Determination of Bile Acids in Canine Biological Samples: Diagnostic Significance.

The comprehensive examination of bile acids is of paramount importance across various fields of health sciences, influencing physiology, microbiology, internal medicine, and pharmacology. While enzymatic reaction-based photometric methods remain fundamental for total BA measurements, there is a burgeoning demand for more sophisticated techniques such as liquid chromatography-tandem mass spectrometry (LC-MS/MS) for comprehensive BA profiling. This evolution reflects a need for nuanced diagnostic assessments in clinical practice. In canines, a BA assessment involves considering factors, such as food composition, transit times, and breed-specific variations. Multiple matrices, including blood, feces, urine, liver tissue, and gallbladder bile, offer insights into BA profiles, yet interpretations remain complex, particularly in fecal analysis due to sampling challenges and breed-specific differences. Despite ongoing efforts, a consensus regarding optimal matrices and diagnostic thresholds remains elusive, highlighting the need for further research. Emphasizing the scarcity of systematic animal studies and underscoring the importance of ap-propriate sampling methodologies, our review advocates for targeted investigations into BA alterations in canine pathology, promising insights into pathomechanisms, early disease detection, and therapeutic avenues.

In Vitro Pharmacokinetic Behavior of Antiviral 3-Amidinophenylalanine Derivatives in Rat, Dog and Monkey Hepatocytes.

Type II transmembrane serine proteases represent pharmacological targets for blocking entry and spread of influenza or coronaviruses. In this study, the depletion rates of the 3-amidinophenylalanine (3-APhA)-derived matriptase/TMPRSS2 inhibitors MI-463, MI-472, MI-485 or MI-1900 were determined by LC-MS/MS measurements over a period of 300 min using suspensions of rat, dog and cynomolgus monkey primary hepatocytes. From these in vitro pharmacokinetic (PK) experiments, intrinsic clearance values (Clint) were evaluated, and in vivo pharmacokinetic parameters (hepatic clearance, hepatic extraction ratio and bioavailability) were predicted. It was found that rat hepatocytes were the most active in the metabolism of 3-APhA derivatives (Clint 31.9-37.8 mL/min/kg), whereas dog and monkey cells displayed somewhat lower clearance of these compounds (Clint 6.6-26.7 mL/min/kg). These data support elucidation of important PK properties of anti-TMPRSS2/anti-matriptase 3-APhAs using mammalian hepatocyte models and thus contribute to the optimization of lead compounds.

Farm to table: Residues of different pesticides in tomato and tomato juice - Food safety aspects.

During plant cultivation, the pesticides can get into the tissue of vegetables due to crop protection processes, and thus into the food chain. Therefore, they constitute a potential risk to the consumer's health. Depletion of pesticides [spirotetramat (Movento), azoxystrobin and difenoconazole (Amistar Top)] was monitored by testing tomatoes treated individually or simultaneously and tomato juices prepared from the treated tomatoes. The investigations aimed to reveal any kinetic interaction between the compounds tested and changes in their elimination, and thus to assess their compliance with the official Maximum Residue Limits (MRLs). The co-presence of pesticides prolonged the elimination of the individual compounds which reached significantly higher residue levels (P < 0.0001) in tomato, especially difenoconazole (45%) and azoxystrobin (50%) on day 8 after treatment that can cause food safety issues to the human consumers. However, the concentrations of pesticides applied alone or simultaneously were found to be below the corresponding MRL values after the withdrawal period in all investigated tomato and tomato juice samples. Accordingly, the investigated pesticides can be safely used simultaneously, their concentrations are in compliance with the legal regulations and thus their concomitant presence does not pose any risk to the consumers' health.

Transfer of certain beta-lactam antibiotics from cow's milk to fresh cheese and whey.

The aim of this study was to investigate the transfer of cephalexin, penicillin-G, and ampicillin & cloxacillin from cow's milk to cheese and whey. For this purpose, raw milk was artificially contaminated to different antibiotic levels and then heat-treated to prepare fresh cheese from it. Antibiotic levels of the milk, whey and cheese were measured with LC-MS/MS. The extent of heat degradation was not sufficient to remove the antibiotic residues from milk. Antibiotic concentrations in whey and fresh cheese were in good accordance with the concentration of the same compound in milk suggesting that contamination of the milk will result in contamination of the product. The investigated antibiotics were transferred less into the cheese curd (1.6-12.5% of the original amount), than into the whey (33.2-74.1%). For penicillin-G even 100% (complete removal) was experienced.

Serum hepcidin measurements in healthy dogs using liquid chromatography/tandem mass spectrometry.

BACKGROUND: Hepcidin is a key regulator of iron homeostasis. The measurement of this hormone is essential for the diagnosis of iron deficiency anemia and might be useful as a prognostic factor in many diseases. Serum hepcidin levels have been infrequently evaluated in dogs. OBJECTIVES: The aim of this study was to measure serum hepcidin in a population of healthy dogs using liquid chromatography/tandem mass spectrometry (LC-MS/MS), considered the gold standard assay for measuring hepcidin like in human medicine. METHODS: Blood samples from 86 healthy dogs of 25 different breeds were measured with the LC-MS/MS method. Synthetic canine hepcidin was used as the standard reagent. Reference values were calculated based on the results. RESULTS: The mean hepcidin concentration of the study population was 16.6 ± 7.7 ng/mL. There reference interval (RI) was defined as 5.3-36.4 ng/mL. No significant difference was found between male and female dogs, or between different age and body weight groups. Hepcidin concentrations did not correlate with red blood cell counts, hemoglobin concentrations, iron levels, iron-binding capacities, and C-reactive protein concentrations in this healthy population. A weak negative correlation was found between hepcidin and the mean corpuscular volume. CONCLUSION: LC-MS/MS proved to be a reliable and time-effective method for the detection of canine hepcidin. The RI was similar but narrower compared with that of human studies.

Heterocyclic Amine Formation in Grilled Chicken Depending on Body Parts and Treatment Conditions.

Heterocyclic amines (HCAs) carcinogenicity is known since the 1970's, but the exact way of their formation is still unclear. During these examinations different body parts (breast filet with and without skin, thigh filet without skin and full wing with skin) of chickens from the same Ross 308 strain were analyzed after grilling with the combination of 3-3 temperature and duration levels (150-180-210 °C and 2.5-5-10 min per side). Five different kinds of heterocyclic amines (HAR, NOR, MeIQx, 4,8-DiMeIQx and PhIP) were detected by HLPC-MS/MS. The results obtained from the present study confirm that, in general, the higher the temperature and longer the duration of the grilling the more HCAs will be generated. Grilling of chicken thigh without bones and skin resulted in lower amounts of HCAs generated in comparison to the grilling of chicken breast without skin. The presence of skin on the chicken breast increased the amounts of HCAs formed, especially if grilling was performed at high temperature for longer duration, especially at 210 °C for 10 min. In case of grilling the chicken wings, the amounts of HCAs formed were lower than observed in the breast.

Predictive Correlation between Apparent Sensory Properties and the Formation of Heterocyclic Amines in Chicken Breast as a Function of Grilling Temperature and Time.

In the present set of experiments, we studied the correlation between the heterocyclic amine (HCA) concentration and the color changes of the chicken breast with or without skin during grilling under open or closed conditions as a function of the applied temperature and time. The concentration of the HCAs formed during grilling was measured by a validated LC-MS/MS method, whereas the color changes were determined either instrumentally or by visual observation. In general, higher temperatures and longer heat treatment times resulted in a more substantial HCA formation, especially on the surface of the samples and in the skin, where the total levels reached 746 ng/g. Results of regression analysis demonstrate a strong correlation (r > 0.7) between the HCA content of the grilled chicken breast and the L* and a* values indicating the significance of brightness and the red parameter of the color scale, respectively. In the case of open grilling, the skinless breast samples showed correlation (r > 0.7) between the HCA content and the color analysis results in both the full sample and the crust, respectively. Breast samples with skin exhibited the same level of correlation when they were grilled closed. In the case of open grilling the breast with skin, and closed-grilling the skinless breast, the linear regression analysis yielded a weaker correlation (0.7 > r > 0.4 or less) between the HCA concentrations and the color. Our results demonstrate that there is a predictive correlation between the color changes perceptible for the consumers and the HCA formation during grilling of chicken breast as a function of time and temperature depending on the type of grilling and the presence of skin.

Heavy metals in seafood purchased from a fishery market in Hungary.

Heavy-metal content (arsenic, cadmium, lead and mercury) was investigated and evaluated in shellfish, oysters and squids. The samples were collected weekly for 20 weeks from a fishery product market in Hungary. The concentration of heavy metals was measured by Inductively Coupled Plasma Optical Emission Spectrometer (ICP-OES) analysis after sample preparation. The results were analysed statistically by one-way ANOVA method. The average concentration of arsenic (3.01 ± 1.46 mg kg-1) in shellfish was significantly higher (p < 0.001) compared to oysters (2.88 ± 1.12 mg kg-1) and squids (1.28 ± 0.52 mg kg-1). The level of mercury was below the limit of detection (0.5 mg kg-1) in each sample and there was no statistical significance in the concentrations of cadmium (p = 0.351) and lead (p = 0.412) in the species investigated. The provisional tolerable intake values were also calculated. Based on the obtained results of the heavy-metal content of the investigated samples, the seafood is considered to be safe for human consumption. However, prolonged ingestion of oysters and squids at these levels may contribute to the consumers' cadmium burden.

LC-MS study of the heat degradation of veterinary antibiotics in raw milk after boiling.

The aim of our study was to examine the degradation of veterinary antibiotics in milk during boiling. Raw cow milk samples were fortified with the target compounds and boiled for various short time-intervals prevailing in household practice. Antibiotic concentrations were determined by LC-MS/MS measurements. Degradation rate constants, half-lives and degradation percentages were calculated. Cefoperazone and cloxacillin proved to be the less and the most heat-stable substance, with 78.3% and 9.6% degradation in 300 s respectively. Aminoglicosides exhibited intermediate (33.8-43.6%), tetracycline (30.4%) and trimethoprim (22.6%) intermediate to high heat stability. The results demonstrate that antibiotic residues possibly present in raw milk exhibit high heat stability when treated for few seconds at around 100 °C. Keeping the milk at this temperature for at least two minutes would allow varying decrease in the amount of some compounds, but does not totally eliminate the potential risks to the consumer's health.

Food safety aspects of primary environmental contaminants in the edible tissues of roe deer (Capreolus capreolus).

The muscle, liver, kidney and fat samples of 20 roe deer of both sexes originating from a hunting area in central Hungary were investigated for the presence of heavy metals such as As, Cd, Hg and Pb, and their contents were evaluated for possible health risk to consumers. Both As and Hg were found at a level below the limit of detection (< 0.5 mg/kg wet weight) in all samples. The median of the measured Cd concentrations was significantly higher in both the kidney and the liver (p = 0.0011) of bucks than of does. In bucks, Cd levels exceeded the respective maximum limits laid down in the European legislation in four kidney and three muscle samples, whereas in does, the measured concentrations were below the respective limits in all samples. The detected amounts of Pb exceeded the maximum limits in the kidney of one buck and eight does, in the liver of two bucks and six does, in the muscle of six bucks and nine does, whereas in all fat tissues of both bucks and does. The concentration of Pb (p = 0.02) was significantly greater in the kidney of does compared to roebucks. Based on data obtained from the present study, the consumption of organs and tissues of the investigated roe deer could be objectionable from food-toxicological point of view and may pose risk to the high consumers of wild game due to their cadmium and lead contents.

Toxic heavy metals in the muscle of roe deer (Capreolus capreolus)--food toxicological significance.

The study was performed on 20 (10 males, 10 females) roe deer (Capreolus capreolus) to investigate the concentration of cadmium, lead, mercury, and arsenic in the muscle tissue. They reside in forest and meadow, about 50 km distance from industrial activities and traffic. Samples were taken from the musculus biceps femoris of each deer without external contamination after shooting during the regular hunting season on a hunting area close to Eger in Hungary. The determination of heavy metal contents was carried out by inductively coupled plasma optical emission spectrometry (ICP-OES). The statistical analysis was performed by statistical package for the social sciences (SPSS) version 11.0. The measured residue concentration of cadmium was below the limit of detection in the roe deer meat indicating no health risk for the consumers. The average lead concentration (0.48 ± 0.21 mg/kg wet weight) exceeded the regulated maximum limit, but its calculated weekly intake was below the provisional tolerable weekly intake (PTWI). The residue level of mercury is not regulated and the average mercury content of roe deer meat (0.87 ± 0.40 mg/kg wet weight) was about half of PTWI, but the consumption of meat with the highest detected concentrations results in higher PTWI than recommended. The measured concentration of arsenic (0.27 ± 0.20 mg/kg wet weight) in the roe deer meat may not pose any health risk for the human consumers according to the PTWI set by the World Health Organization.