Effects of two infectious bursal disease vaccine virus strains on hepatic microsomal enzyme activities in chickens

The influence of two infectious bursal disease vaccines on the activities of hepatic microsomal enzymes aniline hydroxylase, ethylmorphine N-demethylase, NADPH-cytochrome c reductase, aryl sulphotransferase and p-nitrophenol UDP-glucuronyltransferase was investigated in chickens. The vaccines contained attenuated Winterfield 2512 and VMG-91 strains, respectively. The activities of enzymes were determined on postvaccination days 0, 2, 5 and 7. At the same time, post-mitochondrial supernatant, cytosolic and microsomal pellet protein concentrations were determined. As expected, the antibody titres against infectious bursal disease virus in the serum were increased in both tested groups in relation to each administered vaccine. Using RT-PCR, the presence of the VP2 gene fragment of virus in the liver of chicken was demonstrated 4 and 6 h after vaccination. The results of this study suggest that the two commercial vaccines modulate the activities of five enzymes tested, and that the two attenuated vaccines applied triggered induction and/or inhibition of phases I and II of biotransformation enzyme activities.     

Effect of fasting and refeeding on blood glutathione and lipid peroxide concentration of cockerels and pullets

Changes of reduced glutathione (GSH) and TBARS (thiobarbituric acid-reactive products of lipid peroxidation) concentrations and activity of gamma-glutamyltransferase (GGT, EC 2.3.2.2) in the blood of Lohman brown cockerels and pullets in response to 48 hour food deprivation and 24 hour refeeding were examined. The experiment was performed on 61-day-old chickens. Blood samples ware collected from the wing vein (v. brachialis) in heparinized tubes for three times: control sampling before fasting, then after 48 hour food deprivation and after refeeding for 24 hours. Blood GSH concentration after refeeding in cockerels was significantly higher compared with prefasting and fasting values. The concentration of GSH in female chickens was significantly lower after fasting as well as after refeeding compared with control values. In addition to that, in pullets GSH concentration in refeeding was higher than in fasting conditions. The level of TBARS in blood in female and male chickens after fasting and refeeding were significantly lower than the prefasting values. The GGT activity on cockerels after 48 hour food deprivation was significantly higher compared with control sampling and in chickens refeed for 24 hours, whereas in pullets significant difference was exhibit compared only with control values. Concentration of GSH in control sampling in cockerels compared with those in pullets was significantly lower. After 48 hours of fasting, the level of GSH was significantly higher in the cockerels than in the pullets. Results of TBARS concentration in the pullets were higher of control and fasting values than in the cockerels. The GGT activity of control sampling was significantly higher in male chicken. Lipid peroxidation in chickens of both sexes decreased with fasting, but prooxidative-antioxidative processes were more intensive in female chickens, probably because they were not reach sexual maturity.     

Evaluation of nonstarch polysaccharides and oligosaccharide content of different soybean varieties (Glycine max) by near-infrared spectroscopy and proteomics

A total of 832 samples of soybeans were screened by near-infrared (NIR) reflectance spectroscopy, to identify soybean samples with a lower content of oligosaccharides and nonstarch polysaccharides (NSP). Of these, 38 samples were identified on the basis of variation in protein content and agronomic value and submitted to high-resolution NIR spectroscopy. On the basis of the NIR data, 12 samples were further selected for chromatographic characterization of carbohydrate composition (mono-, di-, and oligosaccharides and NSP). Their soluble proteins were separated by two-dimensional gel electrophoresis (2DE). Using partial least-squares regression (PLSR), it was possible to predict the content of total NSP from the high-resolution NIR spectra, suggesting that NIR is a suitable and rapid nondestructive method to determine carbohydrate composition in soybeans. The 2DE analyses showed varying intensities of several proteins, including the glycinin G1 precursor. PLSR analysis showed a negative correlation between this protein and insoluble NSP and total uronic acid (UA).     

Effect of phosphorus concentration of the nutrient solution on the volatile constituents of leaves and bracts of Origanum dictamnus

The chemical composition of the essential oils obtained from the leaves and bracts of hydroponically cultivated Origanum dictamnus were analyzed by GC-MS techniques. Three different concentrations of phosphorus (5, 30, and 60 mg/L) in the nutrient solution were used for the cultivation, using the nutrient film technique (NFT). A total of 46 different compounds were identified and significant differences (qualitative and quantitative) were observed between the samples. Carvacrol and p-cymene were identified as the main compounds in all samples analyzed, whereas thymoquinone was found in higher percentage in the leaves than in bracts. The essential oils were tested for their antibacterial activity against Gram-positive and Gram-negative bacteria. The oils obtained from the bracts were found to be more active. The results obtained from GC-MS analyses were submitted to chemometric analysis.     

3D morphometric study of domestic fowl glomerulus—A scanning electron microscope study of vascular corrosion casts

Microvascularization of domestic fowl kidneys was studied using scanning electron microscopy (SEM) of vascular corrosion casts (VCCs). Two types of nephrons, mammalian-type (MT) and reptilian-type (RT) nephrons and their glomerular structure were analysed quantitatively by 3D morphometry. A significant difference in shape and size between the MT and RT glomeruli was found. The mean diameter of the RT glomeruli was about 56 µm, while that of MT glomeruli was significantly larger, namely about 80 µm. The afferent arterioles in mammalian-type glomeruli usually bifurcated into two lobular branches and formed a complex glomerular capillary network with numerous loops. Reptilian-type glomeruli consisted of a single capillary forming few loops and leaving the glomerulus as efferent arteriole. Diameters of afferent and efferent arteriolar replicas were similar in all three kidney divisions of MT and RT nephrons. The absence of the interconnecting branches between the MT nephron capillaries at the gross inspection suggests that the mammalian-type nephron glomeruli, although more complex than the reptilian type, are not equivalent to those in mammalian kidneys.     

Anatomical variations of the thoracic duct in the dog

Anatomical variations can be frequently found in the lymphatic system, which is also true for the shape and course of the thoracic duct (ductus thoracicus), the biggest lymph vessel in the body. From 2012 to 2019, the thoracic duct was successfully dissected in 43 dog carcasses that were used in the anatomy course at the Faculty of Veterinary Medicine, University of Zagreb. The thoracic duct originated from the cranial border of the cisterna chyli as one lymph vessel in 36 dogs (83.7%), as two vessels in six dogs (14%) and as three vessels in one dog (2.3%). We divided the observed thoracic duct variations into six groups according to their anatomical similarities. Considering the specific embryonic development, we can conclude that all observed variations are the result of minor deviations from the standard ontogenesis. However, the importance of thoracic duct variations is significant in surgical procedures done in the thoracic cavity to prevent or cure the chylothorax. Since this research showed variations in 39 out of 43 dogs (90.7%) throughout the whole course of the thoracic duct, great care must be taken while performing the ligation or embolization of the thoracic duct.     

Thermodynamics and structural features of the yeast Saccharomyces cerevisiae external invertase isoforms in guanidinium-chloride solutions

Recently, four external invertase isoforms (EINV1, EINV2, EINV3, and EINV4) have been isolated from S. cerevisiae. However, there is nothing known about their structural features and thermodynamics of unfolding. Since this information is essential for understanding their functioning at the molecular level as well as applicable in the food industry, we investigated guanidinium-chloride induced structural changes of the isoforms by CD and fluorescence spectroscopy. The resulting unfolding curves measured for each isoform at different temperatures were described simultaneously by a reversible two-state model to obtain the corresponding thermodynamic parameters. Here, we show that they are different for different isoforms and demonstrate that they correlate with the surface charge density of the native isoforms which follows the order EINV1 < EINV2 < EINV3 < EINV4. It appears that at physiological temperatures the thermodynamic stability of the isoforms follows the same order, while above 55 °C, the order is the opposite EINV1 > EINV2 > EINV3 ≈ EINV4. This suggests that increasing the efficiency of the food industry processes involving invertase would require the application of EINV3 and/or EINV4 at physiological temperatures and EINV1 at elevated temperatures.     

Co-oximetry in clinically healthy dogs and effects of time of post sampling on measurements

Objectives : Co‐oximetry is a complex and valuable laboratory method that measures haemoglobin species and oxygenation status by multi‐wavelength spectrophotometry. The purpose of this study was to establish reference intervals for clinically healthy dogs and to determine the effect of time of analyses and sex of animals on the accuracy of results. Methods : Blood was collected from 27 healthy adult dogs of various breeds and sex. Co‐oximetry was performed on a CCX co‐oximeter that measures eight haemoglobin and oxygen transport related parameters: carboxyhaemoglobin (COHb), deoxyhaemoglobin (HHb), oxyhaemoglobin (O₂Hb), methaemoglobin (MetHb), total haemoglobin (tHb), oxygen saturation (SO₂%), oxygen content (O₂Ct) and oxygen capacity (O₂Cap). Results : Results obtained after 2 and 4 hours were not significantly different from those obtained immediately after sampling. But after 48 hours, the results for total haemoglobin, oxygen saturation, oxyhaemoglobin, oxygen content and oxygen capacity were significantly lower, and carboxyhaemoglobin and deoxyhaemoglobin values were significantly higher than determination immediately after sampling. Gender had no significant impact on co‐oximetry values. Clinical Significance : Co‐oximetry offers several advantages compared with other methods, including ease of use, increased accuracy and greater differentiation among haemoglobin species.     

Influence of P-glycoprotein inhibition on secretion of ivermectin and doramectin by milk in lactating sheep

The aim of to the present study was to evaluate the effects of verapamil (VER) on plasma pharmacokinetics of ivermectin (IVM) and doramectin (DOR) in lactating Istrian Pramenka dairy sheep and to investigate the role of P-glycoprotein (P-gp) in transport of avermectins into milk. Pharmacokinetics of IVM and DOR following subcutaneous administration of 0.2mg/kg b.w. was evaluated in four groups of sheep. They were administered either IVM or DOR alone or in combination with verapamil (VER) at a dose of 3.0mg/kg b.w., 3 times at 12h intervals. Blood plasma and milk samples were collected at defined time intervals over 30 days post-treatment to determine IVM and DOR concentration levels. Pharmacokinetic parameters in sheep injected with IVM or DOR alone corresponded to previously published values. Comparison between sheep injected with IVM only, and sheep injected with IVM in combination with VER (IVM+VER) showed significant difference in pharmacokinetic parameters in blood plasma. Area under the concentration-time curve (AUC) truncated at 2 days (AUC(2)) was 15 and 28 μg day/L for group IVM and IVM+VER, respectively. With co-administration of VER, apparent plasma clearance (Cl/F) and mean residence time (MRT) of IVM decreased from 135 to 116 L/day and from 5.8 to 3.8 days, respectively. Similar trends were observed for DOR (AUC(2) 48 vs. 68 μg day/L, Cl/F 61 vs. 46 L/day, and MRT 5.6 vs. 4.4 days for groups DOR and DOR+VER, respectively). This study confirms that co-administration of VER has a significant effect on pharmacokinetic parameters of subcutaneously administered IVM in blood plasma. The influence on DOR pharmacokinetics is much weaker. This could be either due to the difference in lipophilicity or the difference in affinity towards P-gp as a result of structural differences. No significant influence of VER on AUC ratio of IVM and DOR between milk and plasma was observed suggesting that P-gp does not govern transport of avermectins into milk.     

Antimicrobial susceptibility of Clostridium perfringens isolates of bovine, chicken, porcine, and turkey origin from Ontario

Antimicrobial susceptibilities and toxin types were determined for 275 Clostridium perfringens isolates collected in Ontario in the spring of 2005. Minimal inhibitory concentrations (MICs) of C. perfringens isolates for 12 antimicrobials used in therapy, prophylaxis, and/or growth promotion of cattle (n = 40), swine (n = 75), turkeys (n = 50), and chickens (n = 100) were determined using the microbroth dilution method. Statistical analyses and MIC distributions showed reduced susceptibility to bacitracin, clindamycin, erythromycin, florfenicol, and tetracycline for some isolates. Reduced susceptibility to bacitracin was identified in chicken (64%) and turkey (60%) isolates. Swine isolates had predominantly reduced susceptibility to clindamycin (28%) and erythromycin (31%), whereas bovine isolates had reduced susceptibility to clindamycin (10%) and florfenicol (10%). Reduced susceptibility to tetracycline was spread across all species. No clear reduced susceptibility, but elevated MIC(50) for virginiamycin was found in chicken isolates in comparison with isolates from other species. Toxin typing revealed that C. perfringens type A is the dominant toxin type isolated in this study across all 4 host species.