Document Type : Research Paper

Authors

1 College of Veterinary Medicine University of Al-Qadisiyah Iraq

2 Department of Veterinary Microbiology, College of Veterinary Medicine, University of Al-Qadisiyah, Al-Diwaniyah City, Iraq.

10.29079/qjvms.2024.151348.1025

Abstract

Human diarrhea is caused by the zoonotic protozoan Giardia lamblia. This parasite is very important, so it might serve as a study object. The purpose of the work described below was to identify G. lamblia in human stool samples from Baghdad province, Iraq. 140 fecal samples constituted an experiment. The samples were collected throughout the year. PCR, partial gene sequencing, and microscopic analysis were performed. The microscopic approach identified cysts at the highest rate (14/59, 23.7%) in the month of January, and PCR was able to determine that the percentage of humans certified as sick was 21/21 (99.09%). The phylogenetic tree shows that the strains of isolates are very close to the world strains from Poland and Iran. Overall, the work at hand provides substantial pieces of information on the occurrence of G. lamblia in human stool.

Keywords

1.Ankarklev J, Jerlström-Hultqvist J, Ringqvist E, Troell K, Svärd SG. Behind the smile: cell biology and disease mechanisms of Giardia species. Nat Rev Microbiol. 2010;8(6):413-422. doi:10.1038/nrmicro2317
2.Savioli L, Smith H, Thompson A. Giardia and Cryptosporidium join the 'Neglected Diseases Initiative'. Trends Parasitol. 2006;22(5):203-208. doi:10.1016/j.pt.2006.02.015
3.Robertson LJ, Gjerde BK, Furuseth Hansen E, et al. Giardia duodenalis cysts isolated from wild moose and reindeer in Norway: genetic characterization by PCR-RFLP and sequence analysis at two genes. J Parasitol. 2006;92(5):1025-1030. doi:10.7589/0090-3558-43.4.576
4.Adam RD. Biology of Giardia lamblia. Clin Microbiol Rev. 2001;14(3):447-475. doi:10.1017/S1431927604040954
5.Halliez MC, Buret AG. Extra-intestinal and long-term consequences of Giardia duodenalis infections. World J Gastroenterol. 2013;19(47):8974-8985. doi:10.3748/wjg.v19.i47.8974
6.Ryan U, Hijjawi N, Feng Y, Xiao L. Giardia: an under-reported foodborne parasite. Int J Parasitol. 2019;49(1):1-11. doi:10.1016/j.ijpara.2018.07.003
7.Escobedo AA, Almirall P, Robertson LJ, Franco RM. Giardiasis: pharmacotherapy options. Expert Opin Pharmacother. 2019;20(15):1849-1862. doi:10.1517/14656566.8.12.1885
8.World Health Organization. Guidelines for Drinking-Water Quality. 4th ed. World Health Organization; 2011. doi:10.1016/j.pce.2022.103353
9.Adam RD. Biology of Giardia lamblia. Clin Microbiol Rev. 2001 Oct;14(3):447-75. doi: 10.1128/CMR.14.3.447-475.2001.
10.Escobedo AA, Almirall P, Robertson LJ, Franco RM. Giardiasis: the ever-present threat of a neglected disease. Infect Dis Clin North Am. 2010 Jun;24(2):297-316. doi: 10.1016/j.idc.2010.01.009.
11.Hanevik K, Wensaas KA, Rortveit G, Eide GE, Mørch K, Langeland N. Irritable bowel syndrome and chronic fatigue 6 years after Giardia infection: a controlled prospective cohort study. Clin Infect Dis. 2014 Jan;58(3):341-6. doi: 10.1093/cid/cit724.
12.Ryan U, Cacciò SM. Zoonotic potential of Giardia. Int J Parasitol. 2013 Nov;43(12-13):943-56. doi: 10.1016/j.ijpara.2013.06.001.
13.Feng Y, Xiao L. Zoonotic potential and molecular epidemiology of Giardia species and giardiasis. Clin Microbiol Rev. 2011;24:110–40. doi:10.1128/CMR.00033-10
14.Ryan U, Cacci SM. Zoonotic potential of Giardia. Int J Parasitol. 2013;43:943–56. doi: 10.1128/CMR.00033-10
15.Savioli L, Smith H, Thompson A. Giardia and cryptosporidium join the ‘neglected diseases initiative’ Trends Parasitol. 2006;22:203–208. doi:10.1016/j.pt.2006.02.015
16.Smith HV, Mank TG. Diagnosis of human Giardiasis. In: Lujan HD, Svard S, Eds , editors. Giardia a model organism. New York: Springer-Verlag; 2011. pp. 353–74. doi:10.18502/ijpa.v18i2.13180
17.Hooshyar H, Ghafarinasab S, Arbabi M, Delavari M, Rasti S. Genetic variation of Giardia lamblia isolates from food-handlers in Kashan, central Iran. Iran J Parasitol. 2017;12:83–89. PMCID: PMC5522702, PMID: 28761464
18.Stark D, Barratt JL, Van Hal S, Marriott D, Harkness J, Ellis JT. Clinical significance of enteric protozoa in the immunosuppressed human population. Clin Microbiol Rev . 2009;2:634–50. doi:10.1128/CMR.00017-09
19.Daryani A, Sharif M, Meigouni M, Mahmoudi FB, Rafiei A, Gholami S, et al. Prevalence of intestinal parasites and profile of CD4+ counts in HIV+/AIDS people in north of Iran, 2007-2008. Pak J Biol Sci . 2009;12:1277–81. doi:10.3923/pjbs.2009.1277.1281
20.Elmi T, Gholami Sh, Rahimi-Esboei B, Garaili Z, Najm M, Tabatabaie F. Comparison of sensitivity of sucrose gradient, wet mount and formalin – ether in detecting protozoan Giardia lamblia in stool specimens of BALB/c mice. J Pure Applied Microbiol . 2017;11:105–109. doi:10.22207/JPAM.11.1.14
21.Wolfe MS. Giardiasis. Clin Microbiol Rev. 1992;5:93–100. doi:10.1128/CMR.5.1.93
22.Soares R, Tasca T. Giardiasis: an update review on sensitivity and specificity of methods for laboratorial diagnosis. J Microbiol Methods. 2016;129:98–102. doi:10.1016/j.mimet.2016.08.017
23.Gutiérrez-Cisneros MJ, Martínez-Ruiz R, Subirats M, Merino FJ, Millán R, Fuentes I. Assessment of two commercially available immunochromatographic assays for a rapid diagnosis of Giardia duodenalis and crypstosporidium spp. in human fecal specimens. Enferm Inf Microbiol Clin. 2011;29:201–203. doi: 10.1016/j.eimc.2010.09.005
24.Soares R, Tasca T. Giardiasis: an update review on sensitivity and specificity of methods for laboratorial diagnosis. J Microbiol Methods. 2016;129:98–102. doi:10.1016/j.mimet.2016.08.017
25.John DT, Petri WA, Markell EK, Voge M, Eds , editors. Markell and Voge's medical parasitology. New York: Elsevier Health Sciences; 2006. pp. 404–5. doi:10.21125/inted.2023.1933
26.Hiatt RA, Markell EK, Ng E. How many stool examinations are necessary to detect pathogenic intestinal protozoa? Am J Trop Med Hyg. 1995;53:36–9. doi:10.4269/ajtmh.1995.53.36
27.Goka AK, Rolston D, Mathan V, Farthing M. The relative merits of faecal and duodenal juice microscopy in the diagnosis of giardiasis. Trans R Soc Trop Med Hyg. 1990;84:66–7. doi:10.1016/0035-9203(90)90386-S
28.Oguoma VM, Ekwunife CA. The need for a Better Method: Comparison of direct smear and formol-ether concentration techniques in diagnosing intestinal parasites. Int J Trop Med . 2007;3:1–8. doi: 10.4269/ajtmh.16-0436
29.Carvalho GL, Moreira LE, Pena JL, Marinho CC, Bahia MT, Machado-Coelho GL. A comparative study of the TF-Test®, Kato-Katz, Hoffman-Pons-Janer, Willis and Baermann-Moraes coprologic methods for the detection of human parasitosis. Memórias do Ins Oswaldo Cruz. 2012;107:80–84. doi:10.1590/S0074-02762012000100011
30.Engels D, Nahimana S, Gryseels B. Comparison of the direct faecal smear and two thick smear techniques for the diagnosis of intestinal parasitic infections. Trans R Soc Trop Med Hyg. 1996;90:523–5. doi:10.1016/S0035-9203(96)90304-1
31.Babaei Z, Oormazdi H, Rezaie S, Rezaeian M, Razmjou E. Giardia intestinalis: DNA extraction approaches to improve PCR results. Exp Parasitol. 2011;128:159–62. doi:10.1016/j.exppara.2011.02.001
32.Xiao LI, Herd RP. Quantitation of giardia cysts and cryptosporidium oocysts in fecal samples by direct immunofluorescence assay. J Clin Microbiol . 1993;31:2944–6. doi:10.1128/JCM.31.11.2944-2946.1993
33.Mank TG, Zaat JO, Blotkamp J, Polderman AM. Comparison of fresh versus sodium acetate acetic acid formalin preserved stool specimens for diagnosis of intestinal protozoal infections. Eur J Clin Microbiol Infect Dis. 1995;14:1076–81. doi:10.1007/BF01590942 
34.Palmer J. Modified iron hematoxylin/Kinyoun stain. Clin Microbiol News. 1991;13:39–40. doi:10.1177/030098587901600212
35.Garcia LS, editor. Diagnostic medical parasitology. 5th edition. Washington, D.C, USA: ASM Press; 2007. pp. 759–63. doi:10.1016/0196-4399(94)90055-8
36.Bullock WL. The use of the Kohn chlorazol black fixative-stain in an intestinal parasite survey in rural Costa Rica. J Parasitol. 1980;66:811–3. doi:10.2307/3280674
37.Graham CC, Diamond LS. Methods for cultivation of luminal parasitic protists of clinical importance. Clin Microbiol Rev. 2002;15:329–41. doi:10.1128/CMR.15.3.329-341.2002
38.Keister DB. Axenic culture of Giardia lamblia in TYI-S-33 medium supplemented with bile. Trans R Soc Trop Med Hyg. 1983;77:487–8. doi:10.1016/0035-9203(83)90120-7
39.Beal CB, Viens P, Grant RG, Hughes JM. A new technique for sampling duodenal contents. Am J Trop Med Hyg . 1970;19:349–52. doi: 10.4269/ajtmh.1970.19.349
40.Korman SH, Hais ED, Spira DT. Routine in vitro cultivation of Giardia lamblia by using the string test. J Clin Microbiol. 1990;28:368–9. doi: 10.1128/jcm.28.2.368-369.1990
41.Gordts B, Hemelhof W, Van Tilborgh K, Retore P, Cadranel S, Butzler JP. Evaluation of a new method for routine in vitro cultivation of Giardia lamblia from human duodenal fluid. J Clin Microbiol. 1985;22:702–4. doi: 10.1128/jcm.22.5.702-704.1985
42.Gordts B, Retore P, Cadranel S, Hemelhof W, Rahman M, Butzler JP. Routine culture of Giardia lamblia trophozoites from human duodenal aspirates. Lancet. 1984;324:137–8. doi: 10.1016/s0140-6736(84)91050-x
43.Rosenthal P, Liebman WM. Comparative study of stool examinations, duodenal aspiration, and pediatric Entero-Test for giardiasis in children. J pediatrics. 1980;96:278–9. doi: 10.1016/s0022-3476(80)80826-2
44.Hall EJ, Rutgers HC, Batt RM. Evaluation of the peroral string test in the diagnosis of canine giardiasis. J Small Anim Pract. 1988;29:177–83. doi:10.1111/j.1748-5827.1988.tb02275.x
45.Heyworth MF. Diagnostic testing for giardia infections. Trans R Soc Trop Med Hyg. 2014;108:123–5. doi: 10.1093/trstmh/tru005
46.Faubert G. Immune response to Giardia duodenalis. Clin Microbiol Rev. 2000;13:35–54. doi:10.1128/CMR.13.1.35-54.2000
47.Fink , Singer S. The intersection of immune responses, microbiota, and pathogenesis in giardiasis. Trend Parasitol. 2017;33:901–13. doi:10.4049/jimmunol.198.Supp.216.10
48.Adam RD. Biology of Giardia lamblia. Clin Microbiol Rev. 2001;14:447–75. doi:10.1128/CMR.14.3.447-475.2001
49.Abdulla DA. Coccidiosis in domesticated duck in Nineveh governorate. Iraqi J Vet Sci. 2010;24(2):93-97. doi: 10.33899/ijvs.2010.5602
50.Al-Taee AF, Mohammed RG, Mohammed NH. Diagnosis of some helminthic eggs in faces of ducks and geese in Nineveh governorate, Iraq. Iraqi J Vet Sci. 2011;25(1):5-10. doi: 10.33899/ijvs.2011.5696
51.Mohammed NH. Study on the blood protozoa in geese. Iraqi J Vet Sci. 2020;34(1):23-27. doi: 10.33899/ijvs.2019.125499.1028
52.Mohammad Z.A. Some chewing lice (Phthiraptera) species as ectoparasites infested aquatic birds with a new record of three species from Al-Sanaf marsh/ southern Iraq. Iraqi J Vet Sci. 2020;34(1):173-180. doi: 10.33899/ijvs.2019.125721.1139
53.Mohammed NH. Detection of Cryptosporidium spp. In feces of ducks in Nineveh governorate. Iraqi J Vet Sci. 2009;23(1):1-5. doi: 10.33899/ijvs.2009.5689
54.Mohammed NH. Prevalence of Giardia spp. in ducks and geese in Nineveh governorate. Iraqi J Vet Sci. 2009;23(1):1-5. doi: 10. 338999/ijvs.2012.35197
55.Coles, E.H. Veterinary Clinical Pathology. 4th Edition, W.B. Saunders Company, Philadelphia, 1986; 17-19. doi: 10.1111/vcp.13118
56.Feng Y, Xiao L. Zoonotic potential and molecular epidemiology of Giardia species and giardiasis. Clin Microbiol Rev. 2011 Jan;24(1):110-40. doi: 10.1128/CMR.00033-10.
57.Smith HV, Cacciò SM, Cook N, Nichols RA, Tait A. Cryptosporidium and Giardia as foodborne zoonoses. Vet Parasitol. 2007;149(1-2):29-40. doi:10.1016/j.vetpar.2007.07.015
58.Johnson DW, Pieniazek NJ, Griffin DW, Misener L, Rose JB. Development of a PCR protocol for sensitive detection of Giardia intestinalis in wastewater. Water Res. 1995;29(6):1511-1518. doi:10.1016/0043-1354(94)00393-l
59.Adamska M, Leśniańska K, Sawczuk M, et al. Molecular characterization of Giardia intestinalis strains from humans in Poland. Parasitol Res. 2015;114(6):2345-2350. doi:10.1007/s00436-015-4448-5
60.Alfatlawy HH, Alfatlawi MA. Morphological and molecular identification of Parabronema skrjabini of camels (Camelus dromedary) in Najaf province. Iraqi J Vet Sci. 2021;35(3):507-12. doi: 10.33899/ijvs.2020.127101.1459
61.Alfatlawi MSH, Alfatlawi MA. Molecular and phylogenetic study of Theileria spp isolated from ticks in Al-Diwaniyah City, Iraq. Iraqi J Agricul Sci. 2019;50(1):475-79. doi: https://doi.org/10.36103/ijas.v50i1.313
62.Klaif SF, Jassim A, Alfatlawi MA, Ali MA. Major-surface-protein-4-gene-based detection of Anaplasma marginale isolated from sheep in Al-Diwaniyah province, Iraq. Iraqi J Vet Sci. 2022;36(3):85-88. doi:10.33899/ijvs.2021.129230.1635
63.Monis PT, Caccio SM, Thompson RCA, Sundar N. Multilocus genotypic analysis of Giardia intestinalis isolates: evidence of genetic recombination and zoonotic transmission. Int J Parasitol. 1999;29(12):1899-908. doi: 10.1371/journal.pntd.0000558.
64.Sulaiman IM, Fayer R, Bern C, Gilman RH, Trout JM, Schantz PM et al. Triosephosphate isomerase gene characterization and potential zoonotic transmission of Giardia duodenalis. Emerg Infect Dis. 2003;9(11):1444-52. doi: 10.3201/eid0911.030084.
65.Read C, Walters J, Robertson ID. Correlation between genotype of Giardia duodenalis and diarrhoea. Int J Parasitol. 2004;34(6):717-21. doi:10.1016/s0020-7519(01)00340-x
66.Lebbad M, Petersson I, Karlsson L, Botero-Kleiven S, Andersson JO, Svenungsson B et al. Multilocus genotyping of human Giardia isolates suggests limited zoonotic transmission and association between assemblage B and flatulence in children. PLOS Negl Trop Dis. 2011;5(8):e1262. doi: 10.1371/journal.pntd.0001262.