Damaged eggshells result in losses of eggs. The genetic mechanism of variable eggshell strength is still unclear. The current study was conducted to verify whether the eggshell calcification related genes, CALB1, SPP1, DMP4, BMP2 and SLIT2, were associated with eggshell mechanical property. For this purpose quantitative PCR (q-PCR) analysis was performed to detect gene expression between two groups of hens laying strong and weak eggs. The hens were selected from 360 White Leghorn layers at 60 wk to ensure that the strong and weak eggs differed significantly in breaking strength but not in eggshell thickness and weight. Using this special strong/weak eggshell model, we found that the expression of CALB1 in the uterus of strong shell group was about 3-fold higher (P<0.05) than that in weak shell group. The DMP4 expression was significantly higher (2-fold, P<0.05) in the uterus of weak shell group than that in strong shell group. However, no difference was observed for genes of SPP1, SLIT2 and BMP2 between these two groups. The current study provides a new insight to investigate the association of candidate genes with eggshell mechanical property.

Cellular retinoic acid-binding protein 1 (CRABP1) is a well-conserved member of cytosolic lipid-binding protein family. It is an important modulator of retinoic acid signaling. Long serial analysis of gene expression (LongSAGE) analysis suggested that CRABP1 gene was differentially expressed during prenatal skeletal muscle development in porcine. Here, we obtained the full-length coding region sequence and genomic sequence of the porcine CRABP1 gene and analyzed its genomic structures. Subsequently, we examined CRABP1 chromosome assignment using INRA-University of Minnesota 7 000 porcine radiation hybrid panel (IMpRH) and explored its tissue distribution in adult Tongcheng pigs and dynamical expression profiles in prenatal skeletal muscle (33, 65 and 90 days post coitus, dpc) from Landrace (lean-type) (described as L33, L65 and L90) and Tongcheng pigs (obese-type) (described as T33, T65 and T90). The CRABP1 gene was mapped to chromosome 7q11-q23 and closely linked to the microsatellite marker SWR1928. Quantitative real-time PCR showed that CRABP1 mRNA was highly expressed in lung and stomach, moderately expressed in placenta and uterus, and weakly expressed in other tissues. Moreover, CRABP1 gene was down-regulated during prenatal skeletal muscle development in both Landrace and Tongcheng pigs and it was expressed much higher in T33 than L33. Two single-nucleotide polymorphisms (SNPs) were detected by sequencing and mass spectrometry methods, allele frequency analysis indicated that g. 281 (G>A) and g. 2992 (G>A) were deviated from Hardy-Weinberg equilibrium in the Landrace and DLY (Duroc×(Landrace×Yorkshire)) pig breeds.

The present study aims to understand the effects of long-term fertilization on soil organic carbon (SOC), black carbon (BC), enzyme activity, and the relationships among these parameters. Paddy field was continuously fertilized over 30 yr with nine different fertilizer treatments including N, P, K, NP, NK, NPK, 2NPK (two-fold NPK), NPK+manure (NPKM), and CK (no fertilization), N, 90 kg urea-N ha-1 yr-1; P, 45 kg triple superphosphate-P2O5 ha-1 yr-1; K, 75 kg potassium chloride-K2O ha-1 yr-1; and pig manure, 22 500 kg ha-1 yr-1. Soil samples were collected and determined for SOC, BC content, and enzyme activity. The results showed that the SOC in the NPKM treatment was significantly higher than those in the K, P, and CK treatments. The lowest SOC content was found in the CK treatment. SOC content was similar in the N, NP, NK, NPK, 2NPK, and NPKM treatments. There was no significant difference in BC content among different treatments. The BC-to-SOC ratios (BC/SOC) ranged from 0.50 to 0.63, suggesting that BC might originate from the same source. Regarding enzyme activity, NPK treatment had higher urease activity than NPKM treatment. The urease activity of NPKM treatment was significantly higher than that of 2NPK, NP, N, P, K, CK, and NPKM treatment which produced higher activities of acid phosphatase, catalase, and invertase than all other treatments. Our results indicated that long-term fertilization did not significantly affect BC content. Concurrent application of manure and mineral fertilizers increased SOC content and significantly enhanced soil enzyme activities. Correlation analysis showed that catalase activity was significantly associated with invertase activity, but SOC, BC, and enzyme activity levels were not significantly correlated with one another. No significant correlations were observed between BC and soil enzymes. It is unknown whether soil enzymes play a role in the decomposition of BC.

Fadrozole, an aromatase inhibitor, can masculinize genetic female chickens and high-dose decreases the hatchability. Therefore, it is important to study the growth and development of sex-reversed females after hatch. Chick embryos from a population of CAU3 egg-type were treated with different concentrations of Fadrozole prior to the sexual differentiation at E3.0 (st18). At hatch, the phenotypic sex and genetic sex were identified by vent sexing and genetic diagnosis with CHD1, respectively. Body weight and shank length of sex reversal were tested at 8 and 20 wk, respectively. Testicular development, oviduct and ovarian degeneration were observed and serum concentration of estradiol and testosterone were tested with radioimmunoassay (RIA) at 30 wk. The results showed that body weight and shank length of sexreversed females were not significantly different between low-dose groups (0.1, 0.3, and 0.5 mg for F1, F2, and F3, respectively) and high-dose groups (1.0 and 1.3 mg for F4 and F5, respectively) (P>0.05). Left and right testes or ovotestes in F2, F3, F4, and F5 groups were heavier than that of in F1 group (P<0.05). While the gonad weight of treatment groups were less than that in male control (P<0.05), oviduct weight in F2, F3, F4, and F5 groups were significant differences compared with female control and F1 group (P<0.05). Egg number from onset of laying egg to 30 wk in F4 and F5 groups were less than in female control, F1 and F2 groups (P<0.05). Serum testosterone level in F5 group was significant higher compared with female control, F1, F2, F3, and F4 groups (P<0.05), but significant lower compared with male control (P<0.05). While concentration of serum estradiol in F5 group was significant lower compared with female control, F1, F2, and F4 groups (P<0.05). In conclusion, the concentration of Fadrozole do not affect postnatal growth of sex-reversed female chicken and the degree of sex-reversed females elevate with the increase of Fadrozole concentration at sex maturity.