Gülay BAYRAKAL, Emek DÜMEN, Gözde EKİCİ, HAYRETTİN AKKAYA, Funda Hatice SEZGİN

Detection and Molecular Examination of Pathogens in Honey and Bees in the Northern Marmara Region, Turkey

Honey, which has many positive health effects, is fondly consumed in our country and in the world. Although honey is considered to be a micro-organism-free food because of its antimicrobial and bacteriostatic effects, studies refute this idea. In addition to primary contamination, personnel, tools and equipment used in beekeeping and honey production is a potential source of secondary contamination. In addition, honey, which can carry many microorganisms as a result of cross-contamination, is among the important foods and can threat public health. Therefore, it is thought that screening of pathogens that may be present in honey would contribute to the studies. Due to the geographical location, the diversity of climate and vegetation, Turkey is located in the upper row of honey producting countries. In this study, 900 samples examined in Kırklareli province in Northern Marmara Region. Kırklareli region has been chosen as the research area since it is considered as an important province in honey production and is a border province located in the industrial zone. The aim of the study was to investigate the presence of parasitological, bacterial, fungal and viral parameters which are important for the quality of the consumer, bee, larvae, colony and honey. According to the obtained data, positive results were found in many parameters and statistically significant results were obtained.

PDF

___

1. Cengiz MM, Erdoğan Y: Comparison of wintering ability and colony performances of different honeybee (Apis mellifera L.) genotypes in eastern Anatolian/Turkey conditions. Kafkas Univ Vet Fak Derg, 23 (6): 865- 870, 2017. DOI: 10.9775/kvfd.2017.17667

2. Yaman M, Yarılgaç EŞ, Güner BG, Ertürk Ö: Ordu yöresi bal arılarında Nosemosis hastalığının varlığı ve dağılımı. Turkiye Parazitol Derg, 39, 47- 51, 2015. DOI: 10.5152/tpd.2015.3783

3. White JW, Subers MH, Schepartz AI: The identification of inhibine, the antibacterial factor in honey, as hydrogen peroxide and its origin in a honey glucose-oxidase system. Biochim Biophys Acta, 73 (1): 57-70, 1963. DOI: 10.1016/0926-6569(63)90108-1

4. McLoone P, Warnock M, Fyfe L: Honey: A realistic antimicrobial for disorders of the skin. J Microbiol Immunol Infect, 49 (2): 161-167, 2016. DOI: 10.1016/j.jmii.2015.01.009

5. Roy S, Mandal M, Pal NK, Das MK, Halder D, Sircar B, Mandal S: Exploration of antibacterial and antioxidative property of two natural honey samples from Malda district, India. Transl Med (Sunnyvale), 6 (4): 1000187, 2016. DOI: 10.4172/2161-1025.1000187

6. Molan PC: The antibacterial activity of honey: 1. The nature of the anibacterial activity. Bee World, 73 (1): 5-28, 1992. DOI: 10.1080 0005772X.1992.11099109

7. Molan PC: The antibacterial activity of honey: 2. Variation in the potency of the antibacterial activity. Bee World, 73 (2): 59-76, 1992. DOI: 10.1080/0005772X.1992.11099118

8. Almasaudi SB, Al-Nahari AAM, Abd El-Ghany ESM, Barbour E, Al Muhayawi SM, Al-Jaouni S, Azhar E, Qari M, Qari YA, Harakeh S: Antimicrobial effect of different types of honey on Staphylococcus aureus. Saudi J Biol Sci, 24 (6): 1255-1261, 2017. DOI: 10.1016/j.sjbs.2016.08.007

9. Snowdon JA, Cliver DO: Microorganisms in honey. Int J Food Microbiol, 31 (1-3): 1-26, 1996. DOI: 10.1016/0168-1605(96)00970-1

10. Mead PS, Slutsker L, Dietz V, McCaig LF, Bresee JS, Shapiro C, Griffin PM, Tauxe RV: Food-related illness and death in the United States. Emerg Infect Dis, 5 (5): 607-625, 1999. DOI: 10.3201/eid0505.990502

11. Teixeira EW, Chen Y, Message D, Pettis J, Evans JD: Virus infections in Brazilian honey bees. J Invertebr Pathol, 99 (1): 117-119, 2008. DOI: 10.1016/j.jip.2008.03.014

12. Oğuz B, Karapinar Z, Dincer E, Değer MS: Molecular detection of Nosema spp. and black queen-cell virus in honeybees in Van Province, Turkey. Turk J Vet Anim Sci, 41 (2): 221-227, 2017. DOI: 10.3906/vet1604-92

13. Grabensteiner E, Bakonyi T, Ritter W, Pechhacker H, Nowotny N: Development of a multiplex RT-PCR for the simultaneous detection of three viruses of the honeybee (Apis mellifera L.): Acute bee paralysis virus, Black queen cell virus and Sacbrood virus. J Invertebr Pathol, 94 (3): 222- 225, 2007. DOI: 10.1016/j.jip.2006.11.006

14. Bogdanov S, Jurendic T, Sieber R, Gallmann P: Honey for nutrition and health: A review. J Am Coll Nutr, 27 (6): 677-689, 2008. DOI: 10.1080/ 07315724.2008.10719745

15. Maturin L, Peeler JT: Bacteriological analytical manual, Chapter 3: Aerobic Plate Count. U.S. Food and Drug Administration, Silver Spring, MD, 2001. https://www.fda.gov/food/laboratory-methods-food/bam-aerobicplate-count; Accessed date: 05.07.2019

16. Feng P, Weagant SD, Grant MA, Burkhardt W: Bacteriological Analytical Manual, Chapter 4: Enumeration of Escherichia coli and the Coliform Bacteria. U.S. Food and Drug Administration, Silver Spring, MD, 2002. https://www.fda.gov/food/laboratory-methods-food/bam-4-enumerationescherichia-coli-and-coliform-bacteria; Accessed: 05.07.2019

17. ISO 16649-2: E. G. Microbiology of food and animal feeding stuffsHorizontal method for the enumeration of beta‐glucuronidase‐positive Escherichia coli-Part 2: Colony‐count technique at 44 degrees C using 5‐ bromo‐4‐chloro‐3‐indolyl beta‐D‐glucuronide, 2001.

18. Lancette GA, Bennet RW: Staphylococcus aureus and Staphylococcal Enterotoxins. In, Downes FP, Ito K (Eds): Compendium of Methods for the Microbiological Examination of Foods. 4th ed., 387-403, American Public Health Association, Washington, 2001.

19. Solomon HM, Lilly T: Bacteriological Analytical Manual Chapter 17: Clostridium botulinum. U.S. Food and Drug Administration, Silver Spring, MD, 2001. https://www.fda.gov/food/laboratory-methods-food-safety/ bam-clostridium-botulinum; Accessed: 05.07.2019

20. James RR, Skinner JS: PCR diagnostic methods for Ascosphaera infections in bees. J Invertebr Pathol, 90 (2): 98-103, 2005. DOI: 10.1016/j. jip.2005.08.004

21. Latgé JP: Aspergillus fumigatus and aspergillosis. Clin Microbiol Rev, 12 (2): 310-350, 1999. DOI: 10.1128/CMR.12.2.310

22. Horn BW, Greene RL: Vegetative compatibility within populations of Aspergillus flavus, A. parasiticus, and A. tamarii from a peanut field. Mycologia, 87 (3): 324-332, 1995. DOI: 10.1080/00275514.1995.12026537

23. Paxton RJ, Klee J, Korpela S, Fries I: Nosema ceranae has infected Apis mellifera in Europe since at least 1998 and may be more virulent than Nosema apis. Apidologie, 38 (6): 558-565, 2007. DOI: 10.1051/ apido:2007037

24. Tunca RI, Oskay D, Erginoglu S: Monitoring of Nosema infections levels during hygienic honey bee breeding programs in Turkey. Kafkas Univ Vet Fak Derg, 23 (4): 521-526, 2017. DOI: 10.9775/kvfd.2016.17186

25. Goebel B, Wenner T: A survey of Nosema spp. and Varroa destructor mites in honey bee (Apis mellifera) colonies throughout Western Dominica Apiaries. Texas, USA, 2016.

26. Grabensteiner E, Ritter W, Carter MJ, Davison S, Pechhacker J, Kolodziejek J, Boecking O, Derakhshifar I, Moosbeckhofer R, Licek E, Nowotny N: Sacbrood virus of the honeybee (Apis mellifera): Rapid identification and phylogenetic analysis using reverse transcription-PCR. Clin Diagn Lab Immunol, 8 (1): 93-104, 2001. DOI: 10.1128/CDLI.8.1.93- 104.2001

27. Woolson RF, Clarke WR: Tests of Independence and Measures of Association for Two Random Variables. In, Statistical Methods for the Analysis of Biomedical Data. 2nd ed., 263-307, John Wiley & Sons, New York, 2011.

28. Oduwole O, Udoh EE, Oyo-Ita A, Meremikwu MM: Honey for acute cough in children. Cochrane Database Syst Rev, 4, 1-80, 2018. DOI: 10.1002/14651858.CD007094.pub5

29. Simone-Finstrom M, Borba RS, Wilson M, Spivak M: Propolis counteracts some threats to honey bee health. Insects, 8:46, 2017. DOI: 10.3390/insects8020046

30. Simone-Finstrom M, Aronstein K, Goblirsch M, Rinkevich F, de Guzman L: Gamma irradiation inactivates honey bee fungal, microsporidian, and viral pathogens and parasites. J Invertebr Pathol, 153, 57-64, 2018. DOI: 10.1016/j.jip.2018.02.011

31. Leyva-Daniel DE, Escobedo-Avellaneda Z, Villalobos-Castillejos F, Alamilla-Beltrán L, Welti-Chanes J: Effect of high hydrostatic pressure applied to a Mexican honey to increase its microbiological and functional quality. Food Bıoprod Process, 102, 299-306, 2017. DOI: 10.1016/j.fbp. 2017.01.001

32. Vázquez-Quiñones CR, Moreno-Terrazas R, Natividad-Bonifacio I, Quiñones-Ramírez EI, Vázquez-Salinas C: Microbiological assessment of honey in México. Rev Argent Microbiol, 50 (1): 75-80, 2018. DOI: 10.1016/j.ram.2017.04.005

33. Tysset C, Durand C, Taliergio YP: Contribution to the study of the microbial contamination and the hygiene of commercial honey. Rev Med Vet, 146, 1471-1492, 1970.

34. Packer JM, Irısh J, Herbert BR, Hill C, Padula M, Blair SE, Carter DA, Harry EJ: Specific non- peroxide antibacterial effect of manuka honey on the Staphylococcus aureus proteome. Int J Antimicrob Agents, 40 (1): 43-50, 2012. DOI: 10.1016/j.ijantimicag.2012.03.012

35. Dixon B: Bacteria can’t resist honey. Lancet Infect Dis, 3 (2):116, 2003. DOI: 10.1016/S1473-3099(03)00524-3 36. Aureli P, Franciosa G, Fenicia L: Infant botulism and honey in Europe: A commentary. Pediatr Infect Dis J, 21 (9): 866-868, 2002.

37. Chin J, Arnon SS, Midura TF: Food and environmental aspects of infant botulism in California. Rev Infect Dis, 1 (4): 693-697 1979. DOI: 10.1093/clinids/1.4.693

38. Wojtacka J, Wysok B, Kabašinskienė A, Wiszniewska-Łaszczych A, Gomółka-Pawlicka M, Szteyn J, Malakauskas M, Migowska-Calik A: Prevalence of Clostridium botulinum type A, B, E and F isolated from directly sold honey in Lithuania. J Agr Sci Tech, 19 (2): 335-343, 2017.

39. Matović K, Mišić D, Karabasil N, Nedić N, Dmitrić M, Jevtić G, Ćirić J: Clostridium botulinum spores in European honey bees from Serbia. J Apicult Res, 58 (3): 420-426, 2019. DOI: 10.1080/00218839.2018.1560654

40. Özkırım A, Keskin N: Ankara ili ve çevresindeki arılıklarda teşhis edilen başlıca yavru hastalıklarının dağılımı. Mellifera, 2 (4): 8-12, 2002.

41. Jara L, Martínez-López D, Muñoz I, De la Rua P: Epidemiological survey of Ascosphaera apis in small-scale migratory Apis mellifera iberiensis colonies. Sociobiology, 65 (2): 285-290, 2018. DOI: 10.13102/ sociobiology.v65i2.2685

42. Soysal Mİ, Gürcan EK: Tekirdağ ili arı yetiştiriciliği üzerine bir araştırma. Tekirdağ Ziraat Fak Derg, 2 (2): 161-165, 2005.

43. Kırpık MA, Aydoğan MN, Örtücü S, Hasenekoğlu İ: Kafkas arısı (Apis mellifera caucasia Pollman, 1889) (Hymenoptera: Apidea)’nın dış yüzey ve sindirim sistemi mikrofungus florasının belirlenmesi. Kafkas Univ Vet Fak Derg, 16 (Suppl-B): S347-S352, 2010. DOI: 10.9775/kvfd.2010.2589

44. Büyük M, Tunca Rİ, Taşkın A: Determination of Nosema disease in colonies of Kırşehir province. TURJAF, 5 (1): 1-5, 2017. DOI: 10.24925/ turjaf.v5i1.1-5.723

45. Emsen B, Guzman-Novoa E, Hamiduzzaman MM, Eccles L, Lacey B, Ruiz-Pérez RA, Nasr M: Higher prevalence and levels of Nosema ceranae than Nosema apis infections in Canadian honey bee colonies. Parasıtol Res., 115 (1): 175-181, 2016. DOI: 10.1007/s00436-015-4733-3

46. Kar S, Kaya N, Güven E, Karaer Z: Yeni geliştirilen tespit kabı ile ergin arılarda Varroa enfestasyonunun belirlenmesi. Uludag Bee J, 6 (2): 68-73, 2006.

47. Şimşek H: Elazığ yöresi bal arılarında bazı parazit ve mantar hastalıklarının araştırılması. Ankara Üniv Vet Fak Derg, 52, 123-126, 2005.

48. Anido M, Branchiccela B, Castelli L, Harriet J, Campá J, Zunino P, Antúnez K: Prevalence and distribution of honey bee pests and pathogens in Uruguay. J Apicult Res, 54 (5): 532-540, 2015. DOI: 10.1080/00218839.2016.1175731

49. Otim AS, Kajobe R, P’Odyek Abila P, Kasangaki P, Echodu R: Viruses circulating in african honey bees in Uganda. Bee World, 97 (1): 21-25, 2020. DOI: 10.1080/0005772X.2019.1698103

50. Brettell LE, Riegler M, O’Brien C, Cook JM: Occurrence of honey bee-associated pathogens in Varroa-free pollinator communities. J Invertebr Pathol, 171:107344, 2020. DOI: 10.1016/j.jip.2020.107344