The NS3 proteins of global strains of bluetongue virus evolve into regional topotypes through negative (purifying) selection

Comparison of the deduced amino acid sequences of the genes (S10) encoding the NS3 protein of 137 strains of bluetongue virus (BTV) from Africa, the Americas, Asia, Australia and the Mediterranean Basin showed limited variation. Common to all NS3 sequences were potential glycosylation sites at amino acid residues 63 and 150 and a cysteine at residue 137, whereas a cysteine at residue 181 was not conserved. The PPXY and PS/TAP late-domain motifs were conserved in all but three of the viruses. Phylogenetic analyses of these same sequences yielded two principal clades that grouped the viruses irrespective of their serotype or year of isolation (1900-2003). All viruses from Asia and Australia were grouped in one clade, whereas those from the other regions were present in both clades. Each clade segregated into distinct subclades that included viruses from single or multiple regions, and the S10 genes of some field viruses were identical to those of live-attenuated BTV vaccines. There was no evidence of positive selection on the S10 gene as assessed by reconstruction of ancestral codon states on the phylogeny, rather the functional constraints of the NS3 protein are expressed through substantial negative (purifying) selection.     

Identification and validation of housekeeping genes as internal control for gene expression in an intravenous LPS inflammation model in chickens

Real-time PCR has become a powerful tool for the detection of inflammatory parameters, including cytokines. Reference or housekeeping genes are used for the normalization of real-time RT-PCR results. In order to obtain reliable results, the stability of these housekeeping genes needs to be determined. In this study the stability of five genes, including beta-actin, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), hypoxanthine phophoribosyl-transferase (HPRT), ubiquitin (UB) and glucose-6-phosphate dehydrogenase (G6PDH), was determined in a lipopolysaccharide inflammation model in chickens. beta-Actin appeared to be the most stable single gene in our model. Because the use of a single gene for normalization can lead to relatively large errors, the use of the geometric mean of multiple reference genes or normalization factor is preferred. The most stable combination for gene expression analysis in this lipopolysaccharide inflammation model in chickens is G6PDH and UB, since their correlation coefficients were 0.953 and 0.969, respectively (BestKeeper) and an M value of 0.34 and a low V(2/3) value of 0.155 (geNorm) were obtained. The use of HPRT and GAPDH should be avoided. The stable housekeeping genes, G6PDH and UB together, can be used to normalize the expression of pro-inflammatory cytokines in a lipopolysaccharide inflammation model in chickens.     

Effects of α-lipoic acid, vitamins E and C upon the heat stress in Japanese quails

Antioxidant vitamins are commonly used as colorant, preventive, sweetener, nutritive and antioxidant materials in food industry. Thus, many studies have been conducted in recent years. We have performed the present study to understand the effects of α-lipoic acid (ALA), vitamin C and E, added into rations of Japanese quails, on chronic heat stress. This study was carried out in five groups as: control, stress, ALA, vitamin E and vitamin C groups. Heat stress is applied to all groups except the control group. Superficial pectoral muscles tissues were used for biochemical determinations. During the research, it is found that heat stress exerted undesired effects such as increasing lipid peroxidation (LPO) level indicating oxidative stress in Japanese quails. In addition, superoxide dismutase (SOD) and catalase (CAT) enzyme activities and glutathione (GSH) amounts were studied to understand the behaviour of defence mechanism. While stress increased LPO ratio, it was determined that all antioxidant added into the ration decreases LPO significantly (p < 0.05). In addition, it was determined that all of antioxidants added into ration effectively restored SOD activity and LPO ratio which heat stress affected negatively. Interestingly, vitamin C did not adjust GSH ratio in contrast to vitamin E and ALA, where vitamin E and C did not perform any positive effect on heat stress decreased CAT activity. Finally, it can be thought that antioxidant vitamins relatively ameliorated these undesired affects caused by stress factors given.     

First genetic characterization of equine adenovirus type 1 (EAdV-1) in Turkey

Equine adenovirus type 1 (EAdV-1) is a cause of repiratory tract infection in equids. In present study for the first time in Turkey, the prevalence of EAdV-1 in nasal swab samples obtained from horses showing respiratory symptoms was investigated by polymerase chain reaction (PCR), and molecular characterization of the hexon gene detected in the Turkish (TR) strain was performed. Overall, the prevalence of EAdV-1 was found low (1.4%) as indicated by a positive PCR reaction from the nasal swab extracts tested. Phylogenetic analysis based on the partial sequences of the hexon gene of a TR-EAdV-1 strain with those of previously isolated AdVs from different mammals and an EAdV-1 M1 strain showed that the EAdV-1 strains were placed into a unique cluster. Although the TR-EAdV-1 strain was closely related to CAV-1, CAV-2 and bat adenovirus reference strains, larger-scale studies are necessary to better understand the molecular epidemiology and population structure of EAdV-1 in Turkey.     

Species of Staphylococcus isolated from animal infections

One hundred randomly selected clinical strains of staphylococci were identified by species using a commercial micromethod system. Eight species of staphylococci were identified. Staphylococcus intermedius was the most frequent (n = 74) species identified and accounted for 70/74 (94.6%) of the coagulase-positive strains and 70/79 (88.6%) of the total isolates from dogs. Other species identified, in order of their frequency, included S. epidermidis (8), S. aureus (7), S. simulans (4), S. sciuri (2), S. xylosus (2), S. hyicus (2) and S. saprophyticus (1). These results show that at least 8 different species of staphylococci can be recovered from animal infections and that coagulase-positive species such as S. intermedius may be more common than S. aureus. The relative significance of these other species in animal infections needs to be assessed.     

The detection of Salmonella typhimurium varatio copenhagen DT 2 in purebred pigeons

On the occasion of a large exhibition of pure-breed fancy pigeons 398 animals from 49 different dovecotes were examined for Salmonella shedding. Faecal samples were taken after caging of the birds for the exhibition and after 3 days, before the end of the exhibition. Salmonella were detected in faeces of 28 out of 398 pigeons (7.04%). 10 birds were Salmonella positive only after caging for the exhibition, 10 other animals only before the end of the exhibition, and 8 pigeons at both occasions. The Salmonella positive birds originated from 15 different dovecotes, i.e. in ca. 30% of the dovecotes at the exhibition at least 1 Salmonella positive pigeon was identified. The share of positive birds in these dovecotes varied between 5% and 83%. All Salmonella isolates belonged to the serovar Typhimurium variant copenhagen and were of phage type DT 2. The results of this study do not provide complete evidence on the spreading of Salmonella organisms from birds infected at time of caging to other pigeons during the exhibition, however, such transmission cannot be excluded. In only 18 dovecotes pigeons were immunised against Salmonella Typhimurium. However, in these dovecotes all breeder birds but only 13% of the young pigeons had been immunised. Among the vaccinated breeder pigeons the number of Salmonella positive birds was considerably lower (not significant) than among the non-vaccinated breeders. There is epidemiological evidence that vaccination of pigeons has a considerable protective effect against Salmonella exposure. However, in order to effectively reduce Salmonella findings in pure-breed fancy pigeons it is recommended to provide vaccination to pigeons in a greater number of dovecotes and to include the progeny, too.     

Effects of dietary boron and phytase supplementation on growth performance and mineral profile of broiler chickens fed on diets adequate or deficient in calcium and phosphorus

1. Two experiments were designed to determine the effect of dietary boron (B) in broiler chickens. In Experiment 1, a 2 × 4 factorial arrangement of treatments was used to investigate the effect of dietary calcium (Ca) and available phosphorus (aP) (adequate or deficient) and supplemental B (0, 20, 40, and 60 mg/kg diet). In Experiment 2, B, at 20 mg/kg, and phytase (PHY) (500 FTU/kg diet) were incorporated into a basal diet deficient in Ca and aP, either alone or in combination.

2. The parameters that were measured were growth performance indices, serum biochemical activity as well as ash and mineral (i.e. Ca, P, Mg, Fe, Cu and Zn) content of tibia, breast muscle and liver.

3. Results indicated that both supplemental B and dietary Ca and aP had marginal effects on performance indices of chickens grown for 42 d.

4. There were positive correlations (linear effect) between B concentrations of serum, bone, breast muscle and liver and the amount of B consumed.

5. Serum T₃ and T₄ activities increased linearly with higher B supplementation.

6. Increasing supplemental B had significant implications on breast muscle and liver mineral composition. Lowering dietary Ca and aP level increased Cu content in liver and both Fe and Zn retention in breast muscle. Tibia ash content and mineral composition did not respond to dietary modifications with either Ca–aP or B.

7. The results also suggested that dietary contents of Ca and aP do not affect the response to B regarding tissue mineral profile. Dietary combination with B and PHY did not create a synergism with regard to growth performance and bioavailability of the minerals.