Anti-Birnavirus Activity of Methisoprinol – in vitro Study with Infectious Pancreatic Necrosis Virus (IPNV)

This study was conducted to evaluate the influence of methisoprinol, synthetic anti-viral product, on the IPNV replication in vitro by measuring viral RNA synthesis. The monolayers of RTG-2 cells in tissue culture plates (Multiwell, 24 wells, Becton Dickinson, USA) wa

Probiotici i lekovito bilje u uzgoju šarana (cyprinus carpio l.) u zemljanim bazenima - uticaj na prirast ribe, zdravlje i proizvodne rezultate

The aim of the paper is to present the results of feeding carp with traditional grain diet (triticale + wheat) supplemented with probiotics and/or herbs. As probiotics, the EmFarma concentrate, provided by ProBiotics Polska, Poland,was used. This preparation contains consortia of the following microbial bacteria and fungi: Bifidobacterium animalis, Bifidobacterium bifidum, Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus bulgaricus, Lactobacillus casei, Lactobacillus delbrueckii, Lactobacillus plantarum, Lactococcus diacetylactis, Lactococcus lactis, Streptococcus thermophilus, Bacillus subtilis var natto, Saccharomyces cerevisiae and Rhodopseudomonas palustris. The concentrate of probiotic microorganisms was added to the ground corn in an amount of 2 liters of preparation per 1 ton of feed. Prior to adding to the feedstuff, the probiotics were diluted in water. The amount of added water was approximately 10% of feed weight. After mixing probiotics with corn, the feed was left for two hours for swelling. The composition of herbs consisted of powdered Terminalia chebula, Phyllantus emblica, Andrographis paniculata, Tinospora cordifolia and Boerhaavia difusa. The herbal preparation was obtained from the Farmwet company, Poland. The blend of herbs was added in the amount of 3 kg per ton of ground corn feed. The powdered herbs were mixed with grinded corn, moistened with water amounting to approximately 10% of feed dose and left for two hours for swelling. Six feeding groups were examined: - I - natural food only (control group) - II - ground mix of corn - III - ground mix of corn supplemented with probiotics - IV - ground mix of corn supplemented with herbs - V - ground mix of corn supplemented with probiotics and herb - VI - pelleted feed Aller Aqua (referential group) Experimental diets were used for feeding carp fingerlings (C1), two-year restocking material (C2) and consumable carps (C3). Stocking densities of carp, were as follows: - for C1– 20000 ind./ha - for C2– 5000 ind./ha - for C3–1500 ind./ha The following parameters were measured and analyzed: - final body mass (g/ind.) - yield (kg/ha) - survival rate (S) - FCR (kg) - Fulton’s coefficient (F) - number of parasites (Trichodina–Trich., Chilodonella–Chil., Epistylis–Epist., Costia,) - level of lisozyme (mg/l) - level of gamma globulins (g/l)Cilj ovog rada je da prikaže rezultate ishrane šarana tradicionalnom smešom žitarica (tritikale + pšenica) sa dodatkom probiotika i/ili lekovitog bilja. Kao probiotik korišćen je koncentrat EmFarma, koji je obezbedio "ProBiotics Polska" iz Poljske. Ova smesa sadrži skup sledećih mikroorganizama, bakterija i gljivica: Bifidobacterium animalis, Bifidobacterium bifidum, Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus bulgaricus, Lactobacillus casei, Lactobacillus delbrueckii, Lactobacillus plantarum, Lactococcus diacetylactis, Lactococcus lactis, Streptococcus thermophilus, Bacillus subtilis var natto, Saccharomyces cerevisiae and Rhodopseudomonas palustris. Koncentrat probiotskih mikroorganizama dodat je u mleveni kukuruz u količini od 2 litra preparata na 1 tonu hraniva. Pre dodavanja hranivima probiotski preparat je razređen u vodi. Količina dodate vode bila je oko 10% od težine hrane. Posle mešanja probiotika sa kukuruzom, hranivo je ostavljeno dva sata da nabubri. Lekovito bilje sastojalo se od praha Terminalia chebula, Phyllantus emblica, Andrographis paniculata, Tinospora cordifolia i Boerhaavia difusa. Biljni preparat dobijen je od kompanije Farmwet iz Poljske. Mešavina bilja dodata je u količini od 3 kg na tonu kukuruzne prekrupe. Bilje u prahu pomešano je sa prekrupom, zatim navlaženo vodom u količini od oko 10% od doze hrane i ostavljeno dva sata da nabubri. Ispitivanje je vršeno na šest hranidbenih grupa: - I - samo prirodna hrana (kontrolna grupa) - II - smeša sa kukuruznom prekrupom - III - smeša sa kukuruznom prekrupom obogaćena probioticima - IV - smeša sa kukuruznom prekrupom obogaćena lekovitim biljem - V - smeša sa kukuruznom prekrupom obogaćena probioticima i lekovitim biljem - VI - peletirana hrana Aller Aqua (referentna grupa) Eksperimentalne smeše korišćene su za ishranu šaranske mlađi (C1), dvogodišnjaka za dalji uzgoj (C2) i konzumnih šarana (C3). Gustine nasada šarana bile su sledeće: - za C1– 20000 jedinki/ha - za C2– 5000 jedinki/ha - za C3–1500 jedinki/ha Sledeći parametri su mereni i analizirani: - završna telesna masa (g/jedinka) - prinos (kg/ha) - stopa preživljavanja (S) - stopa konverzije hrane, FCR (kg) - Fultonov koeficijent (F) - broj parazita (Trichodina–Trich., Chilodonella–Chil., Epistylis–Epist., Costia,) - nivo lizozima (mg/l) - nivo gama globulina (g/l

Species specific anaesthetics for fish anaesthesia and euthanasia.

There is a need to ensure that the care and welfare for fish maintained in the laboratory are to the highest standards. This extends to the use of anaesthetics for both scientific study, humane killing and euthanasia at end of life. An anaesthetic should not induce negative behaviours and fish should not seek to avoid the anaesthetic. Surprisingly little information is available to facilitate a humane choice of anaesthetic agent for fish despite over 100 years of use and the millions of fish currently held in thousands of laboratories worldwide. Using a chemotaxic choice chamber we found different species specific behavioural responses among four closely related fish species commonly held in the laboratory, exposed to three widely used anaesthetic agents. As previously found for zebrafish (Danio rerio), the use of MS-222 and benzocaine also appears to induce avoidance behaviours in medaka (Oryzias latipes); but etomidate could provide an alternative choice. Carp (Cyprinus carpio), although closely related to zebrafish showed avoidance behaviours to etomidate, but not benzocaine or MS-222; and rainbow trout (Oncorhynchus mykiss) showed no avoidance to the three agents tested. We were unable to ascertain avoidance responses in fathead minnows (Pimephales promelas) and suggest different test paradigms are required for that species

Influence of HMB (β\beta-hydroxy-β\beta-methylbutyrate) on antibody secreting cells (ASC) after in vitro and in vivo immunization with the anti-Yersinia ruckeri vaccine of rainbow trout (Oncorhynchus mykiss)

In practice, protection of fish against disease by immunization is of limited effectiveness. Therefore, research is concentrated on how to improve the potency and efficacy of vaccines and how to optimally activate the cell-mediated immunity and the specific antibody response. In the present study, the influence of HMB ($\beta$-Hydroxy-$\beta$-methylbutyrate) on the antibody secreting cells (ASC) after both in vitro and in vivo immunization of rainbow trout (Oncorhynchus mykiss) with the anti-Yersiniose vaccine was studied. For in vitro immunization, the spleens from 160 fish were sampled and placed each in 35 mm sterile wells with medium containing HMB at concentrations of 0, 0.1, 1, 5, 10, 25, 50 or 100 $\mu$g/mL of medium. The spleens from 80 fish were injected with the vaccine and incubated at 14 °C for 10 days. For the in vivo study, fish were fed pellets containing HMB at doses of 0, 10, 25 and 50 mg/kg bw per day. After 2 weeks of HMB supplementation, the fish were immunized by intraperitoneal injection of the vaccine. At 7, 14, 18, 21, 28 and 35 days after immunization, pronephros were taken from 10 fish in each group for testing. When analyzed by the ELISPOT assay, HMB increased the number of splenic ASC after in vitro immunization at concentrations between 10 and 100 $\mu$g/mL ($P < 0.05$). Dietary HMB also increased the number of total and specific ASC when the fish were vaccinated in vivo. In conclusion, the results of the present study showed that HMB increases the levels of specific ASC after both in vitro and in vivo immunization of rainbow trout with the anti-Yersinia ruckeri vaccine.Influence du HMB sur les cellules productrices d'anticorps après vaccination in vitro et in vivo de la truite arc-en-ciel (Oncorhynchus mykiss) avec le vaccin Anti-Yersiniose. En pisciculture, la vaccination contre les maladies infectieuses ne présente qu'une efficacité limitée. En conséquence, la recherche est orientée vers les moyens d'augmenter l'efficacité des vaccins et de stimuler l'immunité cellulaire et la réponse en anticorps. La présente étude démontre l'influence du $\beta$-hydroxy-$\beta$-méthylbutyrate (HMB) sur les cellules productrices d'anticorps après immunisation in vitro et in vivo de la truite arc-en-ciel (Oncorhynchus mykiss) avec le vaccin "Anti-Yersiniose " . Pour l'étude in vitro, les rates de 160 poissons ont été prélevées et incubées dans du milieu contenant du HMB aux concentrations 0, 0,1, 1, 5, 10, 25, 50 ou 100 $\mu$g/mL de milieu. Les rates de 80 poissons ont été injectées avec le vaccin et incubées à 14 °C pendant 10 jours. Pour l'étude in vivo, des poissons (40-50 g) ont été nourris avec des granulés supplémentés en HMB à raison de 0, 10, 25 et 50 mg/kg de poisson par jour. Au bout de 2 semaines de supplémentation, les poissons ont été vaccinés par injection intra-péritonéale de vaccin. Aux jours 7, 14, 18, 21, 28 et 35 après vaccination, les pronephros ont été prélevés sur 10 poissons par groupe. In vitro, l'analyse ELISPOT a démontré que HMB augmentait le nombre de cellules produisant les anticorps spécifiques aux concentrations comprises entre 10 et 100 $\mu$g/$\mu$L ($P < 0,005$). In vivo, HMB distribué dans l'aliment a augmenté le nombre des cellules productrices d'anticorps spécifiques et non spécifiques après vaccination. En conclusion, in vitro et in vivo, HMB associé à la vaccination augmente, chez la truite arc-en-ciel, le nombre de cellules produisant les anticorps spécifiques