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Determination of steroid esters in hair of slaughter animals by liquid chromatography with tandem mass spectrometry
2019
Matraszek-Żuchowska, Iwona | Woźniak-Sobczak, Barbara | Sielska, Katarzyna | Posyniak, Andrzej
The use of growth promoters in animal husbandry to increase weight gain and efficiency of feed conversion into muscle has been banned in the European Union since 1988, and under Directive 96/23/EC, surveillance for anabolic steroid hormones is obligatory. The hormones present in animal tissues may be of endogenous origin or may result from illegal administration. Steps have been taken to determine selected steroids in the form of esters in the alternative matrix of animal hair. Their detection in biological material is direct proof of the illegal use of anabolics. The procedure for the determination of steroid esters in animal hair, based on digestion, extraction, purification, and liquid chromatography with tandem mass spectrometry was validated under the current regulations. In total, 348 samples of animal hair were examined using this method. Good recoveries and precision values (RSD) were obtained during validation. Decision limits (CCα) and detection capabilities (CCβ) were in the ranges of 2.57–4.18 μg kg⁻¹ and 4.38–7.12 μg kg⁻¹, respectively. The method met the criteria for confirmation techniques with respect to Commission Decision 2002/657/EC. Testing for steroid esters in animal hair was introduced into the National Residue Control Programme in 2017. Steroid esters were not found in any hair samples above the CCα, which indicates that illegal use of anabolics was not confirmed.
Mostrar más [+] Menos [-]Mitochondrial theory of skeletal muscle ageing –new facts, new doubts
2019
Słodki, Sebastian | Bogucka, Joanna
For many years, scientists have been pursuing research on skeletal muscle ageing both in humans and animals. Studies on animal models have extended our knowledge of this mechanism in humans. Most researchers agree that the major processes of muscle ageing occur in the mitochondria as the major energy production centres in muscle cells. It is believed that decisive changes occur at the enzymatic activity level as well as in protein synthesis and turnover ability. Deregulation of ion channels and oxidative stress also play significant roles. In particular, in recent years the free radical theory of ageing has undergone considerable modification; researchers are increasingly highlighting the partly positive effects of free radicals on processes occurring in cells. In addition, the influence of diet and physical activity on the rate of muscle cell ageing is widely debated as well as the possibility of delaying it through appropriate physical exercise and diet programmes. Numerous studies, especially those related to genetic processes, are still being conducted, and in the near future the findings could provide valuable information on muscle ageing. The results of ongoing research could answer the perennial question of whether and how we can influence the rate of ageing both in animals and humans.
Mostrar más [+] Menos [-]Retrospective liver histomorphological analysis in dogs in instances of clinical suspicion of congenital portosystemic shunt
2019
Sobczak-Filipiak, Małgorzata | Szarek, Józef | Badurek, Iwona | Padmanabhan, Jessica | Trębacz, Piotr | Januchta-Kurmin, Monika | Galanty, Marek
Introduction: The clinical symptoms of portosystemic shunts (PSSs) and hepatic microvascular dysplasia (HMD) – portal vein hypoplasia (PVH) in dogs are similar. PSSs are abnormal vascular connections between the portal vein system and systemic veins. HMD is a very rare developmental vascular anomaly, recognisable during histopathological examination. The study aim was to assess the prevalence of HMD–PVH and hepatocellular and vascular pathologies in the liver. Material and Methods: Liver biopsies from 140 dogs (of different breeds and both sexes) arousing clinical suspicion of PSS were examined histopathologically. Results: An initial PSS diagnosis was confirmed in 125 dogs (89.29%). HMD–PVH was found in 12.32% of dogs, as an isolated disease in 9.29%, especially in Yorkshire terriers, and with extrahepatic PSS in 6.67%. Histopathological analysis of muscles around sublobular veins showed that HMD cases presented hypertrophy or hypertrophy with fibrosis. In 2.17% of all dogs with liver vascular developmental disorders calcification was visible around vessels (without correlation by degenerative changes in those vessels), suggesting prior onset of deep metabolic disorders. Clinical suspicion of PSS was also formed upon quite different pathological processes in young dogs. Conclusion: Histopathological findings diagnosed the type of vascular anomalies (PSS or HMD–PVH) or other pathological changes conclusively, therefore detailed hepatic histopathology is an indispensable component of the clinical diagnostic process.
Mostrar más [+] Menos [-]Evaluation of the use of muscle condition score and ultrasonographic measurements for assessment of muscle mass in dogs
2019
Freeman, Lisa M. | Michel, Kathryn E. | Zanghi, Brian M. | Boler, Brittany M Vester | Fages, Julien
OBJECTIVE To evaluate repeatability and reproducibility of muscle condition score (MCS) in dogs with various degrees of muscle loss; to compare MCS, muscle ultrasonographic measurements, and quantitative magnetic resonance (QMR) measurements; and to identify cutoff values for ultrasonographic measurements of muscle that can be used to identify dogs with cachexia and sarcopenia. ANIMALS 40 dogs of various age, body condition score (BCS), and MCS. PROCEDURES A prospective cross-sectional study was conducted. Body weight, BCS, QMR measurements, thoracic radiographic measurements, and muscle ultrasonographic measurements were assessed once in each dog. The MCS for each dog was assessed 3 separate times by 4 separate raters. RESULTS For the MCS, overall κ for interrater agreement was 0.50 and overall κ for intrarater agreement ranged from 0.59 to 0.77. For both interrater and intrarater agreement, κ coefficients were higher for dogs with normal muscle mass and severe muscle loss and lower for dogs with mild and moderate muscle loss. The MCS was significantly correlated with age (r = −0.62), vertebral epaxial muscle score (VEMS; r = 0.71), forelimb epaxial muscle score (FLEMS; r = 0.58), and BCS (r = 0.73), and VEMS was significantly correlated (r = 0.84) with FLEMS. Cutoff values for identification of mild muscle loss determined by use of VEMS and FLEMS were 1.124 and 1.666, respectively. CONCLUSIONS AND CLINICAL RELEVANCE MCS had substantial repeatability and moderate reproducibility for assessment of muscle mass in dogs. Prospective studies of MCS, VEMS, and FLEMS for assessment of muscle mass in dogs are warranted.
Mostrar más [+] Menos [-]Comparison of computed tomographic attenuation values for epaxial muscles in old and young dogs
2019
Sutherland-Smith, James | Hutchinson, Dana | Freeman, Lisa M.
OBJECTIVE To determine whether the degree of CT attenuation of muscle would differ between healthy old and young dogs. ANIMALS 10 healthy old (> 8 years old) and 9 healthy young (1 to 5 years old) Labrador Retrievers with a body condition score of 5 or 6 on a 9-point scale. PROCEDURES CT was performed with the dogs mildly sedated. A freehand closed polygon tool was used to define the outer margin of the left epaxial muscles on each transverse image obtained from the cranial to caudal endplates of T13. The CT attenuation values from every voxel from within these regions of interest were exported from DICOM software as a single dataset in an extensible markup language file. From these data, mean CT attenuation values were calculated for each dog and these mean values were compared between age groups. RESULTS Mean CT attenuation values for the epaxial muscles were significantly lower in old dogs than in young dogs. A significant negative correlation (r = –0.74) was identified between mean CT attenuation values and dog age. CONCLUSIONS AND CLINICAL RELEVANCE In addition to loss of skeletal muscle mass, low muscle CT attenuation values suggested that the old dogs in this study also had greater muscle fat content than did young dogs. Additional studies are warranted to evaluate qualitative and quantitative muscle changes in old dogs.
Mostrar más [+] Menos [-]Acute-phase protein profile in horses subjected to different exercise protocols
2019
Assuncao, P. | Barbosa, T. | Yonezawa, L. | Barbosa, L. | Watanabe, M. | Kohayagawa, A. | Schmidt, E.
High-intensity exercise can be associated with the occurrence of muscle injury, as well as the induction of an acute-phase response (APR). The present study aims to investigate the synthesis and profile of serum proteins in horses before and after participating in 2 different exercise protocols and to relate this profile to the presence or absence of muscular injury caused by exercise. Ten purebred Arabian (n = 5) and Criollo (n = 5) horses were subjected to 2 different tests on a treadmill, one consisting of short-duration and rapid-acceleration training (TRA) that was mostly anerobic and the other of long-duration and slow-acceleration training (TLD) that was predominantly aerobic. Blood samples were obtained before the beginning of exercise (T0) and at 6 post-exercise time points: immediately after (T1) and 30 min (T2), 3 h (T3), 12 h (T4), 24 h (T5), and 48 h (T6) after exercise. Hematocrit was determined by the microhematocrit method. Plasma and serum samples were prepared by centrifugation (1500 × g for 5 min) for plasma concentrations of fibrinogen, total serum proteins (TP), sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and creatine-kinase (CK) serum activity. Total protein concentration and CK serum activity were determined in an automated biochemistry analyzer. Fibrinogen was determined by the heat precipitation method in microhematocrit capillary tubes. Estimated concentrations of haptoglobin (Hp) significantly decreased after TRA and estimated concentrations of alpha-1 acid glycoprotein (AGP) significantly increased after both protocols at T2. Albumin increased after the TLD exercise protocol. Changes in hematocrit, haptoglobin, and albumin concentrations in horses subjected to different treadmill exercise protocols are related to a physiological response to hemoconcentration and hemolysis. Increases of AGP in the TLD protocol suggest the release of catecholamines as a response to avoid oxidative damage to tissue.
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