2个鲤群体(Cyprinus carpio L.)表型生长性状的AI测量与手工测量的相关性分析

鱼类形态是人工育种、功能基因定位以及水产养殖和生态学研究的重要种群资源。鲤(Cyprinus carpio L.)是中国重要的养殖淡水鱼类之一,鲤的自然和人工选择导致不同品种间的表型极具多样性。近年来,随着成像技术、计算能力和硬件设备的飞速发展,基于机器视觉的无损测量方法逐渐成为一种高效、可重复批量检测鱼体的方法。本研究旨在检验AI测量对鱼类表型性状测量的准确性。本文选择了表型指数差异显著的元江鲤(Cyprinus carpio yuankiang)和金背鲤(Cyprinus carpio var.Jinbei)2个群体,共计204尾鱼进行研究。针对全长、体长、体高、体厚、头长、尾柄长、尾柄高、吻长、眼径和眼间距共10种线性、圆形和空间性状进行手工测量和AI测量,并进行了比较分析。其结果显示:(1)元江鲤和金背鲤2个群体中,针对10种表型性状的测量,相较于手工测量,AI测量结果整体偏大;在表型指数中,体宽指数(体厚/体长),头长指数(头长/体长)和尾鳍指数(尾柄长/体长)2种测量方法均无显著差异,而2种方法在体深指数(体高/体长)结果中,呈现显著差异。(2)元江鲤和金背鲤群体中,对2种测量方法进行相关性和一致性分析,发现针对全长、体长、体高、体厚、头长和眼间距共6种性状,2种方法呈现高度一致性(r>0.85);(3)2种测量方法重复性测量的一致性分析中,发现手工测量和AI测量均呈现较好的一致性(r>0.85),但手工测量的一致性要略高于AI测量。综上所述,AI测量准确度的提升能够加快全长等线性表型性状测量的工作效率和选择育种速度,但对于圆形和空间等表型性状测量的准确度,还需进一步提升。

Ontogeny of Immune Related Organs during Early Development of Carp (Cyprinus Carpio L.)

In this paper, ontogeny of immune-related organs during theearly development of carp was studied by histochemical technique-lcineBlue staining under acid conditions and PAS reaction(AB-PAS staining). Thekidney appeared one day before hatching, spleen, liver and pancreas emergedon the same day of hatching, and thymus was not found until the third day afterhatching. Ontogeny of these immune related organs of the carp in thisresearch is earlier than that reported by Botham.

复方中草药对镜鲤(Cyprinus carpio L.)血清转氨酶及红细胞抗氧化酶活性的影响

在基础日粮中添加2%由不同比例的刺五加、枸杞子、金银花和黄芪配伍的复方中草药饲喂镜鲤（Cyprinus carpio L）.,于第14天、第28天及第42天检测红细胞的过氧化氢酶（CAT）、超氧化物岐化酶（SOD）、血清的谷草转氨酶（GOT）以及谷丙转氨酶（GPT）活性,以研究复方中草药对镜鲤的毒副作用。在28及42d时,复方Ⅰ组红细胞CAT活性与对照组差异显著（P〈0.05）;28d时,各复方组红细胞SOD活性均与对照组差异极显著（P〈0.01）;28d时,复方Ⅱ组血清GOT活性及复方Ⅲ组血清GPT活性与对照组差异极显著（P〈0.05）;42d时,各复方组血清GPT活性与对照组差异极显著（P〈0.01）。结果表明,试验复方中药添加到镜鲤的饲料中,可以增强其血清转氨酶及红细胞抗氧化酶活性,其中复方Ⅰ效果最佳,建议用药时间为28d。

不同温度下不同蛋白水平对镜鲤(Cyprinus carpio L.)非特异性免疫的影响

选用德国镜鲤630尾(200±20)g,在18、22和26℃3个温度环境下用30%、33%、36%、39%、42%的五个不同的蛋白水平的饲料投喂,分别测定肝脏、脾脏中的超氧化物歧化酶(SOD)、丙二醛(MDA)、溶菌酶(LYZ)、血清中的碱性磷酸酶(ALP)和补体C3、C4等免疫指标。结果表明,18℃环境下,42%蛋白水平组肝脏SOD活性显著高于其他组(P<0.05);肝脏MDA含量随饲料蛋白水平的增加而降低;30%蛋白水平组脾脏溶菌酶的活性显著高于39%蛋白水平组(P<0.05)。22℃环境下,42%蛋白水平组肝脏SOD活性显著高于其他蛋白水平组(P<0.05);42%蛋白水平组肝脏MDA含量显著低于其他蛋白水平组(P<0.05);42%蛋白水平组血清C3显著高于30%蛋白水平组(P<0.05)。26℃环境下,肝脏SOD活性随饲料蛋白水平的增加而增加,但差异不显著(P>0.05);42%蛋白水平组肝脏MDA含量显著低于其他组(P<0.05)。结果显示,本试验条件下,在18、22和26℃温度下蛋白水平升高可在一定程度上提高鲤鱼的非特异性免疫,主要表现在抗氧化性方面,而对其他的非特异性免疫指标无显著的影响。

鲤鱼(Cyprinus carpio L.)体重和体长QTL的定位

利用265个AFLP标记、127个微卫星分子标记、37个EST-SSR标记和16个RAPD标记(共445个标记)对大头鲤/荷包红鲤抗寒品系的F2代雌核发育群体44个个体进行基因型检测,构建鲤鱼(Cyprinus carpio L.)遗传连锁图谱;利用软件WinQTLCart2.5采用复合区间作图法对体重和体长两个性状进行了QTL定位分析。结果共检测到与两个与体重性状相关的QTL,分别定位到LG24连锁群(qBWh-24-1)和LG20连锁群(qBWh-20-1)上,可解释的表型变异分别为7.98%和20.05%;检测到两个与体长性状相关的QTL,分别位于LG2连锁群(qFS-2-1)和LG20连锁群(qFS-20-1),可解释的表型变异分别为5.69%和12.69%,4个QTL的加性效应均为负值。

The Use of Cyanobacteria Arthrospira platensis and Cladoceran Daphnia magna as Complementary Protein and Lipid Sources in Transitional Diet for Common Carp (Cyprinus carpio L.) Nursery

This study was conducted to evaluate the use of cyanobacteria Arthrospira platensis and cladoceran Daphnia magna biomass as complementary protein and lipid sources in diet supplementation for common carp (Cyprinus carpio L.) nursery. Three experimental diets containing A. platensis and D. magna meal at different concentrations were compared to the commercial (control) diet. Each experimental diet (ED) was set to contain D. magna and A. platensis meal at a specific combination: 2% and 5%, 2% and 7%, and 4% and 5% for ED1, ED2 and ED3, respectively. The protein and lipid content of the experimental diets ranged from 43.20% to 44.60% dry weight (DW) and 10.64% to 13.42% DW, respectively;while the protein and lipid content of the control diet were 43.00% DW and 6.72% DW, respectively. After 20 days of feeding period, ED3 group obtained the highest final body weight (BW) (58.18 ± 35.24 mg), total biomass (1936 ± 1625 mg), food conversion rate (1.34 ± 0.04), and specific growth rate (12.86 ± 0.03% BW day–1) among all treatment groups (P > 0.05);while ED1 group obtained the highest survival (75.5% ± 7.47%) among all treatment groups (P > 0.05). The total bacterial count and total pathogenic Aeromonas sp. in the culture water of the treatment groups (ranged from 2.00 to 2.65 × 105 CFU mL–1 and 3.50 to 4.12 × 103 CFU mL–1, respectively) were lower compared to the water of the control group fed (3.73 × 105 and 4.70 × 103 CFU mL–1, respectively). No significant differences in physicochemical water quality parameters were observed among treatments (P > 0.05). The current study suggests that the combination of A. platensis and D. magna biomass can be used as complementary protein and lipid sources in diet supplementation for common carp larvae and can result in a comparable fish growth, survival and feed utilization in common carp culture at the nursery phase.

Intercross population study reveals that co-mutation of mitfa genes in two subgenomes induces red skin color in common carp(Cyprinus carpio wuyuanensis)

Common carp are among the oldest domesticated fish in the world.As such,there are many food and ornamental carp strains with abundant phenotypic variations due to natural and artificial selection.Hebao red carp(HB,Cyprinus carpio wuyuanensis),an indigenous strain in China,is renowned for its unique body morphology and reddish skin.To reveal the genetic basis underlying the distinct skin color of HB,we constructed an improved highfidelity(HiFi) HB genome with good contiguity,completeness,and correctness.Genome structure comparison was conducted between HB and a representative wild strain,Yellow River carp(YR,C.carpio haematopterus),to identify structural variants and genes under positive selection.Signatures of artificial selection during domestication were identified in HB and YR populations,while phenotype mapping was performed in a segregating population generated by HB×YR crosses.Body color in HB was associated with regions with fixed mutations.The simultaneous mutation and superposition of a pair of homologous genes(mitfa) in chromosomes A06 and B06 conferred the reddish color in domesticated HB.Transcriptome analysis of common carp with different alleles of the mitfa mutation confirmed that gene duplication can buffer the deleterious effects of mutation in allotetraploids.This study provides new insights into genotype-phenotype associations in allotetraploid species and lays a foundation for future breeding of common carp.

Pharmacokinetics of Enrofloxacin and Its Metabolite in Carp (Cyprinus carpio) After a Single Oral Administration in Medicated Feed

A precise and reliable analytical method of high performance liquid chromatography-tandem mass spectrometry(HPLCMS/MS)was developed to measure trace levels of enrofloxacin(ENR)and its major metabolite ciprofloxacin(CIP)in carp tissues.Optimized chromatographic separation was obtained on a Waters Xterra MS C_(18) reversed-phase column using gradient elution with methanol and 0.1%formic acid aqueous solution including 5mmolL^(-1) of ammonium acetate.The established method was applied to study the pharmacokinetics and distribution of ENR and CIP in tissues of carp following a single oral administration in feed at a dosage of 40mgkg^(-1) bw(body weight).Data were analyzed using DAS 2.0 dynamics software,and the experimental results suggest that ENR was rapidly absorbed and extensively distributed in carp tissues through systemic circulation,and the pharmacokinetic characteristics can be described with a two-compartment model.The elimination half-lives(t_(1/2β))from muscle,liver,gill,plasma and skin were 131,160,104,132 and 310 h,respectively.The areas under the drug concentration-time curves(AUC)for these tissues were 491,972,750,249 and 706hmgkg^(-1),respectively.The maximum concentration(C_(max))values were 13,29,37,9 and 5mgkg^(-1) with peak times(t_(max))of 8,4,4,2 and 4 h,respectively.Ciprofloxacin,the active metabolite of ENR,was also detected in carp tissues,indicating that only 1.54%of de-ethylation of ENR occurs in carp.At a water temperature of 18℃,the drug withdrawal time was determined to be no less than 24 d while the carp was fed at a single dosage of 40mgkg^(-1).

Effects of BDE-209 exposure on growth performance, intestinal digestive enzymes, and intestinal microbiome in common carp (Cyprinus carpio L.)

Polybrominated diphenyl ether has become a widespread environmental pollutant in aquatic environments.BDE-209,a prominent PBDEs found in the environment and wildlife,has been proven to cause oxidative stress,immune damage and endocrine system disorders in fish.However,the effects of BDE-209 on the intestinal health of fish remain unclear.This research aimed to determine the effects of BDE-209 on the growth performance,digestive enzymes activities and intestinal microbiota of common carp.After 60-day exposure,the growth performance of common carp were remarkably decreased(P<0.05).Moreover,the levels of protease,amylase and lipase activities remarkably decreased by BDE-209 exposure(P<0.05).In addition,metagenomics analysis revealed that the alpha-diversity values of intestinal microbiota were significantly decreased in BDE-209 exposure groups(P<0.05).The results also indicated that BDE-209 exposure could alter the intestinal microbial compositions.At the phylum level,the proportion of Fusobacteria was markedly reduced(P<0.05),while the proportions of Bacteroidetes and Firmicutes were significantly increased(P<0.05)along with the increasing concentrations of BDE-209.At the genus level,the proportion of Shewanella was higher(P<0.05)in BDE-209 exposure groups than in control group,while the abundance of Cetobacterium showed an opposite trend.Furthermore,the proportions of some aquatic probiotics(such as Bacillus,Rhodopseudomonas and Lactobacillus)were significantly enriched(P<0.05)in control group,and the content of short-chain fatty acid producing bacterium(such as Faecalibacterium,Bacteroides and Clostridium)were decreased along with the increasing BDE-209 concentrations,but no significant differences were found among the four groups(P>0.05).Overall,exposure to BDE-209 could alter the intestinal microbiota of common carp,which may affect their growth and threaten their health.

镜鲤(Cyprinus carpio specularis)肌肉营养成分分析及价值评定

为了研究德国镜鲤肌肉营养成分和营养价值。试验按体重规格40-50g/尾和220-230g/尾随机抽样,各规格抽取8尾。试验结果表明:40-50g与220-230g镜鲤肌肉鲜样中粗蛋白含量分别为16.98%,16.76%,粗脂肪分别为0.60%,1.04%。肌肉干物质中必需氨基酸总量分别为34.34%,36.17%,必需氨基酸与氨基酸总量之比分别为68.86%,71.23%,四种呈味氨基酸总量分别为25.75%,25.46%,呈味氨基酸与氨基酸总量之比分别为37.47%,37.16%,支/芳分别为2.26,2.29,必需氨基酸指数分别为77.44,85.34。基本符合WHO/FAO标准,除了赖氨酸比例偏高,其余氨基酸比例较接近鸡蛋蛋白氨基酸模式。

镉污染对鲤鱼(Cyprinus carpio Linn.)同工酶酶谱的影响

应用聚丙烯酰胺凝胶电泳和酶活性测定技术，比较和分析了饲养于不同浓度镉液中的鲤鱼在镉含量达到积累－释放平衡后，其肌肉和肝脏组织的酯酶同工酶、碱性磷酸酶同工酶及过氧化物酶同工酶酶谱。结果显示，镉的残留量使鲤鱼肌肉和肝脏组织酯酶同工酶和碱性磷酸酶同工酶谱带减少，活性减弱；过氧化物酶同工酶谱带增多，活性增强。

复方中草药对镜鲤(Cyprinus carpio var. specularis)幼鱼非特异性免疫功能的影响

在基础日粮(对照组)中分别添加2%由不同比例的刺五加(Acanthopanax senticosus Harms)、枸杞子(L.chinense Mill.)、金银花(Lonicera japonica Thunb)和黄芪(Astragalus membranaceus)配伍的复方中草药,记为复方Ⅰ、复方Ⅱ、复方Ⅲ、复方Ⅳ,共四个处理组,饲喂镜鲤幼鱼(Cyprinus carpio var.specularis),并分别于第14、28及42天检测其血清中补体C3的含量、血清及免疫器官溶菌酶、NBT阳性细胞及巨噬细胞吞噬能力的活性,以研究本复方中草药对其非特异性免疫功能的影响。结果表明,14d时,复方Ⅰ组血清溶菌酶活性极显著高于对照组(P<0.01);28d时,复方Ⅰ、Ⅲ组显著高于对照组(P<0.05)。14d时,复方Ⅰ-Ⅲ组脾脏溶菌酶活性极显著高于对照组(P<0.01);28d时,复方Ⅱ组显著高于对照组(P<0.05)。28、42d时,各复方组NBT阳性细胞数显著高于对照组(P<0.05)。14和28d时,复方Ⅰ组补体C3含量极显著高于对照组(P<0.01);42d时,各复方组极显著高于对照组(P<0.01)。14d时,复方Ⅰ、Ⅳ组的巨噬细胞吞噬活性极显著高于对照组(P<0.01);28d时,各复方组显著高于对照组(P<0.05);42d时,复方Ⅰ组极显著高于对照组(P<0.01)。试验结果显示本试验中的四组复方中草药分别按质量分数为2%的量加到镜鲤幼鱼饲料中,可不同程度提高镜鲤幼鱼的非特异性免疫能力,其中复方Ⅰ组效果最佳。

Quantitative genetic estimates of growth-related traits in the common carp(Cyprinus carpio L.):A review

The common carp(Cyrpinus carpio L.)is the oldest cultured and the most domesticated fish species,as well as one of the most important freshwater fishes in the world.However,scientific studies on evaluating the growth-related quantitative traits in this fish are limited.Heritability,the most important parameter in selective breeding programs,was extensively studied for the growth-related traits.The values varied widely among the experiments and methods used because of the existence of common environmental,dominance and maternal effects.However,correlations in phenotypic and genetic levels first evaluated several years ago were limited.On the other hand,heterosis was widely reported and easily obtained for growth-related traits in the common carp.Meanwhile,genotype environment interaction and prediction of breeding values have been studied recently,and are very important in conducting selective breeding programs.The developmental quantitative genetics of growth-related traits was first analyzed in the common carp for reasonable selection during ontogeny.It is expected that genetic improvement will be achieved by carrying out direct selective breeding in the common carp.

蓝色鳞鲤(Cyprinus carpio blue var)同工酶分析

采用聚丙烯酰胺凝胶垂直板电泳法对蓝色鳞鲤(Cyprinus carpio blue var)进行同工酶分析。结果表明:在蓝色鳞鲤的眼睛、肌肉、肝脏、心脏、肾脏5种组织中,14种酶(LDH、ADH、GDH、MDH、G-6-PDH、EST、POD、SDH、FDH、SOD、α-AMY、CAT、COX、ME)的同工酶谱均存在明显的组织特异性。14种酶共记录出33个基因位点,其中α-Amy-2、Cox-2、Est-1、Ldh-1、Mdh-1、Mdh-2和Sod-1为多态位点。蓝色鳞鲤群体的多态位点百分数为21.21%(P0.99),平均预期杂合度和平均实际杂合度分别为0.1079和0.2121,遗传偏离指数(d)值为正。平均有效等位基因数(Ne)为1.24。实验表明,目前蓝色鳞鲤群体的种质资源状况尚好,表现出明显的杂交优势。

Preliminarily study on the maximum handling size, prey size and species selectivity of growth hormone transgenic and non-transgenic common carp Cyprinus carpio when foraging on gastropods

The present study preliminarily examined the differences in maximum handling size, prey size and species selectivity of growth hormone transgenic and non-transgenic common carp Cyprinus carpio when foraging on four gastropods species( Bellamya aeruginosa, Radix auricularia, Parafossarulus sinensis and Alocinma longicornis) under laboratory conditions. In the maximum handling size trial, five fish from each age group(1-year-old and 2-year-old) and each genotype(transgenic and non-transgenic) of common carp were individually allowed to feed on B. aeruginosa with wide shell height range. The results showed that maximum handling size increased linearly with fish length, and there was no significant difference in maximum handling size between the two genotypes. In the size selection trial, three pairs of 2-year-old transgenic and non-transgenic carp were individually allowed to feed on three size groups of B. aeruginosa. The results show that the two genotypes of C. carpio favored the small-sized group over the large-sized group. In the species selection trial, three pairs of 2-year-old transgenic and non-transgenic carp were individually allowed to feed on thin-shelled B. aeruginosa and thick-shelled R. auricularia, and five pairs of 2-year-old transgenic and non-transgenic carp were individually allowed to feed on two gastropods species( P. sinensis and A. longicornis) with similar size and shell strength. The results showed that both genotypes preferred thin-shelled Radix auricularia rather than thick-shelled B. aeruginosa, but there were no significant difference in selectivity between the two genotypes when fed on P. sinensis and A. longicornis. The present study indicates that transgenic and non-transgenic C. carpio show similar selectivity of predation on the size-and species-limited gastropods. While this information may be useful for assessing the environmental risk of transgenic carp, it does not necessarily demonstrate that transgenic common carp might have lesser environmental impacts than non-transgenic carp.

蓝色鳞鲤(Cyprinus carpio blue var)的细胞遗传学分析

采用体内注射PHA和秋水仙素,肾细胞短期培养,常规空气干燥法制备蓝色鳞鲤染色体,对100个中期分裂相记数统计,确定蓝色鳞鲤(Cyprinuscarpiobluevar)的染色体数为2n=100。测得核型参数按Levan等的染色体划分标准得出:蓝色鳞鲤有15对中部着丝点染色体(m);13对亚中部着丝点染色体(sm);22对端部和亚端部着丝点染色体(st,t),其染色体总臂数(NF)为156,蓝色鳞鲤核型公式为2n=30m+26sm+44st,t。采用流式细胞分析仪测定了蓝色鳞鲤的DNA含量,与鸡血细胞标准对照相比为1.73±0.10,以鸡红细胞DNA含量2.3pg.N-1计,则蓝色鳞鲤的体细胞DNA含量为3.99pg.N-1。

Effect of Metal Ions on Protease Activities in the Intestines and Hepatopancreas of Red-white Ornamental Carp (Cyprinus carpio L)

[Objective] The aim of this study was to study effects of metal ions on the protease activities in digestive tissues and gland of red-white ornamental carp（Cyprinus carpio L）.[Method] Effects of four kinds of metal ions （K＋,Na＋,Mg2＋ and Ca2＋） on protease activities in hepatopancreas,foregut,midgut,hindgut of red-white ornamental carp were studied by enzyme analysis method.[Result] Effects of four kinds of metal ions on protease activities of red-white ornamental carp were different in the range of experimental concentration from 25 mmol/L to 150 mmol/L.K＋ could promote protease activities in hepatopancreas and hindgut at different levels.Especially,K＋ had the promoting effect at low-concentration level,but the inhibitory effect at high-concentration level in midgut and the inhibitory effect in foregut.Na＋ had the promoting effect on protease activities in hepatopancreas,foregut and hindgut at different levels,but the inhibitory effect in midgut.Mg2＋ and Ca2＋ had the inhibitory effect on protease activities in intestinal and hepatopancreas at different levels.[Conclusion] This study provides basic data and theoretical foundation for researches on the digestive physiology of red-white ornamental carp or the development and optimization of compound feed.

基于线粒体Cyt b基因和D-loop区分析元江鲤和杞麓鲤群体遗传结构

本文利用线粒体Cyt b基因和D-loop区部分序列,分析元江鲤(Cyprinus carpio yuankiang)和杞麓鲤(Cyprinus carpio chilia)群体的遗传结构。结果表明,在两个鲤群体中均获得929 bp的Cyt b基因序列和650 bp的D-Loop区序列。两个群体间平均遗传距离为0.00595(Cyt b)、0.00852(D-loop)。Cyt b基因共检测出元江鲤3种、杞麓鲤15种单倍型,两个群体共享两个单倍型(C-Hap1和C-Hap3);D-loop区序列共检测出元江鲤3种、杞麓鲤6种单倍型,两个群体共享两个单倍型(D-Hap1和D-Hap2)。元江鲤和杞麓鲤群体单倍型多样性(Hd)分别为0.30±0.10(Cyt b)、0.25±0.10(D-loop)和0.89±0.04(Cyt b)、0.73±0.05(D-loop)。元江鲤和杞麓鲤群体核苷酸多样性(π)分别为0.12±0.04(Cyt b)、0.12±0.09(D-loop)和0.77±0.09(Cyt b)、0.87±0.07(D-loop),杞麓鲤遗传多样性水平明显高于元江鲤群体。分子方差分析(AMOVA)显示,两个群体间有明显的遗传分化,且遗传变异主要来源于群体间。系统发育分析显示,元江鲤群体单倍型较少,类型较单一,与杞麓鲤群体差异大,多数元江鲤样本单倍型属于华南鲤(Cyprinus carpio rubrofuscus)类型;多数杞麓鲤样本具特有单倍型,部分属于华南鲤、远东鲤(Cyprinus carpio haematopterus)类型。结果表明,杞麓鲤与元江鲤群体遗传分化明显,元江鲤线粒体遗传多样性较低。

Protective Effect of Bacillus subtilis Peptidoglycan (PG) on β-conglycinin-induced Intestinal Epithelial Cells Damage of Juvenile Carp (Cyprinus carpio)

[Objective] The paper was to investigate the protective effects of different concentrations of Bacillus subtilis peptidoglycan(PG) on β-conglycinin-induced inflammatory injury in intestinal epithelial cells of juvenile carp(Cyprinus carpio).[Method] In 24-cell microplates, the intestinal epithelial cells(IECs) of juvenile carp were primarily cultured for 72 h at 26°C and 6% CO2, and then the IECs were randomly divided into6 groups with 4 replicates per group. One of the six groups was set as negative control group, and the other groups were all supplemented with 1.0 mg/mL β-conglycinin in culture medium to establish inflammatory injury. At 24 h post induction, the culture media were changed into B. subtilis PG culture media with the concentrations of 0(positive control group), 0.15, 0.30, 0.45 and 0.60 mg/mL, respectively. The samples were collected to measure the antioxidant and anti-inflammatory indices at 12, 24 and 36 h post culture.[Result]β-conglycinin exposure significantly decreased the activity of ASA, AHR, SOD, CAT, GPx, and increased the PC content and the mRNA expression of inflammatory cytokines(IL-1β, IL-8, TNF-α1,IL-10 and TGF-β). At 12, 24 and 36 h post PG treatment, the activities of ASA, AHR, SOD, CAT, GPx and the content of PC in cells decreased in a dose-dependent manner;the mRNA levels of IL-1β, IL-8 and TNF-α1 were down-regulated and those of IL-10 and TGF-β were up-regulated.[Conclusion] Different concentrations of B. subtilis PG could protect IECs oxidative damage induced by β-conglycinin and improve the antioxidant capacity of IECs. High concentration of PG could improve the anti-inflammatory ability of IECs by inhibiting inflammatory factors and promoting the gene expression of anti-inflammatory cytokines.

PCR-RFLP analysis of mitochondrial DNA ND5/6 region among 3 subspecies of common carp (Cyprinus carpio L.)and its application to genetic discrimination of subspecies

Mitochondrial DNA ND5/6 region was studied by PCR-RFLP analysis among ten representative strains belonging to three subspecies (Cyprinus carpio carpio, Cypri-nus carpio haematopterus and Cyprinus carpio rubrofuscus) of common carp (Cyprinus carpio L.). A total of 2.4 kb frag-ment was amplified and subjected to restriction endonucle-ase analysis with nine restriction endonucleases subsequently.The results indicated that each subspecies owned one hyplo-type and four restriction enzymes (DdeⅠ, HaeⅢ, TaqⅠand MboⅠ) produced diagnostic restriction sites which could be used for discriminating the three subspecies and as molecu-lar genetic markers for assistant selective breeding of com-mon carp.