Establishment and influencing factors of experimental animal models for intestinal protozoa infection

doi:10.12140/j.issn.1000-7423.2025.01.021

Abstract

Abstract:

Intestinal protozoa are single-celled eukaryotic microorganisms that are widely distributed in ecological environments. Most of the protozoan isolates are derived from excretions or secretions of individuals carrying cysts, which are used for experimental animal infections following purification and culture. Both the limitations of in vitro culture techniques and the host specificity of protozoa may directly affect the difficulty in the establishment of animal models. This review summarizes major in vitro culture techniques and appropriate animal models, and discusses the factors affecting the establishment of animal models. Considering that novel animal models are more effective to simulate the disease progression of human intestinal protozoan diseases, high attention should be paid to the development of experimental animal models of intestinal protozoan infections, notably novel animal models, so as to provide supports for the establishment and research of animal models of intestinal protozoan infections.

Key words: Intestinal protozoa, In vitro culture, Inoculation, Animal infection, Animal model

CLC Number:

R531

ZHANG Xiaocheng, SHEN Yujuan, HU Yuan, JIANG Yanyan, LIU Hua, CAO Jianping. Establishment and influencing factors of experimental animal models for intestinal protozoa infection[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2025, 43(1): 135-139.

Figures/Tables 1

References 42

[1]	Stensvold CR, Berg RPKD, Maloney JG, et al. Molecular characterization of Blastocystis and Entamoeba of muskoxen and sheep in Greenland[J]. Int J Parasitol, 2023, 53(11/12): 673-685.
[2]	金伟. 安徽省肠道原虫病流行病学研究进展[J]. 热带病与寄生虫学, 2019, 17(2): 116-118.
	Jin W. Progress in epidemiological research of intestinal protozoa in Anhui Province[J]. J Trop Dis Parasitol, 2019, 17(2): 116-118. (in Chinese)
[3]	Farid A, Tawfik A, Elsioufy B, et al. In vitro and in vivo anti-Cryptosporidium and anti-inflammatory effects of Aloe vera gel in dexamethasone immunosuppressed mice[J]. Int J Parasitol Drugs Drug Resist, 2021, 17: 156-167.
[4]	Robinson NB, Krieger K, Khan FM, et al. The current state of animal models in research: A review[J]. Int J Surg, 2019, 72: 9-13.
[5]	Fink MY, Shapiro D, Singer SM. Giardia lamblia: Laboratory maintenance, lifecycle induction, and infection of murine models[J]. Curr Protoc Microbiol, 2020, 57(1): e102.
[6]	Al-Hindi AI, El-Khozondar HJ, Tabaza W, et al. Effect of magnetic field on the growth of the cultured Entamoeba histolytica isolated from patients in Palestine[J]. Exp Parasitol, 2021, 226/227: 108126.
[7]	Szpakowska A, Stachowska A, Burzyńska A, et al. Determining the pathogenicity and virulence of parasites on animal models[J]. Przegl Epidemiol, 2023, 77(2): 209-219.
[8]	Taghipour A, Rayatdoost E, Bairami A, et al. Are Blastocystis hominis and Cryptosporidium spp. playing a positive role in colorectal cancer risk? A systematic review and meta-analysis[J]. Infect Agent Cancer, 2022, 17(1): 32.
[9]	Ryan UM, Feng YY, Fayer R, et al. Taxonomy and molecular epidemiology of Cryptosporidium and Giardia: A 50 year perspective (1971-2021)[J]. Int J Parasitol, 2021, 51(13/14): 1099-1119.
[10]	Arrowood MJ. Cryptosporidium oocyst purification using discontinuous gradient centrifugation[J]. Methods Mol Biol, 2020, 2052: 43-59.
[11]	Nourrisson C, Lavergne RA, Moniot M, et al. Enterocytozoon bieneusi, a human pathogen[J]. Emerg Microbes Infect, 2024, 13(1): 2406276.
[12]	Li W, Feng Y, Xiao L. Enterocytozoon bieneusi[J]. Trends Parasitol, 2022, 38(1): 95-96.
[13]	Yu XG, Mu XR, Yuan KJ, et al. Occurrence and genotypic identification of Blastocystis spp. and Enterocytozoon bieneusi in Bamaxiang pigs in Bama Yao Autonomous County of Guangxi Province, China[J]. Animals (Basel), 2024, 14(22): 3344.
[14]	Whelan TA, Fast NM. Microsporidia[J]. Curr Biol, 2023, 33(18): R936-R938.
[15]	Bojko J, Reinke AW, Stentiford GD, et al. Microsporidia: A new taxonomic, evolutionary, and ecological synthesis[J]. Trends Parasitol, 2022, 38(8): 642-659.
[16]	Pan GQ, Bao JL, Ma ZG, et al. Invertebrate host responses to microsporidia infections[J]. Dev Comp Immunol, 2018, 83: 104-113.
[17]	Kuo CJ, Hansen M, Troemel E. Autophagy and innate immunity: Insights from invertebrate model organisms[J]. Autophagy, 2018, 14(2): 233-242.
[18]	Cacciò SM, Lalle M, Sv?rd SG. Host specificity in the Giardia duodenalis species complex[J]. Infect Genet Evol, 2018, 66: 335-345.
[19]	Dayao DA, Sheoran A, Carvalho A, et al. An immunocompetent rat model of infection with Cryptosporidium hominis and Cryptosporidium parvum[J]. Int J Parasitol, 2020, 50(1): 19-22.
[20]	Shiratori M, Patel A, Gerhold RW, et al. Persistent Trichomonas vaginalis infections and the pseudocyst form[J]. Trends Parasitol, 2023, 39(12): 1023-1031.
[21]	Aquino MFK, Hinderfeld AS, Simoes-Barbosa A. Trichomonas vaginalis[J]. Trends Parasitol, 2020, 36(7): 646-647.
[22]	Edwards T, Burke P, Smalley H, et al. Trichomonas vaginalis: clinical relevance, pathogenicity and diagnosis[J]. Crit Rev Microbiol, 2016, 42(3): 406-417.
[23]	Mendoza Cavazos C, Knoll LJ. Entamoeba histolytica: Five facts about modeling a complex human disease in rodents[J]. PLoS Pathog, 2020, 16(11): e1008950.
[24]	刘光英, 陈金富, 温旺荣, 等. 齿龈内阿米巴致病作用的动物实验[J]. 中国寄生虫学与寄生虫病杂志, 2001, 19(4): 229-232.
	Liu GY, Chen JF, Wen WR, et al. Experimental study on the pathogenesis of Entamoeba gingivalis[J]. Chin J Parasitol Parasit Dis, 2001, 19(4): 229-232. (in Chinese)
[25]	陈金富, 刘光英, 温旺荣, 等. 齿龈内阿米巴的连续培养与致病性研究[J]. 中国寄生虫学与寄生虫病杂志, 2000, 18(2): 84-86.
	Chen JF, Liu GY, Wen WR, et al. Studies on the continuous culture and pathogenicity of Entamoeba gingivalis[J]. Chin J Parasitol Parasit Dis, 2000, 18(2): 84-86. (in Chinese)
[26]	Bartelt LA, Bolick DT, Mayneris-Perxachs J, et al. Cross-modulation of pathogen-specific pathways enhances malnutrition during enteric co-infection with Giardia lamblia and enteroaggregative Escherichia coli[J]. PLoS Pathog, 2017, 13(7): e1006471.
[27]	Keselman A, Li EQ, Maloney J, et al. The microbiota contributes to CD8⁺ T cell activation and nutrient malabsorption following intestinal infection with Giardia duodenalis[J]. Infect Immun, 2016, 84(10): 2853-2860.
[28]	Stefano VC, Loiola SH, Dos Santos BM, et al. Effects of hypertonic solutions on two species of human intestinal parasites during fecal examination[J]. Future Microbiol, 2023, 18: 197-203.
[29]	Fitri LE, Candradikusuma D, Setia YD, et al. Diagnostic methods of common intestinal protozoa: Current and future immunological and molecular methods[J]. Trop Med Infect Dis, 2022, 7(10): 253.
[30]	Patel SJ, Pathak P, Thejeswi P, et al. Acute appendicitis secondary to Entamoeba histolytica infestation[J]. Trop Parasitol, 2023, 13(2): 114-116.
[31]	冯霞, 张富强, 徐福阳, 等. 人芽囊原虫感染动物模型构建研究进展[J]. 中国血吸虫病防治杂志, 2021, 33(1): 102-106.
	Feng X, Zhang FQ, Xu FY, et al. Progress of researches on animal models of Blastocystis hominis infections[J]. Chin J Schisto Control, 2021, 33(1): 102-106. (in Chinese)
[32]	Robinson G, Elwin K, Chalmers RM. Cryptosporidium diagnostic assays: Molecular detection[J]. Methods Mol Biol, 2020, 2052: 11-22.
[33]	Xu N, Liu H, Jiang YY, et al. First report of Cryptosporidium viatorum and Cryptosporidium occultus in humans in China, and of the unique novel C. viatorum subtype XVaA3h[J]. BMC Infect Dis, 2020, 20(1): 16.
[34]	DuPont HL. Persistent diarrhea: A clinical review[J]. JAMA, 2016, 315(24): 2712-2723.
[35]	Carrero JC, Reyes-López M, Serrano-Luna J, et al. Intestinal amoebiasis: 160 years of its first detection and still remains as a health problem in developing countries[J]. Int J Med Microbiol, 2020, 310(1): 151358.
[36]	Lange AN, Sch?fer A, Bender A, et al. Galleria mellonella: A novel invertebrate model to distinguish intestinal symbionts from pathobionts[J]. Front Immunol, 2018, 9: 2114.
[37]	Solaymani-Mohammadi S, Petri WA Jr. Zoonotic implications of the swine-transmitted protozoal infections[J]. Vet Parasitol, 2006, 140(3/4): 189-203.
[38]	Vermeire B, Gonzalez LM, Jansens RJJ, et al. Porcine small intestinal organoids as a model to explore ETEC-host interactions in the gut[J]. Vet Res, 2021, 52(1): 94.
[39]	Heo I, Dutta D, Schaefer DA, et al. Modelling Cryptosporidium infection in human small intestinal and lung organoids[J]. Nat Microbiol, 2018, 3(7): 814-823.
[40]	Bhalchandra S, Lamisere H, Ward H. Intestinal organoid/enteroid-based models for Cryptosporidium[J]. Curr Opin Microbiol, 2020, 58: 124-129.
[41]	Guérin A, Striepen B. The biology of the intestinal intracellular parasite Cryptosporidium[J]. Cell Host Microbe, 2020, 28(4): 509-515.
[42]	Kilkenny C, Parsons N, Kadyszewski E, et al. Survey of the quality of experimental design, statistical analysis and reporting of research using animals[J]. PLoS One, 2009, 4(11): e7824.

原虫种类	虫种来源	动物选择	动物接种技术	文献
蓝氏贾第鞭毛虫	自感染贾第鞭毛虫患者或感染动物的粪便中分离培养获得包囊	家犬、家猫、家兔、B细胞缺陷小鼠、C57BL/6N小鼠、C57BL/6J小鼠、长爪沙鼠、蒙古沙鼠等	胃部灌注	[5,18,26 -27]
溶组织内阿米巴	自然感染溶组织内阿米巴患者或感染动物的粪便中分离培养获得虫体	狗、猫、猴、兔、豚鼠、小鼠、金黄地鼠、仓鼠、长爪沙鼠等	盲肠内接种；胃部灌注	[6-7,20⇓⇓-23]
人芽囊原虫	自然感染人芽囊原虫患者或感染动物的粪便中分离培养获得虫体	大鼠、小鼠、豚鼠、猪和兔等	胃部灌注	[8]
隐孢子虫	自然感染隐孢子虫患者或感染动物的粪便中分离培养获得卵囊	大鼠、小鼠、豚鼠、犬、牛、羊、猪、鸡等	小型动物：胃部灌注大型动物：经口感染	[9-10,19]
微孢子虫	自然感染微孢子虫患者或感染动物的粪便中分离培养获得孢子	BALB/c小鼠、C57BL/6J小鼠、SCID小鼠等	腹膜内注射；直肠灌入	[11⇓⇓⇓⇓⇓-17]