Scoring of the extent and intensity of carp (Cyprinus carpio) skin changes made by cormorants (Phalacrocorax carbo sinensis): relationship between morphometric and histological indices

Cormorants cause damage to fish in aquaculture not only by consuming large number of fish but also by wounding them. Quantification of the actual damage in fish induced by wounding is important for reliable estimations of the overall loss in aquaculture caused by cormorants. In this study, specimens of mirror and scaly carp (Cyprinus carpio L.) injured by attacks of great cormorant (Phalacrocorax carbo L.) were collected from the ??E?ka?? fish farm in Serbia. Surfaces of superficial and open subepithelial wounds (expressed as a percent of the total body surface), estimated by computer-assisted image analysis, had mean values 10.66?±?1.86 and 0.85?±?0.90, respectively. Histological evaluation of wounds was performed by two scoring systems: system proposed by Bernet et al. (J Fish Dis 22:25?C34, 1999) and modified system described by Wahli et al. (Aquaculture 225:371?C386, 2003). The most frequent histopathological changes were disturbed epidermal structure and surface cell morphology and increased number of club cells (on more than 75% of specimens); increased number of melanocytes and their distribution in different layers in dermis; and enlarged hypodermis (more than 80% of samples) and myolysis (50% of samples). Comparison of wounds?? surfaces (surface of superficial injures) and histopathological changes, estimated by two scoring systems, revealed significant positive correlation. Bernet??s scoring system showed higher correlation coefficients, when compared with the modified Wahli??s scoring system.     

Nepetalactone content in shoot cultures of three endemic Nepeta species and the evaluation of their antimicrobial activity

Rapid micropropagation of Nepeta rtanjensis, N. sibirica and N. nervosa was performed. Qualitative and quantitative nepetalactone content in methanol extracts of in vitro grown plants was analysed by reverse-phase HPLC coupled with UV and MS detection. Only trans,cis-nepetalactone was detected in shoots of N. rtanjensis, while cis,trans-nepetalactone stereoisomer was present in N. sibirica. No nepetalactone was observed in shoots of N. nervosa. The antimicrobial activity of methanol extracts, against eight bacterial and eight fungal species, was evaluated. All the tested extracts showed significant antibacterial and strong antifungal activity. However, N. rtanjensis extract exhibited the best antimicrobial potential.     

Parthenogenetic Activation of Pig Oocytes Using Pulsatile Treatment with a Nitric Oxide Donor

The nitric oxide donor (+)–S‐nitroso‐N‐acetylpenicillamine (SNAP) is capable of inducing parthenogenetic activation in pig oocytes matured in vitro. However, quite a long exposure to the nitric oxide donor, exceeding 10 h, is necessary for successful oocyte activation. Repeated short‐term treatment with 2 mm SNAP significantly increased the activation rates despite the fact that the overall exposure time to the nitric oxide donor did not exceed 4 h. With regard to the activation rate, 12 repeated treatments lasting 10 min each were found to be the most efficient regimen (63.3%). The continuous exposure to the nitric oxide donor for the same overall time induced parthenogenetic activation in 12.5% oocytes (2‐h continuous treatment with 2 mm SNAP). The development of parthenogenetic embryos increased after repeated short‐term treatment with SNAP. After continuous treatment with 2 mm SNAP for 10 h, only 6.7% of the oocytes cleaved, and none developed beyond the 4‐cell stage. Thirty‐minute treatment repeated four times with 2 mm SNAP induced cleavage in 37.5% of the oocytes, 18.3% developed to the morula stage, and 6.7% reached the blastocyst stage. Based on the results, it is concluded that pulsatile treatment can significantly improve parthenogenetic activation rate when compared with the continuous treatment using nitric oxide donors.     

Distribution of porcine circovirus 2 cap antigen in the lymphoid tissue of pigs affected by postweaning multisystemic wasting syndrome

The lymphatic organs of 50 pigs from a total of eight farms located at different sites in the epizootiological region of North Ba?ka County were studied to obtain data on the prevalence of circoviral infections in Serbia. All of the pigs examined had clinical signs suggestive of postweaning multisystemic wasting syndrome (PMWS). All pigs underwent necropsy and tissue samples were taken for histopathological, immunohistochemical (IHC) and PCR analysis. The presence of porcine circovirus 2 (PCV2) was established by PCR analysis in the organs of the pigs tested. The most frequent histopathological lesions of lymphoid tissue linked with the presence of positive immunostaining for PCV2 Cap antigen confirmed the existence of PMWS in all farms tested in North Ba?ka County. Using PCR, histopathological and IHC techniques, the presence of PMWS was proved in the Republic of Serbia. During necropsy, generalised enlargement of the lymph nodes was evident. The most common histopathological finding was lymphocyte depletion in the follicular and perifollicular areas of lymph nodes. Infiltration by macrophages was also recorded. By IHC analysis, the cytoplasm of macrophages was shown to contain a large amount of the ORF2-coded Cap antigen of PCV2. Lymphocyte depletion and large numbers of macrophages were recorded in the tonsils, spleen, intestinal lymphatic tissue, Peyer's patches and ileocaecal valve. The presence of typical granulomatous lesions with multinuclear giant cells (MGCs) was also recorded in the lymphatic tissue. Cap antigen was shown to be present in macrophages and less often in lymphocytes.     

Biological properties of a naturally attenuated infectious bursal disease virus isolated from a backyard chicken flock

The biological properties of an infectious bursal disease (IBD) virus isolated from bursas collected during an outbreak in a village chicken flock in Macedonia are described. The mortality rate was 50%. Two viruses coexisted in the bursas of infected chickens (IBDVwt and IBDVtc). The virus termed IBDVtc grows on chicken embryo fibroblast (CEF) cells from the first passage. Specific pathogen free chickens inoculated with IBDVtc at passage level 4 did not develop any clinical signs of disease. Some discrete bleeding on the leg muscles was seen and the bursa of Fabricius revealed pathological lesions similar to those caused by classical strains. However, the bursa recovered quickly (bursa lesion score 2) by 14 days post infection (PI). We also found evidence of bursal repopulation by means of perinuclear antigen staining. Strong CD3 influx was evident at 4 days PI, and at 33 days PI the CD3+ cell finding was comparable to the control. The mean antibody titre was 9.2 log 2 at 14 days PI. The amino acid composition of VP2 in IBDVwt (222 Ala, 242 Ile, 253 Gln, 256 Ile, 279 Asp, 284 Ala, 294 Ile and 299 Ser) is described. The same sequence was found in IBDVtc, except for two point mutations, at Gln253→His and Ala284→Thr. Such amino acid substitution is responsible for partial attenuation and the ability of the strain to replicate in cell culture. None of the commercial vaccine viruses has a similar arrangement of amino acids in the variable domain of IBDV. This strongly suggests that IBDVtc originates from a very virulent strain. To the best of our knowledge, this is the first report of a concomitant infection of chickens with highly pathogenic IBDV and its mutant counterpart.     

Effects of soil solarization on nematodes in Croatia

For the first time soil solarization was investigated in Croatia both in the field and in the greenhouse in 1991, 1992, 1993 and 1994. For two months (July and August), the soil was mulched with transparent polyethylene (PE) sheets of 0.015 or 0.050mm thickness. Soil temperatures at depths of 5, 10 and 20cm were recorded daily. In order to assess nematode population densities, soil samples were analysed before mulching and at the end of the mulching treatment. The results of these experiments showed that soil solarization drastically reduced the population of plant-parasitic nematodes (Meloidogyne, Pratylenchus, Paratylenchus, Tylenchus, Tylenchorhynchus spp.) by about 97–100% at a depth of 10cm and 92–97% at a depth of 20cm in the field, while in the greenhouse, the population of plant-parasitic nematodes was reduced by about 89–100% at a depth of 10cm and 98–100% at a depth of 20cm.In the same experiments, the population of saprophytic nematodes in the field was reduced by about 86–90% at a depth of 10cm and 72–89% at a depth of 20cm. In the greenhouse, the population of saprophytic nematodes was reduced by about 87–97% at a depth of 10cm and 87–93% at a depth of 20cm. This data shows that soil solarization was less effective in the control of saprophytic nematodes, which is considered to be an advantage.     

Usninsäure aus jugoslawischen Flechten und ihre antibiotische Wirksamkeit

Zusammenfassung Es wurde L-Usninsäure aus jugoslawischen Flechten in reiner Form isoliert. Der Schmelzpunkt der Substanz (zwei Proben) ist bei . Die Elementarmikroanalyse sowie die qualitativen Reaktionen entsprechen auch der reinen L-Usninsäure. Ein drittes isoliertes Präparat hatte den Schmp. bei 196° und . Das Präparat mit dem Schmp. 204° wurde auf seine bakteriostatische Wirksamkeit verwendet, und zwar in fein pulverisierter Form in einer Blut-Agar-Nährlösung suspendiert (Rinnenmethode), sowie mit der Routinemethode der Diskuse. Ein Lösungsvermittler für die sehr schwer in Wasser lösliche Usninsäure wurde nicht angewendet.Die Methode der Suspendierung in dem Nährmedium ist etwas empfindlicher, aber weniger praktisch. L-Usninsäure wirkt in vitro bakteriostatisch auf Mikroorganismen der Kugelform und zeigt so eine Ähnlichkeit mit Penicillin. Sie entwickelt eine Wirksamkeit gegenüber Streptokokken, Staphylokokken und Pneumokokken. Auf grampositive Bacillen zeigt sie in Suspensions-Konzentrationen von 0,25, 0,5 und 1,0% keine Wirksamkeit, was mit der sehr schweren Löslichkeit der Substanz in Wasser im Zusammenhang stehen wird.Die Wirkungsweise der Usninsäure steht am nächsten dem Penicillin, obwohl dieselbe schwächer ist als beim letzteren.Gegen Penicillin resistente Stämme zeigten sich als empfindlich auf Usninsäure.Die Versuche werden fortgesetzt.

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Summary L-usninic acid was isolated in pure form from lichens of Jugoslavia. The melting point of the substance (two samples) lies at . The elementary microanalysis and the qualitative reactions are in accordance with the pure L-usninic acid. A third isolated preparation showed the melting point 196° C, and . The preparation (melting point 204°) was tested on the bacteriostatic effet, specifically pulverized suspended in a blood-agar-culture solution (Rinnen-method) (channel-method), as well as with the routine method of the discuse. A medium to rise the solubility in water of the very little soluble usninic acid was not used.The method of suspending this acid in the culture solution is somewhat more sensitive, but less usefull. L-usninic acid has in vitro a bacteriostatic effect on spherical microorganisms in resemblance with penicilline. It is effective against streptococci, staphylococci and pneumococci. On Gram-positive bacillae the usninic acid has no effect in concentrations of 0,25 0,5 and 1,0 % of the suspensions, probably in consequence of its very small solubility.The effectiveness of the usninic acid is analogous to that of the penicilline, but weaker than the last-named. Stocks resistant against penicilline showed sensible against usninic acid.The researches will be continued.

     

Peroral Application of Water-soluble Derivative of Propolis (WSDP) and Its Related Polyphenolic Compounds and Their Influence on Immunological and Antitumour Activity

Polyphenolic compounds are widely distributed in the plant kingdom and display a variety of biological activities, including chemoprevention and growth inhibition of tumours. Propolis contains a conglomerate of polyphenolic compounds. We investigated the effect of propolis and polyphenolic compounds, components of propolis, on the growth and metastatic potential of a transplantable mammary carcinoma (MCa) of the mouse. Metastases in the lung were generated by 2 x 10 tumour cells injected intravenously (i.v.). A water-soluble derivative of propolis (WSDP) and the polyphenolic compounds (caffeic acid (CA) and caffeic acid phenethyl ester (CAPE)) were given to mice perorally before or after tumour cell inoculation. WSDP, CA and CAPE reduced the number of metastases in the lung. This implies that the antitumour activities of the compounds used in these studies are mostly related to the immunomodulatory properties of the compounds, their cytotoxicity to tumour cells, and their ability to induce apoptosis and/or necrosis.     

Effects of egg mass and percentage mass loss during incubation on hatchability of eggs of the rock partridge (Alectoris graeca)

1. The effects of initial egg mass (IEM) and percentage mass loss during incubation (%ML) on hatchability of rock partridge eggs were investigated. 2. Eggs at the extremes of IEM had lower fertility and embryonic mortality. 3. Eggs at the extremes of %ML also had low fertility and hatchability was disproportionately reduced in eggs that had lost less mass during incubation. 4. Chick mass was a function of both IEM and mass lost during incubation. 5. In these respects rock partridge eggs are similar to that of other domesticated species of poultry.