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Hypoimmunity and numerous stresses are two major challenges in broiler industry. Nutrient intervention at the specific time of embryonic stage is a feasible way to improve animal performance. This study was conducted to investigate the possible effects of in ovo feeding (IOF) of vitamin C at embryonic age 15th day (E15) on growth performance, antioxidation and immune function of broilers. A total of 240 broiler fertile eggs were randomly divided into two groups (0 and 3 mg injected dose of vitamin C at E15), and new-hatched chicks from each treatment were randomly allocated into six replicates with 10 chicks per replicate after incubation. The results indicated that in ovo vitamin C injection improved the hatchability (P<0.05) and increased immunoglobulin M (IgM) (at the broiler’s age 1st day, D1), IgG and IgM concentrations (D21), as well as lysozyme activity (D21, P<0.05) and total antioxidant capacity (D42, P<0.01) in plasma of broilers. On D21, the splenic expression level of DNA methyltransferase 1 (DNMT1) was up-regulated in vitamin C (VC) group, whereas interleukin (IL)-6, interferon-γ, ten-eleven translocation protein 1 and thymine-DNA glycosylase were down-regulated (P<0.05). On D42, in ovo vitamin C injection up-regulated splenic expression levels of DNMT1, DNA methyltransferase 3B (DNMT3B) and growth arrest and DNA-damage-inducible protein beta (P<0.05), whereas down-regulated splenic expression levels of IL-6, tumour necrosis factor-α and methyl-CpG-binding domain protein 4 (P<0.05). Our findings suggested that IOF of 3 mg vitamin C at E15 could improve, to some extent, the antioxidant activity and immune function in plasma, corresponding with the lower expression of pro-inflammatory cytokines in spleen. However, IOF of vitamin C leading to the changes in the expression of DNA methyltransferases and demethylases may suggest an increased trend of DNA methylation level in spleen and whether DNA methylation variation is associated with the lower expression of pro-inflammatory cytokines in spleen warrants future study.
Hepatitis E is an important zoonosis that is prevalent in China. Hepatitis E virus (HEV) is a pathogen that affects humans and animals and endangers public health in China. In this study, the detection of HEV epidemics in swine in Sichuan Province, China, was carried out by nested real-time PCR. A total of 174 stool samples and 160 bile samples from swine in Sichuan Province were examined. In addition, software was used to analyse the biological evolution of HEV. The results showed that within 2 years of swine HEV (SHEV) infection in China, SHEV was first detected in Sichuan Province. HEV was endemic in Sichuan; the positive rate for pig farms was 11.1%, and the total positive sample rate was 10.5%. The age of swine with the highest positive rate (17.9%) was 5–9 weeks. The examined swine species in order of highest to lowest HEV infection rates were Chenghua pig, Large White, Duroc, Pietrain, Landrace and Hampshire. Nucleotide and amino acid sequence analysis showed that the HEV epidemic in swine in Sichuan Province was related to genotype IV, which had the highest homology to HEV in Beijing. Sichuan strains have greater variation than Chinese representative strains, which may indicate the presence of new HEV strains.
Taihu pig breeds are the most prolific breeds of swine in the world, and they also have superior economic traits, including high resistance to disease, superior meat quality, high resistance to crude feed and a docile temperament. The formation of these phenotypic characteristics is largely a result of long-term artificial or natural selection. Therefore, exploring selection signatures in the genomes of the Taihu pigs will help us to identify porcine genes related to productivity traits, disease and behaviour. In this study, we used both intra-population (Relative Extend Haplotype Homozygosity Test (REHH)) and inter-population (the Cross-Population Extend Haplotype Homozygosity Test (XPEHH); F-STATISTICS, FST) methods to detect genomic regions that might be under selection process in Taihu pig breeds. As a result, we found 282 (REHH) and 112 (XPEHH) selection signature candidate regions corresponding to 159.78 Mb (6.15%) and 62.29 Mb (2.40%) genomic regions, respectively. Further investigations of the selection candidate regions revealed that many genes under these genomic regions were related to reproductive traits (such as the TLR9 gene), coat colour (such as the KIT gene) and fat metabolism (such as the CPT1A and MAML3 genes). Furthermore, gene enrichment analyses showed that genes under the selection candidate regions were significantly over-represented in pathways related to diseases, such as autoimmune thyroid and asthma diseases. In conclusion, several candidate genes potentially under positive selection were involved in characteristics of Taihu pig. These results will further allow us to better understand the mechanisms of selection in pig breeding.
Superior and inferior rice grains have different weights and are located on the upper primary branch and lower secondary branches of the panicle, respectively. To study differences in germination vigour of these two types of grain, a number of factors were investigated from 0 to 48 h of germination. The present study demonstrated that in inferior grains the starch granule structure was looser at 0 h, with full water absorption at 48 h, while in superior grains the structure was tight and dense. Relative water content increased, and dry matter decreased, more rapidly in inferior grains than in superior ones. Abscisic acid and gibberellin levels, as well as α-amylase activity, also changed more rapidly in inferior grains, while soluble sugar content and amylase coding gene expression increased more rapidly in inferior than superior grains during early germination. The expression of OsGAMYB was higher in inferior grains at 24 h but higher in superior grains at 48 h. The phenotypic index of seedlings was higher in seedlings from superior grains at the two-leaf stage. However, the thousand-grain weight and yield per plant in superior and inferior plants showed no significant difference at harvest. The present study indicates that inferior grains germinate faster than superior ones in the early germination stage. Although inferior grains produced weaker seedlings, it is worthwhile using them in rice production due to their comparative yield potential over that of superior grains.
The mammalian target of rapamycin (mTOR) has been shown to be involved in lipopolysaccharide (LPS)-induced immune responses in many mammal cells. Here, we suggest that the mTOR pathway is involved in the intestinal inflammatory responses evoked by LPS treatment in chicken embryos. The intestinal tissue from Specific pathogen free chick embryos was cultured in the presence of LPS for 2 h. Secretory immunoglobulin A (sIgA) concentrations, messenger RNA (mRNA) expression of cytokines, and protein levels of nuclear factor-κB (NF-κB), mitogen-activated protein kinase (MAPK), mTOR and p70 ribosomal S6 kinase (p70S6K) were determined. The results showed that LPS treatment increased sIgA concentrations in a dose-dependent manner. The mRNA levels of interleukine (IL)-6, IL-8, IL-10, tumor necrosis factor-α and Toll-like receptor (TLR) 4 were upregulated by LPS treatment (P<0.05). Lipopolysaccharide increased the phosphorylation of Jun N-terminal kinase (JNK), p38 MAPK and NF-κB (P<0.05) while decreasing the phosphorylation level of mTOR (P<0.05). Supplementation of leucine at doses of 10, 20 and 40 mM dose-dependently decreased sIgA production. Leucine supplementation at 40 mM restored the phosphorylation level of mTOR and p70S6K while suppressing the phosphorylation levels of NF-κB (P<0.05) and partially down-regulating the phosphorylation of p38 MAPK and JNK. The transcription of IL-6 was significantly decreased by leucine supplementation. These results suggested that leucine could alleviate LPS-induced inflammatory responses by down-regulating NF-κB signaling pathway and evoking mTOR/p70S6K signaling pathway, which may involve in the regulation of the intestinal immune system in chicken embryos.
This study aims to investigate the climate–malaria associations in nine cities selected from malaria high-risk areas in China. Daily reports of malaria cases in Anhui, Henan, and Yunnan Provinces for 2005–2012 were obtained from the Chinese Center for Disease Control and Prevention. Generalized estimating equation models were used to quantify the city-specific climate–malaria associations. Multivariate random-effects meta-regression analyses were used to pool the city-specific effects. An inverted-U-shaped curve relationship was observed between temperatures, average relative humidity, and malaria. A 1 °C increase of maximum temperature (Tmax) resulted in 6·7% (95% CI 4·6–8·8%) to 15·8% (95% CI 14·1–17·4%) increase of malaria, with corresponding lags ranging from 7 to 45 days. For minimum temperature (Tmin), the effect estimates peaked at lag 0 to 40 days, ranging from 5·3% (95% CI 4·4–6·2%) to 17·9% (95% CI 15·6–20·1%). Malaria is more sensitive to Tmin in cool climates and Tmax in warm climates. The duration of lag effect in a cool climate zone is longer than that in a warm climate zone. Lagged effects did not vanish after an epidemic season but waned gradually in the following 2–3 warm seasons. A warming climate may potentially increase the risk of malaria resurgence in China.
Information on morbidity burden of seasonal influenza in China is limited. A multiplier model was used to estimate the incidence and number of outpatient visits for seasonal influenza by age group for the 2015–2016 season in Beijing, the capital of China, based on reported numbers of influenza-like illness consultations and proportions of positive cases from influenza surveillance systems in Beijing, general consultation rates and other parameters from previous studies, surveys and surveillance systems. An estimated total of 1 190 200 (95% confidence interval (CI) 830 400–1 549 900) cases of influenza virus infections occurred in Beijing, 2015–2016 season, with an attack rate of 5·5% (95% CI 3·9–7·2%). These infections resulted in an estimated 468 280 (95% CI 70 700–606 800) outpatient visits, with an attack rate of 2·2% (95% CI 0·3–2·8%). The attack rate of influenza virus infections was highest among children aged 0–4 years (31·9% (95% CI 21·9–41·9%)), followed by children aged 5–14 years (18·7% (95% CI 12·9–24·5%)). Our study demonstrated a substantial influenza-related morbidity in Beijing, China, especially among the preschool- and school-aged children. This suggests that development or modification of seasonal influenza targeted vaccination strategies need to recognize that incidence is highest in children.
Diarrhea is a common cause of morbidity and mortality and the incidence of diarrhea in the world has changed little over the past four decades. To assess the prevalence of and healthcare practices for diarrhea, a cross-sectional study was conducted in Pudong, Shanghai, China. In October 2014, a total of 5324 community residents were interviewed. Respondents were asked if they had experienced diarrhea (defined as ⩾3 passages of watery, loose, bloody, or mucoid stools within a 24-h period) in the previous month prior to the interview. The monthly prevalence of diarrhea was 4·1% (95% CI: 3·3–4·8), corresponding to an incidence rate of 0·54 episodes per person-year. The proportion of individuals with diarrhea who sought healthcare was 21·2% (95% CI: 13·4–29·0). Diarrhea continues to impose a considerable burden on the community and healthcare system in Pudong. Young age and travel were identified as predictors of increased diarrhea occurrence.
The E2F transcription factor family is distributed widely in eukaryotes and has been well studied among mammals. In the present study, the E2F transcription factor 4 (E2F4) gene was isolated from fat bodies of Antheraea pernyi and sequenced. E2F4 comprised a 795 bp open reading frame encoding a deduced amino acid sequence of 264 amino acid residues. The recombinant protein was expressed in Escherichia coli (Transetta DE3), and anti-E2F4 antibodies were prepared. The deduced amino acid sequence displayed significant homology to an E2F4-like protein from Bombyx mori L. Quantitative real-time polymerase chain reaction analysis revealed that E2F4 expression was highest in the integument, followed by the fat body, silk glands, and haemocytes. The expression of E2F4 was upregulated in larvae challenged by bacterial (Escherichia coli, Micrococcus luteus), viral (nuclear polyhedrosis virus), and fungal (Beauveria bassiana) pathogens. These observations indicated that E2F4 is an inducible protein in the immune response of A. pernyi and probably in other insects.
Worldwide 350 million people suffer from major depression, with the majority of cases occurring in low- and middle-income countries. We examined the patterns, correlates and care-seeking behaviour of adults suffering from major depressive episode (MDE) in China.
A nationwide study recruited 512 891 adults aged 30–79 years from 10 provinces across China during 2004–2008. The 12-month prevalence of MDE was assessed by the Modified Composite International Diagnostic Interview-short form. Logistic regression yielded adjusted odds ratios (ORs) of MDE associated with socio-economic, lifestyle and health-related factors and major stressful life events.
Overall, 0.7% of participants had MDE and a further 2.4% had major depressive symptoms. Stressful life events were strongly associated with MDE [adjusted OR 14.7, 95% confidence interval (CI) 13.7–15.7], with a dose–response relationship with the number of such events experienced. Family conflict had the highest OR for MDE (18.9, 95% CI 16.8–21.2) among the 10 stressful life events. The risk of MDE was also positively associated with rural residency (OR 1.5, 95% CI 1.4–1.7), low income (OR 2.3, 95% CI 2.1–2.4), living alone (OR 2.6, 95% CI 2.3–3.0), smoking (OR 1.4, 95% CI 1.3–1.6) and certain other mental disorders (e.g. anxiety, phobia). Similar, albeit weaker, associations were observed with depressive symptoms. Among those with MDE, about 15% sought medical help or took psychiatric medication, 15% reported having suicidal ideation and 6% reported attempting suicide.
Among Chinese adults, the patterns and correlates of MDE were generally consistent with those observed in the West. The low rates of seeking professional help and treatment highlight the great gap in mental health services in China.
Pudong White (PW) pigs are distributed in the Taihu region of China and are characterized by their completely white coats. A heated debate concerning this genetic resource and its relationship to Taihu and western pig breeds has arisen, due to the white coat of the animals. To determine whether PW is a unique genetic resource, we performed a detailed assessment of the genetic relationships among PW, six breeds from the Taihu population and three western pig breeds, based on whole-genome single nucleotide polymorphism (SNP) data. A total of 68 102 SNPs were identified in the genomes of the tested populations by next-generation sequencing technology, of which, 64 were determined as the potentially specific to PW breed. The genetic distance between PW pigs and the Taihu population was shorter than that between PW and western breeds. The genetic distance within the PW population was small and neighbour-joining tree analysis revealed that all PW individuals clustered into a separated group, indicating a close genetic relationship among PW individuals which may result from a small effective population size (Ne) and inbreeding. The results of both principal component analysis and evaluation using fastSTRUCTURE demonstrated that PW was clearly differentiated from other breeds. Together, these results indicate that PW is a distinctive genetic resource with a unique genetic structure separate from other Taihu and western pig breeds. Furthermore, this genome-wide comprehensive survey of the relationships among PW, Taihu and western pig breeds, demonstrates the rationality of the current breed classification of PW. The results also provide evidence about the unique genetic resource of PW, based on genome-wide genetic markers. These data will improve our understanding of the genetic structure and current state of PW breed, and facilitate the development of a national project for the conservation and utilization of these pigs.
Association mapping based on linkage disequilibrium is an effective approach for dissecting the inheritance of complex multi-gene traits. In the present study, association mapping was performed for yield traits based on 172 popular Upland cotton (Gossypium hirsutum L.) cultivars in China and 331 polymorphic simple sequence repeat (SSR) markers. The gene diversity index of 331 markers ranged from 0·0387 to 0·7799 with an average of 0·4002, and the polymorphism information content ranged from 0·0379 to 0·7473 with an average of 0·3375. A total of 93 significantly associated markers for seven yield traits were identified across more than one environment, among which 11 were for seed cotton yield, 12 for lint yield, 11 for boll number per plant, 13 for boll weight, 21 for lint percentage, 14 for lint index and 11 for seed index. The corresponding ranges in phenotypic variation explained by markers across four environments for these seven traits were 1·75–10·49, 1·75–9·34, 2·84–11·80, 2·59–9·89, 2·38–13·97, 2·73–14·82 and 2·50–11·88%, respectively. Some of the yield-associated markers detected were found to be linked to or associated with the same traits identified in previous studies. Furthermore, elite alleles for yield traits were also mined. The present study can provide useful information for further understanding the genetic basis of yield traits, and facilitate high-yield breeding by molecular design in Upland cotton.
Twenty Small Tailed Han (STH) and 20 Ujumqin (UJU) ewes naturally infected with gastrointestinal nematodes were randomly assigned to one of four treatments arranged in a 2 × 2 factorial design, receiving anthelmintic treatment (AT) or non-anthelmintic treatment (NonAT) prior to lambing. After lambing, the effects of AT on feed intake, digestion and milk yield in ewes, and the growth rates of lambs fed their mother's milk were assessed for 28 days. Faecal samples were collected to determine faecal egg counts (FECs), milk was collected to measure milk yield and ewes and lambs were weighed to quantify daily body weight change. The results showed that AT significantly increased ewe dry matter intake (2411 g/d for AT and 2209 g/d for NonAT) and decreased FECs (50 eggs/g for AT and 2655 eggs/g for NonAT). All ewes lost weight after lambing, but body weight loss in the AT (43 g/d) was significantly less than in NonAT (84 g/d), and STH ewes (70 g/d) lost more weight than UJU ewes (58 g/d). Anthelmintic-treated ewes produced more milk for their lambs to consume. However, the extent of these positive effects of AT differed between STH and UJU ewes. The average daily body weight gain of lambs in AT was higher than those in NonAT. In conclusion, effective AT in ewes before lambing benefits subsequent lactation in ewes and growth rate in lambs.
Flour whiteness (FW) is an important factor in assessing flour quality and determining the end product quality. It is an integrated sensory indicator reflecting flour colour and is negatively correlated with protein content. In order to dissect the genetic relationship between FW and its five related traits at the quantitative trait locus (QTL)/gene level, a recombinant inbred line population was evaluated under three environments. Quantitative trait loci for FW were analysed by unconditional and conditional QTL mapping. Four unconditional additive QTLs and 16 conditional additive QTLs were detected across the three environments. Of these QTLs, only one major additive QTL (Qfw1D1-1) was consistently identified using both unconditional and conditional QTL analysis. This QTL was independent of flour colour a* (a function of red-green with a positive a* for redness and negative for greenness) and b* (a green-blue value with positive value for yellowness and negative for blueness) and was only slightly affected by flour protein content. A minor additive QTL (Qfw4A-4) was also detected using these two QTL mapping methods, being independent of flour colour a* and b*. Five unconditional and ten conditional epistatic minor QTLs were detected, from which only one pair (Qfw3A-10/Qfw6B-6) was identified by both unconditional and conditional QTL mapping, also independent of flour colour a* and b*. The major QTL (Qfw1D1-1) identified in the current study for the first time can be used for improving wheat FW in marker-assisted breeding.
Feeding a high concentrate (HC) diet is a widely used strategy for supporting high milk yields, yet it may cause certain metabolic disorders. This study aimed to investigate the changes in milk production and hepatic metabolism in goats fed different proportions of concentrate in the diet for 10 weeks. In total, 12 mid-lactating goats were randomly assigned to an HC diet (65% concentrate of dry matter, n=6) or a low concentrate (LC) diet (35% concentrate of dry matter, n=6). Compared with LC, HC goats produced greater amounts of volatile fatty acids and produced more milk and milk lactose, fat and protein (P<0.01). HC goats showed a greater concentration of ATP, NAD, plasma non-esterified fatty acids and hepatic triglycerides than LC goats (P<0.05). Real-time PCR results showed that messenger RNA (mRNA) expression of gluconeogenic genes, namely, glucose-6-phosphatase, pyruvate carboxylase and phosphoenolpyruvate carboxykinase were significantly up-regulated and accompanied greater gluconeogenic enzyme activities in the liver of HC goats. Moreover, the expression of hepatic lipogenic genes including sterol regulatory element-binding protein 1c, fatty acid synthase and diacylglycerol acyltransferase mRNA was also up-regulated by the HC diet (P<0.05). HC goats had greater hepatic phosphorylation of AMP-activated protein kinase than LC (P<0.05). Furthermore, histone-3-lysine-27-acetylation contributed to this elevation of gluconeogenic gene expression. These results indicate that lactating goats fed an HC diet for 10 weeks produced more milk, which was associated with up-regulated gene expression and enzyme activities involved in hepatic gluconeogenesis and lipogenesis.
There are six strains of the complete genomic sequences of black queen cell virus (BQCV) published in the GenBank, including South Africa (AF183905), South Korea (JX149531), Hungary 10 (EF517515), Poland 4 (EF517519), Poland 5 (EF517520) and Poland 6 (EF517521). Based on the six BQCV strains published in the GenBank, ten pairs of primers were designed in the present study using reverse transcription polymerase chain reaction to obtain the first complete genome sequence of a BQCV strain in China, called the BQCV China-JL1 strain (KP119603). A phylogenetic tree was then built to analyse their genetic relationships. The BQCV China-JL1 strain showed 86–93% similarity with the six strains published in the GenBank. The BQCV China-JL1 strain consisted of 8358 nucleotides (nt). The 5′-proximal open reading frame (ORF1) initiated at nt position 546 and terminated at nt position 4676, ORF3 initiated at nt position 4891 and terminated at nt position 5433, and the 3′-proximal ORF (ORF2) was located between nt positions 5750 and 8203.
The objective of this study was to determine if a moderate or high reduction of dietary CP, supplemented with indispensable amino acids (IAA), would affect growth, intestinal morphology and immunological parameters of pigs. A total of 40 barrows (initial BW=13.50±0.50 kg, 45±2 day of age) were used in a completely randomized block design, and allocated to four dietary treatments containing CP levels at 20.00%, 17.16%, 15.30% and 13.90%, respectively. Industrial AA were added to meet the IAA requirements of pigs. After 4-week feeding, blood and tissue samples were obtained from pigs. The results showed that reducing dietary CP level decreased average daily gain, plasma urea nitrogen concentration and relative organ weights of liver and pancreas (P<0.01), and increased feed conversion ratio (P<0.01). Pigs fed the 13.90% CP diet had significantly lower growth performance than that of pigs fed higher CP at 20.00%, 17.16% or 15.30%. Moreover, reducing dietary CP level decreased villous height in duodenum (P<0.01) and crypt depth in duodenum, jejunum and ileum (P<0.01). The reduction in the dietary CP level increased plasma concentrations of methionine, alanine (P<0.01) and lysine (P<0.05), and decreased arginine (P<0.05). Intriguingly, reducing dietary CP level from 20.00% to 13.90% resulted in a significant decrease in plasma concentration of IgG (P<0.05), percentage of CD3+T cells of the peripheral blood (P<0.01), also down-regulated the mRNA abundance of innate immunity-related genes on toll-like receptor 4, myeloid differentiation factor 88 (P<0.01) and nuclear factor kappa B (P<0.05) in the ileum. These results indicate that reducing dietary CP level from 20.00% to 15.30%, supplemented with IAA, had no significant effect on growth performance and had a limited effect on immunological parameters. However, a further reduction of dietary CP level up to 13.90% would lead to poor growth performance and organ development, associated with the modifications of intestinal morphology and immune function.
A fractional model generalized from the Zener model is proposed for the prediction of temperature-dependent free recovery behaviors of amorphous shape memory polymers (SMPs). This model differs from the Zener model in that it involves nonlinear differential equations of fractional, not integer, order. The theoretical solution based on this fractional model is utilized to simulate the isothermal and nonisothermal free recovery of an amorphous SMP compared with the one based on the Zener model. The results show a reasonable improvement in the prediction of the strain recovery response of SMP by the fractional calculus method.
Constitutive models based on fractional calculus are utilized to investigate the viscoelastic response of thermally activated shape memory polymers (SMPs). Fractional calculus-based viscoelastic equations are fitted to experimental data existing in literature compared with traditional viscoelastic models. In addition, a fractional rheology model is applied to simulate the isothermal recovery of an amorphous SMP. The fit results show a significant improvement in the description of the strain recovery response of SMP by the fractional calculus method.