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Isoelectric focusing of proteins in the pH gradient as a tool for identification of species origin of raw meat
2018
Różycki, Mirosław | Chmurzyńska, Ewa | Bilska-Zając, Ewa | Karamon, Jacek | Cencek, Tomasz
Health, religious, and commercial aspects justify the need for meat species identification. The lack of officially approved methods prompts the undertaking of research on validation of isoelectric focusing of proteins (IEF) for official purposes. Samples were prepared from pigs (Sus scrofa ferus domestica), cattle (Bos taurus), and poultry (Gallus gallus domesticus). Meat mixtures were made by blending 50%, 25%, 10%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.2% meat of other species. Samples were examined on ultrathin polyacrylamide gels with pH 3–9 gradient. The results of the study confirmed the stable and reproducible pattern of meat protein bands. The detection limit of raw meat admixtures from pigs, cattle, and poultry mostly ranged from 2% down to 0.2% (0.2% for poultry). However, the IEF method can be used to detect the addition of pig meat to bovine meat in an amount higher than 3%. At the significant mixture level (i.e at least 5% addition of meat of another species) IEF proves itself with 100% specificity, sensitivity, and accuracy. The achieved detection limits provide a basis for recommending the IEF method for routine tests in laboratories detecting the species origin of meat.
Mostrar más [+] Menos [-]Control of residues of thyreostats in slaughter animals in Poland in 2011–2017
2018
Woźniak-Sobczak, Barbara | Matraszek-Żuchowska, Iwona | Sielska, Katarzyna | Witek, Sebastian | Posyniak, Andrzej | Niemczuk, Krzysztof | Żmudzki, Jan
Introduction: In the European Union, the use of thyreostatic drugs for fattening slaughter animals has been banned since 1981 under Council Directive 81/602/EEC. For protection of consumer health against unwanted residues and in compliance with Directive 96/23, each EU country must monitor thyreostats in samples of animal origin. This paper presents the results of research on thyreostatic residues carried out in Poland in 2011–2017. Material and Methods: The material for testing was urine (n = 3,491), drinking water (n = 127), and muscle samples (n = 349) officially collected by Veterinary Sanitary Inspectors in slaughterhouses and farms throughout the country in accordance with the national residue control plan. The samples were examined for the presence of tapazole, thiouracil, methylthiouracil, propylthiouracil, and phenylthiouracil using liquid chromatography tandem mass spectrometry through an accredited method. Results: In four bovine and three porcine urine samples, the permissible thiouracil concentration was exceeded. In one sample of porcine urine, methyl- and propylthiouracil were found. The presence of thiouracil and its derivatives in urine samples is most likely due to feeding animals diet containing cruciferous plants. Conclusions: The results of research indicate that thyreostats are not used for anabolic purposes in slaughter animals in Poland.
Mostrar más [+] Menos [-]Development of a new RT-PCR with multiple primers for detecting Southern African Territories foot-and-mouth disease viruses
2018
Liu, Yali | Ding, Yao-Zhong | Dai, Jun-Fei | Ma, Bing | He, Ji-Jun | Ma, Wei-Min | Lv, Jian-Liang | Ma, Xiao-Yuan | Ou, Yun-Wen | Wang, Jun | Liu, Yong-Sheng | Chang, Hui-Yun | Wang, Yong-Lu | Zhang, Qiang | Liu, Xiang-Tao | Zhang, Yong-Guang | Zhang, Jie
Introduction: The extremely high genetic variation and the continuously emerging variants of foot-and-mouth disease virus (FMDV) of Southern African Territory (SAT) serotypes including SAT1, SAT2, and SAT3 make it necessary to develop a new RT-PCR for general use for monitoring viruses based on the updated genome information. Material and Methods: A FMDV SAT-D8 one-step RT-PCR was established based on the 1D2A2B genes of the SAT serotype viruses with a multiplex primer set. FMDV A, O, C, and Asia 1 serotypes, other vesicular disease viruses, inactivated SAT viruses, and 125 bovine, ovine, caprine and porcine tissue samples collected from the Chinese mainland were included for evaluating the assay. Results: The new RT-PCR was proven to be specific without cross-reactions with Eurasian FMDV, swine vesicular disease virus (SVDV), Seneca valley virus (SVV), or other common viral pathogens of cattle, sheep, goat, and pig. An around 257 bp-sized amplicon clearly appeared when the inactivated SAT viruses were detected. However, all 125 samples collected from FMDV-susceptible animals from the Chinese mainland which has not known SAT epidemics showed negative results. Conclusions: A FMDV SAT-D8 one-step RT-PCR is a promising method for primary screening for FMDV SAT serotypes.
Mostrar más [+] Menos [-]Effects of a surgical sealant on leakage pressure and circumference of fresh canine cadaver small intestinal anastomoses
2018
Mutascio, Liliana M. | Breur, Gert J. | Moore, George E. | Simons, Micha C.
OBJECTIVE To evaluate the effect of a bovine albumin–derivatized glutaraldehyde (BA-DG) biopolymer sealant on leakage pressures of intestinal anastomoses in jejunal tissue collected from fresh canine cadavers and to evaluate changes in circumference and cross-sectional area of the anastomotic site resulting from sealant application. SAMPLE 24 jejunal anastomoses from 4 fresh canine cadavers. PROCEDURES Jejunal tissue specimens were collected, and adjacent segment anastomoses were created within 12 hours after euthanasia of each dog. The tissue constructs were randomly assigned to 1 of 2 groups in which sealant was or was not applied. The outer circumference of all anastomoses in the sealant group was measured before and after application of the sealant; the cross-sectional area at the anastomotic site was then calculated at each time point. Tissue constructs were pressure tested, and leakage pressure and site were recorded. All testing was completed within 24 hours after tissue collection. RESULTS Compared with preapplication findings, there were no significant changes in outer circumference or cross-sectional area at the anastomotic site after sealant application. Leakage pressures in the sealant group were significantly higher than those in the no-sealant group. CONCLUSIONS AND CLINICAL RELEVANCE The use of surgical sealant on fresh canine cadaver jejunal anastomoses resulted in significantly higher leakage pressure at the anastomotic site; no immediate tissue deformation of the outer circumference or cross-sectional area occurred after sealant application. Future in vivo investigations are warranted to evaluate the effects of this sealant and potential benefits for clinical patients undergoing enterectomy.
Mostrar más [+] Menos [-]Comparison of antimicrobial resistance genes in feedlots and urban wastewater
2018
Beukers, A. G. | Zaheer, R. | Cook, S. R. | Chaves, A. V. | Ward, M. P. | Tymensen, L. | Morley, P. S. | Hannon, S. | Booker, C. W. | Read, R. R. | McAllister, T. A.
The use of antibiotics in livestock production in North America and possible association with elevated abundance of detectable antimicrobial resistance genes (ARG) is a growing concern. Real-time, quantitative PCR (RT-qPCR) was used to determine the relative abundance and diversity of ARG in fecal composite and catch basin samples from 4 beef feedlots in Alberta. Samples from a surrounding waterway and municipal wastewater treatment plants were also included to compare the ARG profile of urban environments and fresh water with that of feedlots. The relative abundance of 18 resistance genes across 5 antibiotic families including sulfonamides, tetracyclines, macrolides, fluoroquinolones, and β-lactams was examined. Sulfonamide, fluoroquinolone, and β-lactam resistance genes predominated in wastewater treatment samples, while tetracycline resistance genes predominated in cattle fecal composite samples. These results reflect the types of antibiotic that are used in cattle versus humans, but other factors such as co-selection of ARG and variation in the composition of bacterial communities associated with these samples may also play a role.
Mostrar más [+] Menos [-]Risk of environmental exposure to small coccidia from wild canid feces in rural Ohio
2018
Moreno-Torres, Karla I. | Sinnott, Devinn M. | Wolfe, Barbara A. | Marsh, Antoinette E. | Saville, William J. A. | Moritz, Mark | Garabed, Rebecca B.
OBJECTIVE To determine the extent of environmental exposure to heteroxenous coccidia from wild canid feces in southeastern Ohio. SAMPLE 285 presumed wild canid fecal samples collected across an ecological system in southeastern Ohio. PROCEDURES Morphological classification and molecular analysis were used to determine the canid genus for collected fecal samples. Microscopic and molecular analysis were used to detect coccidian oocysts and DNA. Several variables were analyzed for associations with coccidian DNA detection or prevalence. RESULTS Coccidian DNA was detected in 51 of 285 (17.9%) fecal samples. Of those positive samples, 1% (95% confidence interval, 0.4% to 3%) had positive results for Hammondia heydorni and none had positive results for Neospora caninum, for an estimated environmental N caninum prevalence of 0% (95% confidence interval, 0% to 7%)/1-km2 hexagonal area evaluated. Morphological classification revealed that 78.9% (225/285) of fecal samples were from coyotes and 17.2% (49/285) were from foxes. No difference in proportions of coccidian DNA-positive fecal samples was identified among canid species. Environmental temperature and fecal freshness were associated with coccidian DNA detection. Land use type, relative canid density, and cattle density were not associated with the prevalence of coccidian DNA-positive samples. CONCLUSIONS AND CLINICAL RELEVANCE The low prevalence of coccidia shed in wild canid feces in this study, including the estimated 0% environmental prevalence of N caninum, suggested that the role of the oocyst environmental phase in coccidia transmission to ruminants is likely minor in rural southeastern Ohio.
Mostrar más [+] Menos [-]Intraocular pressure measurements in cattle, sheep, and goats with 2 different types of tonometers
2018
Peche, N. | Eule, J. C.
The aim of this study was to investigate normal intraocular pressure (IOP) values of cattle, sheep, and goats with a rebound tonometer [TonoVet (TV)] and an applanation tonometer [Tono-Pen AVIA (TPA)] and to determine correction functions for the 2 devices. A total of 60 healthy cattle, sheep, and goats (20 of each) underwent slit-lamp biomicroscopy. Intraocular pressure (IOP) readings were taken from both eyes with the 2 different tonometers and statistically analyzed. For calibration purposes, the IOP was preset on each instrument at 5 to 60 mmHg using 5 mmHg increments in 10 bovine, 8 ovine, and 6 caprine freshly enucleated eyes. Readings were taken with both tonometers at each interval and compared to the manometrically controlled IOP (Mann-Whitney U-test, P ≤ 0.05; Bland-Altman plot, and regression analysis). The median IOP measurements (min to max) obtained with the TV were 23 mmHg (12 to 40 mmHg), 11 mmHg (7 to 20 mmHg), and 23 mmHg (9 to 37 mmHg) for cattle, sheep, and goats, respectively. Using the TPA, the median IOP measurements were 16 mmHg (8 to 27 mmHg), 10 mmHg (5 to 18 mmHg), and 13 mmHg (4 to 25 mmHg) for cattle, sheep, and goats, respectively. There were statistically significant differences between the readings taken with the TV and the TPA in all species (Wilcoxon-test, P ≤ 0.05). All measurements obtained with the TV and the TPA during the calibration procedure differed statistically significantly from the manometrically controlled IOP measurements (Mann-Whitney U-test, P ≤ 0.05). For both instruments, regression formulas were calculated to correct the measurements. Both tonometers can be used effectively to assess intraocular pressure in ruminants, using the specific regression formulas.
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