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Determination of veterinary drug residues, mycotoxins, and pesticide residues in bovine milk by liquid chromatography electrospray ionisation –tandem mass spectrometry
2022
Hajrulai-Musliu, Zehra | Uzunov, Risto | Jovanov, Stefan | Kerluku, Maksud | Jankuloski, Dean | Stojkovski, Velimir | Pendovski, Lazo | Sasanya, James Jacob
Multi-class and multi-residue analyses are very complex procedures because of the physico-chemical properties of veterinary drug residues and other contaminants. The purpose of the study was to develop an analytical method for the sensitive determination of 69 analytes in bovine milk by liquid chromatography electrospray ionisation–tandem mass spectrometry. Antimicrobial, anabolic hormone, lactone, β-agonist, mycotoxin and pesticide residues were analysed in 120 raw milk samples from different dairy farms in North Macedonia. Stable isotopically labelled internal standards were used to facilitate effective quantification of the analytes. The linear regression coefficients were higher than 0.99, the limits of detection ranged from 0.0036 to 47.94 μg/L, and the limits of quantification ranged from 0.053 to 59.43 μg/L. The decision limit values ranged from 0.062 to 211.32 μg/L and the detection capability from 0.080 to 233.71 μg/L. Average recoveries of the analytes spiked in raw milk were in the range of 70.83% to 109%, intra-day coefficient of variation (CV) values from 2.41% to 22.29%, and inter-day CV values from 3.48% to 23.91%. The method was successfully applied in the testing of bovine milk samples. In five samples residues were detected. They were sulfadimethoxine (in two samples), enrofloxacin, tetracycline and oxytetracycline and were at concentrations below the EU maximum residue limit. The method is useful for routine testing for this group of chemical hazards in bovine milk.
Show more [+] Less [-]Occurrence and distribution of canine cutaneous mast cell tumour characteristics among predisposed breeds
2019
Smiech, Anna | Łopuszyński, Wojciech | Ślaska, Brygida | Bulak, Kamila | Jasik, Agnieszka
Introduction: Breed predisposition to cutaneous mast cell tumours (MCT) in a population of dogs in Poland affected by various skin tumours was assessed, and the distribution of MCT characteristics such as histological grading, sex, age, and location, in predisposed breeds was evaluated. Material and Methods: The retrospective epidemiological study included 550 dogs affected by cutaneous MCTs with a reference group of 2,557 dogs diagnosed with other skin tumours. Results: A univariable logistic regression analysis was performed to determine the odds ratios (ORs) with 95% confidence intervals. The risk of high-grade MCTs was the highest for Shar-Peis (OR: 26.394) and American Staffordshire Terriers (OR: 2.897). Boxers (OR: 6.619), Labrador Retrievers (OR: 2.630), French Bulldogs (OR: 2.050), Golden Retrievers (OR: 1.949), and American Staffordshire Terriers (OR: 2.592) were mainly affected by low-grade MCTs. The high risk of MCT was calculated to be at the age of 4–6 years for Labrador Retrievers (OR: 2.686) and 7–10 years for Boxers (OR: 2.956) and French Bulldogs (OR: 9.429). MCTs were significantly more often located on the trunk in French Bulldogs (OR: 4.680), American Staffordshire Terriers (OR: 2.520), and Labrador Retrievers (OR: 1.948). There was no statistically significant correlation between gender and the occurrence of MCTs in the breeds. Conclusions: The breed-predicated differences in the clinical course of MCTs suggest a genetic background for the tumours.
Show more [+] Less [-]The impact of selected risk factors on the occurrence of highly pathogenic avian influenza in commercial poultry flocks in Poland
2021
Gierak, Anna | Śmietanka, Krzysztof
Introduction of highly pathogenic avian influenza virus (HPAIV) into a country and its further spread may have a devastating impact on the poultry industry and lead to serious economic consequences. Various risk factors may increase the probability of HPAI outbreak occurrence but their relative influence is often difficult to determine. The study evaluates how the densities of selected poultry species and proximity to the areas inhabited by wild birds impacted HPAI outbreak occurrence during the recently reported epidemics in Poland. The analysis was developed using these risk factors in the locations of affected and randomly chosen unaffected commercial farms. Generalised linear and non-linear models, specifically logistic regression, classification tree and random forest, were used to indicate the most relevant risk factors, to quantify their association with HPAI outbreak occurrence, and to develop a map depicting spatial risk distribution. The most important risk factors comprised the densities of turkeys, geese and ducks. The abundance of these species of poultry in an area increased the probability of HPAI occurrence, and their farming intensity in several areas of central, western, eastern and northern Poland put these areas at the highest risk. The results may improve the targeting of active surveillance, strengthen biosecurity in the areas at risk and contribute to early detection of HPAI in outbreak reoccurrences.
Show more [+] Less [-]Evaluation of real-time PCR based on SYBR Green I fluorescent dye for detection of Bacillus anthracis strains in biological samples
2018
Kędrak-Jabłońska, Agnieszka | Budniak, Sylwia | Szczawińska, Anna | Reksa, Monika | Krupa, Marek | Szulowski, Krzysztof
Introduction: The aim of the study was the application and evaluation of real-time PCRs based on the fluorescence of SYBR Green I intercalating dye for the detection of three Bacillus anthracis genes in contaminated liver and blood samples. The goals for detection were rpoB gene as a chromosomal marker, pag gene located on plasmid pXO1, and capC gene located on plasmid pXO2. Material and Methods: Five B. anthracis strains were used for the experiments. Additionally, single strains of other species of the genus Bacillus, i.e. B. cereus, B. brevis, B. subtilis, and B. megaterium, and strains of six other species were used for evaluation of the specificity of the tests. Three SYBR Green I real-time PCRs were conducted allowing confirmation of B. anthracis in the biological samples. Results: The observation of amplification curves in real-time PCRs enabled the detection of the chromosomally encoded rpoB gene, pag gene, and capC gene of B. anthracis. The specificity of the tests was confirmed by estimation of the melting temperature of the PCR products. The sensitivity and linearity of the reactions were determined using regression coefficients. Strains of other microbial species did not reveal real-time PCR products. Conclusion: All real-time PCRs for the detection of B. anthracis in biological samples demonstrated a significant sensitivity and high specificity.
Show more [+] Less [-]Correlation of Activity Data in Normal Dogs to Distance Traveled
2020
Eskander, Bishoy S. | Barbar, Megan | Evans, Richard B. | Enomoto, Mastaka | Lascelles, B. Duncan X. | Conzemius, Michael G.
The objective of this study was to explore the mathematical relationships between independent variables (patient morphometrics and treadmill speed) and dependent variables (accelerometer or pedometer output) when evaluating data from accelerometers and pedometers in dogs. Twenty dogs took part in 3 randomized activities, consisting of exercise on a treadmill at 1.0, 1.5, and 2.0 m/s for a total distance of 1 km at each speed. Dogs simultaneously wore both an accelerometer and a pedometer. Statistical analysis used multiple regression models to discover the relationships between independent and dependent variables. A formula was developed to predict the distance traveled by a dog based on its morphometrics and activity monitor output. Shoulder height had stronger correlations to accelerometer and pedometer outputs than other morphometric variables. As shoulder height increased, all accelerometer and pedometer outputs decreased. As treadmill speed increased, both accelerometer and pedometer step counts decreased, while accelerometer activity counts increased. According to a formula derived to predict the total distance traveled using patient shoulder height and accelerometer or pedometer output, pedometer steps were the most accurate predictor of distance traveled. Accelerometer steps were less accurate when using the same model. Accelerometer activity counts did not reveal a meaningful predictive formula. The results of this study indicate that patient morphometrics and treadmill speed (as a measure of intensity) influenced accelerometer and pedometer data. The pedometer data more precisely and accurately estimated the distance traveled based on step counts and patient shoulder height. In normal dogs, accelerometer and pedometer steps may reasonably estimate distance traveled.
Show more [+] Less [-]In vitro holding security of four friction knots of monofilament or multifilament suture used as a first throw for vascular ligation
2020
Tremolada, Giovanni | Hazenfield, Kurt M. | Smeak, Daniel D.
OBJECTIVE To evaluate holding security of 4 friction knots created with various monofilament and multifilament sutures in a vascular ligation model. SAMPLE 280 friction knot constructs. PROCEDURES 10 friction knots of 4 types (surgeon's throw, Miller knot, Ashley modification of the Miller knot, and strangle knot) created with 2-0 monofilament (polyglyconate, polydioxanone, poliglecaprone-25, and glycomer-631) and braided multifilament (silk, lactomer, and polyglactin-910) sutures were separately tied on a mock pedicle and pressure tested to the point of leakage. Linear regression analysis was performed to compare leakage pressures among suture materials (within friction knot type) and among knot types (within suture material). RESULTS Mean leakage pressure of surgeon's throws was significantly lower than that of all other knots tested, regardless of the suture material used. All the other knots had mean leakage pressures considered supraphysiological. Significant differences in mean leakage pressure were detected between various friction knots tied with the same type of suture and various suture types used to create a given knot. Variability in leakage pressure among knots other than the surgeon's throw was greatest for poliglecaprone-25 and lowest for polydioxanone. CONCLUSIONS AND CLINICAL RELEVANCE Most differences in knot security, although statistically significant, may not have been clinically relevant. However, results of these in vitro tests suggested the surgeon's throw should be avoided as a first throw for pedicle ligation and that poliglecaprone-25 may be more prone to friction knot slippage than the other suture materials evaluated.
Show more [+] Less [-]Evaluation of relationships between results of electrocardiography and echocardiography in 341 chimpanzees (Pan troglodytes
2020
Drane, Aimee L. | Atencia, Rebeca | Cooper, Stephen-Mark | Feltrer, Yeddra | Calvi, Thalita | Strike, Tai | Palmer, Christopher | Simcox, Sarah | Rodrigues, Pablo | Sánchez, Carlos | Bolhuis, Hester van | Peck, Bruce | Eng, Jaclyn | Moittie, Sophie | Unwin, Steve | Howatson, Glyn | Oxborough, David | Stembridge, Mike R. | Shave, Rob E.
OBJECTIVE To examine potential relationships between ECG characteristics and echocardiographic measures of cardiac structure in chimpanzees (Pan troglodytes). ANIMALS 341 chimpanzees (175 males and 166 females) from 5 sanctuaries and 2 zoological collections. PROCEDURES Chimpanzees were anesthetized for routine health examinations between May 2011 and July 2017 as part of the International Primate Heart Project and, during the same anesthetic events, underwent 12-lead ECG and transthoracic echocardiographic assessments. Relationships between results for ECG and those for echocardiographic measures of atrial areas, left ventricular internal diameter in diastole (LVIDd), and mean left ventricular wall thicknesses (MLVWT) were assessed with correlational analysis, then multiple linear regression analyses were used to create hierarchical models to predict cardiac structure from ECG findings. RESULTS Findings indicated correlations (r = −0.231 to 0.310) between results for ECG variables and echocardiographic measures. The duration and amplitude of P waves in lead II had the strongest correlations with atrial areas. The Sokolow-Lyon criteria, QRS-complex duration, and R-wave amplitude in leads V6 and II had the strongest correlations with MLVWT, whereas the Sokolow-Lyon criteria, QRS-complex duration, and S-wave amplitude in leads V2 and V1 had the strongest correlations with LVIDd. However, the ECG predictive models that were generated only accounted for 17%, 7%, 11%, and 8% of the variance in the right atrial end-systolic area, left atrial end-systolic area, MLVWT, and LVIDd, respectively. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that relationships existed between ECG findings and cardiac morphology in the chimpanzees of the present study; however, further research is required to examine whether the predictive models generated can be modified to improve their clinical utility.
Show more [+] Less [-]Use of three-dimensional time-of-flight magnetic resonance angiography at 1.5 Tesla to evaluate the intracranial arteries of 39 dogs with idiopathic epilepsy
2019
Ishikawa, Chieko | Ito, Daisuke | Tanaka, Natsumi | Kitagawa, Masato
OBJECTIVE To assess visualization of the intracranial arteries and internal carotid artery (ICA) on 3-D time-of-flight (TOF) magnetic resonance angiography (MRA) images obtained at 1.5 T and to investigate factors that affect the image quality of those arteries in dogs. ANIMALS 39 dogs with idiopathic epilepsy. PROCEDURES Each dog underwent 3-D TOF MRA, and 5 pairs of intracranial arteries, the basilar artery, and both ICAs were evaluated. Each artery was assigned an image-quality score on a scale of 0 to 3, where 0 = poor and 3 = excellent. Multivariable regression analysis was used to assess whether age, body weight (BW), serum total cholesterol concentration, intracranial volume (ICV), and mean arterial pressure were significantly associated with the image quality of each vessel. RESULTS In all dogs, the image-quality score was 2 or 3 for the proximal middle cerebral arteries, basilar artery, and caudal aspect of the caudal communicating arteries. In some dogs, the rostral cerebellar arteries, rostral aspect of the caudal communicating arteries, and middle and rostral aspects of the ICA were poorly visualized. For various arteries, image quality was negatively associated with age and positively associated with BW and ICV. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that 3-D TOF MRA images obtained at 1.5 T did not consistently and clearly delineate the ICA and narrow or peripheral intracranial arteries of dogs; therefore, careful attention is required when such images are assessed. Patient age, BW, and ICV can also affect the image quality of some intracranial arteries on 3-D TOF MRA images.
Show more [+] Less [-]Effects of three fentanyl plasma concentrations on the minimum alveolar concentration of isoflurane in Hispaniolan Amazon parrots (Amazona ventralis)
2018
Hawkins, Michelle G. | Pascoe, Peter J. | DiMaio Kynch, Heather K. | Drazenovic, Tracy L. | Kass, Philip H. | Sanchez-Migallon Guzman, David
OBJECTIVE To determine effects of 3 plasma concentrations of fentanyl on the minimum alveolar concentration of isoflurane (MACiso) and cardiovascular variables in Hispaniolan Amazon parrots (Amazona ventralis). ANIMALS 6 adult parrots. PROCEDURES In phase 1, anesthesia was induced and maintained with isoflurane; intermittent positive-pressure ventilation was provided. The MACiso was determined for each bird by use of a bracketing method and supramaximal electrical stimulus. Fentanyl (20 μg/kg) was administered IV, and blood samples were collected over time to measure plasma fentanyl concentrations for pharmacokinetic calculations. In phase 2, pharmacokinetic values for individual birds were used for administration of fentanyl to achieve target plasma concentrations of 8, 16, and 32 ng/mL. At each concentration, MACiso and cardiovascular variables were determined. Data were analyzed with mixed-effects multilevel linear regression analysis. RESULTS Mean ± SD fentanyl plasma concentrations were 0 ng/mL, 5.01 ± 1.53 ng/mL, 12.12 ± 3.58 ng/mL, and 24.93 ± 4.13 ng/mL, and MACiso values were 2.09 ± 0.17%, 1.45 ± 0.32%, 1.34 ± 0.31%, and 0.95 ± 0.14% for fentanyl target concentrations of 0, 8, 16, and 32 ng/mL, respectively. Fentanyl significantly decreased MACiso in a dose-dependent manner. Heart rate and blood pressure significantly decreased at all fentanyl doses, compared with values for MACiso at 0 ng of fentanyl/mL. CONCLUSIONS AND CLINICAL RELEVANCE Fentanyl significantly decreased the MACiso in healthy Hispaniolan Amazon parrots, but this was accompanied by a depressive effect on heart rate and blood pressure that would need to be considered for application of this technique in clinical settings.
Show more [+] Less [-]Associations between dialysate sodium concentration and plasma sodium concentration of dogs receiving intermittent hemodialysis treatments
2018
Foster, Jonathan D. | Drobatz, Kenneth J. | Cowgill, Larry D.
OBJECTIVE To compare dialysate sodium concentration and patient plasma sodium concentration of dogs during intermittent hemodialysis treatments. SAMPLE 211 intermittent hemodialysis treatments performed on 40 client-owned dogs for the management of dialysis-dependent uremia. PROCEDURES Medical records were reviewed to determine the plasma sodium concentration of each dog before and after routine hemodialysis treatments. Associations between detected changes in plasma sodium concentration and dialysate sodium concentration were evaluated by use of Spearman rank correlations and linear regression analysis. RESULTS Significant linear correlations were found between the dialysate sodium concentration and patient sodium concentration. The starting dialysate-to-patient sodium gradient was associated with the strongest correlation to the change in patient sodium concentration at the end of the dialysis session. Modest correlations existed between the dialysate sodium concentration and postdialysis patient sodium concentration as well as between the predialysis dialysate-to-patient sodium gradient and postdialysis dialysate-to-patient sodium gradient. CONCLUSIONS AND CLINICAL RELEVANCE The dialysate sodium concentration was correlated with the patient sodium concentration in dogs, and the dialysate-to-patient sodium gradient could be used to further refine this association to predict the postdialysis patient sodium concentration and potentially manage dysnatremia during hemodialysis. Prospective studies should be performed to determine how these associations can be used to correct aberrations as well as to avoid unwanted alterations in patient sodium concentrations.
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