Neonicotinoid Analysis in Sunflower (Helianthus annuus) Honey Samples Collected around Tekirdag in Turkey.

In recent years, the widespread use of neonicotinoids in agricultural areas has caused environmental pollution due to its lower toxicity to mammals. Honey bees, which are considered as biological indicators of environmental pollution, can carry these pollutants to the hives. Forager bees returning from sunflower crops that have been treated with neonicotinoids treated sunflower fields cause residue accumulation in the hives, which reason colony-level adverse effects. This study analyses neonicotinoid residues in sunflower (Helianthus annuus) honey sampled by beekeepers from Tekirdag province. Honey samples have been subjected to liquid-liquid extraction methods before liquid chromatography-mass spectrometry (LC-MS/MS). The method validation was carried out to fulfill all the necessary requirements of procedures SANCO/12571/2013. Accuracy was in the range of 93.63-108.56%, for recovery in the range of 63.04-103.19%, and for precision in the range 6.03-12.77%. Detection and quantification limits were determined according to the maximum residue limits of each analyte. No neonicotinoid residues were found above the maximum residue limit in the sunflower honey samples analysed.

The investigation of Culicoides (Diptera: Ceratopogonidae) species and Bluetongue virus and Schmallenberg virus in Northwest Türkiye.

Culicoides biting midges (Diptera: Ceratopogonidae) act as mechanical and biological vectors of arboviruses and are crucial in the global spread of these viruses. This study investigated the diversity of distribution of Culicoides species and the presence of Bluetongue virus (BTV) and Schmallenberg virus (SBV) in Tekirdag province in Northwest Türkiye. The fourteen Culicoides species, such as Culicoides newsteadi, Culicoides schultzei, Culicoides nubeculosus comp., Culicoides punctatus, Culicoides circumscriptus, Culicoides obsoletus comp., Culicoides gejgelensis, Culicoides festivipennis, Culicoides longipennis, Culicoides spp., Culicoides pulicaris, Culicoides picturatus, Culicoides odiatus, Culicoides kurensis, and Culicoides flavipulicaris, were detected. Culicoides newsteadi, C. odiatus, and C. pulicaris were the most abundant species. Phylogenetic analyses of Culicoides species' ITS-1 gene region were performed. A pool of C. festivipennis was positive for SBV RNA, while the BTV genomic materials was not found in the qPCR analysis. This is the first report of the presence/detection of SBV in Culicoides species in Türkiye. The survey of bioecological and epizootiological aspects of vector species is essential in implementing effective control measures for arboviral infections.

Presence of Neospora caninum DNA of Wild Birds from Turkey

Objective: Neospora caninum is a protozoon parasite that has a worldwide distribution and mainly causes abortion in cattle and current serological evidence shows that the disease may be also zoonotic. Wild birds play a role as a reservoir of the disease in nature. The study aimed to determine the prensence of N. caninum in wild birds. Methods: In this study, the presence of neosporosis in wild birds (n=55) including 22 different species found in the western side of Turkey, was investigated by polymerase chain reaction (PCR). In addition, PCR positive samples were confirmed by sequencing, BLAST, and phylogenetic analysis using MEGA7. Results: Obtained results showed that the presence of N. caninum DNA was 5.45% (3/55) in brain-heart homogenates wild birds. The bird species which were found positive for N. caninum were little owl (Athene noctua), common buzzard (Buteo buteo), and little tern (Sternula albifrons). According to phylogenetic analysis and BLAST, all samples were compatible with reference N. caninum isolates. Conclusion: To the best of authors' knowledge, this is the first study detecting N. caninum in little tern. In future studies, it may be interesting to investigate the prevalence of N. caninum in other wild animals to elucidate the transmission properties.

Protozoan and Microbial Pathogens of House Cats in the Province of Tekirdag in Western Turkey.

Domestic felines' re-emerging infectious and neglected zoonotic diseases are a significant focus of global "One Health" efforts. This study aimed to rapidly diagnose 14 pathogens, including zoonoses by using PCR primers in 167 client-owned symptomatic cats, routinely accepted to the Veterinary Clinics of Tekirdag. The prevalence of pathogens investigated were as follows: Babesia canis canis (24%), Babesia microti (2.4%), Hepatozoon felis (10.8%), Cytauxzoon felis (6.6%), Bartonella henselae (40.1%), Anaplasma platys (30.5%), Anaplasma phagocytophilum (7.2%), Rickettsia felis (26.3%), Borrelia burgdorferi (21%), and hemotropic Mycoplasma sp. (11.4%). There was a significant difference between the prevalence of the pathogens (χ2 = 152.26, df = 9, p < 0.001). There was also a statistical difference between the gender of the cats in terms of the prevalence of all pathogens considered together (χ2 = 4.80, df = 1, p = 0.028), where the female cats showed a higher prevalence. This was not the case for the different age groups (χ2 = 2.92, df = 1, p = 0.088). The lowest infection was observed for B. microti (p < 0.001), while the highest infection was observed for B. henselae (p < 0.01). Leishmania donovani, Plasmodium spp., Ehrlichia chaffeensis, and Neoehrlichia mikurensis PCR test results were negative in all samples. In conclusion, house cats of Tekirdag are apparently highly susceptible to some neglected zoonoses important for "One Health", and their prevalence in the region is most probably underestimated. Hence, applying PCR tests to assist fast clinic diagnosis in routine, may be an efficient option to protect the public as well as the cats from severe diseases.

The molecular and serological investigation of Feline immunodeficiency virus and Feline leukemia virus in stray cats of Western Turkey.

This study aimed to investigate the Feline immunodeficiency virus (FIV) / Feline leukemia virus (FeLV) infection prevalence among looking healthy stray cats in Western Turkey by serologic and molecular-based tests. A total of 1008 blood samples from the stray cats were used in this study. All samples were tested for FIV antibodies / proviral DNA and FeLV antibodies / antigens / proviral DNA. The genetic characterization and phylogenetic analysis of FeLV and FIV were carried out in this study. These cats also tested for Leishmaniasis and Toxoplasmosis previously. FIV Ab and proviral DNA detected in 25.2 % and 25.5 % of samples, respectively. FeLV Ab, Ag, proviral DNA positivity was in 45.2 %, in 3.3 %, in 69.7 %, respectively. The molecular detection and phylogenetic analysis of the current FeLV pol gene and FIV gag gene performed. The molecular characterization for the pol gene of FeLV (enFeLV and exFeLV) among Turkey's cat population was reported for the first time. The exFeLV pol sequences closer to the FeLV-A genotype, and the enFeLV pol sequences overlapped with other enFeLV. The current FIV gag sequences were clustered within the subtypes A, B, and C. The findings revealed FeLV subtype A and FIV subtype-A, subtype-B, subtype-C circulate among Turkish stray cats. Single and multiple co-infection positivity was found higher compared to previous reports.

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 AND METHODS All experiments were carried out in HEp-2 cell cultures. Proliferation assays were performed in 24-well flat bottom microplates. We inoculated 1x105 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.

Evaluation of cage designs and feeding regimes for honey bee (Hymenoptera: Apidae) laboratory experiments.

The aim of this study was to improve cage systems for maintaining adult honey bee (Apis mellifera L.) workers under in vitro laboratory conditions. To achieve this goal, we experimentally evaluated the impact of different cages, developed by scientists of the international research network COLOSS (Prevention of honey bee COlony LOSSes), on the physiology and survival of honey bees. We identified three cages that promoted good survival of honey bees. The bees from cages that exhibited greater survival had relatively lower titers of deformed wing virus, suggesting that deformed wing virus is a significant marker reflecting stress level and health status of the host. We also determined that a leak- and drip-proof feeder was an integral part of a cage system and a feeder modified from a 20-ml plastic syringe displayed the best result in providing steady food supply to bees. Finally, we also demonstrated that the addition of protein to the bees' diet could significantly increase the level ofvitellogenin gene expression and improve bees' survival. This international collaborative study represents a critical step toward improvement of cage designs and feeding regimes for honey bee laboratory experiments.

GC-MS analysis and antileishmanial activities of two Turkish propolis types.

Propolis is a honeybee product with a very complex chemical composition and various pharmacological properties. This study was aimed to investigate antileishmanial activities of "Bursa" and "Hatay" propolis samples against Leishmania infantum and Leishmania tropica strains. Propolis samples were analysed with the gas chromatography-mass spectrometry technique. Promastigotes were incubated in Roswell Park Memorial Institute culture medium in the absence and presence of several concentrations (50, 100, 250, 500, 750, and 1,000 µg/mL) of each propolis sample. The viability and cell morphology of promastigotes in each concentration were examined after 24, 48, 72, and 96 h of incubation. The growth of leishmania parasites was significantly suppressed in the presence of 500, 750, and 1,000 µg/mL of Hatay propolis. Bursa propolis was found to be efficient in inhibiting the growth of leishmania promastigotes in culture media at these concentrations, 250, 500, 750, and 1,000 µg/mL. Thus, the in vitro results showed that the Hatay and Bursa propolis samples decreased significantly the proliferation of L. infantum and L. tropica parasites (p < 0.001); however, Bursa propolis was found to be more effective than Hatay propolis against leishmania promastigotes. These two natural products may be useful agents in the prevention of leishmanial infections.

[Sudden die-off of honeybee colonies]. / Bal Arilarinda Ani Koloni Sönmesi.

Apis mellifera is used for honeybee keeping all over Turkey. Recently, honeybees have been suddenly disappearing for no apparent reason. It is presumed that some viral and parasitic honeybee pathogens are responsible for this. No medical research has been conducted to determine the pathologic causes of the sudden die-off of the honeybee colonies in Turkey as yet. This is of urgent importance for future of the honeybee industry.

[Treatment of Linognathus vituli and Bovicola bovis infestations of calves with cypermethrin]. / Danalarin Linognathus vituli ve Bovicola bovis enfestasyonlarinin Cypermethrin ile tedavisi.

This study has been conducted to determine the efficacy level of cypermethrin on the 14 calves that were naturally infested by Linognathus vituli and Bovicola bovis. A 10% solution of cypermethrin was diluted to 0.1% and applied to infested animals twice. All L. vituli were found to be dead after the fourth day of the first application and the second day of the second application. After the first day of application, all of the B. bovis were found dead at the end of each treatment. This study confirmed the protective efficacy of cypermethrin against L. vituli and B. bovis showing that infestation is prevented for a period of about 24-35 days following administration of cypermethrin at this dose.

[Investigation of helminth infections of cats and dogs in the Hatay province]. / Hatay ilinde bakisi yapilan kedi ve köpeklerde helmint enfeksiyonlari

In this study autopsies and fecal examinations of 8 cats and 6 dogs were performed. The helminth infection rate was founded to be 50.0% in cats and dogs after the fecal examinations. Trichuris sp. eggs were found in feces of one dog, and Ancylostoma sp. eggs in feces of one cat and one dog. Seven (87.5%) cats and 2 (33.4%) dogs were found to be infected with various helminths according to the results of the autopsies. The helminth species and their rates were Toxocara cati (62.5%), Joyeuxiella pasqualei (50.0%), Hydatigera taeniaformis (25.0%), Mesocestoides sp. (12.5%) and Dipylidium caninum (12.5%) in cats; T. canis (16.7%) and D. caninum (16.7%) in dogs during the autopsies. Numerous tetrathyridium larvae of the Mesocestoides sp. were found in the abdominal cavity of one cat. In conclusion, the high prevalence of ascarid infections in cats and dogs may be important for human health.