeastern Guangdong Province, China, have produced thousands of dinosaur eggs. Macromorphological features （egg diameter, egg shape, outer surface texture, and shell thickness） of 461 eggs were analyzed using non-destructive techniques and subjected to statistical analyses in order to assess their diversity and taxonomic affinities. Three types （1, 2 and 3） of eggs were discerned based on shape and outer surface morphology. Type 1 eggs are spherical to ellipsoidal in shape and have a rough surface. Three subtypes （Type 1-A-l, l-A-2, and l-B） are apparent from scatter plots and cluster analyses of egg diameters and shell thickness. Type 2 eggs are elongate with linear ornamentation on the surface, and are comparable to eggs that belong to the oofamily Elongatoolithidae. Type 3 eggs are elongate with a smooth surface, and are assigned to the oofamily Prismatoolithidae. Macromorphological features of the various egg types suggest that Type 1 could have been laid by ornithischian, sauropod or therizinosaur dinosaurs, Type 2 by oviraptorids, and Type 3 by troodontids. This study represents the first comprehensive statistical analysis of macrofeatures of dinosaur eggs, and reveals taxonomic diversity in the dinosaurs that were laying eggs in the Upper Cretaceous Heyuan area heretofore unrecognized in skeletal remains.

Analysis on the Variation Expression of Difficult-Dissolved-Proteins of Diapause Eggs After Activating by Acid Treatment

Using HCl to activate the diapause eggs is a traditional technique of artificial hatching applied in silkworm egg production. Its mechanism has not yet been clarified. This experiment explored the effect of HCl on the termination of diapause of silkworm eggs cold-stored for 45 days from the point of proteomes. Two-dimensional electrophoresis techniques and ESI- MS-MS were used to compare and analyze the variation expression of difficult-dissolved-proteins of diapause eggs coldstored for 45 days before and after acid treatment. Through analysis on the two-dimensional electrophoretogram, there were 296 dots before acid treatment and 302 after the treatment, respectively. Amongst them 265 dots were matchable. The matchability reached 88.6%. There were 31 specific protein dots before acid treatment and 37 after acid treatment, respectively. ESI-MS-MS analysis was conducted for two specific protein-rich dots which disappeared after acid treatment. The results indicated that the sequence of No. 1 protein dot had 55 amino acids＇ peptide matched with those of chorion protein （Bombyx mori）. While the sequence of No. 2 protein dot had only 15 amino acids＇ peptide matched with those of heat shock protein hsp 19.9 （Bombyx mori）, and it was presumed to be an unknown protein. The difficult-dissolvedproteins of diapause eggs have variation expression after acid treatment. Some proteins before and after acid treatment are changed in MW.

Reduction of hydrocarbon contamination on viability of Acartia pacifica benthic resting eggs

The potential effect of hydrocarbon contamination on the hatching success of benthic resting eggs of Acartia pacifica in Xiamen Bay was investigated experimentally. The number of nauplii emerging from the sediment samples decreased with increasing Fuel Oil #0 concentration. The estimated rate of mortality increased markedly with the increase of Fuel Oil #0 concentration. Successive fuel Oil #0 concentrations from 50 mg/kg to 5000 mg/kg reduced the number of hatched nauplii by 3.8% -100%. The mortality of A. pacifica resting eggs due to Fuel Oil #0 contamination did not significantly increase as time progressed at each concentration level. The LC50 values of resting eggs, changing from 237.12 to 279.59 mg/kg, remained at an almost stable level in two months. The number of A. pacifica nauplii that hatched from the sediment at 10℃ was higher than those from the sediment at 30℃, which indicates that the toxicity of Fuel Oil #0 on A. pacifica resting eggs increases with increasing temperature.

Unhatched and Hatched Eggshells of the Chinese Cobra Naja atra

Changes in structure and composition of the eggshell resulting from embryonic mobilization of minerals from the eggshell are found in all oviparous reptiles studied thus far. In this study, we measured samples of unhatched and hatched eggshells of the Chinese cobra Naja atra to determine the percentage of ash and the phase composition of calcium carbonate. The mean percentage of ash was significantly higher in unhatched eggshells （24.6%） than in hatched eggshells （22.3%）. The dominant phase in unhatched eggshells was the calcite form of calcium carbonate. In addition to the peaks of calcite, a few small peaks were found to be caused by the aragonite and vaterite phases of calcium carbonate, implying that there are small amounts of aragonite and vaterite in the eggshell. The concentration of the various phases calculated from the intensity of the X-ray diffraction spectra allowed the estimation that percentages of calcite, aragonite and vaterite were about 92%, 4% and 4%, respectively. Hatched eggshells produced similar spectral characteristics as unhatched eggshells, with one exception. The dominant phase composition in the hatched eggshell was also calcite, but the amount of the aragonite phase had a marked increase. Our study adds evidence that embryonic mobilization of minerals from the eggshell may result in changes in structure of the eggshell.