Morphological studies in different avian species on artefacts induced by euthanasia with T 61 " or Pentobarbital (Narcoren)

In mammals (e. g. macaques, dogs, cats, rats, sheep) as well as in men (suicides) euthanasia performed by intravenous injection of T 61 leads to serious lesions in lung, kidney or/and liver (endothelial damage, hyperemia, oedema, necrosis). This is caused by the solvent dimethylformamide (DMF). In this study, in contrast, in different species of birds (e. g. blackbird, carrion crow, kestrel, common buzzard, homer pigeon, common wood pigeon, mallard duck) and various modes of applications and dosages T 61, 1.0-3.0 ml/kg body mass, did not induce comparable artefacts in tissues of internal organs in the narcotized animals. Microscopically, only hyperemia and oedema of lung, kidney and/or liver were found. However, milder but similar lesions were detected also in groups of birds euthanized by pentobarbital (200 mg/kg body mass) as well as in control groups (overdosed ketamine intramusculary, 100 mg/kg body mass, and rapid exsanguination). In conclusion, euthanasia of narcotized birds performed by intravenous or intracardial injections ofT 61 seemed to be suitable. The observed lesions could therefore not be interpreted as T61 induced artefacts.     

Reproductive disturbance caused by an S-triazine herbicide in pigs

The aim of the study was to assess the effect of subacute treatment with a low dose of atrazine (1,3,5-triazine-2,4-diamine, 6-chloro-N-ethyl-N'-(1-methyl-ethyl), an s-triazine herbicide, on endocrine oestrus regulation in gilts. A group of nine gilts (F1 generation of Swedish Landrace x Large Yorkshire) were treated with 1 mg atrazine/kg body mass daily, mixed to the feed for 19 days before the onset of expected oestrus. Blood samples were obtained by cranial vena cava puncture three times daily at 3-h intervals on five post-treatment days, i.e. before and during oestrus. The serum concentration of oestradiol-17 beta (E2) was determined by the fluoroimmunochemical method. On Day -2 before the onset of expected oestrus, a significantly lower (P < 0.001) E2 concentration was measured in the serum of treated gilts (31.25 +/- 1.95 and 39.32 +/- 1.38 pg/mL) than in the control pigs (51.43 +/- 1.29 and 68.59 +/- 2.99 pg/mL). In contrast, the E2 concentration measured in the serum of treated animals was significantly higher (P < 0.001) on the day of the expected onset of oestrus and on the subsequent two days (35.43 +/- 1.85, 53.92 +/- 1.98 and 60.32 +/- 2.35 pg/mL, respectively) than in the control animals (13.52 +/- 1.79, 21.53 +/- 1.35 and 20.05 +/- 1.46 pg/mL, respectively). Insufficient serum E2 concentration of the treated gilts resulted in a failure of expected oestrus, as indicated also by the state of dioestrus demonstrated by histopathological examination of the uterus.     

Evaluation of ELISA for detection of Trichinella antibodies in muscle juice samples of naturally infected pigs

The performance characteristics of an ELISA test for trichinellosis in pigs applied to muscle juice was assessed using 314 samples collected from pigs located in endemic areas of Croatia. Peptic digestion was used as the reference method. The diagnostic accuracy of the two compared dilutions (1:10 and 1:100) was considered to be high because the area under the curve (AUC) index was 0.922 and 0.920 for each dilution, respectively. In this study the two graph-receiver operating characteristic (TG-ROC) analysis was used as a tool for selecting cut-off points. Sensitivity, specificity, likelihood ratios, efficiency and Youden's index were used as indices of test accuracy. The cut-off values that minimize overall misclassification cost under an assumption of 3% prevalence were calculated. Our results indicate that the ELISA applied to muscle juice is a highly accurate test and can be adapted to process a large number of samples.     

Levamisole mucosal adjuvant activity for a live attenuated Escherichia coli oral vaccine in weaned pigs

The present study tested the hypothesis that levamisole exerts its immunopotentiating activity in weaned pigs vaccinated against colibacillosis by priming the lymphocytes and macrophages in the mesenteric lymph node (MLN). Ten weaned piglets were used and allocated into two equal groups. The experimental group was intramuscularly primed with levamisole at an immunostimulatory dose of 2.5 mg/kg given daily, in three consecutive days, and controls received saline according to the same schedule. Both groups were orally vaccinated with the vaccinal Escherichia coli strain on day 0 and challenged with the virulent E. coli strain 7 days later. All pigs were killed on postchallenge day 6. Upon virulent challenge the health status of the two groups was evaluated by clinical observations, and expression of CD25, SWC7 and SWC9 activation antigens by MLN and spleen T and B cells and macrophages, respectively, was tested using flow cytometry. Priming by levamisole significantly contributed to the effectiveness of a live attenuated oral vaccine against porcine postweaning colibacillosis, as evidenced by a good health status of primed vaccinated vs. un-primed vaccinated pigs. The CD3+, CD25+ and SWC9+ MLN but not spleen T cells and macrophages increased in experimental vs. control pigs, implying that levamisole exerts its potentiating activity in the MLN by augmenting both recruitment and activation of cells that participate in cell-mediated immunity.     

Proportions and phenotypic expression of peripheral blood leucocytes in pigs vaccinated with an attenuated C strain and a subunit E2 vaccine against classical swine fever

The influence of an attenuated classical swine fever virus C strain vaccine and a subunit E2 vaccine against classical swine fever on the peripheral blood leucocyte proportion and phenotypic expression in 12-week-old pigs was studied. The C strain was amplified in minipig kidney cell culture and final product contained 10(4 +/- 0.15) TCID50/ml, while the subunit vaccine contained 32 microg per dose of gp E2. Haematological findings showed that the vaccines did not cause leucopenia or lymphocytopenia and the number of neutrophils and eosinophils during the observation period was within physiological range. The results of the proportion of CD4a+, CD5a+, CD8a+, wCD21+, CD45RA+, CD45RC+ , non-T non-B, SWC3a+ and CD11b+ cells were gained by single-colour flow cytometry. At the end of the trial a significantly increase of percentage of CD4+, CD5a+, CD8+, wCD21+ cells has been found in pigs that received the subunit vaccine and the percentage of CD4+, CD5a+, CD8+, CD45RA+ and CD45RC+ cells was higher in pigs that received the attenuated vaccine. Twenty-eight days after vaccination the percentage of CD4+, CD45RA+ and CD45RC+ was significantly higher in pigs vaccinated with the C strain than in pigs vaccinated with the subunit vaccine. In contrary, the percentage of the wCD21- cells was higher in pigs that received the subunit vaccine. Statistically higher values of SWC3a+ and lower values of CD11b+ cells was observed in pigs that received the attenuated vaccine than in pigs vaccinated with the subunit vaccine. Taken altogether, our results showed that the subunit vaccine produced a better stimulation of B cells and CD11b+ monocytes/macrophages /granulocytes/NK cells, whereas the attenuated vaccine induced a higher response of Th cells, naive/memory cells and macrophages/neutrophils. Thus, both vaccines were able to influence the porcine immune system, by activating different subsets of the immune effector/accessory cells.     

Characterization of Erwinia amylovora Strains from Croatia

Erwinia amylovora is the causative agent of fire blight, a destructive disease of rosaceous plants. The European population can be divided into several subtypes according to differences in restriction fragment length polymorphism of the XbaI genomic DNA digest analysed with pulsed-field gel electrophoresis. This technique was also used to determine the genetic relatedness of six Croatian isolates to the E. amylovora types found in the countries surrounding Croatia. The isolates belong to the Pt2 pattern type that is characteristic of the East Mediterranean basin. All tested isolates gave essentially the same total cell protein pattern in SDS-polyacrylamide gel electrophoresis. The number of short-sequence DNA repeats in plasmid pEA29 of six isolates was determined by PCR assays and ranged from four to seven. The isolates examined showed high pathogenicity in immature pear fruits. Differences were also revealed in microbiological assays such as amylovoran synthesis, levan formation, siderophore production and colour on coliform medium.     

New aphid species for italy caught by suction trap

Six aphid species new for Italy were caught by suction trap in the Friuli-Venezia Giulia region, Italy. They are:Chaetosiphon chaetosiphon (Nevsky),Illinoia morrisoni (Swain),Myzodium modestum (Hottes),Pseudacaudella rubida (Börner),Schizaphis longicaudata Hille Ris Lambers andTrichosiphonaphis (Xenomyzus)polygonifoliae (Shinji). All species are provided here with characteristic morphological information and line drawings.     

Involution of bursa cloacalis (Fabricii) and thymus in Cyclosporin A-treated chickens

Fifty-day-old Arbor Acres chickens received a daily oral dose of 30 mg/kg Cyclosporin A for 21 consecutive days. This treatment induced the striking morphological changes in the bursa cloacalis (Fabricii). The number of bursal follicles did not change, but they were markedly decreased in size. The pars lymphoreticularis was collapsed and contained smaller accumulations of lymphocytes. The pars lymphoepithelialis was almost totally devoid of lymphocytes and it was composed of an empty network of epithelial cells. Interfollicular epithelium, follicle-associated epithelium, basement-membrane-associated epithelium and bursal epithelial cytoreticulum were not changed. The thymus also displayed substantial structural changes, whereby the thymic medulla was strongly reduced in size.