华中农业大学水产学院/湖北省水生动物病害防控工程技术研究中心/ 农业农村部水生动物疫病专业实验室(华中农业大学),武汉430070
肖杰尹,E-mail:xjy@webmail.hzau.edu.cn
顾泽茂,E-mail:guzemao@mail.hzau.edu.cn
收稿:2025-05-24,
纸质出版:2026-01-15
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肖杰尹,杨浩,顾泽茂.多子小瓜虫掠食体培养基的筛选及添加琼脂糖和EPC细胞对虫体发育的影响[J].华中农业大学学报,2026,45(1):300‐307.
XIAO Jieyin,YANG Hao,GU Zemao.Screening of medium for Ichthyophthirius multifiliis theronts and effect of adding agarose and EPC cells to medium on their development[J].Journal of Huazhong Agricultural University,2026,45(01):300-307.
肖杰尹,杨浩,顾泽茂.多子小瓜虫掠食体培养基的筛选及添加琼脂糖和EPC细胞对虫体发育的影响[J].华中农业大学学报,2026,45(1):300‐307. DOI: 10.13300/j.cnki.hnlkxb.2026.01.027.
XIAO Jieyin,YANG Hao,GU Zemao.Screening of medium for Ichthyophthirius multifiliis theronts and effect of adding agarose and EPC cells to medium on their development[J].Journal of Huazhong Agricultural University,2026,45(01):300-307. DOI: 10.13300/j.cnki.hnlkxb.2026.01.027.
为探究多子小瓜虫(
Ichthyophthirius multifiliis
)掠食体在体外的发育条件,通过比较掠食体在不同培养基中的存活情况获得掠食体培养的最适培养基;并分析在最适培养基中添加鲤上皮瘤(EPC)细胞和琼脂糖(a: 琼脂糖在虫体与细胞混合液下层;b: 琼脂糖与虫体和细胞混合;c: 琼脂糖在虫体和细胞混合液上层)对掠食体发育的影响。结果显示:(1)掠食体在不同培养基中的存活时间为4~6 d,其中在M199培养基中存活时间最长,但培养基中的掠食体无法发育成滋养体;(2)掠食体在添加EPC细胞团的M199培养基中可发育成滋养体,但仅可存活2 d,且滋养体的大小在第2天为(31.32±3.79) µm,无显著生长趋势(
P
>
0.05);(3)掠食体在3种琼脂糖模式中均可发育成滋养体,且存活时间可达3 d。a、b和c 3种模式中滋养体的大小在第3天分别为(37.40±3.99)、(39.51±8.51)和(45.14±10.92) µm,模式c中的滋养体生长最快。结果表明,在M199培养基中添加EPC细胞团和琼脂糖均可促进掠食体发育成滋养体,且在琼脂糖位于虫体和细胞混合液上层的模式下滋养体生长最快。因此,在多子小瓜虫掠食体的体外培养中,建议采用添加EPC细胞团并以琼脂糖覆盖上层的M199培养基,以最优促进其向滋养体发育与生长。
Ichthyophthirius multifiliis
is a ciliated protozoan parasite that infects various freshwater fish, causing ichthyophthiriasis. The life cycle of
I. multifiliis
comprises three stages: theront, trophont, and tomont. Theront, the only infective stage, invades the host and transforms into the trophont. This study first compared the survival of theronts in different cell culture media to identify the most suitable medium for
in vitro
culture. Based on these findings, the effects of adding epithelioma papillosum cyprini (EPC) cells and agarose to the culture medium on theront development were investigated using three models: (a), agarose was positioned in the lower layer beneath the theront and cell mixture; (b), the agarose mixed
directly with theront and cell; and (c),agarose positioned in the upper layer above the theront and cell mixture. The survival rate and size of
I. multifiliis
were calculated for each condition. The results were as follows: (1) The longest survival time for theronts in medium M199 was up to 6 d. However, theronts were unable to transform into trophonts. (2) The addition of EPC cell aggregates to medium M199 enabled theronts to transformed into trophonts, which survived for 2 d. The size of trophonts was (31.32±3.79) µm on d 2, with no significant growth trend observed. (3) In all three agarose models, theronts was transformed into trophonts and survived for 3 d. The diameters of trophonts in models a, b, and c modes were (37.40±3.99) µm, (39.51±8.51) µm, and (45.14±10.92) µm, respectively, on day 3. In conclusion, the present study demonstrated that adding either EPC cell aggregates or agarose to medium M199 promoted the transformation of theronts into trophonts. However, significant trophonts growth occurred only in the model where the medium was supplemented with both agarose and cells.
MATTHEWS R A. Ichthyophthirius multifiliis fouquet and ichthyophthiriosis in freshwater teleosts[J].Advances in parasitology,2005,59:159-241.
王哲,刘春雷,顾泽茂,等.多子小瓜虫的形态发生及其寄生导致翘嘴鲌鳃组织病理变化[J].水生生物学报,2016,40(5):935-941.WANG Z,LIU C L,GU Z M,et al.Morphology and morphogenesis of Ichthyophthirius multifiliis and pathological study of gills of Culter alburnus [J].Acta hydrobiologica sinica,2016,40(5):935-941 (in Chinese with English abstract) .
LI M,BASTOS GOMES G,ZHAO W S,et al.Cultivation of fish ciliate parasites:progress and prospects[J].Reviews in aquaculture,2023,15(1):142-162.
EWING M S,KOCAN K M.Invasion and development strategies of Ichthyophthirius multifiliis ,a parasitic ciliate of fish[J].Parasitology today,1992,8(6):204-208.
YANG H,WANG Z,XIAO J Y,et al.Integrated morphological and transcriptome profiles reveal a highly-developed extrusome system associated to virulence in the notorious fish parasite, Ichthyophthirius multifiliis [J/OL].Virulence,2023,14:2242622[2025-05-24].https://doi.org/10.1080/21505594. 2023. 2242622.
SONG K G,LING F,HUANG A G,et al. In vitro and in vivo assessment of the effect of antiprotozoal compounds isolated from Psoralea corylifolia against Ichthyophthirius multifiliis in fish[J].International journal for parasitology:drugs and drug resistance,2015,5(2):58-64.
LIU Y M,ZHANG Q Z,XU D H,et al.Antiparasitic efficacy of curcumin from Curcuma longa against Ichthyophthirius multifiliis in grass carp[J].Veterinary parasitology,2017,236:128-136.
DICKERSON H W,LOHR A L,GRATZEK J B.Experimental intraperitoneal infection of channel catfish, Ictalurus punctatus (Rafinesque),with Ichthyophthirius multifiliis (Fouquet)[J].Journal of fish diseases,1985,8(1):139-142.
HUTSON K S,CABLE J,GRUTTER A S,et al.Aquatic parasite cultures and their applications[J].Trends in parasitology,2018,34(12):1082-1096.
EKLESS L,MATTHEWS R. Ichthyophthirius multifiliis :axenic isolation and short-term maintenance in selected monophasic media[J].Journal of fish diseases,2006,16(5):437-447.
彭碧琳.小瓜虫体外培养研究及青蒿琥酯等药物对小瓜虫的影响[D].福州:福建农林大学,2007.PENG B L.The study of cultivating Ichthyophthirius multifiliis in vitro and the effect of artesunate on Ichthyophthirius multifiliis [D].Fuzhou:Fujian Agriculture and Forestry University,2007 (in Chinese with English abstract) .
XU D H,KLESIUS P H,SHOEMAKER C A.Evaluation of a cohabitation challenge model in immunization trials for channel catfish Ictalurus punctatus against Ichthyophthirius multifiliis [J].Diseases of aquatic organisms,2007,74(1):49-55.
HU J B,YANG H,TU X,et al.Epicatechin gallate:a promising agent isolated from Sargentodoxa cuneata for the treatment of ichthyophthiriasis in goldfish ( Carassius auratus )[J/OL].Aquaculture,2023,574:739724[2025-05-24]. https://doi.org/10.1016/j.aquaculture.2023.739724.
FU Y W,ZHANG Q Z,XU D H,et al.Cynatratoside-C efficacy against theronts of Ichthyophthirius multifiliis ,and toxicity tests on grass carp and mammal blood cells[J].Diseases of aquatic organisms,2015,117(1):13-20.
TANGE E Ø,MATHIESSEN H,VON GERSDORFF JØRGENSEN L.Effects of pH on free-living stages of a Nordic strain of the economically important freshwater fish parasite Ichthyophthirius multifiliis [J].International journal for parasitology,2020,50(10-11):859-864.
YOSHINAGA T,AKIYAMA K,NISHIDA S,et al. In vitro culture technique for Cryptocaryon irritans ,a parasitic ciliate of marine teleosts[J].Diseases of aquatic organisms,2007,78(2):155-160.
YAO T,ASAYAMA Y.Animal-cell culture media:history,characteristics,and current issues[J].Reproductive medicine and biology,2017,16(2):99-117.
VENTURA M T,PAPERNA I.Histopathology of Ichthyophthirim multifiliis infections in fishes[J].Journal of fish biology,2006,27(2):185-203.
YANG H,TU X,XIAO J Y,et al.Investigations on white spot disease reveal high genetic diversity of the fish parasite, Ichthyophthirius multifiliis (Fouquet,1876) in China[J/OL].Aquaculture,2023,562:738804[2025-05-24]. https://doi.org/10.1016/j.aquaculture.2022.738804.
倪达書,李连祥.多子小瓜虫的形态、生活史及其防治方法和一新种的描述[J].水生生物学集刊,1960(2):197-225.NI D S,LI L X.Studies on the morphology and life cycle of Ichthyophthirius multifiliis and its control,with a description of a new species[J].Acta hydrobiologica sinica,1960(2):197-225 (in Chinese with English abstract) .
卢全章,郭琼林.寄生虫感染草鱼胸腺的观察[J].水生生物学报,1990,14(1):95-96.LU Q Z,GUO Q L.Observation on the infection of the thymus of grass carp ( Ctenopharyngodon idellus ) by parasites[J].Acta hydrobiologica sinica,1990,14(1):95-96 (in Chinese) .
MARGARET H L. Ichthyophthirius multifiliis Fouquet:development and assessment of in vitro systems for long term maintenance[D].Plymouth,South West England,UK:University of Plymouth,1999.
罗思施,汤顺清.琼脂糖在组织工程中的研究进展[J].中国生物工程杂志,2015,35(6):68-74.LUO S S,TANG S Q.Research progress of agarose in tissue engineering[J].China biotechnology,2015,35(6):68-74 (in Chinese with English abstract) .
MUNIZ R S,CAMPBELL P C,SLADEWSKI T E,et al.Revealing spatio-temporal dynamics with long-term trypanosomatid live-cell imaging[J/OL].PLoS pathogens,2022,18(1):e1010218[2025-05-24]. https://doi.org/10.1371/journal.ppat.1010218.
SHAW S,KNÜSEL S,ABBÜHL D,et al.Cyclic AMP signalling and glucose metabolism mediate pH taxis by African trypanosomes[J/OL].Nature communications,2022,13(1):603[2025-05-24].https://doi.org/10.1038/s41467-022-28293-w.
EWING M S,LYNN M E,EWING S A.Critical periods in development of Ichthyophthirius multifiliis (Ciliophora) populations1[J].The journal of protozoology,1986,33(3):388-391.
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