Mustafa Necati Muz

Antiviral Activity of Hatay Propolis Against Replication of Herpes Simplex Virus Type 1 and Type 2

Background Propolis is a bee product widely used in folk medicine and possessing many pharmacological properties. In this study we aimed to investigate: i) the antiviral activities of Hatay propolis samples against HSV-1 and HSV-2 in HEp-2 cell line, and ii) the presence of the synergistic effects of propolis with acyclovir against these viruses. Material/Methods All experiments were carried out in HEp-2 cell cultures. Proliferation assays were performed in 24-well flat bottom microplates. We inoculated 1×105 cells per ml and RPMI 1640 medium with 10% fetal calf serum into each well. Studies to determine cytotoxic effect were performed. To investigate the presence of antiviral activity of propolis samples, different concentrations of propolis (3200, 1600, 800, 400, 200, 100, 75, 50, and 25 μg/mL) were added into the culture medium. The amplifications of HSV-1 and HSV-2 DNA were performed by real-time PCR method. Acyclovir (Sigma, USA) was chosen as a positive control. Cell morphology was evaluated by scanning electron microscopy (SEM). Results The replication of HSV-1 and HSV-2 was significantly suppressed in the presence of 25, 50, and 100 μg/mL of Hatay propolis. We found that propolis began to inhibit HSV-1 replication after 24 h of incubation and propolis activity against HSV-2 was found to start at 48 h following incubation. The activity of propolis against both HSV-1 and HSV-2 was confirmed by a significant decrease in the number of viral copies. Conclusions We determined that Hatay propolis samples have important antiviral effects compared with acyclovir. In particular, the synergy produced by antiviral activity of propolis and acyclovir combined had a stronger effect against HSV-1 and HSV-2 than acyclovir alone.

Molecular detection of Nosema ceranae and Nosema apis infections in Turkish apiaries with collapsed colonies

Extensive losses of honey bees (Apis mellifera L.) have been reported from many countries worldwide in Europe, the USA, and the Middle East, with losses ranging from 1.8% – 85% (Neumann and Carreck, 2010). Similarly, beekeepers in Turkey have experienced extensive colony losses (Giray et al., 2010; Muz, 2008). From the early spring of 2007 to the autumn of 2009, 85 colonies were examined from 21 apiaries in the Hatay wintering region of Turkey where extensive colony losses had occurred, together with 11 samples from the southern Marmara region. All samples that proved positive for Nosema spp. by light microscopy were classified according to their region of origin. Extraction of DNA was then performed as described by Williams et al. (2008) and PCR conditions with specific primers were used as described by Chen et al. (2008). Hungarian Nosema samples were used as a positive control, and water was used as a negative control. The results showed that N. ceranae was the prevalent microsporidian infection compared with N. apis in collapsed colonies of Turkey. Colonies from the Hatay wintering region were found to show infections of 89% N. cerana and 11% N. apis, whilst samples from the southeastern Marmara region were found to show infections of 84% N. cerana and 16% N. apis. Previously, Nosema infections have been reported, this is the first instance where differential molecular diagnosis has been carried out in apiaries showing colony collapse in Turkey. The detection of both Nosema infections has demonstrated the importance of contagious infections in wintering regions as well as those used for honey production. Significantly, the Hatay wintering region borders Syria, so intensive surveys are urgently needed to determine the distribution and prevalence of Nosema species throughout the whole of Turkey. Acknowledgements We are very grateful to Prof. Dr Nowotny and Dr. Bakonyi for help in providing the positive isolates of Nosema spp. from Szent Istvan University, Hungary.

Survey of the occurrence of Deformed Wing Virus and multiple parasites of queens (Apis mellifera L.) in apiaries with collapsed colonies in Hatay, Turkey

Summary Honey bee queens in Hatay, Turkey, were investigated to reveal the status of Deformed Wing Virus (DWV) and parasites. DWV, Nosema sp., Malpighamoeba mellificae, and Varroa destructor were detected among samples. This is the first record of DWV in honey bee queens with multiple parasitic infections in apiaries with collapsed colonies in Turkey.

A molecular epidemiological study of black queen cell virus in honeybees ( Apis mellifera ) of Turkey: the first genetic characterization and phylogenetic analysis of field viruses

Black queen cell virus (BQCV) is one of the most common honeybee pathogens causing queen brood deaths. The 63 apiaries were sampled between 2007 and 2013 from four different ecogeographic regions in Turkey to estimate BQCV molecular structural characteristics. The BQCV positivity was 47.6%. The 25 local Black queen cell viruses (TrBQCVs) were molecularly characterized and investigated for their genetic relationship with previous records. The identity of the helicase gene among the TrBQCVs was 92–98%, whereas the similarity ranged from 37 to 85% in comparison with the intercontinental records. The identity of the partial capsid gene among the TrBQCVs was 91–100%, and the similarity rate varied from 86 to 97, 88–96, 90–97 and 89–99% in comparison with the Asian, African, American and European counterparts, respectively. The four nonsynonymous substitutions on the partial capsid protein suggest a predicted genotype that is specific among TrBQCVs.