Swine Muscle Research Articles

On-line determination of fatty acid composition in intramuscular fat of Iberian pork loin by NIRs with a remote reflectance fibre optic probe

A near infrared spectrometer equipped with a standard 210/210 bundle remote reflectance fibre-optic probe, with a 5 × 5 cm quartz window type, was used for the determination of fatty acids in the Longissimus dorsi muscle of Iberian breed swine. The fatty acids C14:0, C16:0, C16:1, C17:0, C17:1, C18:0, C18:1, C18:2, C18:3, Σpolyunsaturated, Σmonounsaturated and Σsaturated were determined in samples of intramuscular fat from Iberian breed swine by direct application of the fibre-optic probe onto the loin sample, with no treatment or manipulation of the sample. The regression method employed was modified partial least squares. The calibration results using the fibre-optic probe for 74 loin samples had multiple correlation coefficients (RSQ) for C14:0, C16:0, C16:1, C17:0, C17:1, C18:0, C18:1, C18:2, C18:3, Σpolyunsaturated, Σmonounsaturated and Σsaturated acid of 0.785, 0.798, 0.788, 0.825, 0.762, 0.765, 0.696, 0.859, 0.878, 0.807, 0.943, 0.858, respectively, and standard errors of prediction corrected for the same fatty acids (%) of 0.08, 0.63, 0.26, 0.02, 0.02, 0.51, 0,77, 0.64, 0.05, 1.06, 0.34, 0.70, respectively. The robustness of the method was checked by applying the fibre-optic probe to unknown samples of Iberian breed pork loin in a slaughterhouse, using 15 samples for the external validation.

Sensitive PCR analysis of animal tissue samples for fragments of endogenous and transgenic plant DNA.

An optimized DNA extraction protocol for animal tissues coupled with sensitive PCR methods was used to determine whether trace levels of feed-derived DNA fragments, plant and/or transgenic, are detectable in animal tissue samples including dairy milk and samples of muscle (meat) from chickens, swine, and beef steers. Assays were developed to detect DNA fragments of both the high copy number chloroplast-encoded maize rubisco gene (rbcL) and single copy nuclear-encoded transgenic elements (p35S and a MON 810-specific gene fragment). The specificities of the two rbcL PCR assays and two transgenic DNA PCR assays were established by testing against a range of conventional plant species and genetically modified maize crops. The sensitivities of the two rbcL PCR assays (resulting in 173 and 500 bp amplicons) were similar, detecting as little as 0.08 and 0.02 genomic equivalents, respectively. The sensitivities of the p35S and MON 810 PCR assays were approximately 5 and 10 genomic equivalents for 123 bp and 149 bp amplicons, respectively, which were considerably less than the sensitivity of the rbcL assays in terms of plant cell equivalents, but approximately similar when the higher numbers of copies of the chloroplast genome per cell are taken into account. The 173 bp rbcL assay detected the target plant chloroplast DNA fragment in 5%, 15%, and 53% of the muscle samples from beef steers, broiler chickens, and swine, respectively, and in 86% of the milk samples from dairy cows. Reanalysis of new aliquots of 31 of the pork samples that were positive in the 173 bp rbcL PCR showed that 58% of these samples were reproducibly positive in this same PCR assay. The 500 bp rbcL assay detected DNA fragments in 43% of the swine muscle samples and 79% of the milk samples. By comparison, no statistically significant detections of transgenic DNA fragments by the p35S PCR assay occurred with any of these animal tissue samples.

Simultaneous determination of lincomycin and virginiamycin M 1 in swine muscle, liver and kidney by liquid chromatography–electrospray ionization tandem mass spectrometry

A simple, selective and sensitive method for simultaneous determination of lincomycin (LCM) and virginiamycin M 1 (VGM) in swine muscle and organs using liquid chromatography with electrospray ionization tandem mass spectrometry is presented. Acetonitrile was used to extract the analytes from the samples. Lipids in the extracts were removed by liquid–liquid partition with n-hexane. Analytes in the purified acetonitrile extracts were separated by liquid chromatography with a reversed-phase Hypersil BDS C 18 column (250 mm × 2.1 mm, 5 μm) under gradient elution of an ammonium formate buffer (saturated ammonium formate:formic acid:acetonitrile:water, 1:5:50:950, v/v) and 0.1% formic acid in acetonitrile at 0.2 mL min −1. Identification was performed by monitoring two pairs of multiple reaction monitoring ions from the parent ions ( m/ z 407.2→126.1 and 407.2→359.2 for LCM, respectively, and m/ z 526.3→508.3 and 526.3→355.1 for VGM, respectively) at the defined retention time windows and by matching of the specific tolerance of relative abundance of major ions as stated in the European Union Commission Decision 2002/657/EC. Quantification was based on the relative ratios of the sum of peak areas of major ions of the analytes to those of clindamycin (internal standard) with reference to the respective ratios of the calibration standards. The method was validated by determining and evaluating the accuracy and precision of the intra-day and inter-day repeatability at two independent spike levels and the trueness of measurements on analysis of quality check samples. The limits of detection in different matrices were found ranging from 0.6 to 1.5 μg kg −1 and 1.9 to 4.3 μg kg −1 for LCM and VGM, respectively.

Metabolite pattern of Cysticercus cellulosae metacestode from different predilection sites of swine using proton magnetic resonance spectroscopy.

Cysticercosis due to Taenia solium is one of the most common public health problems in various regions of the world. We have performed prolon magnetic resonance spectroscopy (1H MRS) experiments of the fluid aspirated from cysticerci excised from skeletal muscle (n = 16) and brain (n = 9) of infected swine to compare the metabolite pattern of cysticerci in different predilection sites. Perchloric acid extract of cysticercus cysts excised from skeletal muscles (n = 16) was also prepared to ascertain water-soluble, low molecular weight metabolites using 1H MRS. Absolute quantification and statistical analysis of different metabolites was done to look for any significant differences in different locations of cysts. The metabolite pattern of cysticerci was found to be similar in the various predilection sites. Metabolites observed were leucine, valine, alanine, lysine, glycine, lipid contents, lactate, glutamate, acetate, succinate, creatine, choline, and glucose. Concentration of creatine in cysticercus fluid of cysts removed from the muscle was found to be significantly higher (p = 0.001) than the cysts located in the brain. We conclude that the metabolite pattern in the cysticerci is not influenced by the surrounding tissue location; however concentration of certain metabolites may depend upon the tissue location.

ＬＣ／ＭＳによる豚筋肉および肝臓中のカルバドックス代謝体（キノキサリン‐２‐カルボン酸およびデスオキシカルバドックス）の定量

A sensitive and selective method using liquid chromatography-electrospray mass spectrometry (LC-ESI-MS) for the determination of carbadox metabolites, quinoxaline-2-carboxylic acid (QCA) and desoxycarbadox (Desoxy-CDX), in swine muscle and liver has been developed. The LC separation was performed on a Cadenza CD-C18 column (10 cm x 2 mm i.d.) with a gradient system of 0.01% acetic acid-acetonitrile as the mobile phase at a flow rate of 0.2 mL/min. Negative ionization produced the [M-H]- molecular ion of QCA. On the other hand, the positive mode produced the [M+H]+ ion of Desoxy-CDX. The calibration graphs for QCA and Desoxy-CDX were rectilinear from 0.01 to 0.5 ng with selected ion monitoring (SIM). The drugs were extracted with 0.3% metaphosphoric acid-methanol (7:3), and the extracts were cleaned up on an Oasis HLB cartridge (60 mg) and by liquid-liquid extraction. The recoveries of QCA and Desoxy-CDX from swine muscle and liver fortified at 2.5 and 5 ng/g were 70.2-86.3%, and the detection limits were 1 ng/g for both drugs.

ＬＣ／ＭＳ／ＭＳによる食品中のキノロン剤の同時定量

A simple method was developed for the simultaneous determination of seven quinolones (enoxacin, ofloxacin, ciprofloxacin, danofloxacin, lomefloxacin, enrofloxacin and sarafloxacin) in foods using liquid chromatography coupled with electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS). The seven quinolones were extracted with acetonitrile containing 0.2% formic acid, and the extracted solution was cleaned up on a C18 cartridge. The extract was diluted with 5 mmol/L IPCC-MS3 for injection into the LC-ESI-MS/MS. The LC separation was carried out on an ODS column with gradient elution of 5 mmol/L IPCC-MS3-acetonitrile as the mobile phase. Mass spectral acquisition was done in the positive ion mode by applying selected reaction monitoring (SRM). The recoveries of the seven quinolones were mostly greater than 60% from foods fortified at 10 ng/g. The detection limits in foods were 2 ng/g for enoxacin and ciprofloxacin, and 1 ng/g for the other drugs. Twenty cattle muscle, 7 swine muscle, 9 chicken muscle, 16 milk, 19 prawn and 20 broiled eel samples from retail markets were analyzed by this method. Enrofloxacin and its metabolite ciprofloxacin were detected in 9 broiled eel at the level of trace (tr)-34 ng/g and tr-10 ng/g, respectively.

Comparison of a Modified Digestion Assay with Trichinoscopy for the Detection of Trichinella Larvae in Pork

A pepsin-HCl digestion assay and two compressorium techniques (trichinoscopy) for the identification of swine muscle tissue containing low levels of Trichinella larvae were compared as part of the test validation process for quality assurance purposes. Compressoria read with a stereomicroscope detected more larvae (P < 0.0001, n = 57) and more tissues (P =0.0047, n = 57) than did compressoria read with a projection microscope (trichinoscope). The digestion assay evaluated was 3.2 times as likely as the best compressorium technique to identify a positive tissue when these procedures were used to test 1 g of infected muscle (P < 0.001; 95% confidence interval for the odds ratio, 2.0 to 5.4; n = 161 and n = 189, respectively). Detection by trichinoscopy improved as the number of larvae in tissues increased to >2 larvae per g, but trichinoscopy was less sensitive than the digestion assay regardless of the tissue larval load. These data indicate that the quality controlled digestion assay used in this study is more sensitive than trichinoscopic techniques in the detection of tissues containing low levels of Trichinella larvae.

Essentiality of Histidine in Ruminant and Other Animals Including Human Beings

Concept and establishment of essential amino acids in animals and human beings rendered immeasurable contributions to animal production and human health. In ruminant animals, however, essential amino acids have never been completely established. The present review proposes a hypothesis that histidine may not be an essential amino acid for normal growing cattle (Japanese black) at least at the growing stage after about 450 kg of body weight on the basis of the experimental results of histidinol dehydrogenase activities in some tissues of the cattle together with hints from which the hypothesis was derived. At the same time, histidinol dehydrogenase activities in liver, kidney and muscle of swine, mouse, fowl and wild duck will be shown and the essentiality of histidine in these animals will be discussed. Finally, the essentiality of histidine for adult human will briefly be discussed. (Asian-Aust. J. Anim. Sci. 2003. Vol 16, No. 3 : 445-454)

Residue analysis of spectinomycin in tissues of chicken and swine by HPLC

A reversed-phase HPLC method with ultraviolet detection using p-nitrophenyl hydrazine as a pre-column derivatizing reagent was investigated for the determination of the antibiotic spectinomycin (SPCM) in muscle, liver, kidney and fat of chicken and swine. SPCM was extracted from samples with 10% trichloroacetic acid saturated with EDTA-2Na, and then cleaned up with coupled Sep-Pak Plus PS-2 cartridges. The detection limit of SPCM was 0.02 microgram (potency)/g. Recoveries of SPCM ranged from 77.4 to 97.4% for chicken tissues and from 74.5 to 91.8% for swine tissues. The present method was used for the analysis of chicken tissues after the 11th day of withdrawal (SPCM-medicated drinking water: 500 mg (potency)/L, for 7 days), and swine tissues after the 14th day of withdrawal (SPCM-medicated feed: 100 mg (potency)/kg, for 7 days). Results showed that SPCM concentrations were lower than the MRLs in all tissues.

Multilaboratory Trial for Determination of Ceftiofur Residues in Bovine and Swine Kidney and Muscle, and Bovine Milk

A multilaboratory trial for determining ceftiofur-related residues in bovine and swine kidney and muscle, and bovine milk was conducted following regulatory guidelines of the U.S. Food and Drug Administration, Center for Veterinary Medicine. The methods convert all desfuroylceftiofur-related residues containing the intact beta-lactam ring to desfuroylceftiofur acetamide to establish ceftiofur residues in tissues. Four laboratories analyzed 5 sets of samples for each tissue. Each sample set consisted of a control/blank sample and 3 control samples fortified with ceftiofur at 0.5 Rm, Rm, and 2 Rm, respectively, where Rm is the U.S. tolerance assigned for ceftiofur residue in each tissue/matrix: 0.100 microg/mL for milk, 8.0 microg/g for kidney (both species), 1.0 microg/g for bovine muscle, and 2.0 microg/g for swine muscle. Each sample set also contained 2 samples of incurred-residue tissues (one > Rm and one < Rm) from animals treated with ceftiofur hydrochloride. All laboratories completed the method trial after a familiarization phase and test of system suitability in which they demonstrated > 80% recovery in pretrial fortified test samples. Results showed that the methods met all acceptable performance criteria for recovery, accuracy, and precision. Although sample preparation was easy, solid-phase extraction cartridge performance must be carefully evaluated before samples are processed. The liquid chromatography detection system was easily set up; however, the elution profile may require slight modifications. The procedures could clearly differentiate between violative (> Rm) and nonviolative (< Rm) ceftiofur residues. Participating laboratories found the procedures suitable for ceftiofur residue determination.

Demonstration of Scolex in Calcified Cysticercus Lesion using Gradient Echo with or without Corrected Phase Imaging and its Clinical Implications

AIM: This study was performed to determine the magnetic resonance imaging (MRI) sequence that was best suited to demonstrate the scolex in a calcified lesion and to seek the explanation for the appearance of a negative phase in a calcified scolex on corrected gradient refocused echo (GRE) phase imaging. MATERIALS AND METHODS: Forty-nine patients with single/multiple computed tomography (CT) documented homogeneous calcified lesions and/or calcified scolices in cysts were studied with conventional spin echo and corrected GRE phase imaging. Calcium and different paramagnetic substances from cysticerci scolices of a sample of infected swine muscle were quantified. RESULTS: The scolex could be demonstrated in 29/39 patients with single calcified lesion. GRE imaging with an echo time of 35ms was the only sequence that demonstrated scolex in all these 29 cases. 15/29 patients with a single calcified lesion, in all 10 patients with multiple calcified lesions and infected swine muscle with multiple cysts and calcified scolex, corrected GRE phase imaging showed negative phase in all these scolices. Estimation of minerals from the calcified scolices from the swine muscle showed by spectroscopic techniques 41.2% of the total mineral contents as paramagnetic substances. CONCLUSION: We conclude that GRE imaging is the imaging method of choice for demonstration of the scolex in a CT calcified lesion. The negative phase on corrected GRE phase imaging is due to the presence of large amount of paramagnetic substances. Chawla, S. et al. (2002). Clinical Radiology57, 826–834.

Demonstration of Scolex in Calcified Cysticercus Lesion using Gradient Echo with or without Corrected Phase Imaging and its Clinical Implications

AIM: This study was performed to determine the magnetic resonance imaging (MRI) sequence that was best suited to demonstrate the scolex in a calcified lesion and to seek the explanation for the appearance of a negative phase in a calcified scolex on corrected gradient refocused echo (GRE) phase imaging. MATERIALS AND METHODS: Forty-nine patients with single/multiple computed tomography (CT) documented homogeneous calcified lesions and/or calcified scolices in cysts were studied with conventional spin echo and corrected GRE phase imaging. Calcium and different paramagnetic substances from cysticerci scolices of a sample of infected swine muscle were quantified. RESULTS: The scolex could be demonstrated in 29/39 patients with single calcified lesion. GRE imaging with an echo time of 35ms was the only sequence that demonstrated scolex in all these 29 cases. 15/29 patients with a single calcified lesion, in all 10 patients with multiple calcified lesions and infected swine muscle with multiple cysts and calcified scolex, corrected GRE phase imaging showed negative phase in all these scolices. Estimation of minerals from the calcified scolices from the swine muscle showed by spectroscopic techniques 41.2% of the total mineral contents as paramagnetic substances. CONCLUSION: We conclude that GRE imaging is the imaging method of choice for demonstration of the scolex in a CT calcified lesion. The negative phase on corrected GRE phase imaging is due to the presence of large amount of paramagnetic substances. Chawla, S. et al. (2002). Clinical Radiology57, 826–834.

Cyclopiazonic Acid Induce Contractures in Isolated Skeletal Muscles of Malignant Hyperthermia Susceptible Swine

Cyclopiazonic Acid Induce Contractures in Isolated Skeletal Muscles of Malignant Hyperthermia Susceptible Swine

Effect of available dietary carbohydrate on glycolytic potential and meat quality of swine muscles

The aim of this study was to determine whether glycolytic potential (GP) in pork muscle could be modified by the availability of carbohydrates in the diets and, if so, to what extent meat color and drip loss were affected. Biopsy samples of longissimus muscle (LM) from 48 Swiss Large White pigs (25 gilts, 23 barrows) weighing 70 kg were collected, and the GP was determined to vary from 111 to 187 μmol g-1 wet weight. At 90-kg body weight, pigs were moved into individual pens and assigned (blocked by GP and sex) to be fed 2.8 kg of a diet either high (H) or low (L) in available carbohydrate up to 104 kg. Pigs were fasted overnight (15 h) before slaughter. Glycogen, glucose, glucose-6-phosphate and lactic acid content were determined in samples of LM (predominantly glycolytic muscle) collected 30 min and 24 h post-mortem and in samples of the dark part of the semitendinosus (ST, oxidative part of the muscle) 24 h post-mortem. After slaughter, the decline in pH and temperature was recorded in the LM from 30 min to 24 h after bleeding. Regardless of the diet, content of glycolytic intermediates and lactic acid were higher in the LM compared to the ST. Diet did not alter the GP, and did not affect color or drip loss of the LM. However, in gilts fed the H diet muscle pH was lower by 0.2 units from 30 min until 6 h post-mortem than in gilts fed the L diet. These effects were not observed in barrows. ST of gilts fed the H diet had higher levels of glycolytic intermediates and lactic acid and, therefore, higher GP compared to gilts on the L diet, but no dietary effects occurred in barrows. Increased GP resulted in paler color and higher drip loss, whereas ultimate pH was not affected. The GP was positively correlated with L* (0.52), a* (0.49), b* (0.59) and drip loss (0.77) of the ST, whereas poor correlations were observed in the LM. In conclusion, dietary treatment only affected paleness and drip loss of the ST muscle and the effects were more pronounced in gilts than barrows. Increased GP resulted in paler meat with higher drip loss. Key words: Pig, glycolytic potential, meat quality, carbohydrate supply

Abattoir condemnation due to parasitic infections and its economic implications in the region of Trikala, Greece.

The prevalence of parasitic infections responsible for the condemnation of carcasses and viscera during meat inspection, and their economic implication, was estimated in a year long abattoir survey of 10 277 slaughtered farm animals in the region of Trikala, Greece. The organs examined for the presence of parasitic lesions during meat inspection were: liver and lungs of all animals, rumen of cattle, small intestine of lambs and kids, and muscles of cattle and swine. The parasitic lesions observed in the lungs of cattle, sheep and goats were caused only by hydatid cysts. No hydatid cysts were observed in the lungs of swine. The parasitic lesions observed in the liver of cattle, sheep and goats were as a result of hydatid cysts and flukes of Fasciola hepatica and Dicrocoelium dendriticum, while those of swine were due to milk spots only. Moniezia sp. proglottids were found in the small intestine of lambs only. The prevalence of parasites responsible for the condemnation of marketable organs was low (0.26%). Parasites were responsible for 22% of the total of condemned organs, and their annual cost was 99, 00 GDR (approximately 292 Euros). The parasites most contributing to marketable organ condemnation were hydatid cysts (26%) and D. dendriticum flukes (26%).

Characterization of Contractile Function and Expression of Muscarinic Receptors, G Proteins and Adenylate Cyclase in Cultured Tracheal Smooth Muscle of Swine

Smooth muscle cells lose their contractile function and phenotype very rapidly when placed in culture. During organ culture of smooth muscle strips, phenotype is lost more slowly. In the present studies, we established an organ culture model to study contractile function and expression of muscarinic receptors, G proteins and adenylyl cyclase in different serum concentrations in tracheal smooth muscle from swine. The results show that contractile function and the amounts of M₃ receptors, G proteins and adenylyl cyclase were maintained for up to 5 days in culture. The expression of M₂ receptors was significantly decreased in culture when compared to freshly isolated muscles. Maximal isometric tension was significantly increased in cultured muscles compared with freshly isolated muscles. Different serum concentrations did not significantly affect contractile function and expression of muscarinic receptors, G proteins and adenylyl cyclase. In conclusion, our studies suggest that cultured smooth muscle might be used as a model to study the regulation of contractile function of smooth muscle by various signal transduction pathways.

Characterization of contractile function and expression of muscarinic receptors, G proteins and adenylate cyclase in cultured tracheal smooth muscle of Swine.

Smooth muscle cells lose their contractile function and phenotype very rapidly when placed in culture. During organ culture of smooth muscle strips, phenotype is lost more slowly. In the present studies, we established an organ culture model to study contractile function and expression of muscarinic receptors, G proteins and adenylyl cyclase in different serum concentrations in tracheal smooth muscle from swine. The results show that contractile function and the amounts of M(3) receptors, G proteins and adenylyl cyclase were maintained for up to 5 days in culture. The expression of M(2) receptors was significantly decreased in culture when compared to freshly isolated muscles. Maximal isometric tension was significantly increased in cultured muscles compared with freshly isolated muscles. Different serum concentrations did not significantly affect contractile function and expression of muscarinic receptors, G proteins and adenylyl cyclase. In conclusion, our studies suggest that cultured smooth muscle might be used as a model to study the regulation of contractile function of smooth muscle by various signal transduction pathways.

Free-range rearing increases (n-3) polyunsaturated fatty acids of neutral and polar lipids in swine muscles

This study was conducted to determine the effect of rearing pigs on a free-range system, on fatty acid profile of neutral and polar lipids from longissimus dorsi (LD) and masseter (MS) muscles, with special reference to n-3 polyunsaturated fatty acids (PUFA). Food sources for free-reared pigs were basically acorn and pasture, which were high in oleic and linolenic acids, respectively whereas, indoors, animals were fed a concentrate high in oleic acid. The predominantly oxidative MS muscle had lower saturated fatty acid (SFA) and higher PUFA contents than LD in neutral lipids (NL), and higher total n-3 PUFA and arachidonic acid contents and lower linoleic acid content in polar lipids (PL). Rearing the animals outdoors significantly increased total n-3 and n-6 PUFA of NL and total n-3 of PL. All n-3 fatty acids detected in PL were significantly higher in free reared animals, including eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids.

Determination and Confirmation of Chloramphenicol Residues in Swine Muscle and Liver

ABSTRACT: Average high‐performance liquid chromatography (HPLC) recoveries of chloramphenicol (CAP) in swine muscle and liver ranged from 91.3 to 94.2% and 93.1 to 103.7%, respectively, with coefficients of variation ranging from 1.4 to 4.3% and 1.1 to 11.2% for each tissue sample. The method described was repeatable and reproducible in swine muscle and liver, with a limit of quantification of 15 ng/mL and a limit of detection estimated at 5 ng/mL. The limit of identification of CAP was 25 ng/mL, 5ng/mL, and 20 ng/mL for HPLC/PDA, GC/MS selected ion monitoring (SIM), and LC/MS analysis, respectively.

Determination of levamisole in livestock products using high performance liquid chromatography

A method is described for the determination of the anthelmintic levamisole in muscle, liver, kidney and fat of cattle, swine and poultry using high performance liquid chromatography with photodiode array detection. Levamisole was extracted from an alkaline sample with ethyl acetate and back-extracted with 0.1 mol/L hydrochloric acid. The extract was applied to an SCX solid-phase extraction column. The column was washed with water and methanol. Levamisole was eluted with a solution of ammonia in methanol. The eluate was evaporated to dryness and the residue was dissolved in the mobile phase and injected into the HPLC system. Mean recoveries from 0.01-0.10 microgram/g fortified muscle, liver, kidney and fat samples ranged from 78.3 to 99.8%. The detection limit for the assay was 0.005 microgram/g.