The Correlation of Biofilm Formation, Hemolytic and Coagulase Activites and Antifungal Susceptibility among Candida parapsilosis Isolates Recovered from Clinical Speciemens

Öz

Objective: The aim of this study was to investigate the virulence factors such as in vitro biofilm production, hemolytic and coagulase activity of Candida parapsilosis isolates and to compare these factors with antifungal susceptibilities. Methods: Twenty six isolates from Kocaeli University and 14 isolates from Mersin University were included into this study. Haemolytic activity was tested by sheep blood agar with 3% glucose. Coagulase activity was tested by tube test method. Biofilm production was determined by modified Christensen macrotube method and scored as weak, intermediate or strong positive. The MIC values of the strains against fluconazole, caspofungin, micafungin, voriconazole and amphotericin B were measured with the automated system VITEK-2. Results: Study isolates were from peripheral blood 23, blood catheter 7, urine 3, periton 3, nail samples 2, pleura and sputum samples each one. Of 40 isolates, biofilm production was detected in 32 (80%) and scored as 13 (32.5%) weak, 12 (30%) intermediate and 7 (17.5%) strong positive. In 50% of isolates alpha hemolysis and in 32.5% beta hemolysis was detected and 17.5% were nonhemolytic. Coagulase activity was negative in all isolates. All isolates were susceptible to voriconazole and amphotericin B. Fluconazole resistance was determined in two isolates having weak biofilm production. Caspofungin resistance was determined in 10 of isolates and 4 of them were also resistant to micafungin; 7 of them had haemolytic activity and 7 of them produced biofilm as well. Of these resistant 10 isolates 7 were from peripheral blood, 2 were from periton and one was from urine. Conclusion: Biofilm production and haemolytic activity were the main virulence factors of C. parapsilosis and due to these factors it causes candidemia and intravenous catheter related infections. The treatment protocols of these infections may alter in parallel with changing epidemiology or antifungal susceptibility profiles. Studies included more isolates are needed.

Anahtar Kelimeler

• C. parapsilosis
• hemolysis
• biofilm
• coagulase
• VITEK-2 AFT
• caspofungin

Klinik Örneklerden İzole Edilmiş Candi̇da parapsi̇losi̇s Suşlarının Bi̇yofi̇lm Oluşturma, Hemoli̇ti̇k ve Koagülaz Akti̇vi̇teleri̇ ile Anti̇fungal Duyarlılıklarının Karşılaştırılması

Öz

Amaç: Candida parapsilosis suşlarının in vitro biyofilm oluşturma, hemoliz ve koagülaz aktiviteleri gibi virülans faktörlerinin araştırılması ve antifungal duyarlılıklarıyla karşılaştırılması amaçlanmıştır. Yöntem: Kocaeli Üniversitesi’nden 26, Mersin Üniversitesi’nden 14 suş çalışmaya alınmıştır. Hemolitik aktivite %3 glikozlu koyun kanlı agarda 37oC’de 48 saatlik, koagülaz aktivite tüp test metodu ile 37oC’de 4 saatlik, biyofilm oluşumu ise modifiye Christensen makro tüp metodu ile %8 glikozlu Sabouraud Dekstroz sıvı besiyerinde 37oC’de 48-72 saatlik inkübasyon sonrasında değerlendirilerek belirlenmiştir. Flukonazol, kaspofungin, mikafungin, vorikonazol ve amfoterisin B’ye karşı MIK değerleri VİTEK-2 otomatize sistem ile belirlenmiştir. Bulgular: Suşların 23’ü periferik kandan, 7’si damar içi kateterden, 3’er adet periton ve idrardan, 2 adet tırnak örneğinden, 1’er adet plevra ve balgam örneklerinden izole edilmiştir. Biyofilm aktivitesi tüm suşların 32’sinde (%80) saptamış olup, 13’ü (%32,5) zayıf, 12’si (%30) orta, 7’si (%17,5) ise güçlü pozitif olarak skorlanmıştır. Suşların yarısı alfa hemoliz, 13’ü (%32,5) beta hemoliz gösterirken, 7’sinde (%17,5) hemoliz saptanmamıştır. Koagülaz aktivitesi tüm suşlarda negatif bulunmuştur. Tüm suşlar vorikonazol ve amfoterisin B’ye duyarlı saptanmıştır. Biyofilm aktivitesi zayıf olan iki suşta flukonazol direncine rastlanmıştır. Periferik kandan izole edilmiş 7 suş ile 2’si periton ve biri idrar suşu olmak üzere toplam 10 suşta (%25) kaspofungin direnci saptanmıştır. Bu suşların 4’ünde aynı zamanda mikafungin direnci, 7’sinde hemolitik aktivite ve biyofilm oluşumu gözlenmiştir. Antifungal direnci ile virülans faktörleri arasında istatistiksel olarak anlamlı bir ilişki bulunamamıştır. Sonuç: Biyofilm oluşturma ve hemolitik aktivite C. parapsilosis’in başlıca virülans faktörleridir ve sıklıkla bu faktörlere bağlı olarak kandidemi ve kateter enfeksiyonları oluşabilmektedir. Bu enfeksiyonların tedavi protokolleri epidemiyolojik ve antifungal duyarlılık paterni verilerine göre değişim gösterebilir. Daha fazla C. parapsilosis suşu kullanarak ileri çalışmalar yapılması önerilmektedir.

Anahtar Kelimeler

• C. parapsilosis
• hemoliz
• biyofilm
• koagülaz
• VITEK-2 AFT
• kaspofungin

Kaynakça

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