Hakan Bulut

Background: Eggshell microbial contamination of fertile hatching eggs represents a major biosecurity concern in commercial poultry hatcheries because it can compromise embryonic viability, hatchability and chick quality. In commercial hatchery practice, broiler breeder fertile eggs are often stored before incubation to synchronize production schedules. However, prolonged storage may increase microbial contamination of the eggshell, deteriorate internal egg quality and reduce embryonic viability and hatchability. Objectives: This study evaluated thyme essential oil and hyaluronic acid (HA) as natural alternatives to chemical disinfectants for sanitation of broiler breeder fertile eggs. Methods: A total of 1800 Ross 308 hatching eggs were randomly assigned to four main treatments: untreated control, thyme essential oil (1%) emulsified with polysorbate‐20 (PS20), HA (0.1%) and thyme essential oil + HA. Eggs were stored for 0 or 14 days in a 4 × 2 factorial design. A PS20‐only vehicle control was included for selected comparisons. Results: All sanitization treatments significantly reduced eggshell total aerobic bacterial counts compared with the untreated control ( p < 0.001). Storage for 14 days impaired internal egg quality, reducing albumen ratio, albumen index, yolk index and Haugh unit values, while increasing yolk ratio and albumen pH ( p < 0.001). Storage also reduced blastoderm viable cell number and increased dead cell number ( p < 0.001). Thyme essential oil produced the most favourable outcomes after storage, with higher hatchability of set eggs and hatchability of fertile eggs and lower embryonic mortality than HA‐treated eggs ( p < 0.05). Yolk malondialdehyde (MDA) levels were lower in the thyme group than in the control and HA groups after storage ( p < 0.01). Although HA reduced microbial load, it was associated with increased blastoderm dead cell counts and higher embryonic mortality after storage. Conclusions: Thyme essential oil appears to be a promising residue‐free and biologically safe alternative to conventional chemical disinfectants for hatchery sanitation of broiler breeder fertile eggs.

Bovine ephemeral fever (BEF) is an arthropod-borne viral disease characterised by a short-term clinical expression that can lead to significant losses in high-yielding cattle and water buffaloes. In this study, we aimed to generate a recombinant plasmid expressing the glycoprotein (G) of the BEF virus (BEFV) and to stimulate a humoral immune response to this protein in BALB / c mice immunised with the recombinant plasmid. Expression of the encoded protein was demonstrated by western blotting and immunoperoxidase tests. The suitable plasmids were intramuscularly administered to BALB/c mice on days 0, 14 and 21. The antibody response in the immunised mice was measured by a plaque reduction neutralization test (PRNT) and enzyme-linked immunosorbent assay (ELISA). According to BEFV ELISA, only two of the seven animals in these groups exceeded the cut-off value. A significant difference was observed in the mean OD values at 450 nm absorbance in the pcDNA4-G-immunised group when compared with those in the plasmid control group at 30 days (p < 0.05). According to PRNT50 results, a 1:20 (p < 0.05) antibody response was obtained at 30 days in pcDNA4-G (100 mu g)-immunised mice, whereas this ratio was 1:80 (p < 0.001) in BEFV-immunised mice (1,000 PFU/0.5 ml). We conclude that the humoral immune response was stimulated in experimental mice immunised with the recombinant plasmid. However, disappointingly, the antibody response was markedly low in pcDNA4-G-immunised mice.

Background: Extrarenal 1α,25-dihydroxyvitamin D3 (1,25-D) locally produced by immune cells plays crucial roles on the regulation of the immune system. However, in vivo status of extrarenal 1,25-D and 25- hydroxyvitamin D (25-D) in acute inflammatory conditions are unknown. Objective: The aim of this study was to determine extrarenal 1,25-D level in circulation in bilaterally nephrectomized rats induced by low-dose lipopolysaccharide (LPS). Methods: Renal 1,25-D synthesis was terminated through bilateral nephrectomy in rats. The rats received intraperitoneal LPS (50 µg/kg BW) three times and the experiment was ended 24 hours after nephrectomy. Serum 1,25-D, 25-D, calcium, phosphorus, intact parathyroid hormone and calcitonin levels were measured and immunohistochemistry were applied to detect the sources of extrarenal 1,25-D synthesis. Results: Circulatory 1,25-D concentration remarkably increased in the both LPS-treated and non-treated bilaterally nephrectomized rats. Elevated circulatory 1,25-D didn’t have hypercalcemic endocrinal effects. Increased 1,25-D level also resulted in concurrent rapid and dramatic depletion of circulatory 25-D. Conclusions: In conclusion, extrarenal 1,25-D could enter into the systemic circulation and therefore, might have systemic effects besides its autocrin and paracrine functions.

Relatively high prevalence and mortality rates of bovine ephemeral fever (BEF) have been reported in recent epidemics in some countries, including Turkey, when compared with previous outbreaks. A limited number of complete genome sequences of BEF virus (BEFV) are available in the GenBank Database. In this study, the complete genome of highly pathogenic BEFV isolated during an outbreak in Turkey in 2012 was analyzed for genetic characterization. The complete genome of the Turkish BEFV isolate was amplified by reverse transcription-polymerase chain reaction (RT-PCR) and sequenced. It was found that the complete genome of the Turkish BEFV isolate was 14,901 nt in length. The complete genome sequence obtained from the study showed 91-92% identity at nucleotide level to Australian (BB7721) and Chinese (Bovine/China/Henan1/2012) BEFV isolates. Phylogenetic analysis of the glycoprotein gene of the Turkish BEFV isolate also showed that Turkish isolates were closely related to Israeli isolates. Because of the limited number of complete BEFV genome sequences, the results from this study will be useful for understanding the global molecular epidemiology and geodynamics of BEF.

Background: The aim of this study was to compare the effects of lapatinib and trastuzumab on vascular endothelial growth factor on experimental corneal neovascularization.Methods: A total of 35 Wistar albino rats were randomly divided into five groups, each containing seven animals. Corneas of rats in the control group were not cauterized and did not receive any treatment. A silver nitrate pencil was applied on the right corneas of rats in the non‐control groups to induce corneal neovascularization. Rats in the sham, lapatinib, trastuzumab and lapatinib + trastuzumab groups were administered systemic saline, 50 mg/kg lapatinib once a day orally by gavage, 4 mg/kg trastuzumab once a day intraperitoneally, or 50 mg/kg lapatinib once a day orally by gavage together with 4 mg/kg trastuzumab once a day intraperitoneally, respectively, for 7 days. Rats were sacrificed on the eighth day, and corneas were excised using a 4‐mm punch trephine. Vascular endothelial growth factor immunostaining in the corneal epithelial and stromal layers was evaluated. Staining intensities were determined semi‐quantitatively, and corneal vascular endothelial growth factor levels were measured by enzyme‐linked immunosorbent assay.Results: The mean immunostaining intensities of epithelial and stromal vascular endothelial growth factor in the control group were significantly lesser than those in the sham group (P < 0.05). The mean immunostaining intensities of epithelial and stromal vascular endothelial growth factor and vascular endothelial growth factor enzyme‐linked immunosorbent assay levels of corneas in all treatment groups (lapatinib, trastuzumab and lapatinib + trastuzumab groups) were lesser than those in the sham group (P < 0.05); however, it was similar to those in the control group (P > 0.05).Conclusions: It is suggested that systemically administered lapatinib is more effective than systemically administered trastuzumab in preventing corneal neovascularization.

This, partly retrospective study, was designed to determine the seroprevalence of Schmallenberg virus (SBV), a new Orthobunyavirus first reported in Germany in late 2011, in domestic ruminants from the Middle Black Sea, West, and Southeast regions of Turkey. An indirect enzyme-linked immunosorbent assay was used to screen serum samples collected from slaughterhouse animals between 2006 and 2013. The overall seroprevalence was 335/1,362 (24.5 %) with 325/816 (39.8 %), 5/307 (1.6 %), 3/109 (2.8 %), and 2/130 (1.5 %) recorded in cattle, sheep, goats, and Anatolian water buffalo, respectively. This is the first study to demonstrate the presence of antibodies to SBV in Turkish ruminants; it indicates that cattle are more susceptible to infection than sheep, goats, or buffalo and that exposure of domestic ruminants to SBV in Turkey may have occurred up to 5 years prior to the first recorded outbreak of the disease in 2011.