Prevalence of Giardia intestinalis with other co-infecting parasites in Barak Valley, Assam, India: a molecular approach

  • Madhumita Roy
  • Baby SinghaEmail author
  • Debadatta Dhar
  • Shubhadeep Roychoudhury
Original Article


Giardia intestinalis was included in the World Health Organization’s Neglected Disease Initiative in 2004 as it may range from asymptomatic to chronic or severe diarrhoea and chronic disorders post-infection. The present study aimed to find out the rate of sole infection of G. intestinalis and co-infection of this with other protozoan parasites among the inhabitants of Barak Valley region of Southern Assam by conventional and molecular detection. A total of 1168 samples were collected from different groups of individuals, all the collected samples were subjected to microscopy after specific staining by Lugol’s iodine solution, Trichrome staining and modified ZN staining procedures. Microscopically positive samples were further confirmed by PCR using specific primer sets. Of the total no. of samples, 267 (22.85%) were positive by PCR for G. intestinalis with a little higher rate of infection in female (24.06%) (OR = 1.2192, CI = 0.9262 to 1.6049) than male (21.27%). The rate of infection is comparatively higher (25.93%) in the age group of 0-5 years (OR = 1.9149, CI = 1.2558 to 2.9200). In 196 samples G. intestinalis co-existence was observed and detected by PCR with some other protozoan parasites like Entamoeba spp., Cryptosporidium spp. and Blastocystis spp. The rate of infection was higher (31.96%) among the participants who collected water from river. Least of the participants showed diarrhoeal symptoms (18.18%) but majority (28.45%) complained for having abdominal cramps (OR = 1.3402, CI = 0.8815 to 1.7855). Among the human infective assemblages, assemblage specific molecular detection revealed the rate of infection of assemblage B was comparatively higher (60.30%) than assemblage A.


Giardiasis Gastrointestinal Prevalence Assemblages Epidemiology 



The authors want to acknowledge Department of Medicine and Department of Microbiology of Silchar Medical College for providing samples. Authors also greatly acknowledge The Molecular Parasitology Laboratory of Gurucharan College Silchar funded by DBT, DST and UGC for helping in molecular analysis under this study. We are grateful to Prof. Jaishree Paul, School of Life Sciences, JNU, New Delhi for providing all the positive controls used in this study as a kind gift to us. We would also like to convey our sincere gratitude to Prof. Sankar Kumar Ghosh, Vice Chancellor, Kalyani University, West Bengal for helping in the sequencing part in his previous laboratory of Dept. of Biotechnology, Assam University, Silchar. This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

Author’s contribution

The study design, aim and methodology were prepared by Dr. Baby Singha and Dr. Shubhadeep Roychoudhury. All the clinical samples along with patient details were provided by Dr. Debadatta Dhar. The experimentation, data analysis, statistical analysis were done by Dr. Madhumita Roy. The manuscript was prepared primarily by Dr. Madhumita Roy and further checked and modified by all the co-authors.


The research leading to these results has received funding from UGC, DBT and DST grants given to the Molecular Parasitology Laboratory, Department of Zoology, G. C. College, Silchar, Assam.

Compliance with ethical standards

Conflict of interest

All the authors declare that they have no conflict of interest.

Ethics approval

The protocol of the study was reviewed and approved by the Institutional Ethical Committee (Ref. No. GCC/9440) of Gurucharan College, Silchar, Assam, before the commencement of the Study and Silchar Medical College and Hospital, Silchar, Assam.

Informed consent

For the present study all stool samples were collected with the consent of each patient/subject, in the form of questionnaires. All study participants had given written consent before enrolment into the study. On behalf of all the infant participants consent had given by the parents/guardians.


  1. Çelİksöz A, Aciöz M, DeĞerlİ S, Çinar Z, Elaldi N, Erandaç M (2005) Effects of giardiasis on school success, weight and height indices of primary school children in Turkey. Pediatr Int 47:567–571CrossRefGoogle Scholar
  2. Charan J, Biswas T (2013) How to calculate sample size for different study designs in medical research? Indian J Psychol Med 35(2):121CrossRefGoogle Scholar
  3. Dutta G (1965) Cytochemistry and fluorescence microscopy of Giardia intestinalis. Proc Natl Acad Sci India 31:151–165Google Scholar
  4. Easow JM, Mukhopadhyay C, Wilson G, Guha S, Jalan BY, Shivananda PG (2005) Emerging opportunistic protozoa and intestinal pathogenic protozoal infestation profile in children of western Nepal. Nepal Med Coll J 7:134–137Google Scholar
  5. Feng Y, Xiao L (2011) Zoonotic potential and molecular epidemiology of Giardia species and giardiasis. Clin Microbiol Rev 24:110–140CrossRefGoogle Scholar
  6. Flanagan P (1992) Giardia–diagnosis, clinical course and epidemiology. A review. Epidemiol Infect 109:1Google Scholar
  7. Ford BJ (2005) The discovery of Giardia. Microsc Chicago 53:161–167Google Scholar
  8. Ish-horowicz M, Korman SH, Shapiro M, Har-even U, Tamir I, Strauss N, Deckelbaum RJ (1989) Asymptomatic giardiasis in children. Pediatr Infect Dis J 8:773–779CrossRefGoogle Scholar
  9. Kaur R, Rawat D, Kakkar M, Uppal B, Sharma V (2002) Intestinal parasites in children with diarrhea in Delhi, India. Southeast Asian J Trop Med Public Health 33:725Google Scholar
  10. Lasek-Nesselquist E, Welch D, Thompson RCA, Steuart RF, Sogin ML (2009) Genetic exchange within and between assemblages of Giardia duodenalis. J Eukaryot Microbiol 56:504–518CrossRefGoogle Scholar
  11. Lebbad M, Ankarklev J, Tellez A, Leiva B, Andersson JO, Svärd S (2008) Dominance of Giardia assemblage B in Leon, Nicaragua. Acta Trop 106:44–53CrossRefGoogle Scholar
  12. Lwanga SK, Lemeshow S, World Health Organization (1991) Sample size determination in health studies: a practical manual. World Health Organization, GenevaGoogle Scholar
  13. Molina N, Polverino D, Minvielle M, Basualdo J (2007) PCR amplification of triosephosphate isomerase gene of Giardia lamblia in formalin-fixed feces. Rev Latinoam Microbiol 49:6–11Google Scholar
  14. Monis PT (1998) Molecular epidemiology: assumptions and limitations of commonly applied methods. Int J Parasitol 28:981–987CrossRefGoogle Scholar
  15. Parija SC, Khairnar K (2007) Detection of excretory Entamoeba histolytica DNA in the urine, and detection of E. histolytica DNA and lectin antigen in the liver abscess pus for the diagnosis of amoebic liver abscess. BMC Microbiol 7:41CrossRefGoogle Scholar
  16. Potters I, Van Esbroeck M (2010) Negative staining technique of Heine for the detection of Cryptosporidium spp.: a fast and simple screening technique. Open Parasitol J 4:1–4CrossRefGoogle Scholar
  17. Rai K, Sherchand JB, Bhatta DR, Bhattarai NR (2005) Status of Giardia intestinalis infection among the children attending Kanti children hospital, Nepal. Sci. World 3:102–105Google Scholar
  18. Schantz P, Sarti E, Plancarte A, Wilson M, Criales J, Roberts J, Flisser A (1994) Community-based epidemiological investigations of cysticercosis due to Taenia solium: comparison of serological screening tests and clinical findings in two populations in Mexico. Clin Infect Dis 18:879–885CrossRefGoogle Scholar
  19. Sethi S, Sehgal R, Malla N, Mahajan R (1999) Cryptosporidiosis in a tertiary care hospital. Natl Med J India 12:207–209Google Scholar
  20. Sulaiman IM, Fayer R, Bern C, Gilman RH, Trout JM, Schantz PM, Das P, Lal AA, Xiao L (2003) Triosephosphate isomerase gene characterization and potential zoonotic transmission of Giardia duodenalis. Emerg Infect Dis 9:1444CrossRefGoogle Scholar
  21. Vanni I, Cacciò SM, van Lith L, Lebbad M, Svärd SG, Pozio E, Tosini F (2012) Detection of Giardia duodenalis assemblages A and B in human feces by simple, assemblage-specific PCR assays. PLoS Neglect Trop Dis 6:e1776CrossRefGoogle Scholar
  22. Xiao L, Morgan UM, Limor J, Escalante A, Arrowood M, Shulaw W, Thompson R, Fayer R, Lal AA (1999) Genetic diversity within Cryptosporidium parvum and related Cryptosporidium species. Appl Environ Microbiol 65:3386–3391Google Scholar
  23. Yoshikawa H, Abe N, Iwasawa M, Kitano S, Nagano I, Wu Z, Takahashi Y (2000) Genomic analysis of Blastocystis hominisStrains isolated from two long-term health care facilities. J Clin Microbiol 38:1324–1330Google Scholar

Copyright information

© Indian Society for Parasitology 2019

Authors and Affiliations

  1. 1.Department of ZoologyGurucharan CollegeSilcharIndia
  2. 2.Department of Life Science and BioinformaticsAssam UniversitySilcharIndia
  3. 3.Silchar Medical College and HospitalSilcharIndia

Personalised recommendations