Comparison between different methods of Blastocystis hominis detection in stool samples.

Abstract

 Blastocystis hominis is common protozoan in human intestinal tract and can cause so-called

blastocystosis characterized by diarrhea. Its routine identification in clinical laboratories is

made by detection of vacuolar form in stool samples using wet mount smears. The present

study was carried out with the aim of evaluating the effectiveness of different techniques

for diagnosis of  B. hominis in the stool samples from the patients attended Brack Hospital

and Medical Technology Department, Faculty of Engineering and Technology, Brack.

A total of 360 stool samples were collected form randomized patients, presenting different

genders and ages (121 males and 239 females ,and aged from less than one year to 90

years), residing in different localities of Wadi Al-shati province. All specimens were

examined by direct smear microscopy (normal saline, iodine, and eosin stains),

concentration (formalin–ether sedimentation) and two xenic culture systems (Monophasic

Jone’s medium and Diphasic Boeck and Drbohlav's ) for the detection of  B. hominis. The

results highlights the low sensitivity of direct smear microscopy (11.29%) compared to

concentration method (15.5%) and  in-vitro culture methods (22.60%). There was no

significant difference (p>0.05) between direct smear preparations, and concentration

method, meanwhile there was significant difference (p<0.05) between direct smear

preparations and the two xenic culture systems for the detection of  B. hominis. There was

an almost equal numbers of positive samples in both culture techniques (85 samples in

diphasic medium and 78 in monophasic medium), and no significant difference (P>0.05)

was found between the two culture methods. Diphasic Boeck and Drbohlav’s medium

produced highest numbers (7.600±6.379) of  B. hominis cells compared to Monophasic

Jone’s medium (5.051±4.938) after every passage cultures and only the vacuolar

morphologic type of this organism was found in both culture systems. Moreover, a larger

size of vacuolar stage of  B. hominis detected in Diphasic Boeck and Drbohlav's medium,

than in Monophasic Jone's medium.

In vitro cultivation does seem worthwhile in the detection of  B. hominis in diagnostic

laboratories. Of all the diagnostic techniques used, diphasic Boeck and Drbohlav's medium

was the most sensitive method for detecting  B. hominis in stool specimens.

The short-term  in vitro culture methods achieved the best performance with regard to

sensitivity with other studied methods. With the advantages in terms of sensitivity, the  in

vitro culture methods could be applied to identify  B. hominis for both clinical diagnosis and

field study purposes, thus indicating the need to include laboratory techniques that enable

 B. hominis detection on a routine basis.

References

1. Boreham ,P. F .L. and Stenzel, D. J. (1993): Blastocystis in humans and animals: morphology, biology and epizootiology. Adv Parasitol. 32:1-70.

2. Stenzel, D.J. and Boreham, P.F.L. (1996): Blastocystis hominis Revisited. Clin. Microbiol. Rev. 9: 563-584.

3. Suresh, K., and Smith, H. (2004): Comparison of methods for detecting Blastocystis hominis. Eur. J. Microbiol. Infect. Dis. 23: 509-511.

4. Roberts, T., Barrat, J., Harkness, J., Eliss, J. and Stark, D. (2011): Comparison of microscopy, culture and conventional polymerase chain reaction for detection of Blastocystis spp. in clinical stool samples. Am .J. Trop. Med. Hyg. 84:308-312.

5. Zaman, V. and Khan, K. Z. (1994): A comparsion of direct microscopy with culture for dignosis of Blastocystis hominis. Southeast Asian J. Trop. Med. Public Health. 25: 792-793.

6. Leelayoova, S., Taamasri, P., Rangsin, R.; Naaglor, T., Thathaisong. U. and Murgthin, M. (2002): In-vitro cultivation: a sensitive method for detecting Blastocystis hominis. Ann.Trop. Med. Parasitol. 96: 803-807.

7. Amato N, V.; Rodriguezalarcon, R.S.; Gakiya, E. et al (2003). - Blastocistose: controvérsias e indefinições. Rev. Soc. Bras. Med. Trop., 36: 515-517.

8. Al-Fellani, M. A., Abdul rahman, D.M., Khan, A.H. and Abousaif, A.A. (2005): Prevalence of intestinal parasites in Sebha, Libya. Garyounis Med. J. 22:56-60.

9. Awatif Mohammed, Khan, A. H., AbdulQader Ajaili, Gelani, Salem, Mohammed Shabani and Ali Daew. (2008): Prevalence of Blastocystis hominis in Wadi Al-Shati Province. Sebha Med. J. 7:4-10

10. Fathy, F. M. (2011): A study on Blastocystis hominis in food handlers: diagnosis and potential pathogenicity J. Egypt. Soc. Parasitol. 41: 433 – 453.

11. Ruqaia , M .A. (2006): A study of Blastocysis hominis by direct faecal smear and culture method in Sebha. M.Sc. (Desertation). Department of Zoology. Faculty of Science . University of Sebha.

12. Cheesbrough M. (2004): District Laboratory Practice in Tropical Countries. 2ed. ed. Cambridge University Press. 191-199.

13. Cook, G. (1996): Manson's Tropical Diseases. 20th Ed. W.B. Saunders. 1738 – 1739.

14. Jones, W .R. (1946): The experimental infection of rats with Entamoeba histolytica with a method for evaluting the anti-amoebic properties of new compounds. Aun. Trop. Med. Parasitol. 40:130-140.

15. Zierdt, C.H. and Swan, J.C. (1981): Generation time and growth rate of the human intestinal parasite Blastocystis hominis. J. Protozool. 28: 483-485.

16. Pipatsatitpon, D., Rangsin, R., Leelayoova, S., Naaglor, T. and Mungthin, M. (2012). Incidence and risk factors of Blastocystis infection in an orphanage in Bangkok, Thailand. Parasites Vectors. 14:37.

17. Salem, R. A. A., Abdullah, M.E. and Abudl Qader, A.E. (2006): Intestinal Protozoa in Libyan patients in Sirte. Jamahiriya Med. J. 6:59-61.

18. Sadaga G.A. and Kassem, H.H. (2007): Prevalence of Intestinal parasites among School Children in Derna District, Libya. J. Egypt. Soc. Parasitol. 37: 205-214.

19. Saleh, M.M. (2007). Prevalence of human intestinal parasites in Sebha City. M.Sc. Thesis, Sebha university, Seba, Libya.

20. Kassem, H.H. Zaed H.A and Sadoga G. A. (2007). Intestinal parasitic infection among children in Tripoli. J. Egypt. Soc. Parasitol. 36: 107-112.

21. Gelani, S. S., Abdulhafeez K., Abdul Qader, A. Awatif, M. A., Mohammed Al-Shebani and Yosef, K. (2009): A study of prevalence of human intestinal parasites in Wadi Al-Shati region. Sebha Med. J. 2: 3-7.

22. Zierdt, C.H. (1991): Blastocystis hominis – past and future. Clin. Microbiol. Res. 4: 61-79.

23. Zhang, X., Oiao, J.Y., Dong, X. H., Li,Y.Q., Li, X. Q. and Li, C. (2003): Study on morphology of Blastocystis hominis in culture and from diarrhea patients .Zhongguo Ji Sheng Chong Xue Yu J :Sheng Chong Bing Za Zhi .21:116-118.

24. Mohammed, A.A., Khan, A.H., Rugaiya, A., Mubrook, Z.A. and Farjani M. (2007): Prevalence and Clinical features of Blastocystis hominis among patients in Sebha, Libya. Sultan Qaboos Uni. Med. J. 7 : 23-28.

25. Vennila G.D., Suresh K, Khairul A.A., Rajah S., Saminatha R., Sivanandan S. and Ramakrishnan K. (1999): Irregular shedding of Blastocystis hominis. Parasitol. Res. 85(2): 162-164.

26. Suresh K.; Ng, C.G.; Ramachandran, N. P.; Ho, L.C.; Yap, E. H. and Singh, M. (1993): In-vitro encystsment and experimental infections of Blastocystis hominis. Parasitol. Res. 79: 456-460.

27. Yakoob, J., Jafriw, W., Jafri, N., Khan, R., Islam, M., Beg, M. A. and Zaman, V. (2004): Irritable bowel syndrome: In search of an etiology. Role of Blastocystis hominis. Am. J. Trop. Med. Hyg. 70: 383-385.

28. Dogruman Al. F., Simsek, Z., Boorom, K., Edici, E., Sahin, M., Tuncer, C., Kustimur, S. and Altinbas, A. (2010). Comparison of methods for detection of Blastocystis infection in routinely submitted stool samples, and also in IBS/BD patients in Ankara, Turkey. PLOS . one 5: e15484.

29. Moe, K.T., Singh, M., Howe, J., Ho, L.C., Tan, S.W., Chen , X.O. and Yap E.H. (1999): Development of Blastocystis hominis cysyts into vacuolar forms in-vitro. Parasitol. Res. 85: 103-108.

30. Stenzel, D.J. (1995): Ultrastructural and cytochemical studies of Blastocystis spp. Ph.D. thesis. Queensland University of Technology, Brisbane, Australia.

31. Qadri, H.S.M., Al-Okaili, G.A. and Al-Dayel, F. (1989): Clinical significance of Blastocystis hominis. J. Clin. Microbiol. 27: 2407-2409.

32. Adeen, W.E. and Hale, D. (1992): Use of Hemo-De to eliminate toxic agents used for concentration and trichrome staining of intestinal parasites. J. Clin. Microbiol. 30: 1893-1895.

33. Guimaraes, S. and Sogayar, I. L. (1993): Blastocystis homonis occurrence in children and staff member of municipal day-care centers from Botucatu. Sao Paulo state, Brazil. Me. Inst. Oswaldo Cruz Rio J. 88: 427-429.

34. Logar, J., Andlovic, A. and Poljsak-Prijatelj, M. (1994): Incidence of Blastocystis hominis in patients with diarrhea. J. Infect. 28: 151-154.

35. Taamasri, P.; Mungthin, M.; Raugsin, R.; Tongupprakarn, B.; Areekul, W. and Leelayoova, S. (2002): Transmission of blastocystosis related to the quality of drinking water. Southeast Asian J. Trop. Med. Public. Health.31: 112-117.

36. Miller, R. A. and Minshew, B.H. (1988): Blastocystis hominis and organism in search of a disease . Rev. Infect. Dis. 10: 938-950.

37. Tungtrongchitr A., Manatasathi S., Kositchaiwt C., Ongroichanakun J., Munkong N., Chinabutr P., Leelakusolvong S. and Chaicampa W. (2004): Blastocystis hominis infection in irritable bowel syndrome patients. Southeast Asian J. Trop. Med. Public Health. 35: 705-71.

38. Nascimento, S.A. and Moitinho, M.D. (2005): Blastocystis hominis gastroenteritis in a haemophiliac with acquired immune deficiency syndrome. J. Pediatr. Gastroenterol. Nutr. 8: 125-128.

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Published

2018-05-23

How to Cite

Comparison between different methods of Blastocystis hominis detection in stool samples. (2018). Lebda Medical Journal, 3(1), 95–100. Retrieved from https://lebmedj.elmergib.edu.ly/index.php/LMJ/article/view/66

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Lebda Medical Journal Vol. (3) 2017