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Use of a new LC-MS method for the determination of pyrrolizidine alkaloids in feeds Полный текст
2018
Kowalczyk, Ewelina | Kwiatek, Krzysztof
Use of a new LC-MS method for the determination of pyrrolizidine alkaloids in feeds Полный текст
2018
Kowalczyk, Ewelina | Kwiatek, Krzysztof
Pyrrolizidine alkaloids (PAs) are secondary metabolites produced by many plant species. Due to their toxicity PAs can pose a risk to human and animal health. To detect the toxic compounds in feed materials a sensitive method based on liquid chromatography coupled with mass spectrometry has been developed. PAs were extracted with sulphuric acid and purified with cation exchange cartridges. A newly developed solvent mixture consisting of ethyl acetate, methanol, acetonitrile, ammonia, and triethylamine was used to wash alkaloids from the cartridges. After evaporation the residues were reconstituted in water and methanol mixture and subjected to LC-MS analysis. The developed method was validated according to SANTE/11945/2015 guidelines. The recovery was from 84.1% to 112.9%, the repeatability ranged from 3.0% to 13.6%, and the reproducibility was from 4.8% to 18.9%. A sensitive and selective method for determination of PAs in feed materials has been developed and validated. All evaluated validation parameters were in accordance with EU Reference Laboratories document no. SANTE/11945/2015. Almost 41% of the analysed feed samples were positive for the presence of at least one PA.
Показать больше [+] Меньше [-]Use of a new LC-MS method for the determination of pyrrolizidine alkaloids in feeds Полный текст
2018
Kowalczyk Ewelina | Kwiatek Krzysztof
Pyrrolizidine alkaloids (PAs) are secondary metabolites produced by many plant species. Due to their toxicity PAs can pose a risk to human and animal health. To detect the toxic compounds in feed materials a sensitive method based on liquid chromatography coupled with mass spectrometry has been developed.
Показать больше [+] Меньше [-]Coagulation factor activity in units of leukoreduced and nonleukoreduced canine fresh-frozen plasma Полный текст
2019
Foote, Michelle L. | Brooks, Marjory B. | Archer, Todd M. | Willis, Robert W. | Mackin, Andrew J. | Thomason, John M.
OBJECTIVE To evaluate coagulation factors in units of leukoreduced (LR) and nonleukoreduced (non-LR) canine fresh-frozen plasma (cFFP). ANIMALS 8 healthy research dogs. PROCEDURES In a crossover study, dogs were randomly assigned to 1 of 2 groups from which blood was collected and either did or did not undergo leukoreduction. After a recovery period of ≥ 28 days, the dogs were switched between protocols. After each collection, blood samples were centrifuged, and cFFP was stored frozen for later comparative analysis of coagulation factors, antithrombin, and protein C activities (reported as comparative percentages of the corresponding activities determined in a canine pooled plasma standard); prothrombin and activated partial thromboplastin times; and fibrinogen concentration. RESULTS There were no significant differences detected between results for LR cFFP, compared with those for non-LR cFFP. CONCLUSIONS AND CLINICAL RELEVANCE Although there was variation among residual activities of coagulation factors in LR and non-LR cFFP, the variations and differences were considered unlikely to impact the efficacy of LR cFFP transfused for coagulation factor replacement in dogs. However, owing to the small sample size and high variability of results in the present study, additional research with a larger sample size is required for definitive conclusions on the effects of leukoreduction on coagulation factors in cFFP and to develop treatment guidelines for LR cFFP use in dogs with congenital and acquired coagulopathies.
Показать больше [+] Меньше [-]Effect of sensor location in dogs on performance of an interstitial glucose monitor Полный текст
2016
Koenig, Amie | Hoenig, Margarethe E. | Jimenez, David A.
OBJECTIVE To identify variations in glucose values concurrently obtained by use of a continuous glucose monitoring system (CGMS) at the same site, reliability of results for each site, lag time for each site, and influence of site thickness on CGMS accuracy. ANIMALS 8 random-source research dogs. PROCEDURES In experiment 1, 8 CGMS sensors were implanted bilaterally at 1 site (4 sensors/side) in 4 dogs. In experiment 2, 2 CGMS sensors were implanted bilaterally at each of 4 sites (1 sensor/side) in 8 dogs; 4 of those 8 dogs then were subjected to a glycemic clamp technique. The CGMS results were compared among sensors and with criterion-referenced results during periods of euglycemia for all 8 dogs and during hyperglycemia and hypoglycemia for 4 dogs during the glycemic clamp procedure. RESULTS Differences (median, −7 mg/dL; interquartile range [IQR], −18.75 to 3 mg/dL) between CGMS and criterion-referenced glucose concentrations differed significantly among dogs and sites; during euglycemia, they were not different from the expected normal variation between multiple sensors concurrently implanted at the same site. Differences (median, −35 mg/dL; IQR, −74 to −15 mg/dL) between CGMS and criterion-referenced concentrations were greater during changes in glucose concentrations. Thoracic sensors were most accurate but had the shortest mean functional life. CONCLUSIONS AND CLINICAL RELEVANCE Significant differences were detected between CGMS and criterion-referenced glucose concentrations. Overall clinical utility of CGMS was acceptable at all sites, with most of the values from all sensors, sites, and dogs meeting guidelines for point-of-care glucometers.
Показать больше [+] Меньше [-]Comparison of blood pressure measurements of anesthetized dogs obtained noninvasively with a cylindrical blood pressure cuff and an anatomically modified conical blood pressure cuff Полный текст
2016
Ramos, Sara J. | Da Cunha, Anderson F. | Domingues, Michelle | Shelby, Amanda M. | Stout, Rhett W. | Acierno, Mark J.
OBJECTIVE To compare blood pressure measured noninvasively with an oscillometric device that involved use of a novel conical cuff and a traditional cylindrical blood pressure cuff. ANIMALS 17 adult hound-type dogs. PROCEDURES Dogs were anesthetized, and a 20-gauge, 1.5-inch catheter was inserted in the median sacral artery. The catheter was attached to a pressure transducer via fluid-filled noncompliant tubing, and direct blood pressure was recorded with a multifunction monitor. A specially fabricated conical cuff was placed on the antebrachium. Four sets of direct and indirect blood pressure measurements were simultaneously collected every 2 minutes. Four sets of measurements were then obtained by use of a cylindrical cuff. RESULTS The cylindrical cuff met American College of Veterinary Internal Medicine consensus guidelines for validation of indirect blood pressure measurements for mean arterial blood pressure (MAP), systolic arterial blood pressure (SAP), and diastolic arterial blood pressure (DAP). The conical cuff met the consensus guidelines for difference of paired measurements, SD, and percentages of measurements within 10 and 20 mm Hg of the value for the reference method, but it failed a correlation analysis. In addition, although bias for the conical cuff was less than that for the cylindrical cuff for SAP, MAP, and DAP measurements, the limits of agreement for the conical cuff were wider than those for the cylindrical cuff for SAP and MAP measurements. CONCLUSIONS AND CLINICAL RELEVANCE On the basis of results of this study, use of a conical cuff for oscillometric blood pressure measurement cannot be recommended.
Показать больше [+] Меньше [-]Tear film osmolality and electrolyte composition in healthy horses Полный текст
2015
Best, Lori J. | Hendrix, Diane V. H. | Ward, Daniel A.
OBJECTIVE To evaluate the tear film osmolality and electrolyte composition in healthy horses. ANIMALS 15 healthy adult horses. PROCEDURES Each horse was manually restrained, and an ophthalmic examination, which included slit-lamp biomicroscopy, indirect ophthalmoscopy, and a Schirmer tear test, was performed. Tear samples were collected from both eyes with microcapillary tubes 3 times at 5-minute intervals. The tear samples for each horse were pooled, and the osmolality and electrolyte concentrations were measured. The mean (SD) was calculated for each variable to establish preliminary guidelines for tear film osmolality and electrolyte composition in healthy horses. RESULTS The mean (SD) tear film osmolality was 283.51 (9.33) mmol/kg, and the mean (SD) sodium, potassium, magnesium, and calcium concentrations were 134.75 (10), 16.3 (5.77), 3.48 (1.97), and 1.06 (0.42) mmol/L, respectively. The sodium concentration in the tear film was similar to that in serum, whereas the potassium concentration in the tear film was approximately 4.75 times that of serum. CONCLUSIONS AND CLINICAL RELEVANCE Results provided preliminary guidelines with which tear samples obtained from horses with keratopathies can be compared. Measurement of tear film osmolality in these horses was easy and noninvasive. The tear film concentration of divalent cations was greater than expected and was higher than the divalent cation concentrations in the tear films of rabbits and humans. These data may be clinically useful for the diagnosis and monitoring of hyperosmolar ocular surface disease in horses.
Показать больше [+] Меньше [-]Associations between genotypes at codon 171 and 136 of the prion protein gene and production traits in market lambs Полный текст
2007
Evoniuk, Jessica M. | Berg, Paul T. | Johnson, Mary L. | Larson, Daniel M. | Maddock, Travis D. | Stoltenow, Charles L. | Schauer, Chris S. | O'Rourke, Katherin I. | Redmer, Dale A.
Objective—To determine whether selection for the homozygous A136R171 genotype that confers resistance to classic scrapie infection negatively affects production traits in sheep. Animals—996 commercial lambs obtained from 2 flocks at separate locations across 3 consecutive years. Procedures—Genotyping at codon 136 and 171 was performed by use of commercially available testing or a single-nucleotide polymorphism assay. Carcass data were collected without knowledge of genotype approximately 24 hours after slaughter by an experienced grader. The model to analyze associations between prion protein (PRNP) genotype and production traits was based on genotype, breed, or both as fixed effects and days on feed as a covariate. Results—Average daily gain was significantly associated with only combined codons 136 and 171. In flock 1, weaning average daily gain was significantly greater in AA136 sheep than heterozygotes; the difference between QR171 and RR171 sheep, compared with QQ171 sheep, were not significant although QR171 and RR171 sheep had higher values. However, in flock 2, average daily gain was significantly greater in AV136 sheep than AA136 sheep and in QR171 sheep than QQ171 sheep. Conclusions and Clinical Relevance—Findings suggest there is an advantage for average daily gain in lambs with an arginine allele at codon 171, but there were no other genotype effects on production traits. Thus, selection for the resistant arginine allele at codon 171 to comply with USDA scrapie eradication guidelines should not be detrimental to lamb production in commercial flocks. Effects of codon 136 on average daily gain were ambiguous.
Показать больше [+] Меньше [-]Association of environmental air contaminants with disease and productivity in swine
1991
Donham, K.J.
A cross-sectional epidemiologic study associating air quality with swine health was conducted on 28 swine farms in southern Sweden. Correlation of housing air environment to swine diseases and productivity (data collected over the preceding 12 months) were investigated. The most prevalent swine health problems detected at slaughter were pneumonia and pleuritis. In farrowing and nursery operations, the most prevalent problem was neonatal pig mortality. Several air contaminants (dust, ammonia, carbon dioxide, and microbes) were found to be correlated with these swine health problems. Maximal safe concentrations of air contaminants were estimated on the basis of dose-response correlation to swine health or human health problems. Recommended maximal concentrations of contaminants were: dust, 2.4 mg/m3; ammonia, 7 ppm; endotoxin, 0.08 mg/m3; total microbes, 10(5) colony-forming units/m3; and carbon dioxide, 1,540 ppm. The overall quality of the ventilation system was correlated with lower concentration of ammonia, carbon dioxide, microorganisms, and endotoxin, but not with dust concentrations. High animal density was related to high ammonia and air microbe concentrations. Animal density measured as kilograms of swine per cubic meter (compared with kilograms of pig weight or swine per square meter) had the highest correlation to animal health and air contaminants.
Показать больше [+] Меньше [-]Effects of enzyme and cryoprotectant concentrations on yield of equine adipose-derived multipotent stromal cells Полный текст
2018
Duan, Wei | Lopez, Mandi J.
OBJECTIVE To evaluate effects of various concentrations of collagenase and dimethyl sulfoxide (DMSO) on yield of equine adipose-derived multipotent stromal cells (ASCs) before and after cryopreservation. SAMPLE Supragluteal subcutaneous adipose tissue from 7 Thoroughbreds. PROCEDURES Tissues were incubated with digests containing 0.1%, 0.05%, or 0.025% type I collagenase. Part of each resulting stromal vascular fraction was cryopreserved in 80% fetal bovine serum (FBS), 10% DMSO, and 10% Dulbecco modified Eagle medium F-12 and in 95% FBS and 5% DMSO. Half of each fresh and cryopreserved heterogeneous cell population was not immunophenotyped (unsorted) or was immunophenotyped for CD44+, CD105+, and major histocompatability complex class II (MHCII; CD44+-CD105+-MHCII+ cells and CD44+-CD105+-MHCII− cells). Cell proliferation (cell viability assay), plasticity (CFU frequency), and lineage-specific target gene and oncogene expression (reverse transcriptase PCR assays) were determined in passage 1 cells before and after culture in induction media. RESULTS Digestion with 0.1% collagenase yielded the highest number of nucleated cells. Cell surface marker expression and proliferation rate were not affected by collagenase concentration. Cryopreservation reduced cell expansion rate and CD44+-CD105+-MHCII− CFUs; it also reduced osteogenic plasticity of unsorted cells. However, effects appeared to be unrelated to DMSO concentrations. There were also variable effects on primordial gene expression among cell isolates. CONCLUSIONS AND CLINICAL RELEVANCE Results supported the use of 0.1% collagenase in an adipose tissue digest and 5% DMSO in cryopreservation medium for isolation and cryopreservation, respectively, of equine ASCs. These results may be used as guidelines for standardization of isolation and cryopreservation procedures for equine ASCs.
Показать больше [+] Меньше [-]Magnetic resonance imaging measurements of organs within the coelomic cavity of red-eared sliders (Trachemys scripta elegans), yellow-bellied sliders (Trachemys scripta scripta), Coastal plain cooters (Pseudemys concinna floridana), and hieroglyphic river cooters (Pseudemys concinna hieroglyphica) Полный текст
2017
Mathes, Karina A. | Schnack, Marcus | Rohn, Karl | Fehr, Michael
OBJECTIVE To determine anatomic reference points for 4 turtle species and to evaluate data on relative anatomic dimensions, signal intensities (SIs), and position of selected organs within the coelomic cavity by use of MRI. ANIMALS 3 turtle cadavers (1 red-eared slider [Trachemys scripta elegans], 1 yellow-bellied slider [Trachemys scripta scripta], and 1 Coastal plain cooter [Pseudemys concinna floridana]) and 63 live adult turtles (30 red-eared sliders, 20 yellow-bellied sliders, 5 Coastal plain cooters, and 8 hieroglyphic river cooters [Pseudemys concinna hieroglyphica]). PROCEDURES MRI and necropsy were performed on the 3 turtle cadavers. Physical examination, hematologic evaluation, and whole-body radiography were performed on the 63 live turtles. Turtles were sedated, and MRI in transverse, sagittal, and dorsal planes was used to measure organ dimensions, position within the coelomic cavity, and SIs. Body positioning after sedation was standardized with the head, neck, limbs, and tail positioned in maximum extension. RESULTS Measurements of the heart, liver, gallbladder, and kidneys in sagittal, transverse, and dorsal planes; relative position of those organs within the coelom; and SIs of the kidneys and liver were obtained with MRI and provided anatomic data for these 4 turtle species. CONCLUSIONS AND CLINICAL RELEVANCE MRI was a valuable tool for determining the position, dimensions, and SIs of selected organs. Measurement of organs in freshwater chelonians was achievable with MRI. Further studies are needed to establish reference values for anatomic structures in turtles. Results reported here may serve as guidelines and aid in clinical interpretation of MRI images for these 4 species.
Показать больше [+] Меньше [-]Two-dimensional left atrium–to–aorta ratios and left ventricular M-mode transthoracic echocardiographic measurements in clinically normal adult Dachshunds Полный текст
2016
Lim, Chee Kin | Fosgate, Geoffrey T. | Green, Henry W III | Kirberger, Robert M.
OBJECTIVE To estimate the left atrium–to–aorta ratio (LA:Ao) and establish 95% prediction intervals for left ventricular M-mode transthoracic echocardiographic measurements in clinically normal adult Dachshunds. ANIMALS 40 healthy Dachshunds. PROCEDURES For each dog, 3 standard 2-D echocardiographic methods (diameter, circumference, and cross-sectional area) were used to measure the left atrium and aorta and calculate the LA:Ao from right parasternal short axis (RPSA) images obtained at the level of the aortic valve cusps. Left ventricular M-mode measurements were acquired from RPSA images obtained at the chordal level immediately below the mitral valve. Descriptive data were generated, and the 95% prediction intervals were calculated by use of an allometric scaling equation and linear regression and compared with those calculated on the basis of data obtained from dogs of multiple breeds in a previous study.RESULTS The mean (SD) LA:Ao was 1.40 (0.13), 2.09 (0.17), and 2.85 (0.48) for the diameter, circumference, and cross-sectional area methods, respectively. The 95% prediction intervals for the left ventricular M-mode measurements determined by an allometric scaling equation on the basis of Dachshund-specific data were narrower than those determined on the basis of data obtained from dogs of multiple breeds. For that allometric equation, scaling exponents on the basis of Dachshund-specific data ranged from 0.129 to 0.397 and did not absolutely conform to the presumed index for linear measurements (ie, body weight0.333). CONCLUSIONS AND CLINICAL RELEVANCE The LA:Aos and 95% prediction intervals calculated in this study can be used as preliminary guidelines for echocardiographic measurements of clinically normal Dachshunds.
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