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KULLANILMIŞ RUJ VE MASKARALARDA MİKROBİYOLOJİK KONTAMİNASYONUNUN ARAŞTIRILMASI

Year 2023, , 269 - 276, 20.01.2023
https://doi.org/10.33483/jfpau.1209451

Abstract

Amaç: Kozmetik ürünlerin içeriğinde yer alan karbon ve azot kaynakları, mineraller, büyüme faktörleri ve asidik ortam, nem gibi uygun ortam koşulları mikrobiyal üreme için elverişli koşulların oluşmasını sağlar. Bu ürünlerin mikrobiyolojik kontaminasyonu hem tüketici sağlığı açısından risk oluşturması hem de üründe meydana gelebilecek değişiklikler nedeniyle ekonomik kayıplara neden olması açısından önem taşımaktadır. Bu çalışmada tüketiciler tarafından kullanılmış ruj ve rimel örneklerinin mikrobiyolojik kontaminasyon açısından incelenmesi amaçlanmıştır.
Gereç ve Yöntem: Çalışmada tüketiciler tarafından kullanılmış otuz adet ruj ve otuz adet rimel örneği mikrobiyolojik kontaminasyon açısından incelenmiştir. Numuneler, Avrupa Standardı EN ISO 17516'da belirtilen mikrobiyolojik limit değerlerine göre değerlendirilmiştir. Kontaminant bakteriler VITEK® 2 Compact (bioMérieux, Fransa) otomatize sistemi ile tanımlanmıştır.
Sonuç ve Tartışma: Elde edilen sonuçlara göre, test edilen maskara örneklerinden yalnızca birinin, ruj örneklerinden ise dördünün belirtilen mikrobiyolojik limit değerlerine uygun olmadığı saptanmıştır. Maskara örneğinden izole edilen bakteri Staphylococcus sciuri, ruj örneklerinden izole edilen bakteriler ise Kocuria kristinae, Streptococcus mitis, Streptococcus oralis, Streptococcus alactolyticus, Rothia mucilaginosa, Rothia dentocariosa türleri olarak tanımlanmıştır. Kontaminasyona neden olan bakterilerin çoğunluğunun ağız, boğaz, solunum ve cilt mikrobiyotasının üyeleri, Gram pozitif türler olduğu görülmüştür. Test edilen kozmetik ürünlerin bulaşının, tüketicilerin uygun olmayan kullanımlarından, aynı ürünü birden fazla kişinin kullanmasından, tükürük bulaşından, el hijyenine dikkat edilmemesinden kaynaklanmış olabileceği düşünülmüştür. 

References

  • 1. Yavaşal Çarıkçı, A.İ., Uçar, F., Yalçın, H.T. (2008). Kozmetik ürünlerde bakteriyal ve fungal kompozisyonun klasik yöntemler ve PCR yöntemi kullanılarak saptanması. Elektronik Mikrobiyoloji Dergisi TR, 6(1),1-16.
  • 2. Çomoğlu, T. (2012). Kozmetikler. Marmara Pharmaceutical Journal. 16, 1-8. [CrossRef]
  • 3. Azak Sungur, S., Sözen Şahane, B., Yeğenoğlu, S. (2018). Kozmetik ürünlerin tarihçesi, ürün tanıtımlarının tüketici davranışı ve yasal durum açısından değerlendirilmesi. Lokman Hekim Dergisi, 8, 191-197. [CrossRef]
  • 4. The European Parliament and the Council of the European Union Regulation (EC) No. 1223/2009 of the European parliament and of the council of 30 November 2009 on cosmetic products. Official Journal of the European Union, 2009, 342, 59.
  • 5. Kozmetik Kanunu, Resmi Gazete Tarihi: 30.03.2005, Sayı: 25771, Kanun Numarası: 5324.
  • 6. Türkı̇ye İlaç ve Tıbbı̇ Cı̇haz Kurumu Kozmetı̇k Ürünlerde Güvenlı̇lı̇k Değerlendı̇rmesı̇ne İlı̇şkı̇n Kılavuz Sürüm 3.0 (2020). From https://titck.gov.tr/storage/Archive/2020/contentFile/asd_977f480b-4a07-4e0e-bb0e-14db1fcf47d6.pdf. Accessed date: 02.11.2021.
  • 7. Birteksöz-Tan, A.S., Tüysüz, M., Ötük, G. (2013). Investigation of preservative efficacy and microbiological content of some cosmetics found on the market. Pakistan Journal of Pharmaceutical Sciences, 26, 153-157.
  • 8. Bashir, A., Lambert, P. (2019). Microbiological study of used cosmetic products: Highlighting possible impact on consumer health. Journal of Applied Microbiology, 128(2), 598-605. [CrossRef]
  • 9. Lundov, M., Moesby, L., Zachariae, C. and Johansen, J. (2009). Contamination versus preservation of cosmetics: A review on legislation, usage, infections, and contact allergy. Contact Dermatitis, 60, 70-78. [CrossRef]
  • 10. Eldesoukey, R.M.M., Alqhtani, B.S., Alqhtani, A.S., Alqhtani, A.H., Alqhtani, A.M. (2016). Comparative microbiological study between traditional and modern cosmetics in Saudi Arabia. Enzyme Engineering, 5, 2. [CrossRef]
  • 11. Neza, E., Centini, M. (2016). Microbiologically contaminated and over-preserved cosmetic products according rapex 2008-2014. Cosmetics, 3, 3. [CrossRef]
  • 12. Scientific Committee on Consumer Products (2016). The SCCP’s Notes of Guidance for the Testing of Cosmetic Ingredients and Their Safety Evaluation. From https://ec.europa.eu/health/ scientific_committees/consumer_safety/docs/sccs_o_190.pdf. Accessed date: 02.11.2021.
  • 13. Babalola, M., Eze, M. (2018). Microbiological quality and characterization of potential pathogens associated with selected brands of commercial cosmetic products in Nigeria. Microbiology Research Journal International, 9, 1-17. [CrossRef]
  • 14. Michalek, I.M., John, S.M., Caetano dos Santos, F.L. (2019). Microbiological contamination of cosmetic products-observations from Europe, 2005-2018. Journal of European Academy of Dermatology and Venereology, 33, 2151-2157. [CrossRef]
  • 15. FDA (2020a) Microbiological safety and cosmetics. From https://www.fda.gov/cosmetics/cosmetic-products-ingredients/cosmetic-products. Accessed date: 22.11.2022
  • 16. FDA (2020b) ASEAN Guidelines for Cosmetic Good Manufacturing Practice. From https://www.fda.moph.go.th/sites/Cosmetic/Lists/ASEANCH/Attachments/58/Appendix%206%20ASEAN%20Cosmetic%20GMP.pdf. Accessed date: 22.11.2022
  • 17. Halla, N., Fernandes, I.P., Heleno, S.A., Costa, P., Boucherit-Otmani, Z., Boucherit, K., Rodrigues, A.E., Ferreira, I., Barreiro, M.F. (2018). Cosmetics preservation: A review on present strategies. Molecules, 23(7), 1571. [CrossRef]
  • 18. EN ISO 17516 (2014) Cosmetics–Microbiology–Microbiological limits. Beuth, Berlin. From https://www.iso.org/obp/ui/#iso:std:iso:17516:ed-1:v1:en (consulted February 2016). Accessed date: 22.11.2022
  • 19. The Food and Drug Administration (FDA) of the United States (2017). Bacteriological Analytical Manual Microbiological Methods for Cosmetics. Chapter 23. From https://www.fda.gov/food/laboratory-methods-food/bam-chapter-23-methods-cosmetics. Accessed date: 22.11.2022.
  • 20. Kaynak Onurdağ, F., Özgen, S., Abbasoğlu, D. (2010). Microbiological investigation of used cosmetic samples. Hacettepe University Journal of the Faculty of Pharmacy, 30(1), 1-16.
  • 21. European Pharmacopoeia 9.0, 2.6.12. Microbiological examination of non-sterile products: Microbial enumeration tests, 2017.
  • 22. European Pharmacopoeia 9.0, 2.6.13. Microbiological examination of non-sterile products: Microbial enumeration tests, 2017.
  • 23. Hills, S. (1946). The isolation of Cl. tetani from infected talc. New Zeland Medical Journal. 45, 419-423.
  • 24. Jairoun, A.A., Al-Hemyari, S.S., Shahwan, M., Zyoud, S.H. (2020). An investigation into incidences of microbial contamination in cosmeceuticals in the UAE: Imbalances between preservation and microbial contamination. Cosmetics. 7, 92. [CrossRef]
  • 25. Baird, RM. (1998). Contamination of non-steril pharmaceuticals in hospital and community environments. In: W.B. Hugo and A.D. Russell (Eds). Pharmaceutical Microbiology, (pp. 374-384). Blackwell Science, Oxford.
  • 26. Skowron, K., Jakubicz, A., Budzyńska1, A., Kaczmarek, A., Grudlewska, K., Reśliński, A., Gospodarek-Komkowska, E. (2017). Microbiological purity assessment of cosmetics used by one and several persons and cosmetics after their expiry date, national institute of public health-national institute of hygiene. Roczniki Państwowego Zakładu Higieny, 68(2), 191-197.
  • 27. Geis, P.A. (2006). Preservation strategies in: Cosmetic Microbiology, 2nd ed. In: P.A. Geis (Eds). (pp.163-180). New York: Taylor&Francis Group.
  • 28. Giacomel, C., Dartora, G., Dienfethaeler, H. and Haas, S. (2013). Investigation on the use of expired make-up and microbiological contamination of mascaras. International Journal of Cosmetic Science, 35, 375-380. [CrossRef]
  • 29. Kengkoom, K., Ampawong, S. (2017). Staphylococcus sciuri associated to subcutaneous abscess and dermatitis in ICR Mouse. Arquivo Brasileiro de Medicina Veterinária e Zootecnia, 69, 117-122. [CrossRef]
  • 30. Wilson, S.E., Bannan, R.A., Mcdonald, M.B., Kaufman, H.E. (1990). Corneal trauma and infection caused by manipulation of the eyelashes after application of mascara. Cornea. 9, 181-182.
  • 31. Reid, F.R., Wood, T.O. (1979). Pseudomonas corneal ulcer. The causative role of contaminated eye cosmetics. Archives of Ophthalmology, 97(9), 1640-1641. [CrossRef]
  • 32. Dadashi, L., Dehghanzadeh, R. (2016). Investigating incidence of bacterial and fungal contamination in shared cosmetic kits available in the women beauty salons. Health Promotion Perspectives, 6, 159-163. [CrossRef]
  • 33. Vassoler, M., Tonial, F., Fagundes, S.C., Fagundes, M.A., Zortea, N.B., Rossato-Grando, L.G., Bertol, C.D. (2020). Microbiological contamination of in-store lipstick testers available to the consumer. Mundo da Saúde. 44, 261-268. [CrossRef]
  • 34. Jung, I.H., Kim, J.H., Yoo, Y.J., Park, B.Y., Choi, E.S., Noh, H. (2019). A pilot study of occupational exposure to pathogenic microorganisms through lip cosmetics among dental hygienists. Journal of Occupational Health, 61, 297-304. [CrossRef]

INVESTIGATION OF THE MICROBIOLOGICAL CONTAMINATION OF USED LIPSTICKS AND MASCARAS

Year 2023, , 269 - 276, 20.01.2023
https://doi.org/10.33483/jfpau.1209451

Abstract

Objective: Carbon and nitrogen sources, minerals, growth factors in the composition of cosmetic products, and environmental conditions such as convenient acidic environment and humidity constitute the suitable environment for microbial growth. Microbiological contamination of these products is important in terms of both posing a risk to consumer health and causing economic losses due to changes occurring in the product. This study aimed to examine the lipstick and mascara samples used by consumers in terms of microbiological contamination.
Material and Method: Thirty lipsticks and thirty mascaras used by consumers were investigated for microbiological contamination. The samples were evaluated according to the microbiological limits specified in the European Standard EN ISO 17516. Contaminant bacteria were identified by VITEK® 2 Compact (bioMérieux, France).
Result and Discussion: According to the results obtained, it was determined that only one of the tested mascara samples and four of the lipstick samples did not comply with the specified microbiological limit values. Staphylococcus sciuri was the contaminant bacterium determined in the mascara sample. In addition, Kocuria kristinae, Streptococcus mitis, Streptococcus oralis, Streptococcus alactolyticus, Rothia mucilaginosa, Rothia dentocariosa were detected in lipstick samples. All the bacteria detected as contaminants are Gram-positive, most of which are the members of the mouth, throat, respiratory, and skin microbiota. The contamination may have been caused by the inappropriate use of consumers, such as using the same product by more than one person, contamination of saliva, and not paying attention to hand hygiene.

References

  • 1. Yavaşal Çarıkçı, A.İ., Uçar, F., Yalçın, H.T. (2008). Kozmetik ürünlerde bakteriyal ve fungal kompozisyonun klasik yöntemler ve PCR yöntemi kullanılarak saptanması. Elektronik Mikrobiyoloji Dergisi TR, 6(1),1-16.
  • 2. Çomoğlu, T. (2012). Kozmetikler. Marmara Pharmaceutical Journal. 16, 1-8. [CrossRef]
  • 3. Azak Sungur, S., Sözen Şahane, B., Yeğenoğlu, S. (2018). Kozmetik ürünlerin tarihçesi, ürün tanıtımlarının tüketici davranışı ve yasal durum açısından değerlendirilmesi. Lokman Hekim Dergisi, 8, 191-197. [CrossRef]
  • 4. The European Parliament and the Council of the European Union Regulation (EC) No. 1223/2009 of the European parliament and of the council of 30 November 2009 on cosmetic products. Official Journal of the European Union, 2009, 342, 59.
  • 5. Kozmetik Kanunu, Resmi Gazete Tarihi: 30.03.2005, Sayı: 25771, Kanun Numarası: 5324.
  • 6. Türkı̇ye İlaç ve Tıbbı̇ Cı̇haz Kurumu Kozmetı̇k Ürünlerde Güvenlı̇lı̇k Değerlendı̇rmesı̇ne İlı̇şkı̇n Kılavuz Sürüm 3.0 (2020). From https://titck.gov.tr/storage/Archive/2020/contentFile/asd_977f480b-4a07-4e0e-bb0e-14db1fcf47d6.pdf. Accessed date: 02.11.2021.
  • 7. Birteksöz-Tan, A.S., Tüysüz, M., Ötük, G. (2013). Investigation of preservative efficacy and microbiological content of some cosmetics found on the market. Pakistan Journal of Pharmaceutical Sciences, 26, 153-157.
  • 8. Bashir, A., Lambert, P. (2019). Microbiological study of used cosmetic products: Highlighting possible impact on consumer health. Journal of Applied Microbiology, 128(2), 598-605. [CrossRef]
  • 9. Lundov, M., Moesby, L., Zachariae, C. and Johansen, J. (2009). Contamination versus preservation of cosmetics: A review on legislation, usage, infections, and contact allergy. Contact Dermatitis, 60, 70-78. [CrossRef]
  • 10. Eldesoukey, R.M.M., Alqhtani, B.S., Alqhtani, A.S., Alqhtani, A.H., Alqhtani, A.M. (2016). Comparative microbiological study between traditional and modern cosmetics in Saudi Arabia. Enzyme Engineering, 5, 2. [CrossRef]
  • 11. Neza, E., Centini, M. (2016). Microbiologically contaminated and over-preserved cosmetic products according rapex 2008-2014. Cosmetics, 3, 3. [CrossRef]
  • 12. Scientific Committee on Consumer Products (2016). The SCCP’s Notes of Guidance for the Testing of Cosmetic Ingredients and Their Safety Evaluation. From https://ec.europa.eu/health/ scientific_committees/consumer_safety/docs/sccs_o_190.pdf. Accessed date: 02.11.2021.
  • 13. Babalola, M., Eze, M. (2018). Microbiological quality and characterization of potential pathogens associated with selected brands of commercial cosmetic products in Nigeria. Microbiology Research Journal International, 9, 1-17. [CrossRef]
  • 14. Michalek, I.M., John, S.M., Caetano dos Santos, F.L. (2019). Microbiological contamination of cosmetic products-observations from Europe, 2005-2018. Journal of European Academy of Dermatology and Venereology, 33, 2151-2157. [CrossRef]
  • 15. FDA (2020a) Microbiological safety and cosmetics. From https://www.fda.gov/cosmetics/cosmetic-products-ingredients/cosmetic-products. Accessed date: 22.11.2022
  • 16. FDA (2020b) ASEAN Guidelines for Cosmetic Good Manufacturing Practice. From https://www.fda.moph.go.th/sites/Cosmetic/Lists/ASEANCH/Attachments/58/Appendix%206%20ASEAN%20Cosmetic%20GMP.pdf. Accessed date: 22.11.2022
  • 17. Halla, N., Fernandes, I.P., Heleno, S.A., Costa, P., Boucherit-Otmani, Z., Boucherit, K., Rodrigues, A.E., Ferreira, I., Barreiro, M.F. (2018). Cosmetics preservation: A review on present strategies. Molecules, 23(7), 1571. [CrossRef]
  • 18. EN ISO 17516 (2014) Cosmetics–Microbiology–Microbiological limits. Beuth, Berlin. From https://www.iso.org/obp/ui/#iso:std:iso:17516:ed-1:v1:en (consulted February 2016). Accessed date: 22.11.2022
  • 19. The Food and Drug Administration (FDA) of the United States (2017). Bacteriological Analytical Manual Microbiological Methods for Cosmetics. Chapter 23. From https://www.fda.gov/food/laboratory-methods-food/bam-chapter-23-methods-cosmetics. Accessed date: 22.11.2022.
  • 20. Kaynak Onurdağ, F., Özgen, S., Abbasoğlu, D. (2010). Microbiological investigation of used cosmetic samples. Hacettepe University Journal of the Faculty of Pharmacy, 30(1), 1-16.
  • 21. European Pharmacopoeia 9.0, 2.6.12. Microbiological examination of non-sterile products: Microbial enumeration tests, 2017.
  • 22. European Pharmacopoeia 9.0, 2.6.13. Microbiological examination of non-sterile products: Microbial enumeration tests, 2017.
  • 23. Hills, S. (1946). The isolation of Cl. tetani from infected talc. New Zeland Medical Journal. 45, 419-423.
  • 24. Jairoun, A.A., Al-Hemyari, S.S., Shahwan, M., Zyoud, S.H. (2020). An investigation into incidences of microbial contamination in cosmeceuticals in the UAE: Imbalances between preservation and microbial contamination. Cosmetics. 7, 92. [CrossRef]
  • 25. Baird, RM. (1998). Contamination of non-steril pharmaceuticals in hospital and community environments. In: W.B. Hugo and A.D. Russell (Eds). Pharmaceutical Microbiology, (pp. 374-384). Blackwell Science, Oxford.
  • 26. Skowron, K., Jakubicz, A., Budzyńska1, A., Kaczmarek, A., Grudlewska, K., Reśliński, A., Gospodarek-Komkowska, E. (2017). Microbiological purity assessment of cosmetics used by one and several persons and cosmetics after their expiry date, national institute of public health-national institute of hygiene. Roczniki Państwowego Zakładu Higieny, 68(2), 191-197.
  • 27. Geis, P.A. (2006). Preservation strategies in: Cosmetic Microbiology, 2nd ed. In: P.A. Geis (Eds). (pp.163-180). New York: Taylor&Francis Group.
  • 28. Giacomel, C., Dartora, G., Dienfethaeler, H. and Haas, S. (2013). Investigation on the use of expired make-up and microbiological contamination of mascaras. International Journal of Cosmetic Science, 35, 375-380. [CrossRef]
  • 29. Kengkoom, K., Ampawong, S. (2017). Staphylococcus sciuri associated to subcutaneous abscess and dermatitis in ICR Mouse. Arquivo Brasileiro de Medicina Veterinária e Zootecnia, 69, 117-122. [CrossRef]
  • 30. Wilson, S.E., Bannan, R.A., Mcdonald, M.B., Kaufman, H.E. (1990). Corneal trauma and infection caused by manipulation of the eyelashes after application of mascara. Cornea. 9, 181-182.
  • 31. Reid, F.R., Wood, T.O. (1979). Pseudomonas corneal ulcer. The causative role of contaminated eye cosmetics. Archives of Ophthalmology, 97(9), 1640-1641. [CrossRef]
  • 32. Dadashi, L., Dehghanzadeh, R. (2016). Investigating incidence of bacterial and fungal contamination in shared cosmetic kits available in the women beauty salons. Health Promotion Perspectives, 6, 159-163. [CrossRef]
  • 33. Vassoler, M., Tonial, F., Fagundes, S.C., Fagundes, M.A., Zortea, N.B., Rossato-Grando, L.G., Bertol, C.D. (2020). Microbiological contamination of in-store lipstick testers available to the consumer. Mundo da Saúde. 44, 261-268. [CrossRef]
  • 34. Jung, I.H., Kim, J.H., Yoo, Y.J., Park, B.Y., Choi, E.S., Noh, H. (2019). A pilot study of occupational exposure to pathogenic microorganisms through lip cosmetics among dental hygienists. Journal of Occupational Health, 61, 297-304. [CrossRef]
There are 34 citations in total.

Details

Primary Language English
Subjects Pharmacology and Pharmaceutical Sciences
Journal Section Research Article
Authors

Kübra Yazıcı 0000-0002-2597-9097

Müjde Eryılmaz 0000-0003-3760-1996

Publication Date January 20, 2023
Submission Date November 24, 2022
Acceptance Date December 28, 2022
Published in Issue Year 2023

Cite

APA Yazıcı, K., & Eryılmaz, M. (2023). INVESTIGATION OF THE MICROBIOLOGICAL CONTAMINATION OF USED LIPSTICKS AND MASCARAS. Journal of Faculty of Pharmacy of Ankara University, 47(1), 269-276. https://doi.org/10.33483/jfpau.1209451
AMA Yazıcı K, Eryılmaz M. INVESTIGATION OF THE MICROBIOLOGICAL CONTAMINATION OF USED LIPSTICKS AND MASCARAS. Ankara Ecz. Fak. Derg. January 2023;47(1):269-276. doi:10.33483/jfpau.1209451
Chicago Yazıcı, Kübra, and Müjde Eryılmaz. “INVESTIGATION OF THE MICROBIOLOGICAL CONTAMINATION OF USED LIPSTICKS AND MASCARAS”. Journal of Faculty of Pharmacy of Ankara University 47, no. 1 (January 2023): 269-76. https://doi.org/10.33483/jfpau.1209451.
EndNote Yazıcı K, Eryılmaz M (January 1, 2023) INVESTIGATION OF THE MICROBIOLOGICAL CONTAMINATION OF USED LIPSTICKS AND MASCARAS. Journal of Faculty of Pharmacy of Ankara University 47 1 269–276.
IEEE K. Yazıcı and M. Eryılmaz, “INVESTIGATION OF THE MICROBIOLOGICAL CONTAMINATION OF USED LIPSTICKS AND MASCARAS”, Ankara Ecz. Fak. Derg., vol. 47, no. 1, pp. 269–276, 2023, doi: 10.33483/jfpau.1209451.
ISNAD Yazıcı, Kübra - Eryılmaz, Müjde. “INVESTIGATION OF THE MICROBIOLOGICAL CONTAMINATION OF USED LIPSTICKS AND MASCARAS”. Journal of Faculty of Pharmacy of Ankara University 47/1 (January 2023), 269-276. https://doi.org/10.33483/jfpau.1209451.
JAMA Yazıcı K, Eryılmaz M. INVESTIGATION OF THE MICROBIOLOGICAL CONTAMINATION OF USED LIPSTICKS AND MASCARAS. Ankara Ecz. Fak. Derg. 2023;47:269–276.
MLA Yazıcı, Kübra and Müjde Eryılmaz. “INVESTIGATION OF THE MICROBIOLOGICAL CONTAMINATION OF USED LIPSTICKS AND MASCARAS”. Journal of Faculty of Pharmacy of Ankara University, vol. 47, no. 1, 2023, pp. 269-76, doi:10.33483/jfpau.1209451.
Vancouver Yazıcı K, Eryılmaz M. INVESTIGATION OF THE MICROBIOLOGICAL CONTAMINATION OF USED LIPSTICKS AND MASCARAS. Ankara Ecz. Fak. Derg. 2023;47(1):269-76.

Kapsam ve Amaç

Ankara Üniversitesi Eczacılık Fakültesi Dergisi, açık erişim, hakemli bir dergi olup Türkçe veya İngilizce olarak farmasötik bilimler alanındaki önemli gelişmeleri içeren orijinal araştırmalar, derlemeler ve kısa bildiriler için uluslararası bir yayım ortamıdır. Bilimsel toplantılarda sunulan bildiriler supleman özel sayısı olarak dergide yayımlanabilir. Ayrıca, tüm farmasötik alandaki gelecek ve önceki ulusal ve uluslararası bilimsel toplantılar ile sosyal aktiviteleri içerir.