Milan Pogačnik

Ivermectin pharmacokinetics in lactating sheep.

Ivermectin (IVM) concentrations in plasma and milk were studied in six Istrian Pramenka dairy sheep after a single subcutaneous dose of 0.2 mg/kg b.w. of IVM in the early lactation period to describe IVM disposition in milk and to evaluate the transfer of IVM residues via milk to suckling lambs. Large inter-animal in concentration variability of IVM in both matrices was observed. The highest overall concentration was found in the same animal: 21.7 microg/l of H(2)B(1a) in plasma on the second day and 44.9 microg/kg of H(2)B(1a) in milk on the first day after the drug was administered. The mean time in which IVM concentrations fell below the limit of detection for the whole ewe group was 22 and 23 days for plasma and milk, respectively. Time course of IVM concentration in milk was following the time course of IVM concentration in plasma, with an overall mean+/-S.D. of milk/plasma ratio of 1.67+/-0.50 for the first 7 days of the experiment. A mean of 0.7% of the dose was excreted through milk. Individual pharmacokinetic parameters were determined by fitting a one-compartment model to the milk and plasma concentration-time profiles. Mean t(max), c(max), t(1/2k(e)) and AUC values for plasma data were: 1.70+/-0.65 days, 11.88+/-6.96 microg/l, 2.85+/-1.97 days and 63.99+/-28.34 microg day/l, respectively, and for milk: 1.28+/-1.07 days, 22.67+/-18.27 microg/l, 3.56+/-2.01 days and 114.60+/-60.41 microg day/l, respectively. The highest level of concentration in suckling lamb plasma, 0.36 microg/l of H(2)B(1a), was slightly above the limit of determination. The mean lamb to ewe ratio of areas under the plasma concentration-time curve for the first 5 days was 0.02. On the basis of obtained results, it can therefore be claimed that indirect IVM exposure of the suckling lambs via milk was negligible.

Linearity of eprinomectin pharmacokinetics in lactating dairy sheep following pour-on administration : Excretion in milk and exposure of suckling lambs

Pharmacokinetics of eprinomectin (EPR) were studied in blood plasma and milk in two groups of six Istrian Pramenka dairy sheep and their suckling lambs following pour-on administration of EPR to ewes at dose levels of 0.5 and 1mg/kg. Maximum concentration in plasma was 2.22 and 5.25 microg/l, and AUC was 13.6 and 33.7 microg day/l for the 0.5 and 1.0mg/kg dose, respectively. These results indicate that drug exposure with a dose of 0.5mg/kg, which is commonly used in cattle, may be subtherapeutic. The concentration time course in milk paralleled plasma concentrations. In the dose range studied, linear pharmacokinetics of EPR were demonstrated. Milk-to-plasma AUC ratio was 0.79+/-0.12 and 1.12+/-0.43; the fraction of dose recovered in milk was 0.037+/-0.011 and 0.058+/-0.027% for the low and high dose, respectively. Maximum residual levels in milk were below the maximum acceptable level of 20 microg/kg; however, EPR was detected in all samples investigated. Despite low permeability in milk, AUC in plasma of suckling lambs was between 20 and 30% of the AUC in plasma of ewes.

Uterine heterologous malignant mixed Müllerian tumor in a dwarf rabbit (Oryctolagus cuniculus)

Malignant mixed Müllerian tumor (MMMT) is a rare neoplasm of the female genital system. A case of MMMT in the uterus of an 8-year-old female dwarf rabbit, which died with clinical signs associated with severe acute dyspnea and anorexia, is described. At necropsy, an oval, firm tumor was found in each of the 2 uterine horns. Numerous metastases were scattered throughout the mediastinum, thoracic diaphragm, and all pulmonary lobes. Microscopically, the tumors consisted of a poorly demarcated, unencapsulated neoplasm, composed of closely associated carcinomatous and sarcomatous components and areas of osteosarcomatous differentiation. Metastases were composed entirely of the sarcomatous component with osteosarcomatous differentiation. Immunohistochemically, the neoplastic epithelial component was positive for cytokeratin and negative for α-smooth muscle actin (α-SMA), vimentin, and desmin. The sarcomatous component was diffusely and strongly positive for vimentin, focally positive for α-SMA (<20% of cells positive), and negative for desmin. The neoplasm was diagnosed as a heterologous MMMT with metastases to the lung, mediastinum, and thoracic diaphragm.

Factors influencing dissipation of avermectins in sheep faeces

Factors influencing fate of avermectins (abamectin, doramectin) in faeces of treated sheep were investigated under different experimental conditions. In the laboratory, concentrations of both avermectins were declined in homogenised faeces of treated animals until day 14 of exposure, regardless of experimental conditions. After that day, no significant decrease in concentrations was observed till the end of the experiment. Established DT(50) did not exceed 9 days. In the karst pasture, an average DT(50) of 27 days was established for abamectin and 23 days for doramectin in natural faeces of treated sheep. In the compost mixture, doramectin concentration was decreased by 38.9+/-2.6% during 21 days of the thermophilic phase of composting. Therefore, DT(50) was not established. A possible influence of moisture content of sheep faeces on concentrations of avermectins was observed.

Subchronic liver injuries caused by microcystins.

Abstract The subchronic effects of cyanobacterial lyophilizate (CL) containing microcystins on liver were investigated in female New Zealand rabbits. Sterilised CL containing microcystins was injected i.p. Liver toxicity was assessed by histological examination of liver samples. Non-invasive magnetic resonance imaging (MRI) of liver was also performed in order to assess changes in the homogeneity of liver tissue. Subchronical intoxication with microcystins caused morphological changes of liver tissue that were also detected by use of MRI. Histological analysis showed that changes seen on MRI represent liver injury characterised with fatty infiltration and periportal fibrosis. This demonstrates that subchronic exposure to microcystins can lead to liver degeneration, which can easily be detected in vivo by use of MRI.

Haemocytic neoplasia in Mediterranean mussels (Mytilus galloprovincialis) in the Slovene Adriatic Sea

The health status of cultured and wild Mediterranean mussels in the Slovene Sea has so far been unexplored. Initially, 1280 adult Mediterranean mussels (Mytilus galloprovincialis), 960 from a shellfish farm and 320 from natural beds, were collected over a one-year period. Water temperature, oxygenation and salinity were measured at each sampling. Mussels were measured and weighted to calculate the condition index and microscopically examined for the presence of haemocytic neoplasia. Haemocytic neoplasia was detected in 14 mussels (1.1%) with a higher prevalence in cultured mussels. Neoplastic cells singularly infiltrated the connective tissue, in small foci or diffusely. Necrosis and multifocal atrophy of digestive tubules were noticed in mussels with diffuse neoplasia, whereas severe haemocytic infiltration of connective tissue was seen in mussels with single neoplastic cells. Haemocytic neoplasia was more frequently observed in spring and autumn. The average condition index of mussels with haemocytic neoplasia was slightly higher than in healthy ones. This is the first report of haemocytic neoplasia in Mediterranean mussels in the Northern Adriatic Sea. The disease occurs only sporadically and to date no significant impact on the mussel population has been noted.

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.

Quill injury - cause of death in a captive Indian crested porcupine (Hystrix indica, Kerr, 1792).

Abstract Indian crested porcupine (Hystrix indica) is a member of the family of Old World porcupines (Hystricidae). Its body is covered with multiple layers of quills, which serve for warning and attack if animal is threatened. However, the literature data on injuries caused by Indian crested porcupine are absent. We describe pathomorphological lesions in an Indian crested porcupine from the Ljubljana Zoo, which died after a fight with a younger male that caused a perforative quill injury of the thoracic wall, followed by septicaemia. Macroscopic, microscopic and bacteriological findings were detailed.

Epidemiologic effects on helicobacter spp. infections of dogs in Slovenia

The aims of our study were to determinate the prevalence of helicobacter in a population of Slovenian dogs, to characterize the intensity of infection and to find out if the epidemiological parameters i. e. age, feeding regimen, gender, breed, location and indoor/outdoor living conditions have any influence on infection and the intensity of infection. A total of 185 randomly chosen dogs from all parts of Slovenia, at ages from 9 days to 15 years, of both genders and 44 different breeds, without any gastrointestinal disorders, were included in our study. Helicobacter were detected in stomachs of 92.4% of dogs. We determined a mild infection in 17.3% of dogs, a moderate infection in 48.1%, and a strong infection in 27% of dogs. Studying epidemiological parameters and their effects on infection we concluded that age and feeding regimen affect the infection and the intensity of infection, whereas gender, breed, location and indoor/ outdoor living conditions do not.

Morphologic and morphometric analysis of adrenal gland cortex of sheep grazing on pastures with the ground of electrofilter ash

Electrofilter ash is a side product of power plants using coal as a fuel. Trials for transforming ash covered areas into cultivated landscape are ongoing e.g., efforts to form grasslands that would be usable for animal breeding and production. However the question of how safe grazing is on such surfaces in relation to animal health and the use of animal products, produced on such surfaces, for human consumption has not been truly resolved yet. The main goal of our work is a systematic search for the possible negative effects on animals grazing on such surfaces. In this work we present the results of morphologic and morphometric analyses that were made to evaluate the potential toxic or some other impact of electrofilter ash constituents on the adrenal gland cortex. No conclusive evidence of such effects was determined in this study.