Effect of relative humidity at chronic temperature on growth performance,glucose consumption,and mitochondrial ATP production of broilers

An experiment was conducted to investigate the effect of relative humidity(RH) at chronic temperature on growth performance, glucose consumption, and mitochondrial adenosine triphosphate(ATP) production of broilers. A total of 180 28-day-old Arbor Acres broilers(half males and half females) were randomly allocated to three treatments, each containing six replicates of 10 birds per treatment, using a completely randomized design. Birds were reared at 35, 60 or 85% RH at 32°C for 15 days(temperature increased by 3°C every 3 days from 20 to 32°C within 15 days: 20–23–26–29–32°C). RH affected(P0.05) average daily feed intake(ADFI), average daily gain(ADG), average daily water consumption(ADWC), blood glucose concentrations, muscle glycogen levels, avian uncoupling protein(av UCP) mRNA expression, and cytochrome c oxidase(CCO) activity in liver of broilers at 42 days of age. The 85% RH decreased(P0.05) ADFI, ADG and ADWC; 35% RH decreased(P0.02) ADG. Both 85 and 35% RH increased(P0.01) blood glucose and decreased(P0.05) muscle glycogen. Both 85 and 35% RH increased(P0.05) av UCP mRNA expression. 35% RH decreased(P0.05) CCO activity. In conclusion, both high and low RH inceased glucose consumption and reduced mitochondrial ATP poduction, leading to a decline in growth rate.     

Effects of dietary forage to concentrate ratio and wildrye length on nutrient intake,digestibility, plasma metabolites,ruminal fermentation and fecal microflora of male Chinese Holstein calves

Twenty-eight male, weaned Chinese Holstein calves((156.8±33.4) kg) were used to investigate the effects of dietary forage to concentrate ratio(F:C) and forage length on nutrient digestibility, plasma metabolites, ruminal fermentation, and fecal microflora. Animals were randomly allocated to four treatments in a 2×2 factorial arrangement: whole-length forage(WL) with low F:C(50:50); WL with high F:C(65:35); short-length forage(SL) with high F:C(65:35); and SL with low F:C(50:50). Chinese wildrye was used as the only forage source in this trial. The grass in the SL treatments was chopped using a chaff cutter to achieve small particle size(~50% particles 19 mm). Dry matter intake(DMI) and organic matter(OM) intake was increased by increasing both F:C(P0.01) and forage length(FL)(P0.05), while acid detergent fiber(ADF) and neutral detergent fiber(NDF) intakes were only increased by increasing the F:C(P0.01). The digestibility of NDF was increased as the FL increased(P0.01), and it was also affected by interaction between F:C and FL(P0.05). Cholesterol(CHO)(P0.01), leptin(LP)(P0.05), and growth hormone(GH)(P0.01) concentrations in plasma were increased as dietary F:C increased. A significant increase in plasma triglyceride(TG)(P0.01), insulin(INS)(P0.05), and GH(P0.01) levels was observed with decreasing dietary FL. Ruminal p H values of calves fed with low F:C diets were significantly lower than those in high F:C treatment(P0.05). Increasing the F:C enhanced ruminal acetic acid(P0.05) and acetic acid/propionic acid(P0.01). Fecal Lactobacillus content was significantly higher, while Escherichia coli and Salmonella contents were significantly lower in WL and high F:C groups(P0.05). Lower fecal scores(higher diarrhea rate) were observed in calves fed with SL hay compared to WL hay(P0.05). Denatured gradient gel electrophoresis(DGGE) bands and richness index(S) were significantly affected by the interaction between F:C and FL(P0.05), under high F:C, band numbers and richness index from WL group were higher than that from SL group(P0.05), whereas there were no differences between WL andSL groups under low F:C(P0.05). Microflora similarity was 50–73% among the different treatments. It is concluded that the WL with high F:C(65:35) diet is suitable for weaned calves.     

Grain dehydration rate is related to post-silking thermal time and ear characters in different maize hybrids

Mechanized grain harvest of maize becomes increasingly important with growing land plot size in Northeast China. Grain moisture is an important factor affecting the performance of mechanized grain harvest. However, it remains unclear what influences grain dehydration rate. In this study, maize grain dehydrating process was investigated in a two-year field experiment with five hybrids under two planting densities in 2017 and 2018. It was found that damaged-grain ratio was the main factor affecting mechanized harvest quality, and this ratio was positively correlated with grain moisture content at harvest (R²=0.6372, P<0.01). To fulfill the national standard of <5% damaged-grain ratio for mechanized grain harvest, the optimal maize grain moisture content was 22.3%. From silking to physiological maturity, grain dehydrating process was mostly dependent on the thermal time (growing degree days, GDDs) (r=–0.9412, P<0.01). The average grain moisture content at physiological maturity was 29.4%. Thereafter, the linear relationship between GDDs and grain moisture still existed, but the correlation coefficient became smaller (r=–0.8267, P<0.01). At this stage, grain dehydrating process was greatly affected by genotypes. Grain dehydrated faster when a hybrid has a smaller husk area (r=0.6591, P<0.05), larger ear angle (r=–0.7582, P<0.05), longer ear peduncle (r=–0.9356, P<0.01) and finer ear (r=0.9369, P<0.01). These parameters can be used for breeders and farmers to select hybrids suitable for mechanized grain harvest.     

Effects of Dietary Protein and Temperature on Growth and Flesh Quality of Songpu Mirror Carp

The study was conducted to demonstrate the effects of dietary protein and water temperature on growth and flesh quality of Songpu mirror carp （Cyprinus carpio） at an initial weight （165.24±5.08） g. Practical diets were formulated to contain five protein levels （29.12%, 31.46%, 34.49%, 38.17%, and 40.13%）, and each diet was randomly assigned triplicate groups of 15 fishes at three temperatures （18℃, 22℃, and 26℃） in the recirculation system. Fishes were fed twice daily to apparent satiation for 56 days. Results indicated that fishes had higher weight gain rate at 22℃ and 26℃ than that at 18℃ （P〈0.05）, but there were no significant differences between 22℃ and 26℃ （P〉0.05）. Based on the weight gain rate, dietary protein level 29.12% could meet the requirement of the body at 18℃, 22℃, or 26℃. Crude protein, crude lipid, moisture and ash of muscle had no significant differences among those treatments （P〉0.05）. pH （after 24 h） of muscle was the highest at 18℃ and the lowest at 22℃ （P〈0.05）, but no differences were observed among different protein level groups at each temperature （P〉0.05）. No significant differences on shear force, water holding capacity, collagen, glycogen and lactate among all the treatments were found （P〉0.05）. It was concluded that when C. carpio fed to apparent satiation, the growth mainly depended upon temperature. Dietary protein could not significantly affect flesh quality, but temperature significantly affected pH of muscles. A dietary protein level 29.12% could meet the requirement of the body at 18℃, 22℃, or 26℃.     

Effect of dioscorea opposite waste on growth performance,blood parameters,rumen fermentation and rumen bacterial community in weaned lambs

This study investigated the effects of dioscorea opposite waste (DOW) on the growth performance, blood parameters, rumen fermentation and rumen microbiota of weaned lambs. Sixty healthy weaned Small-Tailed Han lambs (male, (22.68±2.56) kg initially) were used as the experimental animals. Four levels of concentrate: 0 (control, CON), 10% (DOW1), 15% (DOW2) and 20% (DOW3), were replaced with DOW in the basal diet as experimental treatments. The results showed that lambs fed the DOW2 diet had a higher (P<0.05) dry matter intake (DMI) than the other groups. There was no significant difference (P>0.05) among DOW groups in average daily weight gain (ADG), and replacing concentrate with DOW linearly or quadratically increased (P<0.05) the ADG, while lambs fed the DOW2 diet showed greater (P<0.05) ADG than the CON group. The relative plasma concentration of growth hormone (GH), insulin like growth factor-1 (IGF-1) and insulin were affected by DOW, replacing concentrate with DOW linearly or quadratically (P<0.05) enhanced the plasma concentration of GH, IGF-1 and insulin, which was significantly higher (P<0.05) in the DOW2 group than in the CON, DOW1 and DOW3 groups. In addition, the DOW treatment showed a lower (P<0.05) concentration of blood urea nitrogen (BUN) than the CON group. Replacing concentrate with DOW quadratically decreased (P<0.05) the ruminal ammonia nitrogen (NH₃-N) and increased (P<0.05) the total of volatile fatty acids (TVFAs) at 0 and 4 h after feeding as well as linearly decreased (P<0.05) the NH₃-N at 8 h after feeding. Replacing concentrate with DOW linearly decreased (P<0.05) the propionate and increased the aceate before feeding, and linearly decreased (P<0.05) propionate and quadratically increased (P<0.05) the aceate at 4 and 8 h after feeding. Lambs fed the DOW2 diet increased the phylum Firmicutes and genera Succiniclasticum and Ruminococcus_1 groups, whereas decreased (P<0.05) the relative abundance of phylum Deferribacteres and genera intestinimonas and Ruminiclostridium. In summary, replacing the concentrate with 15% DOW was beneficial for improving the rumen fermentation and ADG by increasing the DMI and modulating the rumen microbial community.     

Effect of dietary supplementation with mulberry (Morus alba L.) leaves on the growth performance,meat quality and antioxidative capacity of finishing pigs

The present study was conducted to evaluate the effect of dietary mulberry(Morus alba L.) leaves powder(MLP) supplementation on meat quality of finishing pigs.A total of 40 Duroc×Landrace×Yorkshire pigs(initial body weight of(40.5±0.63) kg) were randomly allotted into two treatments,fed either with control diet or 15% MLP diet for 85 d.The results showed that MLP diet decreased(P≤0.05) average daily gain(ADG) and increased(P0.05) feed/gain ratio(F/G) in the finishing and whole period.MLP diet also decreased(P0.05) carcass weight,dressing percentage,last rib and average backfat depth.However,MLP diet increased(P0.05) intramuscular fat(IMF) content,decreased(P0.05) shear force,cooking loss and drip loss.In addition,MLP diet increased(P0.05) total antioxidative capacity,glutathione peroxidase and tended(P0.10) to increase total superoxide dismutase in serum.In longissimus thoracis,myosin heavy chain(MyHC) I and IIa mRNA levels were increased(P≤0.05) for MLP diet.In conclusion,15% MLP supplementation reduced the growth performance and carcass traits,but improved meat quality of finishing pigs possibly through the change of myofiber characteristics,enhancement of antioxidative capacity and increase of IMF.     

大豆异黄酮和抗生素对文昌鸡生长性能、肉品质和血浆抗氧化指标的影响

【目的】对比大豆异黄酮和抗生素对文昌鸡生长性能、肉品质及血浆抗氧化指标的影响,为大豆异黄酮在文昌鸡生产中的实际应用提供参考。【方法】选用1日龄文昌鸡母雏540只。试验分3组,包括空白对照组、抗生素组和大豆异黄酮组,每组4个重复,每个重复45羽。分小鸡、中鸡和大鸡3个阶段进行试验。【结果】生长性能结果显示,1~30日龄小鸡各组之间的平均日增质量、平均日采食量和料重比均无显著差异(P0.05);31~80日龄阶段,大豆异黄酮组的平均日增质量显著高于对照组(P0.05);81~120日龄阶段,大豆异黄酮组的平均日采食量显著高于对照组(P0.05)。肉品质结果显示,各组的肉色L*、a*、b*和pH差异均不显著(P0.05);与对照组相比,大豆异黄酮组的熟肉率显著提高(P0.05)。血浆抗氧化指标结果显示,31~80日龄阶段,大豆异黄酮组的总抗氧化能力(Total antioxidant capacity,T-AOC)活性显著高于抗生素组(P0.05);81~120日龄阶段,大豆异黄酮组的肌酸激酶(Creatine kinase,CK)活性显著低于抗生素组,谷胱甘肽(Glutathione,GSH)含量显著高于抗生素组,抗生素组和大豆异黄酮组的T-AOC活性显著高于对照组(P0.05)。【结论】中鸡阶段饲粮中添加20 mg·kg~(–1)大豆异黄酮可以提高鸡日增质量和抗氧化水平;大鸡阶段饲粮中添加15 mg·kg~(–1)大豆异黄酮能够提高鸡采食量、熟肉率和抗氧化水平。     

Characterization of T-complex polypeptide 1 (TCP-1) from the Chilo suppressalis HSP60 family and its expression in response to temperature stress

Many proteins require assistance from molecular chaperones at various stages to attain correctly folded states and functional conformations during protein synthesis. In this study, the gene encoding T-complex polypeptide 1(TCP-1), which belongs to the heat shock protein 60(HSP60) family, was isolated and characterized from the rice stem borer, Chilo suppressalis, by RACE and q PCR, respectively. The full-length c DNA of Tcp-1 was 2 144 bp and encoded a 1 635-bp ORF; the deduced translational product contained 545 amino acids with 5′-and 3′-UTRs and an isoelectric point of 5.29. Cluster analysis confirmed that the deduced amino acid sequence shared high identity(60–99%) with TCP-1 from other insects. To investigate Tcp-1 expression in response to abiotic stress, q PCR was used to analyze expression levels of Tcp-1 m RNA in C. suppressalis larvae exposed to temperatures ranging from –11 to 43°C. With respect to heat shock, Tcp-1 expression was higher than the control after a 2-h exposure to 30 and 36°C and declined at 39 and 43°C. Difference in Tcp-1 expression was observed at temperatures ranging from –11 to 27°C. q PCR analyses revealed that Tcp-1 expression was the highest in hindgut tissue as compared to heads, epidermis, fat body, foregut, midgut, and malpighian tubules. Our results indicated that Tcp-1 expression was differentially expressed in C. suppressalis tissues, and was impacted by temperature stress.     

Cold plasma promotes Sertoli cell proliferation via AMPK–mTOR signaling pathway

This study investigated cold plasmas for multiple biological applications. Our previous work has found dielectric barrier discharge plasma improves chicken sperm quality. The number of Sertoli cells (SCs) decides spermatogenesis. However, whether cold plasma can regulate SC proliferation remains unclear. This study explored the effects of cold plasma on immature chicken SC proliferation and the regulation mechanism. Results showed that cold plasma exposure at 2.4 W for 30 s twice with an interval of 6 h produced (P<0.05) the maximum SC viability, cell growth, and cell cycle progression. SC proliferation-promoting effect of cold plasma treatment was regulated by increasing (P<0.05) the adenosine triphosphate production and the respiratory enzyme activity in the mitochondria. This process was potentially mediated by the adenosine monophosphate-activated protein kinase (AMPK)–mammalian target of rapamycin (mTOR) signaling pathway, which was regulated by the microRNA (miRNA) targeting regulation directly and by the intracellular reactive oxygen species homeostasis indirectly. The cold plasma treatment increased (P<0.01) the miR-7450-5p expression and led to a decreased (P<0.01) AMPKα1 level. On the other hand, miR-100-5p expression was reduced (P<0.05) and led to an increased (P<0.05) mTOR level in SCs. A single-stranded synthetic miR-7450-5p antagomir and a double-stranded synthetic miR-100-5p agomir reduced (P<0.05) the SC proliferation. However, this could be ameliorated (P<0.05) by the cold plasma treatment. Our findings suggest that appropriate cold plasma treatment provides a safe strategy to improve SC proliferation, which is beneficial to elevating male chicken reproductive capacity.     

Analysis of DNA methylation of CD79B in MDV-infected chicken spleen

Marek's disease (MD), an immunosuppressive disease induced by Marek's disease virus (MDV), provides an ideal model for studying diseases caused by a carcinogenic virus. CD79B is a B-cell antigen receptor complex-associated protein β-chain precursor which is involved in the activation, proliferation, differentiation of B-cell and the transmission of downstream signals. This study analyzed CD79B gene mRNA expression and methylation by two schemes #20 (5´ flanking to intron 1) and #27 (intron 2 to intron 3), between MDV-infected tumorous spleens (TS) and non-infected spleens (NS). Results showed that average methylation levels of CpGs in #20 and #27 were higher in TS than in NS (P<0.05), while, CD79B mRNA expression was lower in TS than in NS (P<0.01). Six of 40 CpG sites showed significantly (P<0.05) different methylation levels between TS and NS. Correlation analysis showed that the average methylation level rather than a single site methylation level in #20 affected (P<0.05) mRNA expression. Collectively, it was found that the change of CD79B gene expression after MDV infection might be partly explained by modification of DNA methylation.     

Effects of Bupleurum extract on blood metabolism,antioxidant status and immune function in heat-stressed dairy cows

This experiment was conducted to evaluate the effects of Bupleurum extract(BE) on blood metabolites, antioxidant status, and immune function in dairy cows under heat stress. Forty lactating Holstein cows were randomly assigned to 1 of 4 treatments. The treatments consisted of 0, 0.25, 0.5, and 1.0 g of BE kg–1 dry matter. Supplementation with BE decreased(P0.05) blood urea nitrogen(BUN) contents and increased blood total protein(TP) and albumin(ALB) levels compared with control cows, but it had no effects(P0.05) on blood glucose(GLU), nonesterified fatty acid(NEFA), total triglyceride(TG), low-density lipoprotein cholesterol(LDL-C), and high density lipoprotein cholesterol(HDL-C). Compared with control cows, cows fed BE had higher(P0.05) superoxide dismutase(SOD) and glutathione peroxidase(GSH-Px) activity. However, supplementation with BE had no effect(P0.05) on total antioxidant capacity(T-AOC) or malondialdehyde(MDA) levels. The immunoglobulin(Ig) A and G contents increased(P0.05) in cows fed 0.25 or 0.5 g of BE kg–1. Interleukin(IL)-2 and IL-4 levels were higher(P0.05) in cows fed 0.5 and 1.0 g of BE kg–1, and IL-6 was significantly elevated(P0.05) in cows fed 0.5 g of BE kg–1. There were no treatment effects(P0.05) on the CD4+ and CD8+ T lymphocyte ratios, CD4+/CD8+ ratio, or tumor necrosis factor-α(TNF-α) level among the groups. These findings suggest that BE supplementation may improve protein metabolism, in addition to enhancing antioxidant activity and immune function in heat-stressed dairy cows.     

The toxicological effect of dietary excess of saccharicterpenin,the extract of camellia seed meal,in piglets

Recently, saccharicterpenin extracted from Camellia oleifera seed meal has become a widely used feed supplement in animal husbandry. In order to assess its safety, this study was designed to investigate the toxicity and histopathological effects of saccharicterpenin on piglets. One hundred-fifty weaned pigs((Yorkshire×Landrace)×Duroc), 75 males and 75 females with body weight(BW) of(7.35±0.29) kg, were randomly allotted to groups receiving diets supplemented with 0, 500, 1 000, 2 500 or 5 000 mg kg~(–1) saccharicterpenin for 70 d. The diet with 500 mg kg~(–1) saccharicterpenin supplementation improved liver glutathione peroxidase(GSH-Px) activity, and the diet with 1 000 mg kg~(–1) saccharicterpenin supplementation improved liver glutathione S-transferase(GSH-S) activity in piglets on d 70(P0.05). At 2 500 mg kg~(–1), saccharicterpenin in the diets reduced average daily feed intake(ADFI) and average daily gain(ADG) of piglets from d 1 to 35, damaged the cardiac tissue and liver on d 35, and decreased white blood cell counts(WBC), activities of catalase(CAT) and GSH-Px, and concentrations of glucose(GLU) and urea nitrogen(BUN) in the blood of piglets on d 70(P0.05). In addition, diets with 5 000 mg kg~(–1) saccharicterpenin supplementation reduced ADFI, ADG and increased diarrhea rates of piglets from d 36 to 70, and decreased hemoglobin(HGB) concentration and activity of CAT in the blood of piglets on d 70(P0.05). Moreover, at a rate of 5 000 mg kg~(–1), saccharicterpenin supplementation increased pancreas index on d 35 and hepatic index on d 70, and damaged cardiac tissue, liver and spleen during the whole experimental period(P0.05). These results suggested that dietary 500 mg kg~(–1) saccharicterpenin supplementation had beneficial effects on piglets, but excessive supplementation(2 500 or 5 000 mg kg~(–1)) of saccharicterpenin in the diets could lead to growth retardation, hematological abnormalities and organ injuries.     

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 m L 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(P0.05) the embryonic mortalities, and decreased(P0.05) hatchabilities compared to the non-injected control, but no differences(P0.05) 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 Zn SO4·7 H2 O in a 0.1-m L solution, or non-injection(the positive control), respectively at E9–10. The results from the experiment 2 demonstrated that no differences(P0.05) 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(P0.05) the embryonic mortality, and injections of 150 and 200 μg Zn/egg decreased(P0.05) hatchabilities compared with the controls. The embryonic tibia Zn contents at E20 were increased(P0.05) by injections of 150, 200 and 250 μg Zn/egg. Zinc injection did not affect(P0.05) malonaldehyde(MDA) contents, copper-and Zncontaining superoxide dismutase(Cu Zn SOD) activities and m RNA 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(P0.05) the metallothionein(MT) mR NA 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 m RNA expression levels in the embryonic liver at E20, however, injections of 150–200 μg Zn/egg were harmful to the embryonic development.     

Knockdown of the Meq gene in Marek's disease tumor cell line MSB1 might induce cell apoptosis and inhibit cell proliferation and invasion

Marek's disease (MD), a highly cell-associated and contagious disease of chickens caused by Marek's disease virus (MDV) can result in neural lesions, immunosuppression and neoplasia in chicken. The Meq gene is an important oncogene in the MDV genome, and it is expressed highly in MD tumor tissues and MD T-lymphoblastoid cell lines. An experiment was conducted to elucidate the role of Meq in MD tumor transformation. RNA interference technology was used to block its expression, and then analyzed the biological effects of Meq knockdown on the MD tumor cell line MSB1. A small interfering RNA with an interference efficiency of 70% (P<0.01) was transfected into MSB1 cells to knock down the expression of Meq gene. The cell proliferation, cycle and apoptosis were detected post-Meq knockdown. The results showed that MSB1 cell proliferation was downregulated remarkably at 48 h (P<0.01), 60 h (P<0.05) and 72 h (P<0.01) post-Meq knockdown. The cell cycle was unaffected (P>0.05). B-cell lymphoma 2 gene (BCL2) was anti-apoptotic and caspase-6 was the effector in the apoptosis pathway. The activity of caspase-6 was upregulated (P<0.05) significantly and BCL2 gene expression was downregulated (P<0.05) significantly post-Meq knockdown, suggesting cell apoptosis might be induced. MSB1 cell migration did not exhibit any obvious change (P>0.05) post-Meq knockdown, but the expression of two genes (matrix metalloproteinase 2 (MMP2) and MMP9) that are correlated closely to cell invasion was downregulated (P<0.05) remarkably post-Meq knockdown. The Meq knockdown might affect the main features of tumorous cells, including proliferation, apoptosis, and invasion, suggesting that the Meq gene might play a crucial role in interfering with lymphomatous cell transformation.     

The comparative analysis and identification of secondary metabolites between Tibet wild and cultivated pomegranates (Punica granatum L.) in China

Secondary metabolites are closely related to the nutritional quality and health functions of plants. We investigated the secondary metabolites of both wild (n=23) and cultivated (n=27) pomegranate plants (Punica granatum L.) growing in China. The total flavonoid (TF) and tannin (TT) contents from the peel and juice were determined and the secondary metabolites in the peel (ZLP) and juice (ZLZ) of ‘Zela 4’ were identified using liquid chromatography–electrospray ionization tandem mass spectrometry (LC–ESI-MS/MS). Analysis of variance (P<0.05) showed that there were significant differences in the TF content of peel (TF (P)) and juice (TF (J)), and the TT content of peel (TT (P)) and juice (TT (J)) among different pomegranate accessions. Pearson correlation analysis showed that latitude and altitude might be the main environmental factors affecting TF and TT contents in pomegranates. In this study, 279 secondary metabolites were identified in the ZLP and ZLZ. In addition, we report for the first time 227 secondary metabolites in pomegranates. Using orthogonal partial least squares discriminant analysis, 90 differential metabolites were identified in ZLP and ZLZ. In addition, we screened eight specific germplasms (high-TF (P), ‘Junyong 3’; low-TF (P), ‘Yanzhihong’; high-TF(J), ‘Zela 4’; low-TF (J), ‘Yudazi’, high-TT (P), ‘Junyong 4’; low-TT (P), ‘Anba 1’; high-TT(J), ‘Yeba 1’; and low-TT (J), ‘Baihuayushizi’). The results of our study provide a reference for the development and utilization of wild pomegranate resources and pomegranate breeding in China.     

miR-34c inhibits proliferation and enhances apoptosis in immature porcine Sertoli cells by targeting the SMAD7 gene

MicroRNAs(miRNAs) are implicated in swine spermatogenesis via their regulations of cell proliferation, apoptosis, and differentiation. Recent studies indicated that miR-34 c is indispensable in the late steps of spermatogenesis. However, whether miR-34 c plays similar important roles in immature porcine Sertoli cells remain unknown. In the present study, we conducted two experiments using a completely randomised design to study the function roles of miR-34 c. The results from experiment I demonstrated that the relative expression level of miR-34 c in swine testicular tissues increased(P=0.0017) quadratically with increasing age, while the relative expression level of SMAD family member 7(SMAD7) decreased(P=0.0009) with curve. Furthermore, miR-34 c expression levels showed a significant negative correlation(P=0.013) with SMAD7 gene expression levels. The results from experiment II indicated that miR-34 c directly targets the SMAD7 gene using a luciferase reporter assay, and suppresses(P0.05) SMAD7 mRNA and protein expressions in immature porcine Sertoli cells. Overexpression of miR-34 c inhibited(P0.05) proliferation and enhanced(P0.05) apoptosis in the immature porcine Sertoli cells, which was supported by the results from the Cell Counting Kit-8(CCK-8) assay, the 5-Ethynyl-2′-deoxyuridine(EdU) assay, and the Annexin V-FITC/PI staining assay. Furthermore, knockdown of SMAD7 via small interfering RNA(siR NA) gave a similar result. It is concluded that miR-34 c inhibits proliferation and enhances apoptosis in immature porcine Sertoli cells by targeting the SMAD7 gene.     

Evaluation of a new method for quantification of heat tolerance in different wheat cultivars

Heat stress seriously affects wheat production in many regions of the world. At present, heat tolerance research remains one of the least understood fields in wheat genetics and breeding and there is a lack of effective methods to quantify heat stress and heat tolerance in different wheat cultivars. The objective of this study was to use various wheat cultivars to evaluate stress intensity(δ) and a new method for quantification of heat tolerance and compare this technique with three other currently utilized methods. This new parameter for heat tolerance quantification is referred to as the heat tolerance index(HTI) and is an indicator of both yield potential and yield stability. Heat treatments were applied in a controlled setting when anthesis had been reached for 80% of the wheat. The stress intensity evaluation indicated heat shock was the main factor associated with kernel weight reduction while grain yield reduction was mainly associated with chronic high temperature. The methods evaluation showed that a temperature difference of 5°C from natural temperatures was a suitable heat treatment to compare to the untreated controls. HTI was positively correlated with yield under heat stress(r=0.8657, δ_(2010)=0.15, in 2009–2010; r=0.8418, δ_(2011)=0.20, in 2010–2011; P0.01), and negatively correlated with yield reduction rate(r=–0.8344, in 2009–2010; r=–0.7158, in 2010–2011; P0.01). The results of this study validated the use of HTI and temperature difference control for quantifying wheat heat tolerance that included the yield potential and the stability of different wheat cultivars under heat stress. Additionally, 10 wheat cultivars showed high HTI and should be further tested for their heat confirming characteristics for use in wheat heat tolerance breeding.     

Comparison of forage yield,silage fermentative quality,anthocyanin stability,antioxidant activity,and in vitro rumen fermentation of anthocyanin-rich purple corn (Zea mays L.) stover and sticky corn stover

The objective of this study was to observe the forage yield, silage fermentative quality, anthocyanin stability, and antioxidant activity during the storage period and in vitro rumen fermentation of anthocyanin-rich purple corn (Zea mays L.) stover (PS) and sticky corn stover (SS). Forage yield of corn stover was weighed and ensiled with two treatments: (1) hybrid sticky waxy corn stover (control), and (2) hybrid purple waxy corn stover (treatment). Samples were stored in mini-silos for periods of 0, 7, 14, 21, 42, 63, 84, and 105 d. The results showed that PS had significantly higher (P<0.05) yields of dry matter (DM), organic matter (OM), gross energy (GE), crude protein (CP), neutral detergent fiber (NDF), acid detergent fiber (ADF), and total anthocyanins than that of the SS. Anthocyanin-rich purple corn stover silage (PSS) showed higher (P<0.05) levels of DM and CP relative to the sticky corn stover silage (SSS). Although anthocyanin-rich PSS displayed a lower (P<0.05) level of pelargonidin-3-glucoside (P3G), it had higher (P<0.05) levels of peonidin (Peo) and pelargonidin (Pel) compared to the control. Delphinidin (Del) and malvidin (Mal) were not detected in SSS during the ensilage period; in PSS, Del was no longer detected after 7 d of ensilage. Specifically, total anthocyanins in anthocyanin-rich PSS decreased rapidly (P<0.05) prior to 7 d of ensilage, and then remained at relatively stable (P>0.05) constants. Compared to the anthocyanin-rich PSS, SSS displayed significantly higher (P<0.05) pH value and ammonia nitrogen (NH₃-N) content. Propionic acid (PA) at 0 d and butyric acid (BA) during the entire study period were not detected, whereas anthocyanin-rich PSS showed a higher (P<0.05) level of lactic acid (LA) than that of the SSS. Compared with the SSS extract, anthocyanin-rich PSS extract showed a higher (P<0.05) level of 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity and displayed a lower (P<0.05) half maximal inhibitory concentration (IC₅₀) value. Moreover, anthocyanin-rich PSS reduced (P<0.05) gas production (GP), and displayed lower levels of immediately soluble fraction and ratio of acetic acid (AA) to PA at 12 h, but the other parameters were unaffected (P>0.05) relative to the control. Taken together, the results indicated that: (1) anthocyanins could be stable in silage; (2) anthocyanin-rich PSS showed better silage fermentative quality and stronger antioxidant activity; and (3) anthocyanin-rich PSS had no negative effect on rumen fermentation parameters.