Asphyxial cardiac arrest, resuscitation and neurological outcome in a Landrace/Large-White swine model

The vast majority of laboratory studies on animals have focused on ventricular fibrillation (VF) and not on cardiac arrest (CA) resulting from asphyxia. The aim of this study was to develop a clinically relevant animal model in Landrace/Large-White swine of asphyxial CA resuscitated using the European Resuscitation Council guidelines. Survival and 24 h neurological outcome in terms of functional deficit were also evaluated. Asphyxial arrest was induced by clamping the endotracheal tube (ETT) in 10 Landrace/Large-White piglets. After 4 min of untreated arrest, resuscitation was initiated by unclamping the ETT, 100% oxygen mechanical ventilation, 2 min chest compressions and epinephrine administration. Advanced Life Support algorithm was followed. In case of restoration of spontaneous circulation, the animals were supported for one hour and then observed for 23 h. Coronary perfusion pressure was significantly higher in surviving animals (P < 0.001) during cardiopulmonary resuscitation. End-tidal CO(2) was significantly higher in the animals that survived than in non-surviving animals (P = 0.001). All of the animals were severely neurologically impaired 24 h after CA. This refined model of asphyxia CA is easily reproducible and may be used for pharmacological studies in CA.     

Cardiopulmonary resuscitation in the mouse.

We sought to develop a model of cardiac arrest and resuscitation on mice that would be comparable to that of large mammals and would allow for more fundamental investigations on cardiopulmonary arrest and cardiac resuscitation. A model of cardiopulmonary resuscitation previously developed by our group on rats was adapted to anesthetized, mechanically ventilated adult male Institute of Cancer Research mice that weighed 46 +/- 3 g. The trachea was intubated through the mouth, and end-tidal PCO(2) (PET(CO(2))) was measured with a microcapnometer. Catheters were advanced into the aorta and into the right atrium, and coronary perfusion pressure (CPP) was computed. A 1.5-mA alternating current was delivered to the right ventricular endocardium, which produced ventricular fibrillation or a pulseless rhythm. Precordial compression was begun 4 min later. Ten sequential studies were performed, during which five animals were successfully resuscitated and five failed resuscitation efforts. Successful resuscitation was contingent on the restoration of threshold levels of CPP and PET(CO(2)) during chest compression. As in rats, swine, and human patients, threshold levels of mean aortic pressure, CPP, and PET(CO(2)) were critical determinates of resuscitability in this murine model of threshold level of cardiac arrest and resuscitation.     

Cardiopulmonary resuscitation in the rat

A standardized method of cardiopulmonary resuscitation in rodents has been developed for anesthetized, mechanically ventilated rats. Ventricular fibrillation was induced and maintained by an alternating current delivered to the right ventricular endocardium. After 4 min of ventricular fibrillation, the chest was compressed with a pneumatic piston device. Eight of 14 animals were successfully resuscitated with DC countershock after 6 min of cardiac arrest. In confirmation of earlier studies from our laboratories in dogs, pigs, and human patients, this rodent model of cardiopulmonary resuscitation demonstrated large venoarterial [H+] and PCO2 gradients associated with reduced pulmonary excretion of CO2 during the low-flow state. Mean aortic pressure, coronary perfusion pressure, and end-tidal CO2 during chest compression were predictive of successful resuscitation.     

DNA microsatellite analysis for parentage control in Austrian pigs.

We present an efficient parentage control for pigs based on ten polymorphic microsatellite markers analyzed in a single PCR reaction. Assuming one known parent ("paternity control"), combined exclusion probabilities (CEPs) ranged from 99.18% (Landrace), 99.74% (Piétrain) to 99.76% (Large White) for the most important Austrian breeds. Assuming a known parent-pair ("parentage control", e.g. a substituted offspring), the CEP of the 10-plex PCR increased to 99.97% (Landrace) and 99.99% (Piétrain and Large White). We developed an additional standby battery of 5 markers, which might be applied in those cases, where the CEP of the 10-plex PCR is not sufficient. Therefore an automated, cost and time reduced genotype analysis for pigs is available.     

用焦磷酸测序技术研究猪线粒体细胞色素B基因单倍型

选择43头大白猪,79头长白猪,66头皮特兰猪和60头清平猪作试验材料,采用焦磷酸测序技术分析猪线粒体细胞色素b(CytB)基因单倍型。研究结果显示CytB基因可分为4种单倍型E1,E2,A1和A2。清平猪仅存在于A1单倍型(100%),大白猪和长白猪存在于E1(49.19%,79.25%)和A1(55.81%,20.25%)单倍型,皮特兰则存在于E1(57.58%)和A2(42.42%)单倍型。 Abstract：To detect porcine mitochondrial cytochrome b (CytB) gene haplotypes, Pyrosequencing, which is a novel DNA sequencing method, has been used to analyze SNPs selected Large White, Landrace, Pietrain and Qingping pigs. The pyrosequencing analysis of CytB gene displayed four distinct haplotypes E1, E2, A1 and A2 respectively. Qingping pigs are only present in haplotype A1, Large White and Landrace pigs are present in haplotype E1 and A1, and Pietrain pigs are present in haplotupe E1 and A2.     

Bi-PASA genotyping of a new polymorphism in the APOB gene shows no evidence for an association with fatness in pigs

Sequence analysis of PCR products of a 343-bp fragment from exon 29 of the porcine APOB gene of four Erhualian and four Landrace pigs revealed a missense G/C substitution at position of 6117 in this gene. Two allele-specific primers were designed to genotype this polymorphism using the Bi-PASA technique. Genotyping of 146 animals from Erhualian, Hampshire, Large White, Landrace and Duroc breeds revealed large breed differences in allele frequency. No association with fatness was observed within each of the four European breeds, where animals had been selected to be those with the highest and those with the lowest backfat depth at 100 kg liveweight from a large carcass dissection project.     

Trends in economic traits, halothane sensitivity, blood group and enzyme systems of Swiss Landrace and Large White pigs

Pigs deriving from 150 breeding centres constituting a representative section of elite breeding herds (2496 Swiss Landrace pigs, 587 Swiss Large White pigs) were subjected to blood typing during the period 1981 to 1984. Production traits such as daily gain, feed conversion ratio, lean meat content and meat quality score were available to show the trend in these performance traits since 1978. Field data on the halothane reaction of 14 270 Swiss Landrace (SL) pigs were used to assess the porcine stress syndrome during the period 1978–1983.
In SL pigs the frequency of the alleles H^a, Phi^B and Ada^A decreased significantly, and that of the H^c and Phi^A increased during the period of the study. The frequency of the H^a allele dropped from 0.36 in 1981 to 0.20 in 1984, whereas the H^c allele rose from 0.22 to 0.37. In Swiss Large White (SLW) pigs, on the other hand, the frequency of the H^a allele increased constantly from 0.31 to 0.37 during this period.
An initial frequency of 17.7 % (1978) halothane reactors in SL pigs was lowered to 0.7 % (1982) after five years of halothane testing.
In SL pigs the meat quality scores improved regularly, whereas in SLW pigs it did not change very much. The percentage of PSE animals in the SL breed was reduced from 32.7 % in 1978 to 7.1 % in 1983.
Because the Hal locus is associated with production traits such as meat quality, linkage disequilibria could explain the observed associations between the H and Phi types and production traits.     

Unexpectedly high allelic diversity at the KIT locus causing dominant white color in the domestic pig.

Mutations in KIT encoding the mast/stem cell growth factor receptor (MGF) are responsible for coat color variation in domestic pigs. The dominant white phenotype is caused by two mutations, a gene duplication and a splice mutation in one of the copies leading to skipping of exon 17. Here we applied minisequencing and pyrosequencing for quantitative analysis of the number of copies with the splice form. An unexpectedly high genetic diversity was revealed in white pigs. We found four different KIT alleles in a small sample of eight Large White females used as founder animals in a wild boar intercross. A similar number of KIT alleles was found in commercial populations of white Landrace and Large White pigs. We provide evidence for at least two new KIT alleles in pigs, both with a triplication of the gene. The results imply that KIT alleles with the duplication are genetically unstable and new alleles are most likely generated by unequal crossing over. This study provides an improved method for genotyping the complicated Dominant white/KIT locus in pigs. The results also suggest that some alleles may be associated with negative pleiotropic effects on other traits.     

猪CAST基因的单核苷酸多态性及其对肉质性状的效应

CAST基因作为肉质性状的主要候选基因.以80头外来猪和190头地方猪为材料,在CAST基因内含子24上检测到两个多态性位点(A916G 和C1633G ).在916位点上,长白猪和大白猪以A基因为优势基因,其频率分别为0.88和1.00;莱芜猪,大薄莲猪,沂蒙黑猪和里岔黑猪以B基因为优势基因,其频率分别为0.93, 0.97, 0.78和0.68.在1633位点上,长白猪和大白猪以C基因为优势基因,其频率分别为0.82和0.79:莱芜猪,大薄莲猪,沂蒙黑猪和里岔黑猪以D基因为优势基因,其频率分别为1.00, 1.00, 0.88, 0.78.在试验猪种中,共检测到6种单倍型(AACC,AACD,AADD,ABCC,BBCC,BBDD).单倍型分布的多重比较结果表明,外来猪种(长白猪和大白猪)与地方猪种(莱芜猪,大薄莲猪,沂蒙黑猪和里岔黑猪)比较差异极显著(P < 0.01).固定效应模型分析结果表明,嫩度,屠宰45 min后pH值和滴水损失单倍型间差异显著(P < 0.05).最小二乘分析结果表明,外来猪种与地方猪种在嫩度,屠宰45 min后pH值和滴水损失间差异显著(P < 0.05).BBDD单倍型个体与其它单倍型个体比较,嫩度及滴水损失差异显著(P < 0.05);AADD,BBCC,BBDD单倍型个体与其它单倍型个体比较,屠宰45 min后pH值差异显著(P < 0.05).因此,在育种过程中将CAST基因应用于标记辅助选择,将有利于改善猪肉品质,加快育种进程.     

猪ATF4基因多态性与生产性状的关联及基因表达分析

为进一步了解和认识ATF4基因的功能,揭示ATF4对猪脂肪代谢的影响,寻找与肉质性状相关联的分子标记,文章采用PCR方法扩增了ATF4基因部分序列,通过序列比对发现在翻译起始密码子ATG下游159 bp处存在A159G转换,通过PCR-AluⅠ-RFLP对大白猪、长白猪、梅山猪和通城猪进行酶切分型,发现在大白猪和长白猪中均为AA基因型,在梅山猪和通城猪中均为GG基因型。进一步对大白猪×梅山F2群体资源家系进行了酶切分型,并分析该位点的多态性与生产性状的关系。结果表明,ATF4的多态性与臀部平均膘厚存在极显著相关(P<0.01),与胸腰椎间膘厚、平均膘厚、眼肌高、眼肌面积存在显著相关(P<0.05)。采用Real-time PCR分析了ATF4基因在大白猪与梅山猪背最长肌不同发育阶段的表达模式。结果表明,ATF4基因在大白猪和梅山猪胚胎期65 d和出生后3 d中的表达水平相对都比较低,且在两品种间无明显差异;而在出生后60 d和120 d,ATF4基因在大白猪中与梅山猪均出现了上调表达,并且在梅山猪中的相对表达水平要显著高于大白猪。研究结果为进一步深入研究猪ATF4基因在脂肪代谢中的分子机理奠定了基础。     

A Genome-Wide Association Study in Large White and Landrace Pig Populations for Number Piglets Born Alive

The number of piglets born alive (NBA) per litter is one of the most important traits in pig breeding due to its influence on production efficiency. It is difficult to improve NBA because the heritability of the trait is low and it is governed by a high number of loci with low to moderate effects. To clarify the biological and genetic background of NBA, genome-wide association studies (GWAS) were performed using 4,012 Large White and Landrace pigs from herdbook and commercial breeding companies in Germany (3), Austria (1) and Switzerland (1). The animals were genotyped with the Illumina PorcineSNP60 BeadChip. Because of population stratifications within and between breeds, clusters were formed using the genetic distances between the populations. Five clusters for each breed were formed and analysed by GWAS approaches. In total, 17 different significant markers affecting NBA were found in regions with known effects on female reproduction. No overlapping significant chromosome areas or QTL between Large White and Landrace breed were detected.     

Trends in economic traits, halothane sensitivity, blood group and enzyme systems of Swiss Landrace and Large White pigs

Pigs deriving from 150 breeding centres constituting a representative section of elite breeding herds (2496 Swiss Landrace pigs, 587 Swiss Large White pigs) were subjected to blood typing during the period 1981 to 1984. Production traits such as daily gain, feed conversion ratio, lean meat content and meat quality score were available to show the trend in these performance traits since 1978. Field data on the halothane reaction of 14 270 Swiss Landrace (SL) pigs were used to assess the porcine stress syndrome during the period 1978-1983. In SL pigs the frequency of the alleles Ha, PhiB and AdaA decreased significantly, and that of the Hc and PhiA increased during the period of the study. The frequency of the Ha allele dropped from 0.36 in 1981 to 0.20 in 1984, whereas the Hc allele rose from 0.22 to 0.37. In Swiss Large White (SLW) pigs, on the other hand, the frequency of the Ha allele increased constantly from 0.31 to 0.37 during this period. An initial frequency of 17.7% (1978) halothane reactors in SL pigs was lowered to 0.7% (1982) after five years of halothane testing. In SL pigs the meat quality scores improved regularly, whereas in SLW pigs it did not change very much. The percentage of PSE animals in the SL breed was reduced from 32.7% in 1978 to 7.1% in 1983. Because the Hal locus is associated with production traits such as meat quality, linkage disequilibria could explain the observed associations between the H and Phi types and production traits.     

西部地区主要猪种H-FABP基因多态性,IMF含量及不同基因型脂肪细胞脂滴量的关系

H-FABP是FABP家族成员之一,在长链脂肪酸的吸收和代谢平衡中发挥关键作用,但它对猪IMF含量的影响还知之甚少,对中国西部地区的猪种更是如此.文章利用PCR-RFLP（HinfⅠ、HaeⅢ和Msp Ⅰ 3种限制性内切酶）分子标记技术,检测了杜洛克猪、长白猪、大白猪、内江猪、荣昌猪、汉江黑猪、汉中白猪、八眉猪和野猪共计265头猪H-FABP基因5＇-上游区和第二内含子区的遗传变异,利用最小二乘模型分析了H-FABP基因对猪肌内脂肪含量的遗传效应,并运用猪脂肪细胞培养,油红O染色和TG测定等技术检测了H-FABP基因不同基因型脂肪细胞内脂滴的形态和沉积的量.结果表明：（1）在HinfⅠ-RFLP位点上,上述品种和野猪均存在多态性,其中大白猪、八眉猪、汉江黑猪、汉中白猪和野猪表现为低度多态,杜洛克、长白猪、内江猪和荣昌猪为中度多态;除汉江黑猪（P＜0.05）和野猪（P＜0.01）外,其他猪种基因频率和基因型频率都处于Hardy-Weinderg平衡状态（P＞0.05）;而在HaeⅢ-RFLP和Msp Ⅰ-RFLP位点上,仅内江猪、荣昌猪、汉江黑猪和八眉猪为单态;（2）9种基因型对肌内脂肪（IMF）含量的影响,HH＞Hh＞hh,DD＜Dd＜dd,AA＜Aa＜aa,遗传效应值分别为：3.89,3.42,3.17,2.27,2.49,2.91,2.28,2.70,2.95,H-FABP基因可显著地提高IMF含量（P＜0.05）;（3）aaddHH型的脂肪细胞脂滴的形态,密度和含量与其他基因型细胞差异显著（P＜0.05）.结果提示：可通过提高＂aaddHH＂基因型的频率来增加IMF含量,达到改善猪肉质的目的.     

The swine histo compatibility system SLA: serological studies in the common Swiss and Danish swine breeds

Alloantisera were produced in common Swiss and Danish swine breeds by immunization with leucocytes or skin-grafts. Out of the 126 antisera, 66 were chosen for further study based on titrations employing lymphocytes from unrelated pigs typed with French SLA antisera (Vaiman, Chardon & Renard, 1979). The 66 selected antisera and the French reagents defining 26 SLA specificities were used to type lymphocytes from 595 unrelated pigs of the common Swiss and Danish breeds. The reaction-patterns of the French, Swiss and Danish antisera were adequately correlated for the French SLA specificities Nos FJ 2, 3, 6, 7, 8, 9, 11, 14, 19, 20 and 24 and for the haplotypes FJ 15.1.18 and FJ 5.4. In addition, a cluster of correlated Danish and Swiss antisera characterized a new specificity, provisionally designated CPH 31. This specificity was frequent in the Danish Landrace pigs.
Using the reagents identified in this report, the segregation of SLA markers was studied. Back-cross families demonstrated segregation of 15 distinct SLA haplotypes of which 14 are common in French Landrace or Large White. Differences were found in haplotype frequencies in both the Swiss and the Danish Landrace and Large White breeds.     

The swine histocompatibility system SLA: serological studies in the common Swiss and Danish swine breeds

Alloantisera were produced in common Swiss and Danish swine breeds by immunization with leucocytes or skin-grafts. Out of the 126 antisera, 66 were chosen for further study based on titrations employing lymphocytes from unrelated pigs typed with French SLA antisera (Vaiman, Chardon & Renard, 1979). The 66 selected antisera and the French reagents defining 26 SLA specificities were used to type lymphocytes from 595 unrelated pigs of the common Swiss and Danish breeds. The reaction-patterns of the French, Swiss and Danish antisera were adequately correlated for the French SLA specificities Nos FJ 2, 3, 6, 7, 8, 9, 11, 14, 19, 20 and 24 and for the haplotypes FJ 15.1.18 and FJ 5.4. In addition, a cluster of correlated Danish and Swiss antisera characterized a new specificity, provisionally designated CPH 31. This specificity was frequent in the Danish Landrace pigs. Using the reagents identified in this report, the segregation of SLA markers was studied. Back-cross families demonstrated segregation of 15 distinct SLA haplotypes of which 14 are common in French Landrace or Large White. Differences were found in haplotype frequencies in both the Swiss and the Danish Landrace and Large White breeds.     

A two-nucleotide deletion renders the mannose-binding lectin 2 (MBL2) gene nonfunctional in Danish Landrace and Duroc pigs

The mannose-binding lectins (MBLs) are central components of innate immunity, facilitating phagocytosis and inducing the lectin activation pathway of the complement system. Previously, it has been found that certain single-nucleotide polymorphisms (SNPs) in porcine MBL1 and MBL2 (pMBL1, pMBL2) affect mRNA expression, serum concentration, and susceptibility to disease, but the combinatory effect of pMBL1 and pMBL2 genotypes needs further elucidation. In the present study, pMBL1 and pMBL2 alleles, combined pMBL haplotypes, and MBL-A concentration in serum were analyzed in purebred Landrace (N?=?30) and Duroc (N?=?10) pigs. Furthermore, the combined pMBL haplotypes of 89 Piètrain × (Large White × Landrace) crossbred pigs were studied, and the genotypes of 67 crossbreds challenged with Escherichia coli were compared to their individual disease records. In the purebred animals, three non-synonymous SNPs and a two-nucleotide deletion were detected in the coding sequence of pMBL2. The two-nucleotide deletion was present at a frequency of 0.88 in the Landrace pigs and 0.90 in the Duroc pigs, respectively. In the crossbreds, the T allele of the SNP G949T in pMBL1—previously shown to have profound effect on MBL-A concentration even in the heterozygote condition—was detected in 47 % of the animals. Finally, an association was found between low-producing MBL genotypes and low body weight on the day of weaning in the same animals.     

Quantitative Analysis of Striped Coat-Color Patterns in Large White→Duroc Chimeric Pigs With Special Reference to the Genetic Control Mechanisms of the Dominant Black-Eyed White Phenotype

Coat colors of four chimeric pigs produced by the microinjection of dissociated blastomeres of (Landrace × Large White) blastocysts to the blastocyst cavity of (Duroc × Duroc) blastocysts (Kashiwazaki et al., 1992) exhibited characteristic horizontal stripe-patterns. We carried out quantitative analysis of those patterns in order to derive information concerning the genetic regulatory mechanisms of the dominant black-eyed white phenotypes in the pig. In the four chimeras, the theoretical mean widths of the single-clone stripe calculated from the estimated widths of minimal recognizable stripe (MRS) (Tachi, 1988) were 2.1 ± 0.1, 2.23 ± 0.15,1.89 ± 0.06, and 1.93 ± 0.28 cm respectively. The estimated number of single-clone stripes in the thoracico-lumbar region of those animals were 42.3, 40.7, 46.3, 44.2, and about twice the mean number of vertebrae in the same region (Duroc, 20 or 21; Large White, 21 or 22). Furthermore, the mean length of thoracico-lumbar vertebrate in two of the chimeric pigs, as measured on X-ray radiographs, was approximately twice the mean single-clone stripe width. It was concluded that the stripe-patterns of the chimeric pigs probably represented the dermatome patterns of epidermis; and in the pig, a single somite was likely to be derived from the clones of two primordial cells, as originally proposed by Gearhart & Mintz (1972) in the mouse. It was suggested, furthermore, that in the Large White→Duroc chimeric pigs, melanocytes that migrated into the region of skin formed by a Large White dermatome could not survive, thus creating a clearly demarcated white stripe. Possible involvement of KL or c-kit in the dominant black-eyed white phenotype of the pig is discussed.     

H2AFZ,RNF4 and NR4A1 loci are associated with boar semen quality by population association studies

Abstract

Spermatogenesis is a complex process regulated by many genes. In this study, H2AFZ, RNF4 and NR4A1 genes were selected as candidate genes for boar semen quality traits based on their functions during spermatogenesis, and the associations of three loci (H2AFZ c.192?+?210–192?+?213delCGAT, RNF4 c.374?+?358 T?>?C and NR4A1 c.956?+?796 A?>?G) with sperm quality traits were analyzed in Duroc (n?=?185), Large White (n?=?87) and Landrace (n?=?49) pig populations. The results showed H2AFZ c.192?+?210–192?+?213delCGAT AA boars produced 1.52% lower abnormal sperm rate (ASR) than AB boars in Landrace pigs (p?<?0.05); RNF4 c.374?+?358?TC boars produced 0.31?×?10⁸/ml higher sperm concentration (SCON) than CC boars (p?<?0.05) in Large White pigs; NR4A1 c.956?+?796 A?>?G was associated with ASR in Duroc and Large White pigs and was associated with sperm motility (MOT) in Large White and Landrace pigs. This study indicated the H2AFZ, RNF4 and NR4A1 loci were the potential molecular markers for improving the semen quality traits in boars.     

The effects of air dried poultry manure on performance,organ weights and haematological measurements of weaner pigs reared in the tropics

Sixty-three Large White and Large White × Landrace pigs, initially weighing approximately 10 kg, were allocated to 9 groups of 7 pigs each with equal constant sex and breed ratios. These were fed on one of 9 diets containing 0, 5 and 10% dried poultry manure (DPM) at three levels of protein (17, 19 and 21%) for 16 weeks. Growth rate, feed intake and nutrient digestibility were measured. At the end, all animals were slaughtered, blood samples collected for haematological studies, fresh organ weights recorded and tissue and organs examined for evidence of pathological conditions. Growth rate was not significantly influenced by DPM levels but was significantly (P < 0.05) influenced by protein levels. Apparent digestibility of nutrients was not significantly influenced by protein or DPM levels. Increasing DPM levels significantly increased feed consumption and worsened feed/gain ratio. Organ weights were not significantly influenced by treatments but total white blood cells, percentage banded neutrophils, segmented neutrophils and eosinophils differed significantly (P < 0.05) among groups. The banded and segmented neutrophils were higher, while the lymphocytes were lower, than the normal values; this suggests the presence of pathogenic organisms in pigs fed on these DPM diets.