嘉兴长白猪氟烷基因频率检测初报

氟烷基因是控制猪应激综合征(porcine stress syndrome,PSS)的主效基因,可造成个体突然死亡或宰后产生PSE肉,同时也对猪的繁殖性有一定的负面影响,但它对猪产肉力有增效作用,隐性纯合体表现对氟烷麻醉敏感因而称氟烷基因。Knudson等(1990)发现兰尼啶受体(Rynodine receptor,RYR1)的胰蛋白酶消化图谱MH敏感猪与MH正常猪不同,建议RYR1基因作为氟烷基因的主要候选基因。Fujii(1991)克隆和测序了皮特兰猪(MH敏感)和大约克猪(MH正常)的全长RYR1cDNA序列分析表明有18处发生突变唯有C1843突变为T1843引起arg615→cys615,从而确认了猪…     

猪RYR1基因三种PCR-RFLPs检测方法的比较

猪应激综合症是猪在应激因子 (如运输、转栏、高温、预防注射和配种等 )的作用下诱发的一种综合症 ,其发生主要是由于RYR1基因的点突变造成的。本研究对RYR1基因的三种PCR -RFLPs检测方法进行了比较 ,分别利用三种方法对 60头猪的检测结果证明 :三者具有相同的准确性。但是三引物法和四引物法较二引物法所需的时间更少 ,检测成本更低     

江口萝卜猪和梅山猪的猪应急综合征主效基因RYR1的PCR-RFLP检测

猪兰尼定受体1(Ryanodine Receptor1,RYR1)基因是导致猪应激综合征、影响猪肉质的主效基因.试验采用RFLP方法,以大白猪、2个杂交猪群(杜洛克×长白猪×大白猪和长白猪×大白猪)为对照,检测了江口萝卜猪与引入的梅山猪群RYR1基因的分布类型.结果表明,对照组中,外三元、外二元杂交猪和大白猪中均检测到...     

七个贵州地方猪种RYR1基因突变的分布

兰尼定受体1(Ryanodine Receptor1,RYR1)基因中的单个氨基酸(R615C)是导致猪应激综合征、影响肉质的重要原因。采用RFLP方法,以大白猪、二个杂交猪群(杜洛克×长白猪×大白猪和长白猪×大白猪)为对照,研究了糯谷猪、萝卜猪、柯乐猪、宗地花猪、香猪、关岭猪和黔南黑猪7个贵州地方猪种RYR1基因突变的分布类型。结果表明,从柯乐猪中检测到一例杂合基因型RYR1Nn,杂合基因型的发生频率为2.08%,其余6个贵州地方猪种糯谷猪、萝卜猪、宗地花猪、香猪、关岭猪和黔南黑猪均为正常的基因型RYR1NN,对照组中,外三元、外二元杂交猪和大白猪中均检测到RYR1Nn基因型,杂合基因型的发生频率分别为11.11%、12.12%和5.77%,说明贵州地方猪种中RYR1基因的突变频率低于商品猪,但也存在一定的比例,应加强对地方猪种的利用和保护。     

PCR一步法检测猪RYR1基因

在传统的PCR加酶切检测猪的RYR1基因基础上，设计能特异地扩增突变和正常序列的4条引物，根据扩增片段大小和条带多少可直接鉴别不同的RYR1基因型，从而建立了简单、廉价、准确地PCR一步检测猪应激综合征的新方法。     

猪氟烷基因的研究进展

猪应激综合征(PSS)是产生PSE和DFD肉的主要原因,控制PSS的基因称为氟烷基因(Ha1)或兰尼受体基因(RYR1)。本文综述了猪氟烷基因的研究进展。相关的研究表明,氟烷基因杂合子猪具有较高的生长速度、瘦肉率及较好的肉品质、高的繁殖性能。总体而言,完全清除猪氟烷基因对畜牧业是不利的。     

利用三个基因标记选育大白核心群后备种猪初报

猪氟烷基因(RYR1)c.1843C>T突变位点是造成猪应激综合征的主效基因位点.硬脂酰辅酶A去不饱和酶基因(SCD)是控制单不饱和脂肪酸合成的关键酶,其启动子区域-233上T>C突变位点对肥胖和背膘厚有重要影响.α1岩藻糖基转移酶基因(FUT1)基因是ETEC F18受体蛋白基因,其开放阅读框M307的G>A突变位点影响仔猪断乳后水肿和腹泻的发生.该研究采用PCR-RFLP方法,检测了福建仁锋种猪有限公司大约克核心群60头后备种猪个体在这3个基因相应突变位点的基因型.结果表明,该群体已完全淘汰了应激敏感型n等位基因,建立了氟烷应激抵抗系:且保持着高频率(90.8%)的SCD基因、有利降低背膘沉积、提高瘦肉率的T等位基因;但FUT1的ECF18抗性等位基因A的频率相对较低(26.7%),易感等位基因G的频率较高(73.3%).该研究结果结合该场的性能测定结果运用于指导该大白核心群后备种猪的选种选育实践,可望提高核心群内种猪产肉量性状、抗应激能力和降低仔猪断乳后腹泻抗性发生.     

三江白猪DNA的提取及PCR反应条件的优化

猪应激综合征(Porcine Stress Syndrome，PSS)由一对氟烷基因(Hahn、Haln)控制。试验采取组织DNA和血液DNA的提取，并对氟烷基因的PCR反应条件进行优化，为三江白猪的氟烷基因从分子角度上进行检测提供科学的理论依据。     

猪应激综合征(PSS)〔综述〕

一、一般概念猪应激综合征(PSS,Porcine Stress Syndrome)是在现代生产条件下,各国致力改良猪种,寻找高能饲料,发展集约化饲养方式,提高经济效益,促进养猪业发展,多产肉、产好肉的同时带来的一个问题,即猪的某些生物学特性受到了破坏,易于发生各种应激综合征.     

新民猪RYR1和MC4R基因的多态性检测分析

为了调查兰尼定1型受体(RYR1)基因和黑素皮质素受体4(MC4R)基因[其目前公认的数量性状基因座(QTN)]在新组建的民猪群体内的分布情况,试验采用PCR-RFLP的方法,对黑龙江省农业科学院畜牧研究所收集整理的23头民猪的RYR1基因和MC4R基因进行检测,并与其他已报道的猪种进行比较。结果表明:RYR1基因的N等位基因频率为86.96%,n等位基因频率为13.04%;MC4R的A等位基因频率为76.19%,G等位基因频率为23.81%,与其他猪种相比,n等位基因频率偏高。     

Halothane Sensitivity,The H Blood Group System and Phosphohexose Isomerase (PHI) in Pigs: A Linkage Study Of Physiological Importance

Previous investigations have clearly shown the existence of associations between halothane sensitivity, the H blood group system and the PHI enzyme system in pigs (Rasmusen & Christian 1976, Jørgensen et al. 1976). These associations which have considerable practical interest are most probably linkage phenomenons (Jørgensen 1977, Andresen & Jensen 1977). The major recessive locus for halothane sensitivity (HAL) comprises the two alleles N and n, n being responsible for halothane sensitivity. The distances between this locus and the loci for H and PHI are still not known exactly. This communication aims at clarifying these problems.     

Halothane anesthesia of several hours' duration in swine of the German Landrace breed, selected for resistance to malignant hyperthermia

The malignant hyperthermia syndrome is widely spread in pigs of the German Landrace. This has precluded the use of halothane anesthesia. The advent of homozygous halothane-negative strains stimulated us to test whether such animals were suitable for longduration halothane anesthesia. Nine sows of the German Landrace (n = 4, homozygous halothane-positive (H+); n = 5, homozygous halothane-negative (-)) were used. Anesthesia was induced by Azaperon/Metomidat-HCL followed by intubation and exposure to halothane. All 4 halothane-positive animals died within 45-90 min following the onset of halothane administration, while all 5 halothane-negative animals survived a 4-5 h deep anesthesia without problems. Thus, homozygous hylothane-positive strains of German Landrace pigs may be surgically anesthetized for long durations with halothane anesthesia.     

Endocrine and metabolic responses to halothane and pentobarbitone anaesthesia in sheep

Some metabolic and endocrine responses to anaesthesia in sheep were studied. Adult sheep were anaesthetised with thiopentone and halothane (n=9), acepromazine, thiopentone and halothane (n=8) and pentobarbitone (n=10) on separate occasions. Routine cardiovascular monitoring was carried out and blood samples were taken for assay of cortisol, adrenocorticotrophic hormone (ACTH), arginine vasopressin (AVP), glucose and lactate. Halothane anaesthesia induced hypotension, hypercapnia and respiratory acidosis. Sheep anaesthetised with pentobarbitone were also hypercapnic and acidotic but did not develop hypotension. Plasma cortisol, ACTH and AVP (mean maximum values: cortisol: 83 ng/ml, ACTH 278 ng/ml, AVP 135 pg/ml), increased during halothane anaesthesia but did not change significantly from control values during pentobarbitone anaesthesia (mean maximum values: cortisol: 30 ng/ml, ACTH 71 ng/ml, AVP 7.8 pg/ml). Glucose tended to increase during both halothane and pentobarbitone anaesthesia but lactate decreased. It is not clear what facet of halothane anaesthesia evokes the stress response but it may be associated with cardiovascular depression.     

The effects of halothane sensitivity on carcass composition and meat quality in HAL-1843-normal pigs

Objectives of this study were to determine the incidence of halothane sensitivity in pigs that are homozygous normal at the ryanodine receptor nucleotide 1843 (HAL-1843-normal) and the relationships between halothane sensitivity and carcass composition or meat quality. In Exp. 1, piglets (Lines A, B, C, and D; n = 168, 170, 168, and 169, respectively) were obtained from mating a HAL-1843-normal sire line to four HAL-1843-normal dam lines. In Exp. 2, piglets from Lines A and B (n = 87 and 90, respectively) were included with piglets (Lines E, F, G, and H; n = 94, 92, 89, and 89, respectively) obtained from mating four HAL-1843-normal sire lines to a single HAL-1843-normal dam line. Pigs were subjected to 3% halothane at approximately 9 wk of age. In Exp. 1, limb rigidity, blotching of the skin, and muscle tremors were visually assessed, and based on these criteria, halothane sensitivity (HS) was observed in 48% of the pigs. To better characterize this response, a scoring system was developed and used in Exp. 2. Using this system, 25, 42, and 33% of the pigs in E and 40, 33, and 27% of the pigs in Line G were categorized as HS-low (HS-L), HS-intermediate (HS-I), and HS-high (HS-H), respectively. In Lines F and H, 13 and 18% of the pigs were HS-I, and 0 and 2% were HS-H, respectively. No consistent effects due to HS were observed in carcass composition or meat quality; however, when a subset of pigs from Exp. 2 were subjected to more extensive handling and transportation before slaughter, ultimate pH was lower and drip loss was higher in LM from HS-H compared with HS-L pigs (P < 0.05; n = 71). These results demonstrate that some pigs are sensitive to halothane anesthesia even in the absence of the known HAL-1843 polymorphism. Additionally, halothane sensitivity may be associated with inferior pork quality under adverse antemortem conditions.     

Atypical reactions to halothane in a subgroup of homozygous malignant hyperthermia(MH)-susceptible pigs: indication of a heterogenous genetic basis for the porcine syndrome.

Malignant hyperthermia (MH) is a pharmacogenetic disorder of skeletal muscle. In genetically susceptible pigs, MH can be induced by volatile, halogenated anaesthetics such as halothane. Within a series of pharmacological investigations, a fulminant MH could be induced in 59 of 66 homozygous halothane-susceptible pigs by a challenge with 3% halothane for 15 minutes. The typical MH was characterized by sudden appearance of tachycardia, muscle rigidity with typical extension of the hindlimbs, increase of body temperature, acidosis-caused by rapid increase of CO2 and lactate production-, hyperkalaemia and increased activity of creatine kinase (CK) and aspartate transaminase (AST). In seven homozygous MH-susceptible pigs, this typical MH could not be induced by halothane. These animals responded with sudden appearance of bradyarrhythmia and decrease of arterial pressure. In these MH-atypical pigs (MHA) neither the typical extension of hindlimbs nor a hyperthermia occurred. Compared to a group of 6 MH-susceptible pigs with typical reactions to halothane (MHS), the biochemical alterations were significantly retarded in MHA-pigs. These atypical reactions to halothane could be the effect of decreased cardiac output. Concerning the atypical reactions, we observed a familiar predisposition in MH-susceptible pigs. Although atypical reactions were not found in a group of homozygous halothane-nonsusceptible pigs (MHN), a possible explanation for atypical reactions could be a MH-independent halothane-susceptibility of the myocardium+ in MHA-pigs. On the other side the data may indicate that a primary defect in both the skeletal muscle and also the myocardium is involved in MH. The different reactions to halothane in MH-susceptible pigs could point to a genetic heterogeneity.     

Inheritance of Sensitivity to Halothane in Pigs

The inheritance of sensitivity to halothane was studied in experimental families of four generations. In a five minutes' halothane test, with evaluation of the signs in animals born from variously combined makings of sensitive pigs (Belgian Landrace) and resistant inbred animals (Canadian Landrace), sensitivity to halothane was found to be inherited as a single-gene autonomic recessive trait with "high" or complete penetrance, but with variable expressivity .     

Identification of halothane gene carriers by use of in vivo 31P nuclear magnetic resonance spectroscopy in pigs.

In vivo muscle 31P nuclear magnetic resonance spectroscopy was performed on 12 homozygous halothane-nonsensitive female pigs and 13 female pigs heterozygous with respect to the halothane gene. Fifteen female pigs of a third line, consisting of heterozygotes and halothane-nonsensitive homozygotes, were also available. Body weight ranged from 12 to 18 kg. Mean decrease in phosphocreatine concentration in the biceps femoris of anesthetized pigs was significantly lower for heterozygous vs homozygous pigs (3.46% vs 5.94%, P less than 0.01) after 40 minutes of halothane exposure (3%; oxygen flow, 3 L/min). Also, a statistically significant difference, with respect to the initial (7.21 vs 7.11, P less than 0.008) and end muscle pH values (7.18 vs 7.06, P less than 0.0002), was observed for homozygous vs heterozygous pigs. By means of canonical discriminant analysis, it was possible to distinguish nonsensitive homozygotes from heterozygotes (P less than 0.0001). When applying this classification method to pigs of the same strain, 2 populations (nonsensitive homozygotes, heterozygotes) emerged, with a proportion of pigs corresponding to the expected value on the basis of breeding records. In contrast to the phenotypic expression of muscular rigidity related to the malignant hyperthermia syndrome, the expression of metabolic variables (phosphocreatine, pH) was shown to be dominant.     

Carrier rate of Factor XI deficiency in stunted Japanese black cattle

Blood examinations and genotyping of Factor XI (F11) were performed in growth retardation Japanese Black cattle and their dams. Genotyping of F11 revealed that the recessive homozygous and heterozygous genotype frequencies were 5.2% and 50.0% in the Claudin-16 (CL-16) deficiency group (n=58), 0% and 14.2% in the renal dysplasia group (n=7), 0% and 26.1% in the non-CL-16 deficiency nephritis group (n=23), 8.9% and 46.7% in the hypogenesis syndrome group (n=45), 6.2% and 25.0% in the neonatal weak calf syndrome group (n=32), 9.1% and 38.6% in the respective dams group (n=44), 0% and 23.1% in the normal cattle group (n=13), and 5.9% and 38.2% in total (n=222), respectively. These results showed that the carrier rate of F11 deficiency was high in Japanese Black cattle, and that the CL-16 deficiency, hypogenesis syndrome, neonatal weak calf syndrome, and dams groups had a large amount of recessive homozygous genotype than the other groups. No abnormal bleeding was observed clinically in the present study, and 4 of the recessive homozygous dams showed normal growth and parturition.     

Serum pyruvate kinase and its relation to stress susceptibility in pigs

SUMMARY: Four blood tests, complementary to the halothane test, have been screened for their capacity to discriminate between homozygous and heterozygous stress resistant and stress susceptible pigs: TBARS in plasma, lipid peroxides from erythrocytes and plasma creatine and pyruvate kinase activity. Blood samples were collected from homozygous (NN) and heterozygous (Nn) stress resistant and homozygous stress susceptible (nn) pigs, while subjected to the halothane test. Serum creatine kinase and pyruvate kinase tests were retained for a more elaborated study. Pyruvate kinase appears to be a more reliable indicator of stress susceptibility than the well-known creatine kinase. However both parameters display a considerable standard deviation. Consecutive assays show the pyruvate kinase activity to be dependent on age (weight), which accounts for part of the variation; other sources of variation are the conditions of the exeriment and the way the imposed stress is perceived by the animal. During this experiment a rapid and accurate DNA-test for HAL-genotype detection has been developed (Fujii et al. 1991; Otsu et al. 1991; Coppieters et al. 1992). In spite of this, the pyruvate kinase parameter is a much more discriminating and remarkable indicator for stress susceptibility than the, in previous years, widely used creatine kinase test. ZUSAMMENFASSUNG: Serum Pyruvat Kinase und seine Relation mit Streβanf?lligkeit bei Schweinen Vier Blutteste, komplement?r dem HAL-Test, wurden in einem Screening-Verfahren untersucht um homozygote und heterozygote stre?resistente und stre?anf?llige Schweine zu differenzieren: TBARS in Plasma, Lipid Peroxyde von Erythrocyten und Plasma Kreatine und Pyruvat Kinase Aktivit?t. W?hrend des Halothantestes wurde Blut entnommen von homozygoten (NN) und heterozygoten (Nn) stre?resistenten und homozygoten stre?anf?lligen Schweinen (nn). Mit Serum Kreatine Kinase und Pyruvat Kinase Aktivit?t Tests wurden eine umfassende überprüfung durchgeführt. Letztere zeigt sich als ein zuverl?ssiger Indikator der Stre?anf?lligkeit als die wohl bekanntere Kreatine Kinase. Mehrmalige Analysen zeigen, da? Pyruvat Kinase Aktivit?t vom Alter (Gewicht) abh?ngig ist, was teilweise die Variabilit?t verursacht; andere Quellen der Variabilit?t sind die experimentellen Umst?nde und die Weise, wie der auferlegte Stre? durch das Tier erfahren wird. W?hrend der Durchführung dieses Experimentes wurde ein schneller und akkurater DNA-Test für HAL-Genotyp Detektion entwickelt (Fujii et al. 1991; Otsu et al. 1991; Coppieters et al. 1992). Im Vergleich mit dem, in vorgehenden Jahren, h?ufig angewendeten Kreatine Kinase Test, ist der Pyruvat Kinase Parameter trotzdem ein bemerkenswerter und diskriminierender Indikator für Stre?anf?lligkeit.