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Partial purification and characterization of bovine fibroblast interferon.
1988
Allen G.K. | Grothaus G.D. | Rosenquist B.D.
Bovine fibroblast interferon (BoF-IFN), produced in primary bovine embryonic kidney cell cultures after priming and infection with bluetongue virus, was purified by controlled pore glass (CPG) chromatography to a specific activity of 10(6) U/mg of protein, with 40% recovery of the original activity. The crude IFN was concentrated more than sevenfold during purification. This proved to be a relatively simple, practical method of obtaining sufficient quantities of partially purified natural BoF-IFN for further studies. The CPG-purified BoF-IFN was further concentrated by sequential ultrafiltration and was analyzed by sodium dodecyl sulfate/polyacrylamide-gel electrophoresis (SDS-PAGE). Interferon, recovered from denaturing conditions either by dialysis against phosphate-buffered saline solution or by dilution in cell culture medium containing 10% fetal bovine serum, migrated as a single stainable protein with molecular weight of 21,000 on analytic SDS-PAGE gels. Recovered IFN activity from preparative SDS-PAGE totalled 8.7% of that applied. Attempts to further purify CPG-purified BoF-IFN by zinc chelate affinity chromatography were unsuccessful.
Mostrar más [+] Menos [-]Liquid chromatography–mass spectrometry analysis of carvacrol in chicken tissues
2022
Śniegocki, Tomasz
Carvacrol is an essential oil derived from oregano that is used as a natural additive to improve the efficiency of livestock nutrition. Residues of natural additives such as carvacrol should be monitored in food of animal origin to ensure consumer safety. The aim of this study was to appraise the quick, easy, cheap, effective, rugged and safe (QuEChERS) approach coupled with liquid chromatography and mass spectrometry as a means of carvacrol analysis in chicken tissue. A 5 ± 0.05 g portion of plasma, lung, muscle and liver was mixed for 15 min with 5 mL of 1-butanol and 20 mL of water, then centrifuged. A 0.5 mL volume from the top layer was transferred, then 60 mg of octadecylsilane sorbent, 30 mg of primary and secondary amine and 200 mg of MgSO₄ were added. The extract was mixed and centrifuged. The top layer was filtered and then transferred to an autosampler vial for analysis by high-performance liquid chromatography–tandem mass spectrometry. The limit of detection was calculated at 0.06 μg g⁻¹ and the limit of quantification was 0.2 μg g⁻¹, with relative standard deviation repeatability and reproducibility below <20%. The validation results showed that this method could be a good alternative to determination of carvacrol by gas chromatography and is suitable for carvacrol analysis in different matrices.
Mostrar más [+] Menos [-]Environmental contamination of free-range hen with dioxin
2021
Mikołajczyk, Szczepan | Pajurek, Marek | Warenik-Bany, Małgorzata | Maszewski, Sebastian
The transfer of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) and polychlorinated biphenyls (PCBs) from a contaminated environment into the food chain is a serious consumer safety problem. As part of the Polish National Surveillance Program of PCDD/Fs and PCBs in food of animal origin, a concentration of PCDD/Fs of 4.61 ± 0.75 pg WHO-TEQ/g fat was determined in a sample of free-range eggs, which exceeded the permitted limit of 2.5 pg WHO-TEQ/g. The aim of the study was to investigate the source of the egg contamination and the risk for the eggs’ consumers. Eggs, muscles, feed and soil from the place where backyard waste burning had been carried out in the past and ash from a household stove tipped onto the paddock were analysed using the isotope dilution technique with high-resolution gas chromatography coupled with high-resolution mass spectrometry. The concentration in ash was low at 0.20 pg WHO-TEQ/g and the congener profile did not indicate the source of contamination. The dioxin content in soil from the backyard waste-burning site was 2.53 pg WHO-TEQ/g dry matter (d.m.) and the soil’s profile of PCDD/F congeners matched the profile of the contaminated eggs. By reason of the congener profile similarity, the investigation concluded, that the cause of the contamination was the backyard waste-burning site soil which the animals had access to. Frequent consumption of contaminated eggs from the analysed farm could pose a health risk due to chronic exposure, especially for vulnerable consumers.
Mostrar más [+] Menos [-]Farmed fish as a source of dioxins and PCBs for Polish consumers
2020
Mikołajczyk, Szczepan | Warenik-Bany, Małgorzata | Maszewski, Sebastian | Pajurek, Marek
This paper reports polychlorinated dibenzo-p-dioxin (PCDD), polychlorinated dibenzofuran (PCDF), and polychlorinated biphenyl (PCB) concentrations in fish collected from Polish and Vietnamese farms and the related risk for consumers. Altogether, 160 samples were analysed using an isotope dilution technique with high-resolution gas chromatography coupled with high-resolution mass spectrometry (HRGC-HRMS). To characterise the potential health risk associated with PCDD/F and dioxin-like polychlorinated biphenyl (DL-PCB) intake, doses ingested in two 100 g portions of fish by adults and children were calculated and expressed as the percentage of the tolerable weekly intake (TWI) newly established by the EFSA in November 2018 at 2 pg WHO-TEQ kg⁻¹ b.w. Generally, levels in fish muscles were low in relation to maximum limits (4), being in the range of 0.02–3.98 pg WHO-TEQ g⁻¹ wet weight (w.w.) for PCDD/F/DL-PCBs and 0.05–24.94 ng g⁻¹ w.w. for NDL-PCBs. The highest concentration was found in eel muscles. The least polluted were pangas and zanders and the levels were at the limits of quantification. Consumption of two portions of fish per week results in intakes of 9– 866% TWI by children and 4–286% TWI by adults. Frequent consumption of some species (for example eel and bream) can pose a health risk to vulnerable consumers and especially children and pregnant women.
Mostrar más [+] Menos [-]Fatty acid profile in edible eggs of snails from the Cornu genus
2020
Maćkowiak-Dryka, Monika | Szkucik, Krzysztof | Ziomek, Monika | Klimek, Kamila
The aim of this study was to determine the content of fatty acids in eggs harvested from two edible subspecies of Polish-bred common garden snail from the Cornu genus, as well as this content in the retail-ready product obtained from these eggs. Material for the study consisted of eggs from two subspecies of edible snails: the small (Cornu aspersum aspersum), and large (Cornu aspersum maxima) common garden snails. The eggs studied were in two forms, the first of which had undergone initial processing to the half-product stage and the second of which was the final product available on the Polish market under the name “Snail Eggs”. The gas chromatography method was used to determine the content of fatty acids. More than 75% of the studied fats were saturated fatty acids, dominated by palmitic and stearic acids. The average content of polyunsaturated fatty acids was 0.37%, and it was a combination of two acids: linoleic (C18:2n6c), and its trans isomer (C18:2n6t). No significant differences were found comparing individual fatty acids content between the two species’ eggs as half-products, or between the half-products and the final product. The fat in raw and processed eggs of common garden snails holds low nutritional value, and the processing did not affect the content of fatty acids.
Mostrar más [+] Menos [-]Levels of the natural hormones 17β-oestradiol and testosterone in serum of cattle: results from population studies in Poland
2016
Woźniak-Sobczak, Barbara | Witek, Sebastian | Matraszek-Żuchowska, Iwona | Kłopot, Alicja | Posyniak, Andrzej
Introduction: The present study is a comprehensive overview of the natural occurrence of 17β-oestradiol and testosterone in serum of cattle in Poland. Material and Methods: The serum samples (n = 826) were collected from cattle within five years. The samples were examined for the presence of oestradiol and testosterone using ELISA or gas chromatography with mass spectrometry. Results: In 98 samples (24%) 17β-oestradiol was detected above decision limits of applied methods, including five samples over the recommended concentration of 0.1 μg L⁻¹. Of the serum samples taken from cows (≤18 months of age), 95 and 99 percentiles of the animals had 17β-oestradiol concentration below 0.027 and 0.086 μg L⁻¹ and of samples from cows over 18 months of age - below 0.059 and 0.125 μg L⁻¹ respectively. Calculated values for bulls (≤18 months of age) were 0.025 and 0.034 μg L⁻¹ and for the animals older than 18 months of age - 0.035 and 0.041 μg L⁻¹. The natural presence of testosterone was detected in 201 serum samples (48.7%). According to the obtained data, 95% and 99% of cows (≤18 months of age) serum samples had testosterone concentration below 0.05 and 0.23 μg L⁻¹ and the animals over 18 months of age - 0.30 and 0.49 μg L⁻¹, respectively. For bulls these values did not depend on the age of the animals and were in the ranges of 5 - 6.3 μg L⁻¹ (95%) and 11.4 - 12.1 μg L⁻¹ (99%). Conclusion: Our study showed that the threshold values for these hormones in plasma of cattle designated years ago are correct, but they need to be supplemented for animals older than 18 months.
Mostrar más [+] Menos [-]Desflurane and sevoflurane elimination kinetics and recovery quality in horses
2015
OBJECTIVE To evaluate pharmacokinetics, recovery times, and recovery quality in horses anesthetized with 1.2 times the minimum alveolar concentration of sevoflurane or desflurane. ANIMALS 6 healthy adult horses. PROCEDURES Anesthesia was maintained with sevoflurane or desflurane for 2 hours at 1.2 times the minimum alveolar concentration. Horses recovered without assistance. During recovery, end-tidal gas samples were collected until horses spontaneously moved. Anesthetic concentrations were measured by use of gas chromatography. After a 1-week washout period, horses were anesthetized with the other inhalation agent. Video recordings of anesthetic recovery were evaluated for recovery quality on the basis of a visual analogue scale by investigators who were unaware of the anesthetic administered. Anesthetic washout curves were fit to a 2-compartment kinetic model with multivariate nonlinear regression. Normally distributed interval data were analyzed by means of paired Student t tests; ordinal or nonnormally distributed data were analyzed by means of Wilcoxon signed rank tests. RESULTS Horses recovered from both anesthetics without major injuries. Results for subjective recovery evaluations did not differ between anesthetics. Area under the elimination curve was significantly smaller and time to standing recovery was significantly less for desflurane than for sevoflurane, although distribution and elimination constants did not differ significantly between anesthetics. CONCLUSIONS AND CLINICAL RELEVANCE Differences in area under elimination the curve between anesthetics indicated more rapid clearance for desflurane than for sevoflurane in horses, as predicted by anesthetic blood solubility differences in this species. More rapid elimination kinetics was associated with faster recovery times, but no association with improved subjective recovery quality was detected.
Mostrar más [+] Menos [-]Comparison of inferred fractions of n-3 and n-6 polyunsaturated fatty acids in feral domestic cat diets with those in commercial feline extruded diets
2013
Backus, Robert C. | Thomas, David G. | Fritsche, Kevin L.
Objective-To compare presumed fatty acid content in natural diets of feral domestic cats (inferred from body fat polyunsatrated fatty acids content) with polyunsaturated fatty acid content of commercial feline extruded diets. Sample-Subcutaneous and intra-abdominal adipose tissue samples (approx 1 g) from previously frozen cadavers of 7 adult feral domestic cats trapped in habitats remote from human activity and triplicate samples (200 g each) of 7 commercial extruded diets representing 68% of market share obtained from retail stores. Procedures-Lipid, triacylglycerol, and phospholipid fractions in adipose tissue samples and ether extracts of diet samples were determined by gas chromatography of methyl esters. Triacylglycerol and phospholipid fractions in the adipose tissue were isolated by thin-layer chromatography. Diet samples were also analyzed for proximate contents. Results-For the adipose tissue samples, with few exceptions, fatty acids fractions varied only moderately with lipid fraction and site from which tissue samples were obtained. Linoleic, α-linolenic, arachidonic, eicosapentaenoic, and docosahexaenoic acid fractions were 15.0% to 28.2%, 4.5% to 18.7%, 0.9% to 5.0%, < 0.1% to 0.2%, and 0.6% to 1.7%, respectively. As inferred from the adipose findings, dietary fractions of docosahexaenoic and α-linolenic acid were significantly greater than those in the commercial feline diets, but those for linoleic and eicosapentaenoic acids were not significantly different. Conclusions and Clinical Relevance-The fatty acid content of commercial extruded feline diets differed from the inferred content of natural feral cat diets, in which dietary n-3 and possibly n-6 polyunsaturated fatty acids were more abundant. The impact of this difference on the health of pet cats is not known.
Mostrar más [+] Menos [-]Partial purification and characterization of bovine fibroblast interferon
1988
Allen, G.K. | Grothaus, G.D. | Rosenquist, B.D.
Bovine fibroblast interferon (BoF-IFN), produced in primary bovine embryonic kidney cell cultures after priming and infection with bluetongue virus, was purified by controlled pore glass (CPG) chromatography to a specific activity of 10(6) U/mg of protein, with 40% recovery of the original activity. The crude IFN was concentrated more than sevenfold during purification. This proved to be a relatively simple, practical method of obtaining sufficient quantities of partially purified natural BoF-IFN for further studies. The CPG-purified BoF-IFN was further concentrated by sequential ultrafiltration and was analyzed by sodium dodecyl sulfate/polyacrylamide-gel electrophoresis (SDS-PAGE). Interferon, recovered from denaturing conditions either by dialysis against phosphate-buffered saline solution or by dilution in cell culture medium containing 10% fetal bovine serum, migrated as a single stainable protein with molecular weight of 21,000 on analytic SDS-PAGE gels. Recovered IFN activity from preparative SDS-PAGE totalled 8.7% of that applied. Attempts to further pruify CPG-purified BoF-IFN by zinc chelate affinity chromatography were unsuccessful.
Mostrar más [+] Menos [-]Rapid presumptive diagnosis of anaerobic infections in animals by gas-liquid chromatography
1989
Bogaard, A.E.J.M. van den | Hazen, J. | Maes, J.H.
The detection of volatile fatty acids (VFA) by gas chromatography of 85 purulent specimens from abscesses or pyogenic infections in cats, dogs, rodents, and ruminants was compared with the results of bacteriologic culturing, and proved to be a rapid means of presumptively diagnosing anaerobic infections. Of 83 bacteriologically positive specimens, 52 (61%) yielded obligate anaerobes and in 50 specimens, 1 or more VFA (butyric acid, isobutyric acid, valeric acid, isovaleric acid, caproic acid, or isocaproic acid) was detected. Forty-six specimens were positive for culturing of anaerobes and for detection of 1 or more of these VFA. By contrast, pus from infections caused by (facultative) aerobic microorganisms contained no VFA or only acetic and/or propionic acid.
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