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.     

Molecular epidemiology of the rabies virus in Slovenia 1994-2010

A molecular epidemiology study was performed on a selection of 30 rabies-positive brain samples collected between 1994 and 2010 in Slovenia and originating from the red fox (n=19), badger (n=3), cattle (n=3), dog (n=2), cat (n=1), marten (n=1) and horse (n=1). Based on the comparison of 1092 and 672 nucleotide sequences of nucleoprotein (N) and partial glycoprotein (G) gene regions, a low genetic diversity of the circulating strains was detected, but both phylogenetic trees were consistent with the topology where partial nucleoprotein or glycoprotein genes were used. A high sequence identity in the N and G gene to rabies virus isolates from neighbouring countries was found. The Slovenian strains were clearly different from the vaccine strains SAD B19 and SAD Bern, which have been used in Slovenia since 1988.     

Cytokine profiles and phenotype regulation of antigen presenting cells by genotype-I porcine reproductive and respiratory syndrome virus isolates

ABSTRACT: The present study examined the immunological response of antigen presenting cells (APC) to genotype-I isolates of porcine reproductive and respiratory syndrome virus (PRRSV) infection by analysing the cytokine profile induced and evaluating the changes taking place upon infection on immunologically relevant cell markers (MHCI, MHCII, CD80/86, CD14, CD16, CD163, CD172a, SWC9). Several types of APC were infected with 39 PRRSV isolates. The results show that different isolates were able to induce different patterns of IL-10 and TNF-α. The four possible phenotypes based on the ability to induce IL-10 and/or TNF-α were observed, although different cell types seemed to have different capabilities. In addition, isolates inducing different cytokine-release profiles on APC could induce different expression of cell markers.     

Effects of two probiotic additives containing Bacillus spores on carcass characteristics, blood lipids and cecal volatile fatty acids in meat type chickens

The objective of this study was to evaluate effects of two commercially available probiotic additives, containing Bacillus spores, on carcass and meat characteristics, serum lipids and concentration of cecal volatile fatty acids of meat type chickens. Birds were fed regular corn-soy meal based feed (control), supplemented with additive A, containing 1.6 × 10(6) spores per gram of feed of Bacillus subtilis and Bacillus licheniformis (group A) or additive B, containing the same concentration of Bacillus cereus var. toyoi spores (group B). One hundred and twenty birds (20 per replicate) were slaughtered at the age of 55 days. Results showed that birds in group B had higher (p < 0.05) final body weight compared to birds from group A and higher carcass weights and yield percentages compared with control. Breasts and whole legs were also heavier in group B, compared to control, but not the yield. Group A had higher yield of wings and lower abdominal fat weight compared to group B (p< 0.05), but not compared with control. Total cholesterol was not affected by the dietary treatment, on contrary both probiotics elevated the LDL (p < 0.05) and lowered HDL cholesterol, thus unfavourably changed animal's blood serum cholesterol profile. Both probiotics influenced the cecal fermentation, which was observed as decrease in cecal concentrations of propionic, butyric, n-butyric and n-valeric acids, but the differences compared to control group were statistically significant for group A only. It was established that probiotic additive B was more effective regarding carcass and meat part weights than additive A, however the animals from group B also had more abdominal fat and their meat had significantly higher conductivity than control group, which is not considered as beneficial.     

Using Leaf Mass per Area as predictor of light interception and absorption in crop/weed monoculture or mixed stands

The partitioning of light absorption is difficult to assess in natural or cultivated mixed vegetation canopies. Previous research has shown that it is possible to estimate light absorption in monospecific canopies from the Leaf Mass per Area (M_A), which is closely related to the prevailing light conditions experienced by the leaf during its development.The effectiveness of this approach in assessing light partitioning at individual plant level was tested on monospecific and mixed crop/weed plots. Canopy absorbed PAR (APAR) was estimated in monocultures or 1:1 mixed stands of Abutilon theophrasti Medicus and Glycine max L. (soybean) using a relationship between intercepted PAR (IPAR) and M_A, calibrated at individual leaf level against incident light measurements. The accuracy of estimates was evaluated through comparison with APAR calculated from incident light measurements taken independently at various heights at whole canopy level.The use of the experimentally assessed relationship of IPAR vs. M_A provided acceptable absorption estimates. The average departure of estimates from measurements, expressed as Root Mean Squared Error (RMSE), was 10.2%. By comparison, a numerically optimized version of the model that excluded the effects of experimental errors in single-leaf PAR measurements, yielded estimates with an average RMSE of 4.8%. The lower accuracy of the estimates based on the experimental IPAR-M_A relationship was due to experimental error but also to a high sensitivity of the M_A-based estimates to the model coefficients.M_A variability in monoculture or mixed canopies was confirmed to be highly dependent on the PAR distribution, so M_A is potentially suitable for use as a predictor of light absorption. However, due to the sensitivity of the model, a high experimental accuracy in IPAR-M_A assessment as well as in M_A and LAI distribution has to be guaranteed in order to perform reliable estimates by this method.     

Molecular evidence of genetic diversity changes in pea (<Emphasis Type="Italic">Pisum sativum</Emphasis> L.) germplasm after long-term maintenance

Maintaining germplasm genetic integrity is a key objective of long-term ex situ conservation. Periodic regeneration, performed on limited plots with small number of individuals, increases the risk of genetic drift and genetic diversity changes. In this study, six accessions of white flowered, dry seed pea varieties (Pisum sativum L. subsp. sativum var. sativum): Bohatyr, Klatovsky zeleny, Hanák, Moravsky hrotovicky krajovy, Raman and Viktoria-75 and four accessions of colour flowered, fodder pea (P. sativum subsp. sativum var. arvense (L.) Poiret: Arvika, Česky banán, Moravská krajová and Niké, representing Czechoslovak varieties and landraces, bred over the last 40–80 years, were analyzed using ten microsatellite locus specific markers. Each accession was represented by 20 individual seeds of two temporally different samples, spanning the period of 20 or 40 years. Together with intra-accession variation (except of cv. Hanák), evidence of genetic changes, e.g. differences in allele frequencies as well as genetic composition of sample, was detected in six out of ten accessions (Arvika, Bohatyr, Česky banán, Moravsky hrotovicky krajovy, Moravská krajová and Raman). Evidence of genetic erosion was found in three accessions (Česky banán, Moravsky hrotovicky krajovy and Raman), while in another three (Arvika, Bohatyr and Moravská krajová) the level of diversity was found to have increased. Moreover in three samples of Bohatyr (2004) and Klatovsky zeleny (1963 and 2004), low levels of heterozygosity was detected. These results demonstrate that in pea, a self-pollinating and highly homozygous plant, the danger of the loss of genetic integrity exists. These findings are significant for long-term ex situ germplasm management.     

Comparison of real-time PCR with conventional PCR and culture to assess the efficacy of a live attenuated Salmonella enterica serovar Typhimurium vaccine against Salmonella enterica serovar Enteritidis in commercial leghorn chicks vaccinated under field and laboratory conditions

The efficacy of a live attenuated Salmonella Typhimurium Megan Vac 1 vaccine (MV1) was evaluated against Salmonella Enteritidis in chicken pullets with the use of PCR and culture methods. Two hundred Hyline W-32 white leghorn chicks were obtained from a local hatchery and divided into four treatment groups. Two of the groups served as positive and negative controls. The MV1 vaccine was administered to the chicks in the remaining two groups at 1 and 35 days old by either the coarse spray (field) or the oral route (laboratory) method. The chicks were challenged with a high dose of a Salmonella Enteritidis strain at 10 wk old and euthanatized 3 days postinoculation. Samples for PCR analysis were collected prior to enrichment, after pre-enrichment in buffered peptone water (BPW) and after primary enrichment from the ceca, liver, and spleen. None of the samples tested yielded positive results for the Salmonella Typhimurium vaccine strain by either the culture or PCR methods. Results from the standard culture method showed that vaccinating the birds with MV1 reduced the counts of Salmonella Enteritidis recovered from the challenged birds. In addition, fewer pre-enriched samples tested positive for Salmonella Enteritidis among the challenged groups that were vaccinated when compared to the unvaccinated challenged group. Under the conditions of this study, MV1 was unable to prevent colonization of other internal organs such as the liver and spleen. Real-time PCR was significantly more sensitive than conventional PCR (C-PCR) prior to enrichment, but after enrichment the sensitivities of the two methods were similar. Enrichment significantly increased the sensitivity of both PCR methods for the detection of Salmonella Enteritidis in cecal samples, but did not significantly increase the sensitivity for detection of Salmonella Enteritidis in liver and spleen samples that were pre-enriched in BPW. There was no significant difference between the laboratory or field vaccination methods with respect to either the prevalence of Salmonella Enteritidis isolation or the bacterial loads in culture-positive samples. Collectively, the data suggest that MV1 offered some protection against Salmonella Enteritidis in commercial layer chick pullets under the conditions of this study. Given the labor and time required to perform the C-PCR and culture methods, the real-time PCR method may prove to be a more useful method to use in diagnostics.     

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.     

Effects of castration-induced visceral obesity and antioxidant treatment on lipid profile and insulin sensitivity in New Zealand white rabbits

Molecular mechanisms, responsible for the impaired insulin-sensitivity state due to the obesity are not fully understood in both humans and animals. The purpose of this study was to investigate the effects of castration-induced visceral obesity and the influence of two antioxidants on constituents of blood lipid profile and insulin sensitivity in New Zealand white rabbits. Twenty-six clinically healthy male New Zealand white rabbits were used in the experiment and were divided into 3 groups: first group (CI, n = 7) - castrated-obese and treated with antioxidants “Immunoprotect” for 2 months; second group (CO, n = 7) - castrated-obese; third group (NC, n = 12) - control group (non-castrated, non-obese). At the end of the follow-up period of 2 months after castration an intravenous glucose tolerance test (IVGTT) was performed after a 12-h fasting period as the blood samples for determination of glucose and insulin and their kinetic parameters were obtained at 5 and 0 min before and at 5, 10, 30, 60 and 120 min after the infusion of the glucose. The constituents of lipid profile, triglycerides (TG), total cholesterol (TC) and HDL-cholesterol (HDL-C) were also assessed in the overnight fasting blood samples. The body weight (BW), body mass index (BMI), amount of the visceral fat (VF) and VF/BW ratio were both measured and calculated before the IVGTT and at the end of the experimental period. All measured markers of obesity (BW, BMI, VF, VF/BW) were significantly higher in both groups of castrated rabbits than in the control group. Apart HDL-C, the plasma concentrations of all constituents of lipid profile (TG, TC, HDL-C) were the highest in CO group. There were generally no differences between CI and NC groups for the same traits. After glucose injection blood glucose concentrations and glucose and insulin kinetic parameters were considerably higher (except of glucose elimination rate) in CO rabbits than in NC ones. Castrated rabbits treated with “Immunoprotect” showed lower fasting plasma insulin and improved glucose kinetics dynamics than CO rabbits, but commensurable values of glucose and insulin kinetics parameters than NC group. The results of the current study clearly indicated that castration-induced visceral obesity affected negatively the lipid profile and insulin sensitivity and/or responsiveness. Treatment with antioxidant supplementation, consisted of d-limonene and vitamin E, improved blood lipid profile, fatty liver, glucose homeostasis and insulin sensitivity in obese rabbits. In addition, based on our results we may suggest that castrated male New Zealand white rabbits might be considered as an appropriate animal model to study various metabolic abnormalities related to visceral obesity, such as dyslipidemia and impaired insulin sensitivity.