Myxosporeans and myxosporidiosis of allogynogenetic gibel carp (Carassius auratus gibelio Bloch) in China

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1 Myxosporeans and myxosporidiosis of allogynogenetic gibel carp (Carassius auratus gibelio Bloch) in China Zhang Jinyong Laboratory of Fish Diseases; Institute of Hydrobiology (IHB), Chinese Academy of Sciences (CAS) Szarvas, Hungary May. 21.

2 Brief history of fish myxosporean research in China IV developmental stages I. Initial stage: ; description solely based on morphology of myxospores Prof. Chen Prof. Ma II. Developmental stage: ; traditional taxonomy, epidemiology, histopathology Representative work: Publication of Fauna Sinica, Myxozoa, Myxosporea in 1998 III. Mature stage: ; combined morphological and molecular characteristics, host, tissue tropism; molecular epidemiology; life cycle, ecology, host-parasite relationships; IV. Progressive stage: 2013 afterwards; Genomic era; evolution; phylogenomic anlysis; functional genomics Representative work: available of full genome data of Thelohanellus kitauei; ongoing genomic project of Myxobolus honghuensis Recorded 575 species in 23 genera

3 Allogynogenetic gibel carp Introduction Dual reproduction (sexual reproduction and gynogynesis) Rapid growth 500g/year, third-fifth freshwater cultured species (after grass carp, common carp, silver carp and bighead) above 2 million ton/year and replacing crucian carp in Chinese aquculture.. Certificate of New variety of 3 rd gibel carp bred by IHB, CAS Cited from Prof. Gui (IHB, CAS)

4 Diversity of myxosporean species from gibel carp: Morphological and molecular characterization since 2001; (About 100 myxosporean infecting gold fish and gibel carp belong to 15 genera recorded in Fauna Sinica, Myxozoa, Myxosporea by Chen & Ma (1998) although most only simply described with line drawing and spore morphological data.) 1. Validation of described or recorded species; 2. Description of new species Outline Life cycles of fish myxosporean in China since 2009; no one species with known full life cycle before 2009; Myxosporidiosis of gibel carp since 2001; 3 species (Thelohanellus wangi, T. wuhanensis and Myxobolus honghuensis) severely affected the sustainable development of gibel carp culture industry

5 Myxobolus pyramidis Redescription M.pyramidis Chen, 1958; Infection site: gill lamellae Molecular data: HQ μ m Showing plasmodium locating at the tip of gill filaments and spore in frontal and sutural view Cluster in capillary network of gill lamellae-infecting clade Showing the location of plasmodium and no inflammatory responses besides mechanistic extrusion

6 Myxobolus hearti Redescription Infection site: myocardium Molecular data: GU normal round plasmodium Unnormal plasmodium Cluster with species with round spore

7 Myxobolus ampullicapsulatus n. sp. Infection site: gill filaments Molecular data: DQ339482, KJ725082, KC M.honghuensis M. ampullicapsulatus Showing tiny morphological and molecuar difference between M. ampullicapsulatus and M. honghuensis (molecular similarity is above 99.2% based on comparsion of SSU rdna sequence, and morpholgocially almost undistinguished )

8 Henneguya doneci n. sp. Redescription M.doneci Schulman, 1962; Infection site: gill filaments Molecular data: EU ; HM146129; LC011456, JQ Gill-infecting cllade Cited from Ye et al. 2012

9 Henneguya globulata n.sp. Infection site: gill filaments Molecular data: KJ725074; JF Myxobolus nielii (Nie et Li, 1973) Landsberg et Lom, 1991 Infection site: gill filaments Molecular data: KJ725074; JF Phylogenetic relationship consistent with spore morphology (round spore body)

10 Myxobolus oralis n. sp. Infection site: buccal cavity Molecular data: KC Cluster with species with round spore Cited from Liu et al. 2014

11 Myxobolus turpisrotundus n.sp. Correction of M. rotundus recorded in China; Infection site: connective tissue of gill arch, fins, intestine and body surface Molecular data: EF690299, GU Plasmodium being going to detach from the dorsal fins Showing location of plasmodia and no severe host inflammatory response

12 Myxobolus wulii Synonyms: Myxosoma magna wu & Li, 1986; Myxobolus quanqiaoensis Wu & Wang, 1997; Infection site: hepatopancreas Molecular data: EF Honeycomb-like hepatocyte completely replaced by mature spores Very pathogenic sp., but the infection prevalence is low (below 10%)

13 Thelohanellus testudineus n. sp. Infection site: skin Molecular data: KC Showing plasmodium developing in the dermis of the skin and distinct inflammatory infiltration, Cluster in a Thelohanellus clade Cited from Liu et al. 2013

14 Myxobolus honghuensis n.sp. Synonyms: Myxosoma pharynae Xiu & Lu 2013;M. ampullicapsulatus Jsa.a Infection site: phyarynx Molecular data: KJ725074; JF Tofukasu-like cysts Many small plasmodia forming a big cyst Cluster with species with elongated spore

15 Current severe myxosporidiosis of gibel carp in China 1. Pharynx myxosporidiosis caused by Myxobolus honghuensis Infection prevalence: 70%; Mortality rate: up to 100%, especially in Northern Jiangsu Province; Infection season: from early May to late October, especially summer; Economic losses: 20 billion Chinese yuan per year; Control strategy: blocking the transmission and inactivating the infective actinospores by Ino-org (inosine-arginine);

16 Thelohanellus wangi n.sp. Infection site: gill filaments Molecular data: JX Cluster in a Thelohanellus clade Obvious epithelial hyperphasia at the anterior part of infected gill filaments and a thin connective tissue membrane delimited the plasmodium from cartilage cells of gill filaments

17 2. Gill myxosporidiosis caused by Thelohanellus wangi Infection prevalence: 60%; Mortality rate: up to 100%, especially under anoxia, crowded or other stress; Infection season: only from early April to late June on fry fish below 7cm in body length; Economic losses: 10 million Chinese yuan per year; Control strategy: blocking the transmission and inactivating the infective actinospores by Ino-org (inosine-arginine);

18 Thelohanellus wuhanensis n. sp. Infection site: skin Molecular data: JQ968687; JQ088179; HQ Membranous sheath surrounding spore cited from Liu et al plasmodium Raised plasmodium Showing plasmodium developing in the stratum spongiosum of dermis and many melanocytes gathering together around the plasmodia

19 3. Skin myxosporidiosis caused by Thelohanellus wuhanensis Infection prevalence: 70%; Mortality rate: no directly lethal,but up to 100% under anoxia, crowded or other stress condition; Infection season: only from early April to late June on fry fish below 7cm in body length; Economic losses: 10 million Chinese yuan per year; Control strategy: blocking the transmission and inactivating the infective actinospores by Ino-org (inosine-arginine);

20 Myxozoan life cycle studies in China Firstly reported Triactinomyxon type from Branchiura sowerbyi (Wang et al. 2000) no molecuar data Secondly Triactinomyxon type from Branchiura sowerbyi (Zhai et al. 2012) molecuar data: HM Short spore axis

21 Firstly Raabeia type from Branchiura sowerbyi (Zhai et al. 2013; Xi et al. 2013) molecular data:ab121146; HQ Firstly Aurantiactinomyxon type from Branchiura sowerbyi (Xi et al. 2013); molecular HQ613406

22 Firstly Guyenotia type from Branchiura sowerbyi (Xi et al. 2013); molecular data unavailable Secondly Raabeia type from Branchiura sowerbyi molecular data: HQ613410

23 Firstly Echinactionmyxon type from Branchiura sowerbyi; molecular data: EF branches at the end of caudal processes Secondly Echinactionmyxon type from Branchiura sowerbyi; molecular data: unavailable many branches at the end of caudal processes

24 Thirdly Triactinomyxon from Branchiura molecuar data: JX Firstly Neoactinomyxum type from Branchiura sowerbyi; molecuar data: KP642135, JX458816

25 Molecular data match Experimental infection and molecular match Molecular data match Molecular data match

26 Acknowledgements: Dr. Gu ZM and Dr. Liu Y from Huazhong Agricultural University, Wuhan, China; Prof. Csaba Székely and Prof. Molnár Kálmán from Institute for Veterinary Medical Research, Centre for Agricultural Research, HAS; Dr. Zhai and Dr. Ye LT from Institute of Hydrobiology, Chinese Academy of Sciences; Thank you for your attention