Frequency of the Rendement Napole RN- allele in a population of American Hampshire pigs

A total of 204 purebred Hampshire pigs were obtained from 23 breeders. These animals were the progeny of 41 sires and 123 dams. A sample of purebred Yorkshire (n = 24) pigs were also used in the study. Animals were classified by glycolytic potential determined on a live-animal longissimus muscle biopsy sample. Hampshire pigs (n = 176) with glycolytic potential greater than 185 micromol/g (x = 238.8 micromol/g; SD = 29.54) were classified as heterozygous or homozygous for the dominant RN- allele (RN-rn+ or RN-RN-, respectively), whereas cohorts (n = 28) with glycolytic potential less than 185 micromol/g (x = 141.3 micromol/g; SD = 24.48) were considered as homozygous normal (rn+rn+). All Yorkshire pigs (n = 24) had a mean glycolytic potential level of 146.1 micromol/g (SD = 20.18) and were considered as homozygous normal (rn+rn+). The Hardy-Weinberg equilibrium yielded frequencies of .630 and .370 for the dominant RN- allele and normal rn+ alleles in the Hampshire population, respectively, and genotypic frequencies of .397 (RN-RN-), .466 (RN-rn+), and .137 (rn+rn+). Hampshires with glycolytic potential > or = 185 micromol/g had significantly lower longissimus muscle ultimate pH, intramuscular fat, subjective marbling scores, and percentage of protein (P < .001) and had greater longissimus muscle percentage of moisture (P < .001), drip loss (P < .01), and cooking loss (P < .001) than rn+rn+ Hampshires and Yorkshires. These data suggest the RN- allele exists at a high frequency within the American Hampshire breed. Higher glycolytic potential levels, which accompany the allele, may cause decreased meat quality.     

Effect on Rendement Napole genotype on metabolic markers in Ossabaw pigs fed different levels of fat

We investigated effects of Rendement Napole (RN ) genotype on metabolic markers in Ossabaw pigs fed diets with different levels of dietary fat. Thirty‐two pigs, belonging to either the wild‐type (WT , rn⁺/rn⁺) or carrier (CAR , RN ^?/rn⁺) genotypes (n  = 16/genotype), were divided into two dietary groups, (high fat [HF ] or low fat [LF ]) diets, for 12 weeks (n  = 8 pigs/genotype/diet) after which pigs were killed for gene expression analysis by RT ‐PCR . Feeding HF diet caused increased daily gain (ADG , p  <  .05) and final body weight (BW ) (p  <  .05) in comparison with the LF diet (p  <  .05). Feed efficiency (gain:feed) was higher (p  <  .05) in pigs on the HF and was higher (p  <  .05) in CAR pigs compared to WT . There was genotype × diet interaction (p  =  .05) on final BW such that CAR animals on LF diet had the same final BW as animals of both genotypes on HF diet. Carrier pigs on LF diet had higher (p  <  .05) average daily gain and gain:feed than WT pigs. There was a trend (p  <  .08) for a higher feed consumption in pigs on the LF diet. Backfat thickness was higher (p  <  .01) in pigs on the HF diet. Serum triglyceride was higher (0.62 vs. 0.33 mg/dl, p  <  .01) in pigs on HF diet. Serum insulin was higher (p  <  .05) in CAR versus WT pigs (0.40 vs. 0.015 μg/ml). Pigs on the HF diet had a higher (p  <  .05) serum insulin compared to those on the LF diet (0.032 vs. 0.023 μg/ml). Carnitine palmitoyl transferase 1‐alpha was higher (p  <  .05) in the longissimus dorsi and semitendinosus muscles of pigs on HF diet. Acyl‐CoA oxidase I was elevated (p  <  .05) in the liver of pigs on HF diet. Fatty acid synthase was lower in the longissimus dorsi muscle, liver and mesenteric fat (p  <  .05) of carrier pigs. The RN gene regulates specific metabolic markers in the Ossabaw pigs.     

The effect of the Halothane and Rendement Napole genes on carcass and meat quality characteristics of pigs

The objective of this study was to determine the effects of the Halothane (N) and Rendement Napole (RN) genes on carcass and meat quality characteristics in pigs. Halothane and RN carrier (Nn/RN- rn+) Hampshire boars (n = 4) were mated to dams that were homozygous for the normal allele of both genes (NN/rn+ rn+) to produce progeny of four genotypes: 1, NN/rn+ rn+ (n = 31); 2, Nn/rn+ rn+ (n = 27); 3, NN/RN- rn+) (n = 30); and 4, Nn/RN- rn+ (n = 23). A DNA test was used to determine Halothane genotype, and longissimus glycolytic potential was used to predict the RN genotype. Pigs were reared under standard conditions to approximately 120 kg live weight and slaughtered at a commercial plant, and carcass characteristics and meat quality were evaluated. Halothane carriers (Nn/ _ _), in comparison to Halothane normal (NN/_ _) pigs, had shorter carcasses, lower longissimus ultimate pH, higher Minolta L* and b* values, and greater drip loss. Rendement Napole gene carriers (_ _/RN- rn+) had higher L* and b* values and drip and cooking loss and lower longissimus ultimate pH than homozygous recessive animals (_ _/rn+ rn+). There were Halothane x RN genotype interactions (P < 0.05) for subjective color, firmness, and marbling scores, and for shear force. Animals that were normal for both genes (NN/rn+ rn+) had the highest subjective scores for color (2.60, 1.88, 1.85, and 1.95, SE = 0.181, P < 0.05), firmness (2.53, 2.03, 2.10, and 1.89, SE = 0.182, P < 0 .05), and marbling (2.11, 1.44, 1.53, and 1.55, SE = 0.153, P < 0 .05) for genotypes 1, 2, 3, and 4, respectively, suggesting darker, firmer muscle with a higher level of marbling for this genotype. Shear force was highest for Nn/rn+ rn+ animals (3.83, 4.41, 3.79, and 3.70, respectively, SE = 0.172, P < 0.05). Gilts had less s.c. backfat thickness, greater longissimus muscle area, and lower subjective marbling scores than barrows. There was no effect (P > 0.05) of gender on other meat quality traits. This study illustrates the negative effects of the Halothane and RN genes on fresh pork quality and suggests that in combination the detrimental effects of the two genes are additive for ultimate pH, objective color, and water-holding capacity.     

Effect of exercise on proglycogen and macroglycogen content in skeletal muscles of pigs with the Rendement Napole mutation

OBJECTIVE: To investigate influence of the Rendement Napole (RN-) mutation on proglycogen (PG) and macroglycogen (MG) content in skeletal muscles before and after exercise and evaluate glycogen concentrations within various muscle fiber types. ANIMALS: 5 pigs with the RN- mutation and 3 noncarrier pigs. PROCEDURE: Pigs performed 2 exercise tests on a treadmill. In the first, pigs (mean body weight, 27 kg) ran a distance of approximately 800 m. In the second, pigs (mean body weight, 63 kg) ran until fatigued. Biopsy specimens (biceps femoris muscle) for determination of PG and MG contents were obtained before and after exercise, 24 hours after the first test, and 3 hours after the second test. Histochemical analysis was performed on specimens obtained before and after the second test. RESULTS: Before exercise, PG stores did not differ markedly between groups, but MG stores were twice as high in pigs with the RN- mutation, compared with noncarrier pigs. The MG content decreased to a similar extent in both groups after exercise. Resynthesis of MG was greater in pigs with the RN- mutation than in noncarrier pigs by 3 hours after exercise. A low glycogen content after exercise was observed in many type I and type IIA fibers and in some type lIB fibers. CONCLUSIONS AND CLINICAL RELEVANCE: The RN- mutation was associated with high MG stores in skeletal muscle that did not influence exercise performance. The RN- mutation did not impair glycogenolysis during exercise but may induce faster resynthesis of MG after exercise.     

Growth performance, carcass composition, quality, and enhancement treatment of fresh pork identified through deoxyribonucleic acid marker-assisted selection for the Rendement Napole gene

Progeny (n = 70) from unrelated, DNA tested, Rendement Napole carrier (RN-/rn+) Hampshire sires, and DNA tested, Rendement Napole normal (rn+/rn+) Yorkshire dams were genotyped for the segregating RN- allele via DNA marker-assisted methodology. Six slaughter groups ensued, with littermates all being represented within the same slaughter group. Boneless pork loins were removed from right carcass sides after a 48-h chill at 2 degrees C. The anterior portions of the loins were not enhanced, whereas the posterior sections were enhanced with a solution containing 0.5% sodium chloride and 0.5% sodium tripolyphosphate to 110% of their initial weight. Carcasses of carrier pigs had less (P < 0.05) 10th rib fat depth and a greater (P < 0.01) percentage carcass lean than carcasses of normal pigs. Postmortem LM pH of carrier pigs was lower (P < 0.002) at 3, 6, 12, and 24 h, and tended to be lower (P = 0.062) at 48 h compared with that of normal animals. Samples of LM from carrier pigs had greater (P < 0.01) glycolytic potential values, drip loss percentages, and a* values, and lower pH values at fabrication than LM from normal pigs. No genotype differences (P > 0.05) were found for LM lactate, L*, or b* values. Nonenhanced semimembranosus samples from carrier pigs exhibited greater (P < 0.05) purge loss percentages and L* values, and lower (P < 0.01) pH values than samples from normal pigs. Enhanced LM samples exhibited greater (P < 0.05) drip and purge loss percentages, greater pH, and lower L* values at fabrication, regardless of Napole status. These findings suggest that the Napole gene has a positive influence on carcass leanness but detrimental effects for lean quality, which were often further compounded when meat was subjected to enhancement treatment.     

Screening 52 single nucleotide polymorphisms for extreme value of glycolytic potential and drip loss in pigs

Low heritability of meat quality traits and the lack of their systematic registration in breeding programs have encouraged the search for single nucleotide polymorphisms (SNPs) located within genes coding the proteins involved in muscle and fat metabolism. In this report, a panel of 52 SNPs was used to find which alleles and genotypes are more/less frequent in groups of pigs differentiated by extreme value of glycolytic potential (GP) and drip loss (DL). The analysis was carried out in 52 fatteners (chosen from 246 pigs), of which 28 were Landrace and 27 Landrace × Yorkshire. Two designs were performed: I, fatteners were divided into two groups showing extreme value of GP (<125 versus >145), II, fatteners were divided into two groups showing extreme value of DL (<6.0 versus >6.0). Allele frequency differences between the phenotypic groups of extreme GL or DL were not influenced by the breed. The frequency of 52 SNPs alleles for each of group was calculated and a chi‐squared test was used to estimate the significance of differences in allele frequencies between alternative groups in each experimental design. Three SNPs (DECR1, PPARGC1, MC4R) and another two (CYP21, SFRS1) showed significant differences between groups of extreme GP and DL, respectively. To exemplify and validate potential associations of candidate SNPs for GP and DL, 293 fatteners representing three commercial breeds/crosses (95 Landrace, 66 Landrace × Yorkshire and 132 Landrace × Yorkshire × Duroc were genotyped for DECR1 and CYP21 by PCR‐RFLP assays. DECR1 showed significant associations with GP in Landrace and Landrace × Yorkshire × Duroc fatteners. CYP21 showed significant associations with DL in all breeds/crosses. Interestingly, the CYP21 polymorphism revealed adverse associations trend in Landrace × Yorkshire × Duroc pigs in comparison to Landrace and Landrace × Yorkshire fatteners.     

军牧1号白猪、杜洛克猪和藏猪的氟烷基因和酸肉基因多态性分布

为了研究军牧1号白猪、杜洛克猪和藏猪氟烷基因和酸肉基因的多态性分布情况,试验采用PCR-RFLP方法对61头军牧1号白猪、51头杜洛克猪和51头藏猪的氟烷基因和酸肉基因多态性进行了检测。结果表明:军牧1号白猪的氟烷基因表现为单一的NN型;杜洛克猪的氟烷基因表现为多态性,等位基因N的频率为0.196 1,基因型分布符合哈代-温伯格平衡(χ2=3.033 9,P=0.081 5);藏猪的氟烷基因表现为单一的nn型。3个猪种的酸肉基因均表现为单一的rn/rn型。     

Effects of multiple concurrent stressors on rectal temperature, blood acid-base status, and longissimus muscle glycolytic potential in market-weight pigs

Sixty-four market-weight (130.0 +/- 0.65 kg) barrows (n = 16) and gilts (n = 48) were used in a split-plot design with a 2 x 2 x 2 factorial arrangement of treatments: 1) handling intensity (gentle vs. aggressive), 2) transport floor space (0.39 vs. 0.49 m(2)/pig), and 3) distance moved during handling (25 vs. 125 m) to determine the effects of multiple concurrent stressors on metabolic responses. For the handling intensity treatment, pigs were moved individually approximately 50 m through a handling course with either 0 (gentle) or 8 (aggressive) shocks from an electric goad. Pigs were loaded onto a trailer and transported for approximately 1 h at floor spaces of either 0.39 or 0.49 m(2)/pig. After transport, pigs were unloaded, and the distance moved treatment was applied; pigs were moved 25 or 125 m through a handling course using livestock paddles. Rectal temperature was measured, and blood samples (to measure blood acid-base status) were collected 2 h before the handling intensity treatment was applied and immediately after the distance moved treatment was applied. A LM sample to measure glycolytic potential was collected after the distance moved treatments on a subset of 32 pigs. There were handling intensity x distance moved interactions (P < 0.05) for several blood acid-base measurements. In general, there was no effect of distance moved on these traits when pigs were previously handled gently. However, when pigs were previously handled aggressively, pigs moved 125 compared with 25 m had greater (P < 0.05) blood lactate and less (P < 0.05) blood pH, bicarbonate, and base-excess. Pigs transported at 0.39 compared with 0.49 m(2)/pig had a greater (P < 0.01) increase in creatine kinase values; however, transport floor space did not affect any other measurements. Data were analyzed by the number of stressors (the aggressive handling, restricted transport floor space, and 125-m distance moved treatments) experienced by each pig (0, 1, 2, or 3). As the number of stressors experienced by the pig increased, rectal temperature, blood lactate, and LM lactate increased linearly (P 相似文献   

Effect of vitamin E on improving fresh pork quality in Berkshire- and Hampshire-sired pigs

This study was designed to evaluate the effects of vitamin E supplementation on pork quality of two genotypes with distinct differences in pork quality traits. Pigs (n = 240; BW = 87 +/- 0.35 kg) were allotted by weight to one of 20 treatments (4 pens/treatment, 3 pigs/pen) in a 2 x 2 x 5 factorial randomized complete block design. Factors included 1) genotype (Berkshire or Hampshire sired), 2) sex (gilts or barrows), and 3) vitamin E level (12.1, 54.7, 98.8, 174.0, and 350.6 IU of vitamin E/kg diet). Hampshire-sired pigs had greater average daily gain (1.05 vs 0.98 kg) and gain:feed (0.30 vs 0.27) and less average daily feed intake (ADFI) (3.46 vs 3.62 kg) than Berkshire-sired pigs (P < 0.001) for the 6-wk study. Hampshire-sired barrows consumed more feed (3.54 vs 3.38 kg/d) and were less efficient (0.29 vs 0.31) than Hampshire-sired gilts (P < 0.05), but this sex difference was not observed in Berkshire-sired pigs (interaction, P < 0.05). Berkshire-sired pigs had greater backfat (34.1 vs 21.1 mm; P < 0.001), reduced longissimus muscle area (37.6 vs 46.3 cm2; P < 0.001), reduced lean percentage (53.0 vs 55.8; P < 0.001), and a greater head-on yield (79.8 vs 79.2; P < 0.05). Vitamin E increased (P < 0.05) ADFI linearly (P < 0.05), but had no effects on carcass composition. Loin chops from Hampshire-sired pigs had reduced ultimate pH (5.64 vs 5.91), greater drip loss (92.2 vs 66.3 mg), and increased Minolta L* (52.6 vs 48.6), a* (8.9 vs 7.5), and b* (6.9 vs 5.2) values compared to Berkshire-sired pigs (P < 0.001). Vitamin E had no effect on pH, temperature, drip loss, and L* or a* values, but tended (P < 0.07) to increase b* values linearly (P < 0.06). Oxidation as indicated by thiobarbituric acid reactive substances (TBARS) was greatest in Hampshire-sired gilts at the lowest level of vitamin E, and decreased linearly (P < 0.001) with additional vitamin E. However, TBARS responded in a cubic fashion (P < 0.05) to vitamin E in Hampshire-sired barrows and were not affected in Berkshire-sired gilts or barrows (three-way interaction, P < 0.02). Hampshire-sired pigs had greater TBARS than Berkshire-sired pigs (0.053 vs 0.047 mg malondialdehyde equivalents/kg). Vitamin E supplementation increased serum concentrations of vitamin E on d 21 (1.06 to 4.79 microg/mL) and d 42 (1.02 to 2.82 microg/mL) and increased tissue concentrations of vitamin E (1.99 to 4.83 microg/g) linearly (P < 0.001). Vitamin E supplementation was not effective in improving fresh meat quality in genotypes with poor or superior meat quality traits.     

Investigations on potential dietetic effects of lactulose in pigs

Effects of lactulose (lac) as feed additive on chyme and feces quality in pigs (reared piglets: n = 10; fattening pigs: n = 20) were tested. Furthermore, fattening pigs were experimentally infected with Salmonella Derby (orally 2.6 × 10⁸ cfu/pig, single dose infection on 1 day) to examine potential prophylactical effects. Pigs were fed a conventional diet ad libitum (in the control group without lac, in the experimental group including lac: 28.6/26.9 g lac/kg dry matter). In piglets and fattening pigs lac intake did not alter fecal dry matter (DM) content or feces pH. Contrary to findings in human beings only small amounts of lac reached the hindgut, although higher lac concentrations were measured in ileal chyme (in piglets: 10–40 g, in fattening pigs 40–70 g lac/kg DM of chyme). Lac ingestion led neither to increased concentrations of lactic acid or volatile fatty acids (vfa) nor to reduced pH data in chyme of small intestine, but in cecum and colon contents higher vfa concentrations (significant effects in piglets) were measured. Furthermore, lac ingestion resulted in reduced ammonia levels in small and large intestine contents. But no protective effects of lac ingestion were recognized when fattening pigs were infected orally with Salmonella Derby. Due to the fact that only parts of lac reach the hindgut there are doubts, whether lac can lead to comparable dietetic effects in pigs as known from human beings (laxation, forced elimination of bacteria a.s.o.). The obtained results in pigs underline the necessity of experimental studies in the target species before substances are used and recommended for dietetic purposes in feeding of pigs.     

Effect of feed withdrawal and handling intensity on longissimus muscle glycolytic potential and blood measurements in slaughter weight pigs

This study was carried out to evaluate the effect of feed withdrawal and handling intensity on blood acid-base responses and muscle glycolytic potential in slaughter-weight pigs. Sixty crossbred pigs (BW = 107.7 +/- 0.56 kg; 44 barrows and 16 gilts) were used in a randomized complete block design with a 2 x 2 factorial arrangement of treatments: 1) feed withdrawal (0 vs. 24 h), and 2) handling intensity (low vs. high). The high-intensity handling treatment consisted of moving the pigs through a passage (12.2 m long x 0.91 m wide) for eight laps using an electric goad two times per lap. Pigs in the low-intensity handling treatment were moved at their own pace through the passage for eight laps using a livestock panel and paddle. Biopsy samples were collected from the LM at the beginning of feed withdrawal, at the end of the handling procedure, and 4 h after handling. Blood samples were collected 2 h before and immediately after the handling procedure. There were no interactions between feed withdrawal and handling intensity for any of the variables measured. Feed withdrawal decreased (P < 0.05) baseline and posthandling body temperature (38.85 vs. 38.65 degrees C; SEM = 0.060 and 39.70 vs. 39.37 degrees C; SEM = 0.04, respectively) and blood glucose, lowered (P < 0.05) baseline partial pressure of oxygen and partial pressure of carbon dioxide, and increased (P < 0.01) baseline and posthandling plasma free fatty acid concentrations. High-intensity handling produced higher (P < 0.01) posthandling lactate and glucose, and lower (P < 0.01) posthandling blood pH (7.33 vs. 7.18 +/- 0.02, respectively), bicarbonate, base excess, and total carbon dioxide than low-intensity handling. Longissimus muscle glycolytic potential of fasted pigs was lower (P < 0.01) than in fed pigs at the end of the handling procedure (177.2 vs. 137.0 micromol/g of wet tissue; SEM = 10.08, respectively). There was no effect of handling intensity on longissimus muscle glycolytic potential. Feed withdrawal did not attenuate the blood acid-base changes caused by handling; however, the combination of feed withdrawal and handling decreased muscle glycolytic potential.     

藏香猪毕氏肠微孢子虫感染情况和基因分型

采集了青海省和陕西省部分地区共计450份藏香猪新鲜粪便样品,基于小亚基核糖体RNA基因(small-subunit ribosomal RNA,SSU rRNA)的巢式PCR检测样品中毕氏肠微孢子虫(Enterocytozoon bieneusi)的感染情况,并分析其遗传多样性。结果显示,218份样品感染了毕氏肠微孢子虫,总感染率为48.4%(218/450);青海省所调查藏香猪的毕氏肠微孢子虫感染率为65.8%(154/234),显著高于陕西省的感染率29.6%(64/216)(P0.001);4个年龄段间藏香猪毕氏肠微孢子虫感染率差异显著(P0.001),其中哺乳仔猪感染率最高(92.7%),而成年猪感染率最低(9.2%)。序列分析发现,藏香猪存在5个SSU rRNA基因型(Type 1～Type 5),其中Type 1为优势基因型。对218份毕氏肠微孢子虫阳性分离株的多位点序列分型发现,分别有59,26,9和99份样品在MS1、MS3、MS4和MS7基因位点成功扩增,呈现18,4,4和4种单倍型,基于MS1、MS3和MS4等3个基因位点共形成6种多位点基因型(multi-locus genotypes,MLGs)。     

Pulsed-field gel electrophoresis genotyping of Salmonella gallinarum and comparison with random amplified polymorphic DNA

Salmonella gallinarum is gram-negative bacteria that cause fowl typhoid (FT) in chickens. Since the first outbreak of FT reported in 1992 in Korea, it has widely spread throughout the country. Today, FT is one of the most devastating diseases of poultry. The aim of the present study was to ascertain a genetic relationship among S. gallinarum isolates collected from different regions of Korea over a 10-year period. We examined a total of 38 isolates of S. gallinarum obtained in 29 regions of Korea from 1992 to 2001 including the 9R vaccine strain and the standard strain of S. gallinarum (ATCC 9184). The PFGE profiles produced 12 different patterns with the XbaI-digestion and 11 different patterns with the SpeI-digestion. The RAPD using URP-6 primers showed eight different genotypes with the same Salmonella isolates. The PFGE patterns of the 9R vaccine strain and ATCC 9184 of S. gallinarum were different from the identical type A, the most common genotype among field isolates in our study. In conclusion, a low genetic heterogeneity was observed among Korean S. gallinarum isolates. In addition, PFGE appeared to be a more accurate and reproducible method for genotyping of S. gallinarum isolates than RAPD.     

Effect of feed withdrawal and dietary energy source on muscle glycolytic potential and blood acid-base responses to handling in slaughter-weight pigs

The effects of dietary energy source and feed withdrawal on muscle glycolytic potential (GP) and blood acid-base responses to handling were investigated in slaughter-weight pigs (initial BW 94.7 ± 1.01 kg). Crossbred pigs (n = 96; 48 barrows, 48 gilts) were used in a randomized complete block design with a 4 × 2 × 2 factorial arrangement of treatments: l) diet [control, high fat (10% supplemental fat), low-digestible carbohydrate (20% total starch), and high-fat/low-digestible carbohydrate (10% supplemental fat and 20% total starch)]; 2) feed withdrawal (0 and 36 h); and 3) sex (barrow and gilt). Diets were fed for 28 d before the feed withdrawal treatment was applied, at the end of which all pigs were individually moved through a 12.20-m-long × 0.91-m-wide passageway for 16 laps (195 m total distance), with the assistance of an electric goad (2 times per lap). Longissimus muscle biopsies were collected at the beginning of the feeding and feed withdrawal periods and immediately after and 4 h after the handling procedure. Venous blood was collected 2 h before and immediately after the handling procedure to measure acid-base responses. At the end of the feeding period, pigs fed the control and high-fat diets were heavier (P < 0.001) than those on the low-digestible carbohydrate and the high-fat/low-digestible carbohydrate diets (129.8, 130.9, 114.0, and 122.1 kg, respectively; SEM 1.91). Diet, feed withdrawal, and sex did not affect (P > 0.05) blood acid-base responses to handling. Muscle GP at the end of the feeding period and 4 h posthandling was least (P < 0.05) for pigs fed the high-fat diet and similar for the other 3 diet treatments. Pigs subjected to 36 h compared with 0 h of feed withdrawal had less GP (P < 0.05) immediately after and 4 h after the handling procedure. There was an interaction between diet and feed withdrawal treatments for changes in GP from the start of feed withdrawal to 4 h posthandling. The reduction in GP was greater (P < 0.05) for fasted than for fed pigs receiving the control and high-fat diets, but was similar (P > 0.05) for fasted and fed pigs receiving the 2 low-digestible carbohydrate diets. In conclusion, neither dietary energy source nor fasting affected blood acid-base responses to handling; however, fasting-induced changes in LM GP were diet dependent.     

Preslaughter handling effects on pork quality and glycolytic potential in two muscles differing in fiber type composition

The objective of the present experiment was to investigate the effects of transportation, lairage, and preslaughter stressor treatment on glycolytic potential and pork quality of the glycolytic longissimus and the oxidative supraspinatus (SSP) or serratus ventralis (SV) muscles. In a 2 x 2 x 2 factorial design, 384 pigs were assigned randomly either to short (50 min) and smooth or long (3 h) and rough transport, long (3 h) or short (< 45 min) lairage, and minimal or high preslaughter stress. Muscle samples were taken from the LM at 135 min and from the SSP at 160 min postmortem for determination of the glycolytic potential and rate of glycolysis. At 23 h postmortem, pork quality was assessed in the LM and the SV. Effects of transport and lairage conditions were similar in both muscle types. Long transport increased (P < 0.01) the glycolytic potential and muscle lactate concentrations compared with short transport. Both long transportation and short lairage decreased (P < 0.01) redness (a* values) and yellowness (b* values) of the LM and SV. In combination with short lairage, long transport decreased (P < 0.05) pork lightness (lower L* values), and electrical conductivity was increased (P < 0.05) after long transport. Several interactions between stress level and muscle type (P < 0.001) were observed. High preslaughter stress decreased (P < 0.001) muscle glycogen in both the LM and SSP, but this decrease was greater in the LM. Lactate concentrations were increased (P < 0.001) only in the LM by high preslaughter stress. Increases in ultimate pH (P < 0.001) and decreases in a* values (P < 0.01) were greatest in the SV, whereas increases in electrical conductivity (P < 0.001) were greatest in the LM. The lack of interactions among transportation, lairage, and muscle type was attributed to the relatively minor differences in stress among treatments. It was concluded that, in glycolytic muscle types such as the LM, the high physical and psychological stress levels associated with stress in the immediate preslaughter period have a greater effect on the water-holding capacity of the meat and may promote PSE development. Conversely, oxidative muscle types tend to have higher ultimate pH values and produce DFD pork in response to intense physical activity and/or high psychological stress levels preslaughter.     

Plasma cell-free DNA in canine lymphoma patients as a novel material for genotyping

Canine lymphoma is a disease with high morbidity and poor long-term prognosis, despite a high response rate to chemotherapy. In this study, we focused on liquid biopsy, in which small amounts of substances from body fluids were analysed, to determine whether cell-free DNA (cfDNA) in the plasma can be used as a biomarker for lymphoma in dogs. We found that 23 patients with lymphoma had significantly higher cfDNA concentrations than the 12 healthy dogs (median 2360 ng/mL versus 299 ng/mL, p < .0001). Polymerase chain reaction for antigen receptor rearrangement (PARR) was also employed using cfDNA from the lymphoma group to investigate whether cfDNA could be used for the detection of genetic clonality of lymphomas, as well as the genomic DNA (gDNA) extracted from an original lesion in each case. The correlation of the PARR results between cfDNA and gDNA was observed in 100% of B-cell lymphomas (10/10), 77.8% of T-cell lymphomas (7/9), and 100% of other types of lymphomas (4/4), respectively. These results indicate that plasma cfDNA levels are increasing in canine lymphoma patients, that cfDNA concentration can be a novel diagnostic tool, and that it can be used as a diagnostic tool for PARR.     

Detection and genotyping of porcine circovirus in naturally infected pigs by oligo-microarray

A rapid and reliable method for the identification of porcine circovirus (PCV) genotypes based on oligonucleotide microarray hybridization has been developed. The genotype-specific oligonucleotides (22-30 mer) immobilized on the surface of glass slides were selected to bind to the multiple target sites within the replication gene that are conserved among individual PCV genotypes. Cy5-labeled DNA targets were amplified in a PCR with primers common to both genotypes. The identification of PCV genotype was based on hybridization with several individual genotype-specific oligonucleotides. This approach combines the high sensitivity of PCR with the selectivity of DNA-DNA hybridization. The utility and feasibility of oligonucleotide microarray hybridization was evaluated by testing standard and 87 clinical isolates. Analysis of the specimens showed that this microarray-based method is capable of unambiguous identification of both genotypes and fivefold more sensitive than gel electrophoresis. Our results indicated that the oligonucleotide array is useful for the identification and discrimination of PCV from clinical isolates and specimens in a clinical laboratory.     

猪圆环病毒检测与分型寡核苷酸芯片的建立及应用

本研究应用寡聚核苷酸基因杂交技术建立了一种快速可靠的检测猪圆环病毒(PCV)基因型的方法.在PCV保守序列复制酶基因内设计了2对特异性引物,在此物之间设计了2种基因型特异的核苷酸探针(22～30 mer).通过PCR扩增Cy5标记的DNA片段与固定在玻片表面的探针杂交进行PCV基因分型.该技术结合了PCR方法的高度敏感性和DNA-DNA杂交技术的选择特异性.利用该方法对58倒临床样品进行了检测鉴定.结果显示,该技术能准确鉴别PCV病毒基因型.且其灵敏度是凝胶电泳的5倍.试验结果表明寡核苷酸基因芯片技术可有效地应用于PCV临床诊断和分子流行病学调查.