Hakan Bulut

A Large-Scale Outbreak of Bovine Ephemeral Fever in Turkey, 2012

ABSTRACT Regional cases of bovine ephemeral fever (BEF) were documented previously in Turkey. Previous cases were confirmed in South-East Turkey with low cow mortality. Recent BEF-suspected outbreaks with high mortality were documented in many regions of Turkey in 2012. The aim of study was the epidemiological examination of the outbreak and molecular characterization of the viruses detected from the outbreak. For this reason, blood samples were collected from BEF-suspected outbreak regions. From the results of RT-PCR, high rate of BEF-suspected samples (48/60 or 80%) was found positive for BEF virus (BEFV) RNA. The nucleotide sequences of the G1 region of G gene of BEFV in the current study during the 2012 outbreak were grouped into cluster II of BEFV. It was suggested that BEFV may be spread out to other neighbor countries in the future years.

Detection of avian metapneumovirus subtypes in turkeys using RT-PCR.

This study investigated the prevalence of avian metapneumovirus (aMPV) and the detection of molecular subtypes of field strains of the virus using RT-PCR in clinically healthy turkeys and those showing signs of respiratory disease. In the RT-PCR examination of 624 tracheal tissue samples collected from a local turkey abattoir, 2.9 per cent (18/624) of samples tested positive. In the examination of tracheal swab samples collected from flocks with respiratory problems, 18 of 20 samples tested positive. When the results were assessed at flock level, aMPV infection was detected in only one of the 23 clinically healthy turkey flocks, whereas all four flocks with respiratory problems were infected. Molecular typing using primers specific to the attachment glycoprotein (G) gene showed that all 36 positive samples belonged to subtype B. Partial sequence analysis of DNA samples showed 95 per cent homology between the field types and the reference strain aMPV subtype B. Whereas clinically healthy turkeys had been vaccinated with a subtype A virus vaccine, the flocks with respiratory problems had been vaccinated with a subtype B virus vaccine. Despite four blind passages of RT-PCRpositive samples on Vero and chicken embryo fibroblast cells, no cytopathic effect was detected by microscopic examination.

Effects of lapatinib and trastuzumab on vascular endothelial growth factor in experimental corneal neovascularization.

The aim of this study was to compare the effects of lapatinib and trastuzumab on vascular endothelial growth factor on experimental corneal neovascularization.

Use of cauda epididymis extract as immunocontraceptive

The objective of this study was to evaluate use of a cauda epididymis extract (CEE) as an immunocontraceptive in rats. Twenty-two rats in puberty or 19 rats that gave birth once (primipara) were immunized with intraperitoneal (IP) injection of CEE. Rats in puberty received one or two injections of CEE containing 1.5 x 10(9) sperm/mL, while primipara rats received injections of CEE containing 3 x 10(6) sperm/mL up to three times. Animals were tested for the presence and concentration of anti-CEE antibody by enzyme-linked immunosorbent assay (ELISA) and monitored for pregnancy following natural insemination. Results revealed that 38 (92.6%) of the 41 rats were positive for anti-CEE antibodies, regardless of animal type or immunization procedure. However, there was no relation between pregnancy rates and concentration of anti-CEE antibody in rats immunized with CEE. These results indicate that immune response against CEE may not play a major role in contraception in rats.

Prevalence of Rota-and Reoviruses in Turkey Enteritis in Turkey

The objective of this study was to compare the presence of rotavirus (TRotV) and reovirus (TReoV) in clinically healthy turkey flocks and in those with poult enteritis complex (PEC) using reverse transcription-polymerase chain reaction (RT-PCR). TRotV and TReoV were detected in 2.6% (6/230) of the birds each and in 26.08% (6/23) and 13.04% (3/23) of the flocks, respectively. Mixed infection with both agents was found in one sample. None of these two viruses were detected in turkeys originating from clinically healthy flocks.

Outbreak of clinical infectious laryngotracheitis in Turkey

FIG 1: Agarose gel electrophoresis of PCR and digestion products. Lane 1 Positive control (A96 strain of infectious laryngotracheitis virus [ILTV]), Lane 2 Tracheal sample showing the presence of a 588 base pair (bp) fragment by direct PCR, Lane 3 Negative control (distilled water added to PCR reaction as a template), Lane 4 Negative tissue sample, Lane 5 BamHI cut PCR product amplified from the A96 strain of ILTV, Lane 6 BamHI cut PCR product amplified from tracheal sample (fragments approximately 149 and 439 bp size), Lane M 100 bp DNA ladder (Promega) M 1 2 3 4 5 6

The effects of Kisspeptin antibodies on delayed estrus in rats

In recent years, numerous studies have been conducted on the effects of Kisspeptin/GPR54 system on sexual cycle, which proved that this system regulated gonadotropin release through GnRH. This study aims to determine the effects of hyperimmune serum containing antibodies produced against Kisspeptin on the sexual cycle and GnRH receptors in rat pituitaries. To this end, five Wistar female rats were passively immunised using a hyperimmune serum obtained from two Wistar female rats against Kisspeptin 10. Another five rats were selected as the control group. Anti-Kisspeptin antibodies of the hyperimmunised rats in the serum were identified by ELISA. The sexual cycles of the rats were followed by the measurements of vaginal irrigations and the estradiol concentrations in their blood samples. The number of follicles and corpora lutea in their ovaries was determined through histopathological tests, and the GnRH receptors in their pituitaries were identified by immunohistochemistry. Consequently, strong seropositivity was detected in all passively immunised rats and the hyperimmune serum. However, no difference was found between the groups with regard to the number of estrous cycles observed, the interval between estrous periods, estradiol concentrations, the number of follicles and corpora lutea and immunohistochemical results.

Complete genome analysis of highly pathogenic bovine ephemeral fever virus isolated in Turkey in 2012

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.