Diversity of currently circulating rabies virus strains in Croatia

Sylvatic rabies has been present in Croatia for more than three decades, with the red fox (Vulpes vulpes) as the main reservoir. The present epidemic of sylvatic rabies in Croatia started already in 1977 and in the past ten years the disease has become enzootic in the entire country and thus represents a considerable veterinary and public health threat. A genetic characterization and phylogenetic analysis of rabies virus isolates (RABV) from Croatia was performed using panel of 32 selected rabies-positive brain samples from domestic and wild animals collected between 2008 and 2010. Based on the comparison of 367-nucleotide sequences of a conserved region of the nucleoprotein (N) gene (nucleotides 75-441), the phylogenetic analysis revealed a low genetic diversity of currently circulating RABV strains in Croatia. 18 RABV isolates mainly originating from Eastern Croatia clustered with the formerly established Eastern European (EE) lineage, and the rest (14) were identical with the West European (WE) group. Both phylogenetic groups seem to coincide in central regions on both sides along the Save River. A high sequence identity in the N gene of the RABV isolates from neighbouring countries was found.     

The effectiveness of selected feed and water additives for reducing Salmonella spp. of public health importance in broiler chickens: A systematic review, meta-analysis, and meta-regression approach

Eating inappropriately prepared poultry meat is a major cause of foodborne salmonellosis. Our objectives were to determine the efficacy of feed and water additives (other than competitive exclusion and antimicrobials) on reducing Salmonella prevalence or concentration in broiler chickens using systematic review-meta-analysis and to explore sources of heterogeneity found in the meta-analysis through meta-regression. Six electronic databases were searched (Current Contents (1999-2009), Agricola (1924-2009), MEDLINE (1860-2009), Scopus (1960-2009), Centre for Agricultural Bioscience (CAB) (1913-2009), and CAB Global Health (1971-2009)), five topic experts were contacted, and the bibliographies of review articles and a topic-relevant textbook were manually searched to identify all relevant research. Study inclusion criteria comprised: English-language primary research investigating the effects of feed and water additives on the Salmonella prevalence or concentration in broiler chickens. Data extraction and study methodological assessment were conducted by two reviewers independently using pretested forms. Seventy challenge studies (n=910 unique treatment-control comparisons), seven controlled studies (n=154), and one quasi-experiment (n=1) met the inclusion criteria. Compared to an assumed control group prevalence of 44 of 1000 broilers, random-effects meta-analysis indicated that the Salmonella cecal colonization in groups with prebiotics (fructooligosaccharide, lactose, whey, dried milk, lactulose, lactosucrose, sucrose, maltose, mannanoligosaccharide) added to feed or water was 15 out of 1000 broilers; with lactose added to feed or water it was 10 out of 1000 broilers; with experimental chlorate product (ECP) added to feed or water it was 21 out of 1000. For ECP the concentration of Salmonella in the ceca was decreased by 0.61log(10)cfu/g in the treated group compared to the control group. Significant heterogeneity (Cochran's Q-statistic p≤0.10) was observed among studies examining all organic acids (controlled or challenge experiments), butyric acid, formic acid, a formic/propionic acid mixture, fermented liquid feed, and D-mannose. Meta-regressions were conducted to examine the source of heterogeneity among studies. For prevalence outcomes, 36% and 60% of the total variance was within and between studies, respectively. For concentration outcomes, 39% and 33% of the total variance was within and between studies, respectively. Inadequate blinding and randomization was common, and no studies undergoing meta-analysis or meta-regression were conducted on a commercial farm. The strength of evidence of the effect of these additives was very low. Studies conducted under commercial conditions are needed to understand the potential benefit of these interventions for the poultry industry and to improve the strength of evidence of the effectiveness of these additives.     

Mass spectrometric dereplication of nitrogen-containing constituents of black cohosh (Cimicifuga racemosa L.)

Black cohosh preparations are popular dietary supplements among women seeking alternative treatments for menopausal complaints. For decades, triterpene glycosides and phenolic acids have dominated the phytochemical and biomedical research on this plant. In this study, we provide evidence that black cohosh contains an unexpected and highly diverse group of secondary nitrogenous metabolites previously unknown to exist in this plant. Using a dereplication approach that combines accurate mass measurements, database searches and general knowledge of biosynthetic pathways of natural products, we identified or tentatively identified 73 nitrogen-containing metabolites, many of which are new natural products. The identified compounds belong to several structural groups including alkaloids, amides or esters of hydroxycinnamic acids and betains. Among the alkaloids, several classes such as guanidino alkaloids, isoquinolines and β-carbolines were identified. Fragmentation patterns for major compound classes are discussed, which provides a framework for the discovery of these compounds from other sources. Identification of alkaloids as a well-known group of bioactive natural products represents an important advance in better understanding of the pharmacological profile of black cohosh.     

Effects of two infectious bursal disease vaccine virus strains on hepatic microsomal enzyme activities in chickens

The influence of two infectious bursal disease vaccines on the activities of hepatic microsomal enzymes aniline hydroxylase, ethylmorphine N-demethylase, NADPH-cytochrome c reductase, aryl sulphotransferase and p-nitrophenol UDP-glucuronyltransferase was investigated in chickens. The vaccines contained attenuated Winterfield 2512 and VMG-91 strains, respectively. The activities of enzymes were determined on postvaccination days 0, 2, 5 and 7. At the same time, post-mitochondrial supernatant, cytosolic and microsomal pellet protein concentrations were determined. As expected, the antibody titres against infectious bursal disease virus in the serum were increased in both tested groups in relation to each administered vaccine. Using RT-PCR, the presence of the VP2 gene fragment of virus in the liver of chicken was demonstrated 4 and 6 h after vaccination. The results of this study suggest that the two commercial vaccines modulate the activities of five enzymes tested, and that the two attenuated vaccines applied triggered induction and/or inhibition of phases I and II of biotransformation enzyme activities.     

Effect of fasting and refeeding on blood glutathione and lipid peroxide concentration of cockerels and pullets

Changes of reduced glutathione (GSH) and TBARS (thiobarbituric acid-reactive products of lipid peroxidation) concentrations and activity of gamma-glutamyltransferase (GGT, EC 2.3.2.2) in the blood of Lohman brown cockerels and pullets in response to 48 hour food deprivation and 24 hour refeeding were examined. The experiment was performed on 61-day-old chickens. Blood samples ware collected from the wing vein (v. brachialis) in heparinized tubes for three times: control sampling before fasting, then after 48 hour food deprivation and after refeeding for 24 hours. Blood GSH concentration after refeeding in cockerels was significantly higher compared with prefasting and fasting values. The concentration of GSH in female chickens was significantly lower after fasting as well as after refeeding compared with control values. In addition to that, in pullets GSH concentration in refeeding was higher than in fasting conditions. The level of TBARS in blood in female and male chickens after fasting and refeeding were significantly lower than the prefasting values. The GGT activity on cockerels after 48 hour food deprivation was significantly higher compared with control sampling and in chickens refeed for 24 hours, whereas in pullets significant difference was exhibit compared only with control values. Concentration of GSH in control sampling in cockerels compared with those in pullets was significantly lower. After 48 hours of fasting, the level of GSH was significantly higher in the cockerels than in the pullets. Results of TBARS concentration in the pullets were higher of control and fasting values than in the cockerels. The GGT activity of control sampling was significantly higher in male chicken. Lipid peroxidation in chickens of both sexes decreased with fasting, but prooxidative-antioxidative processes were more intensive in female chickens, probably because they were not reach sexual maturity.     

Influence of dietary enzyme complex on the performance of broilers fed on diets with and without antibiotic supplementation

1. The aim of the experiment was to test the possible interactions of an enzyme complex and a food antibiotic on the growth and metabolism, carcase yield, whole body composition and nutrient deposition in broilers. The basal diet contained 400 g/kg barley. The four treatments were as follows: O—without supplements (control), E—enzyme complex Roxazyme® G, 200 mg/kg, A—antibiotic avoparcin, 10 mg/kg, EA—Roxazyme G, 200 mg/kg plus avoparcin, 10 mg/kg.

2. Roxazyme G positively influenced weight gain, food conversion efficiency, energy metabolisability, fat and nitrogen utilisation and the dry matter content of droppings. Fat and energy deposition in the whole body were also increased, whereas protein deposition and carcase yield were not influenced.

3. Avoparcin increased energy metabolisability and fat utilisation, but had no influence on nitrogen utilisation. No significant improvements from avoparcin were seen in growth or in nutrient deposition in the body. The fibre degradability (NDF and ADF fraction) was significantly depressed by antibiotic supplementation.

4. The inclusion of both supplements to the diet did not have a fully additive effect on growth, energy metabolisability, or fat and nitrogen utilisation. The interaction between enzyme and antibiotic for food conversion efficiency during the first experimental period (7 to 21 d) was nearly significant (P= 0.053). Except for fibre degradability (P ≤ 0.01), no other significant interactions between enzyme and antibiotic were found.     

Aphid parasitoids (Hymenoptera: Braconidae: Aphidiinae) on citrus: Seasonal abundance, association with the species of host plant, and sampling indices

Samples were collected from southern Greece during 1996–2000 in order to investigate the presence of parasitoids onAphis gossypii Glover infesting several citrus species. The species of aphidiines found to have a significantly different preference forA. gossypii wereAphidius colemani Viereck,Aphidius matricariae Haliday,Diaeretiella rapae (M’Intosh),Ephedrus persicae Froggat,Lysiphlebus testaceipes (Cresson),Binodoxys acalephae (Marshall) andBinodoxys angelicae (Haliday). In another sampling experiment, the relative abundance of aphidiine parasitoids on aphids infesting orange and tangerine trees was studied in southern Greece (Nea Kios) in 1996 and 1997.A. gossypii constituted the largest part of the aphid population and was the only species parasitized.B. angelicae andA. colemani were the most abundant parasitoid species. The parasitization rate differed among the parasitoid species.B. angelicae had the highest colonization rate in centrally located and large host (A. gossypii) groups, whereasA. colemani was found in more isolated and relatively small host groups. The percentage of parasitism byB. angelicae was high mainly in large host groups, whenB. angelicae was the only parasitoid present. However, in cases of coexistence ofB. angelicae withA. colemani with hyperparasitoids, in the same sampling unit, the percentage of parasitism was relatively low. http://www.phytoparasitica.org     

Effect on neonatal oestrogen and hCG treatment on the genital organs of adult female rats

The ovaries and the uterine as well as vaginal mucous membranes of 80-, 180- and 365-day-old intact female rats and females neonatally treated with a single dose of oestrogen and repeated doses of human chorionic gonadotropin (hCG) were studied. Numerous follicles, interstitial cells and corpora lutea (CL) were present on the ovary of intact females from 80 up to 365 days of age. The number of primary and secondary follicles decreased in the intact female rats between Day 180 and Day 365 of life. On the ovaries of 180- and 365-day-old female rats neonatally treated with oestrogen, interstitial glandular cells and cystic follicles predominated. No CL were present on these ovaries. The height of epithelial cells of the uterine and vaginal mucous membranes increased in intact female rats from 80 to 365 days of age, whereas in oestrogen-treated females the height of epithelium decreased. From 80 up to 365 days of age, the height of epithelial cells of uterine and vaginal mucous membranes of rats neonatally treated with repeated doses of hCG was similar to that in the corresponding control animals.