Taxonomy, diversity and diagnosis of Tetrahymenosis, and its recent identification measures in aquaculture inferred from infections in Northeastern China

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Increasing knowledge and identification of Tetrahymenosis are important. Methods Four parasitic Tetrahymena species were collected from eight commercially farmed fishes in Harbin, northeastern China. Specific oligonucleotide probe and fluorescence in situ hybridization staining was designed and tested. Hoechst33342 staining methods, gene sequencing and phylogenetic analyses were also conducted. Results Tetrahymena pyriformis, T. vorax, T. chironomi and T. bergeri are the four species responsible for Tetrahymenosis in various fishes in Harbin. Taxonomy, diagnosis, diversity, pathogenicity, and histopathology of Tetrahymenosis were supplemented, analyzed and summaried, based on the present and previous work. The term Tetrahymenosis is diagnosised as diseases affecting a number of fishes, crustaceans, mollus, beetle, dragonfly, salmon, slug, midge larvae and freshwater mussel species that caused by ciliates of the genus Tetrahymena which constitutes an abundant group inhabit various aquaculture and natural habitats. Improved classification of parasitic Tetrahymena was provided. Phylogeny of parasitic Tetrahymena species was studied, and an SSU-rDNA targeted oligonucleotide probe labeled with a fluorochrome was designed, and the FISH protocol was optimized for identification of Tetrahymena species, Conclusions The manifestations of histopathology in fish typically include lesions on the body surface, and affected organs include the skin, musculature, viscera, eye socket and spinal cord; masses of ciliates can be detected in copious amount of mucus and between spaces in the damaged tissues. Improved classification of parasitic Tetrahymena is provided: facultatively parasitic forms ( T. pyriformis , T. rostrate , T. bergeri and T. vorax ), facultatively free-living forms ( T. chironomi , T. corlissi, T. rotunda, T. glochidiophila and T. papula ) and parasitic forms ( T. stegomyiae and T. limacis ).The phylogenic results indicate that Tetrahymena spp. belonging to ‘borealis’ group have a greater probability to become parasitism. the method of which can be used for quick and early detection of Tetrahymenosis. Parasitology Aquaculture and Mariculture Tetrahymenosis facultative parasitic ciliates Simultaneous fluorescence in situ hybridization diversity histopathology Figures Figure 1 Figure 2 Figure 3 Figure 4 Background Tetrahymena , assigned to class Oligohymenophorea and family Tetrahymenidae, is a pyriform-shaped ciliate, with buccal cavity equipped with one paroral membrane and three serially arranged membranelles [ 1 – 15 ]. Occasionally, this microorganism can become parasitic and affect a wide range of hosts, including ornamental fish, edible fish, beetle, dragonfly, salmon, slug, midge larvae and freshwater mussel [ 16 – 21 ]. Pathogenic characteristics of infected fishes by Tetrahymena spp. are similar: possessing typical whitish lesions on the body surface, and affected organs include the skin, musculature, viscera, eye socket and spinal cord; masses of ciliates can be detected in copious amount of mucus and between spaces in the damaged tissues [ 16 – 18 , 22 ]. Some of them are facultative parasites while others might be obligate parasites [ 5 ]. The most pathogenic species of this genus, according to published literature, is T. corlissi , which often leads to a systemic infection and well-known as “guppy killer disease” [ 23 – 25 ]. They are responsible for serious economic losses in commercial fish farms around the world. While morphological method is the routine way to identify Tetrahymena spp., it has the limitations as follows: 1) the commonly used silver staining methods for reliable identification need considerable experience and staining failure is often the case, making species identification difficult, 2) parasitic Tetrahymena species are hard to control when massive infection occurs. Thus, early detection, quantification, and control of these infectious pathogens are important in aquaculture [ 1 , 26 ]. FISH with specific fluorochrome-labeled oligonucleotide probes is a staining technique that allows molecular identification of targeted organisms in mixed assemblages by means of a fluorescence microscopy, this method can overcome the drawbacks of the morphological method and yield prompt detection, and has been applied successfully to pathogenic ciliates (e. g. Pseudocohnilembus persalinus, Boveria labialis and B. subcylindrica ) [ 27 – 30 ]. However, to date, there are no general fluorochrome-labeled oligonucleotide probes for parasitic Tetrahymena spp. Methods Ciliates isolation, cultivation and morphological identification . Tetrahymena pyriformis, T. vorax, T. chironomi and T. bergeri were collected from commercially farmed fishes e. g. Malapterurus electricus , Tetraodon palembangensis , Poecilia latipinna , Channa aurantimaculata , Carassius auratus , zebrafish, Phoxinus lagowskii, Pelteobagrus fulvidrac , from fish farms or markets in Harbin, northeastern China between September 2017 and November 2019. Mucus or tissue were spread on glass slides using glass slides and the fresh mounts examined under a light microscope (20X and 40X objective) to confirm infection. Slides containing ciliates were selected for silver staining and morphological description. Cells were cultured at 25 °C and maintained in a culture medium (sterilized distilled water boiled with fish meat, stored at -20 ° C). Live observations were made under a microscope with phase-contrast illumination. Silver carbonate [ 31 ] and protargol [ 32 ] staining methods were used to reveal the infraciliature. All measurements of stained specimens were made under oil immersion (1250 x). Design and test of the specific oligonucleotide probe and FISH staining . Probe (nucleotides, 19; GC contents, 47.37%; nucleotide-nucleotide Tm, 53.1 o C) was designed using the probe design tool as implemented in the ARB software package for the SSU-rDNA sequence of the present Tetrahymena pyriformis, T. vorax, T. chironomi and T. bergeri , and other Tetrahymena species from GenBank [ 30 ]. Generated probes were checked against the GenBank sequence collection by a standard nucleotide-nucleotide BLAST search [ 33 ]. FISH was used to visualize Tetrahymena spp. above both in field samples and a mixture of species as well as Pseudocohnilembus persalinus and Uronema marinum that frequently occurred in the same hosts as the negative controls. Cells were fixed with 50% Bouin’s solution and filtered onto a 2-µm-pore-size cellulose nitrate membrane (25 mm in diameter) using low under pressure. The membrane was then washed five times with 2 ml of filtered sterile water. The basic hybridization followed the protocol of Stoeck et al. (2003) and Zhan et al. (2014). Hoechst33342 staining methods . Cells of parasitic Tetrahymena species were stained and fixed in 4% PFA for 20 min at room temperature, then were washed with PBS for 5 min at room temperature. Cells were then incubated with Hoechst 33342 (100 ng/mL in PBS) in the dark for 15 min at room temperature, then cells were washed with PBS for 5 min at room temperature. At last, cells were viewed using a fluorescence microscope with emission 350 nm and a blue/cyan emission filter (460 nm) (Fig. 3 ) [ 34 ]. Gene sequencing and Phylogenetic analyses . Single cells of Tetrahymena pyriformis, T. vorax, T. chironomi and T. bergeri were washed individually with distilled water. Genomic DNA was extracted from five cells using a DNeasy Blood & Tissue Kit (Qiagen, Hilden, Germany), following the manufacturer’s instructions. The SSU rRNA gene was amplified with the primers 82F- (5’-GAA ACT GCG AAT GGC TC-3’) and 18 s-R (5’-TGA TCC TTC TGC AGG TTC ACC TAC-3’) [ 35 ]. Bidirectional sequencing was performed by the Shanghai Sunny Biotechnology Company (Shanghai, China). The SSU rRNA gene sequences of T. pyriformis, T. vorax, T. chironomi and T. bergeri were aligned with sequences of other related taxa on the European Bioinformatics Institute web server (URL: http://www.ebi.ac.uk/Tools/msa/muscle/ ) using the MUSCLE package. Resulting alignments were refined by trimming both ends using BioEdit 7.0.5.2 [ 36 ]. Bayesian inference (BI) analysis was carried out with MrBayes on XSEDE v3.2.6 (Ronquist and Huelsenbeck 2003) on CIPRES Science Gateway [ 37 ] using the GTR + I + G evolutionary model as the best-fit model selected by MrModeltest v.2 [ 38 ] according to the Akaike Information Criterion (AIC). A Maximum Likelihood (ML) tree was constructed using RAxML-HPC2 v. 8.2.10 [ 39 ] on the CIPRES Science Gateway with the optimal model GTR + I + G evolutionary model as the best model according to the AIC criterion selected by the program Modeltest v.3.4 [ 40 ]. Node support came from 1000 bootstrap replicates. TreeView v.1.6.6 [ 41 ] and MEGA v5 were used to visualize tree topologies. Results Taxonomy The term Tetrahymenosis covers diseases affecting a number of fishes, crustaceans, mollus, beetle, dragonfly, salmon, slug, midge larvae and freshwater mussel species that caused by ciliates of the genus Tetrahymena which constitutes an abundant group inhabit various aquaculture and natural habitats (Table 1). Tetrahymena belongs to the phylum Ciliophora, Class Oligohymenophorea, order Tetrahymenida and family Tetrahymenidae [4]. Currently, the genus Tetrahymena Furgason, 1940 (Ciliophora, Hymenostomatida) is comprised of over 40 species [2–5, 13, 19]. Tetrahymena is generally pyriform-shaped, uniformly covered with meridional kineties, the buccal cavity is equipped with one curved undulating membrane with paired basal bodies organized in zigzag pattern (i.e. stichodyad), located on right edge of the buccal cavity, and three parallel membranelles. Within species, size variations depend on culture medium, physiological state and life stage (Corliss, 1973). The life cycle of Tetrahymena species is typified by the following developmental stages: theront, trophont, tomont and tomite. Diversity of parasitic Tetrahymena Tetrahymena species commonly exhibits a whole range of existence from the completely free-living state through stages of facultative parasitism to obligate endoparasitism [6]. Corliss (1971a) classified Tetrahymena parasitism into three types: facultatively parasitic forms (generally found free-living in nature but for which a parasitic existence is not uncommon), facultatively free-living forms (typically found in association with a host but capable in nature as well as experimentally) and parasitic forms (as endoparasites associated with specific hosts). To date, about ten Tetrahymena species ( T. corlissi, T. pyriformis, T. stegomyiae , T. chironomi, T. vorax, T. rostrata, T. limacis, T. rotunda, T. glochidiophila and T. papula ; Table 1) have been generally reported as pathogenicity to various invertebrates and vertebrates [16–18, 22]. In the current work, T. pyriformis, T. vorax, T. chironomi and T. bergeri are the four species isolated from fish farms or markets in Harbin. The cells commonly showed a high densities and vitalities when just isolated from the various fishes (Figs 1, 2). The detailed morphology of the four species are reported before, and the Harbin populations of the four species are similar with those previous descriptions (Figure 2) [1, 3, 7, 42]. The topologies of BI trees are similar to that of ML trees, so only the ML trees are shown in Figure 4 with branch support values for both analyses. ­­­­In the consensus topology, all the parasitic Tetrahymena spp. (Table 1) having sequence data are selected to constructed trees. As is shown, all the parasitic forms cluster in the ‘borealis’ group. The clustering pattern indicates that ‘borealis” Tetrahymena spp. might have a greater probability to become parasitism [13, 43]. In addition, Harbin population of T. pyriformis, T. vorax, and T. bergeri cluster with their previous populations, respectively, which also supports the morphological identifications. Diagnostic features: Pathogenicity/Histopathology. As pathogenic parasites, histopathology of parasitic Tetrahymena spp. have been studied in detail and reported many times [16, 18, 22, 44–48]: possessing typical whitish lesions on the body surface, and affected organs include the skin, musculature, viscera, eye socket and spinal cord; masses of ciliates can be detected in copious amount of mucus and between spaces in the damaged tissues. In fishes, extensive studies reporting the organ-specific pathological changes when infected with Tetrahymenosis are available, and typical gross lesions of Tetrahymena infection are characterized by protrusion of scales, swelling, ulcerative wounds on the skin, blindness, and protrusion of eyes [47, 48, 22, 18]. In our recent study, four Tetrahymena species were isolated Malapterurus electricus , Tetraodon palembangensis , Poecilia latipinna , Channa aurantimaculata , Carassius auratus , zebrafish, Phoxinus lagowskii, Pelteobagrus fulvidrac from farms in Harbin . At arrival, fish did not have any apparent gross signs of Tetrahymena infection. Nonetheless, the parasite was observed in fresh-mount preparations on the skin, in the gills and in internal organs (Figure 2). Noticeable, many cells of Tetrahymena bergeri were observed in the skin mucus of Phoxinus lagowskii , individuals of P. lagowskii exhibited symptoms that no feeding, staying upside down, gasping attack edges and secreting a lot of mucus (Figure 3G, H, arrowhead). Tetrahymena pyriformis were observed from guts of diseased zebrafish (Figure 3F, arrowhead). Pelteobagrus fulvidraco had many large, round deep skin lesions all over the body with hyperaemic margins exposing the skeletal muscle, and scrapings revealed the presence of T. vorax (Figure 2G, arrowheads). Gills of all diseased fish were shriveling, dark and anaemic, gills of several fish had dark-red ulceration (Figure 2I; arrowhead). Some cells of T. vorax were also detected in gaps among the gill filaments. Skin lesions or mucus of fishes were directly stained in vivo by Hoechst33342 staining method, and Figure 3A–C show the process of facultatively parasitic Tetrahymena sp. eating bacteria around the cell (Figure 3B), and Tetrahymena sp. eating tissues (skin or gill) as well as bacteria of host (Figure 3C). Cytoplasm of Tetrahymena sp. always contain many (approximately 2–5 μm in diameter) bacteria-or fish tissue- filled food vacuoles and variable-sized granules, distributed randomly (Figure 3C, arrowhead). The result indicates that facultatively parasitic Tetrahymena sp. can physically devour tissue, not only by producing enzyme [49]. Detection and identification using FISH The probe Tetr2020 evaluated with the probe match tool of the ARB software package and the GenBank BLAST tool showed that they are specific to Tetrahymena spp. The probe Tetr2020 (5 ’ - TGTAGTAGCCGTTTCTCAG-3 ’ ) had at least two mismatches to other closely related species like Uronema spp., Pseudocohnilembus spp. and Cyclidium spp. FISH with the probes Tetr2020 resulted in the presence of a red fluorescence signal each for T. pyriformis, T. vorax, T. chironomi and T. bergeri (Figure 3D, E), clearly distinguishable from the faint autofluorescence signal achieved with negative-control hybridizations using the Tetr2020 probe to hybridize the untargeted ciliates Pseudocohnilembus persalinus , Uronema marinum and isolated from the same hosts (Figure 3F). Thus, the probes Tetr2020 is also a general probe suitable for widely and rapidly detecting Tetrahymenosis. Discussion Corliss (1971a) classified Tetrahymena parasitism into facultatively parasitic forms ( T. pyriformis and T. rostrata ), facultatively free-living forms ( T. chironomi and T. corlissi ) and parasitic forms ( T. stegomyiae and T. limacis ). However, based on later and the present works (Tables 1, 2), supplementary classification is as follows: facultatively parasitic forms ( T. pyriformis , T. rostrate , T. bergeri and T. vorax ), facultatively free-living forms ( T. chironomi , T. corlissi, T. rotunda, T. glochidiophila and T. papula ) and parasitic forms ( T. stegomyiae and T. limacis ). and the morphology of pathogens and the hosts and data sources are compared in Tables 1 and 2. Tetrahymena bergeri is reported in the present work as facultatively parasitic forms in Channa aurantimaculata for the first time (free-living reported before), the reason possibly is that high latitude cold areas may have multiple life-styles when cold weather, due to the slow degradations of microorganisms and insufficiency of food [50]. Except for the ten species showed above, many unidentified Tetrahymena sp. conducted infections were also recorded, e. g. Stolk (1960) isolated Tetrahymena sp. from the central nervous system of larvae of Cyprinous carpio , Abramis brama and Ameiurus sp., Kim et al., (2002) examined the fish imported from Southeast Asian countries into Korea and reported of infection from the consignment of ornamental fish, Laoprasert et al., (2001) reported of tetrahymenosis in guppies from local fish farms, appearing as a white ulcerative disease. Thilakaratne et al., (2003) reported of Tetrahymena sp. infection in guppies from commercial ornamental fish farms in Sri Lanka, along with other parasites in different ornamental fish species. Pimenta-Leibowitz et al., (2005) reported that poor environmental and adverse physiological conditions pre-dispose to infections with Tetrahymena sp. As a histophagous parasite, Tetrahymena disintegrates host tissue and feeds on cell debris. Infection is showed by whitish patches due to the masses of ciliates in copious amount of mucus, patches usually located on the dorso-lateral skin around the abdomen (Johnson, 1978). Typical gross lesions of Tetrahymena infection are characterized by protrusion of scales, swelling, ulcerative wounds on the skin, blindness, and protrusion of eyes (Johnson, 1978). Numerous Tetrahymena spp. may form a rim around the eye orbit (spectacled eye). The parasites route of entry into the body has not been reported. In histopathological analyses, the ciliates were found to be distributed between spaces in the damaged tissues, and extensive necrotic changes occur in the muscle and subdermal tissue. Necrosis of the epithelial cells, extending to the musculature, and disfigured dermal and subcutaneous tissue with edema and hemorrhage was also reported [24, 51]. Compared with Scuticociliatosis, Tetrahymenosis can be found in a more wide-ranging hosts (e. g. slugs, hick embryos, dragonfly, helgramite, roach, cockroach and caterpillar), and Tetrahymenosis is easier to discovered in freshwater fish (vs. mostly in marine habitat fishes for Scuticociliatosis) (24, 46, 48, 52). However, their major clinico-pathological manifestations are similar: anemia, weight loss, dark coloration, enteritis, excessive body mucus, yellowish intestinal mucus, loss of scales, hemorrhagic and/or bleached spots on the skin, and dermal necrotic lesions that finally destroy tissues lead to high mortalities [8, 16, 50, 52]. As a test case using probes for the identification of pathogenic ciliates in aquaculture, the present study utilized the probe Tetr2020 to unambiguously detect Tetrahymenosis (Figure 3, D). The general identification by strong fluorescence signals from the oligonucleotide probe can be used for quick and early detection of Tetrahymenosis infections on both invertebrates and vertebrates, especially fish. The FISH probe designed here has the potential to be used for confirming the geographical distribution of parasitic Tetrahymena spp. and detecting the possible dispersal of this facultative pathogen [27–30]. The FISH approach also provided some morphological information such as body shape, macro and micronuclei shape and number of micronuclei of parasitic Tetrahymena spp. Conclusions The manifestations of histopathology in fish typically include lesions on the body surface, and affected organs include the skin, musculature, viscera, eye socket and spinal cord; masses of ciliates can be detected in copious amount of mucus and between spaces in the damaged tissues. Improved classification of parasitic Tetrahymena is provided: facultatively parasitic forms ( T. pyriformis , T. rostrate , T. bergeri and T. vorax ), facultatively free-living forms ( T. chironomi , T. corlissi, T. rotunda, T. glochidiophila and T. papula ) and parasitic forms ( T. stegomyiae and T. limacis ).The phylogenic results indicate that Tetrahymena spp. belonging to ‘borealis’ group have a greater probability to become parasitism. the method of which can be used for quick and early detection of Tetrahymenosis. List of Abbreviations Fluorescence in situ hybridization FISH Declarations Ethics approval and consent to participate Not applicable. Consent for publication Availability of data and materials The datasets supporting the conclusions of this article are included within the article. Representative sequences are submitted to the GenBank database (accession numbers are provided in Figure 4). Competing interests PXM and CY work at Harbin Normal University, Harbin, China. The study was conducted as part of a research program to investigate Tetrahymenosis in aquaculture in Northeastern China. Funding Natural Science Foundation of China (project numbers: 31970498), Heilongjiang Province Science Foundation for Youths (project number: QC2017017). Authors' contributions The study design, protocol and were prepared by ZSX, PMM, LWY, SM, WCN, LJL, WX and ZL, and report of the study and reviewed by PXM and CY. PXM and his team at Harbin Normal University were responsible for the animal phase and data collection. PXM were responsible for compiling the first draft of the manuscript. All authors revised and approved the final version. Acknowledgements This work was supported by the Natural Science Foundation of China (project numbers: 31970498), Heilongjiang Province Science Foundation for Youths (project number: QC2017017). References Corliss JO. Tetrahymena chironomi nov., a ciliate from midge larvae, and the current status of facultative parasitism in the genus Tetrahymena . Parasitology. 1960;50:111–153. Corliss JO. Facultative parasitism in Protozoa and its possible evolutionary significance. (Abstr.) Parasitology. 1962;52: Corliss JO. Tetrahymena , a ciliate genus of unusual importance in modern biological research. Acta Protozool. 1965;3:1– Corliss JO. The value of ontogenetic data in reconstructing protozoan phylogenies. Trans Amer Micros Soc. 1968;87:1– Corliss JO. 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Investigations on ciliates living in lamelli branchiates of small waterbodies. Bull Acad Polonaise Sci. 1958;6:113– Hoffman GL, Landolt M, Camper JE, Coats DW, Stookey JL, Burek JD. A disease of freshwater fishes caused by Tetrahymena corliss Thompson, 1955, and a key for identification of holotrich ciliates of freshwater fishes. J Parasitol. 1975;6:217– Johnson SK. Tet diseases of tropical fishes and an evaluation of correction techniques. Fish Disease Diagnostic Laboratory F12, Texas A & M University, Austin. 1978. Kazubski SL. Trichia lubomirskii (Helicidae), a new host of Tetrahymena limacis (Warren, 1932) Kozloff, 1946 (Ciliata) and Zonitoides nitidus Mull. (Zonitidae), a new host of T. rostrata (Kahl, 1926) Corliss, 1952, in Poland. Bull Acad Polonaise Sci. 1958;6:247–52. Kim JH, Hayward CJ, Joh SJ, Heo GJ. Parasitic infections in live freshwater tropical fishes imported to Korea. Dis Aquat Organ. 2002;52:169– Kozloff EN. The morphology and systematic position of a holotrichous ciliate parasitizing Deroceras agreste (L.). J. Morph. 1946;79:445–65 Kozloff EN. A comparison of the parasitic phase of Tetrahymena limacis (Warren) with clones in culture, with particular reference to variability in the number of primary ciliary meridians. J Protozool. 1956a;3:20– Kozloff EN. Tetrahymena limacis from the pulmonate gastropods Monadenia fidelis and Prophysaon andersoni. J Protozool. 1956b;3:204– Kozloff EN. A species of Tetrahymena parasitic in the renal organ of the slug Derocercas reticidatum. J Protozool. 1957;4:75– Laird M. Parasites of Singapore mosquitoes, with particular reference to the significance of larval epibionts as an index of habitat pollution. Ecology. 1959;40:206– Laoprasert T, Khaisri C, Chinabut S. Study on white ulcerative disease in guppy ( Poecilia reticulata ). Proceeding of the Seminar on Fisheries 2001. The Aquatic Animal Health Research Institute (AAHRI) newsletter, abstracts from Volume 10 No.2. Bangkok, Thailand. 2001. Lynn DH, Molloy D, LeBrun R. Tetrahymena rotunda sp. (Hymenostomatida: Tetrahymenidae), a ciliate parasite of the hemolymph of Simulium (Diptera: Simuliidae). Trans Am Microsc Soc. 1981;134–141. Michelson EH. Distribution and pathogenicity of Tetrahymena limacis in the slug Deroceras reticulatum . Parasitology. 1971;62:125– Ronquist F, Huelsenbeck J. MrBayes 3: Bayesian phylogenetic inference under mixed models. Bioinformatics. 2003;19:1572–1574. Seaman GR, Tosney T, Berglund R, Goldberg G. Infectivity and recovery of Tetrahymena pyriformis strain S from adult female cockroaches ( Periplaneta americana ). J Protozool. 1972;19:644– Stout JD. The ecology, life history and parasitism of Tetrahymena [ Paraglaucoma ] rostrata (Kahl) Corliss. J Protozool.1954;1:211– Wakita K, Imai S, Hatai K. "Histopathological study on Tetrahymena infection in Dwarf Gourami (Colisa lalia)." 5th Symposium on Diseases in Asian Aquaculture. 2002. Wang Q, Luo S, Ghonimy A, Chen Y, Guo Z, Liu H, Zhang D. Effect of dietary l-carnitine on growth performance and antioxidant response in Amur minnow ( Phoxinus lagowskii Dybowskii ). Aquacult Nutr. 2019;25:749–760. Williams NE. The polymorphic life history of Tetrahymena patula . J Protozool. 1960; 7:10– Wilson MJ, Coyne C, Glen DM. Low temperatures suppress growth of the ciliate parasite, Tetrahymena rostrata , and pathogenicity to field slugs, Deroceras reticulatum . Biocontrol. Sci. Techn. 1998;8:181– Wright JF. Tetrahymena pyriformis (Ehrenberg) and corlissi Thompson parasitic in stream dwelling triclads (Platyhelminthes: Turbellaria). J Parasitol. 1981;67:131–133. Xue Z, Zhang Y, Lin M, Sun S, Gao W, Wang W. Effects of habitat fragmentation on the population genetic diversity of the Amur minnow ( Phoxinus lagowskii ), Mitochondrial DNA Part B 2017;2:331–336. Lwoff, A. Infection experimentale a Glaucoma piriformis chez Qalleria mellonella (Lepidoptere). C.R. Acad Sci Paris. 1924;178:1106– Windsor DA. Colpoda steinii and Tetrahymena limacis from terrestrial pulmonated gastropods collected in Illinois. (Abstr.) J Protozool. 1959;6:33. Janda V, JÍRovEC O. Uber kunstlieh hervorgerufenen Parasitismus eines freilebenden Ciliaten Glaucoma piriformis und Infektionsversuch mit Euglena gracilis and Spirochaeta biftexa. Vestnik Ceskoslov. Zool. spolec. Praze. (Mem. Soc. zool. tchicosl.) 1937;5:34– Corliss JO. 1951. Morphology and systematic status of pure culture strains in the Colpidium-Glaucoma-Leucophrys-Tetrahymena (Abstr.) Proc Amer Soc Protozool. 2:11. Tables Table 1. Morphological identification characteristics of parasitic Tetrahymena spp. Species Size in vivo Body shape Caudal cilia SK No. Mi No. CVP No. & position References T. corlissi an average 52 × 32 μm elongate, pointed anteriorly and rounded posteriorly present 25-31 1 2 or 3; SK6-10 16 T. pyriformis ca. 50 × 30 μm pear-shaped absent 15-25 N/A 2; SK5, 6 5 T. stegomyiae 60-100 μm pear-shaped absent 25-30 1 - 1 T. chironomi ca. 40 × 23 μm elongate absent 23-28 1 3; SK6-9 1 T. vorax < 150 μm posterior end is drawn out into a tail absent 18-23 0 2-6; SK5-10 7 T. rostrata 50-60 × 30-40 μm Vary present 36-58 1 3 or 4; SK6-12 5,7 T. limacis 40-55 × 30-40 μm Cucumber-shaped with strongly apiculate anterior end absent 26-46 1 2 or 3; 5-8 58 T. rotunda 40-90 μm (after staining) bluntly ovoid to globose present 50-66; -- not observed 64 T. papula 80-100 μm piriform, bag-shaped absent 33-45 1 2-6; SK9-16 78 Table 2. Reports of parasitic Tetrahymena species and their hosts . Parasite Host Data source T. bergeri Channa aurantimaculata From the present work T. chironomi Fourth-instar larva ( Chironomus plumosus ) 1 T. chironomi Phoxinus lagowskii From the present work T. corlissi Beetle, Dragonfly, Guppies, Helgramite, Tadpoles, Tent caterpillar 16 T. corlissi Guppies ( Poecilia reticulatus ) 54 T. corlissi Freshwater triclads 73 T. corlissi Atlantic Salmon Salmon salar 23 T. corlissi Fishes 46 T. corlissi Fresh fish 17 T. corlissi Guppies ( Poecilia reticulata ) 24 T. corlissi Guppies 25 T. corlissi Guppies ( Poecilia reticulata ) 22 T. corlissi Dwarf Gourami ( Colisa lalia ) 69 T. corlissi Freshwater ornamental fish 48 T. corlissi Guppies ( Poecilia reticulata ) 18 T. corlissi Golden perch ( Macquaria ambiqua ) 50 T. glochidiophila Freshwater mussel 21 T. limacis European slug ( Deroceras retioulatum) 58 T. limacis European slug ( Deroceras reticulatum ) , native slug ( Prophysaon andersoni ) , native snail ( Monadenia fidelis ) 59, 60 T. limacis Fresh-water clams 53 T. limacis Land snail ( Trichia lubomirskii ) 56 T. limacis European slug ( Deroceras reticulatum ) 76 T. limacis Slug ( Deroceras reticulatum ) 65 T. papula Tent caterpillar ( Malacosoma americana larva) 16 T. pyriformis Insects 75 T. pyriformis Insects 77 T. pyriformis Clams 53 T. pyriformis Beetle, Chick embryos, Dragonfly, Helgramite, Roach, cockroach, Tent caterpillar 16 T. pyriformis Oligochaete, Allolobophora caliginosa var. trapezoides 17 T. pyriformis Mosquitoes 62 T. pyriformis Adult female cockroaches ( Periplaneta americana ) 67 T. pyriformis Channel catfish ( Ictalurus punctatus ) 44 T. pyriformis Freshwater triclads 73 T. pyriformis Fishes 45 T. pyriformis Freshwater crayfish ( Cherax quadricarinatus ) 47 T. pyriformis Ornamental fishes 51 T. pyriformis Freshwater ornamental fish 48 T. pyriformis Phenacogrammus interruptus From the present work T. rostrata European slug ( Deroceras reticulatum ) 61 T. rostrata Land snail ( Zonitoides nitidua ) 56 T. rostrata European slug ( Deroceras reticulatum ) 16 T. rostrata Field slugs ( Deroceras reticulatum ) 72 T. rotunda Simulium 9 T. stegomyiae Mosquito 1 T. vorax Chick embryos, Cockroach, Dragonfly, Helgramite, Tent caterpillar 16 T. vorax Pelteobagrus fulvidraco From the present work Supplementary Files GAJFP.jpg Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-15239","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Short report","associatedPublications":[],"authors":[{"id":371262,"identity":"7b5de5a7-35e0-4209-8161-e6933f5e9316","order_by":1,"name":"Xuming Pan","email":"","orcid":"https://orcid.org/0000-0002-7071-1751","institution":"Harbin Normal University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Xuming","middleName":"","lastName":"Pan","suffix":""},{"id":371263,"identity":"e5ad7baf-2eb6-4709-93dd-d947db20f620","order_by":2,"name":"shuixian Zhang","email":"","orcid":"","institution":"Harbin Normal University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"shuixian","middleName":"","lastName":"Zhang","suffix":""},{"id":371264,"identity":"6a86cdc8-08f5-4b54-8c7e-7309f6cd2fe3","order_by":3,"name":"mengmeng Pan","email":"","orcid":"","institution":"Harbin Normal University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"mengmeng","middleName":"","lastName":"Pan","suffix":""},{"id":371265,"identity":"20966f61-b621-4e6d-80e9-08256a96b4ae","order_by":4,"name":"Wenyu Li","email":"","orcid":"","institution":"Harbin normal University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Wenyu","middleName":"","lastName":"Li","suffix":""},{"id":371266,"identity":"3479adfe-da58-4319-8aec-a3262ffffbfe","order_by":5,"name":"meng Sun","email":"","orcid":"","institution":"Harbin Normal University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"meng","middleName":"","lastName":"Sun","suffix":""},{"id":371267,"identity":"e0872704-a5fd-406b-93d2-be6b261cc264","order_by":6,"name":"chongnv Wang","email":"","orcid":"","institution":"Harbin Normal University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"chongnv","middleName":"","lastName":"Wang","suffix":""},{"id":371268,"identity":"1428d0d3-4a17-48be-bffb-37e086a58115","order_by":7,"name":"Jiale Liang","email":"","orcid":"","institution":"Harbin Normal University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Jiale","middleName":"","lastName":"Liang","suffix":""},{"id":371269,"identity":"1725794d-1d94-46ee-b7d8-b14a6becea24","order_by":8,"name":"xiang Wang","email":"","orcid":"","institution":"Harbin Normal University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"xiang","middleName":"","lastName":"Wang","suffix":""},{"id":371270,"identity":"fc37c642-0c2a-4f1a-9da6-8569417bb2eb","order_by":9,"name":"lei Zhu","email":"","orcid":"","institution":"Harbin Normal University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"lei","middleName":"","lastName":"Zhu","suffix":""},{"id":371271,"identity":"98f98b81-738d-4533-82c8-0abaa622e90d","order_by":10,"name":"Ying Chen","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAyElEQVRIiWNgGAWjYBACxmbmgw8+VPyX42dvIFILcztbsuGMM8zGkj0HiNTC3s+jJs3bxpxocCOBSC28zTzMhjPY2BIMbj7eeIOhxiaaoBbJZl6gX3h48iRvpxVbMBxLy20gpMWwmQ/oFwmJYr7bOWYSjA2HCWuxP8xjJs1jYJDYcPMMkVoYm0FaEhISJ9zgIVoLKJAPHAAGMtAvCcT4hbH/8MEHH/8dAEbl4Y03PtTYENaCDAwkEkhRDtFCqo5RMApGwSgYGQAApyFBWTsjlVEAAAAASUVORK5CYII=","orcid":"","institution":"Harbin Normal University","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Ying","middleName":"","lastName":"Chen","suffix":""}],"badges":[],"createdAt":"2020-02-25 13:52:48","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-15239/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-15239/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":595051,"identity":"eeff2954-f86f-4454-8a30-68acf4264c5f","added_by":"auto","created_at":"2020-03-04 17:11:51","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":5465709,"visible":true,"origin":"","legend":"Diseased fishes isolated from farms or aquaculture markets of Harbin, northeastern China infected by Tetrahymena spp. A. Malapterurus electricus; B. Tetraodon palembangensis; C, Poecilia latipinna; D, Channa aurantimaculata; E, Carassius auratus; F, zebrafish; G, H, Phoxinus lagowskii; I. J. Pelteobagrus fulvidraco. Scale bars: A. 10 cm (A, D); 4 cm (B, E, G, H, J).","description":"","filename":"Fig.1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-15239/v1/Fig. 1.jpg"},{"id":595052,"identity":"3a107a98-fbc8-437e-82c8-441bd2ed30c7","added_by":"auto","created_at":"2020-03-04 17:11:51","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":8308483,"visible":true,"origin":"","legend":"Photomicrographs of parasitic Tetrahymena species isolated from Harbin in vivo (A–D, F, H, K) and after silver carbonate staining (E, G, J, L). (A, B) Views of numerous parasitic Tetrahymena sp. in low magnification. (C–E) Tetrahymena pyriformis. (F, G) Tetrahymena vorax. (H–J) Tetrahymena chironomi. (K, L) Tetrahymena bergeri, arrowhead shows caudal cilium. Scale bars: 100 μm (A, B), 25 μm (C, D, F–L), 10 μm (E).","description":"","filename":"Fig.2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-15239/v1/Fig.2.jpg"},{"id":595053,"identity":"47ee587f-6424-4e0e-b15e-c44ee7795aaf","added_by":"auto","created_at":"2020-03-04 17:11:51","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":8329560,"visible":true,"origin":"","legend":"Simultaneous fluorescence in situ hybridization (FISH; D–F) and Hoechst33342 (A–C) staining of parasitic Tetrahymena species (FISH; A–D) and other test ciliates (F). (A) Views of numerous cells of parasitic Tetrahymena sp. in low magnification. (B) To show bacteria (arrowhead) around Tetrahymena sp. (C) To show the process that Tetrahymena sp. (arrow) eating tissue (arrowhead) as well as bacteria of host. (D, E) Presence of red fluorescence signals for T. pyriformis, T. vorax, T. chironomi and T. bergeri (arrowhead, signals of four species were similar, so only that of T. pyriformis are showed). (F) Other test ciliates (not assigned to Tetrahymena) after simultaneous fluorescence in situ hybridization. Scale bars: 50 μm (A, D, E), 20 μm (B, C).","description":"","filename":"Fig.3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-15239/v1/Fig.3.jpg"},{"id":595054,"identity":"2ef45e53-0956-470c-8a09-41c688114daa","added_by":"auto","created_at":"2020-03-04 17:11:52","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":3211697,"visible":true,"origin":"","legend":"Maximum likelihood tree inferred from SSU rDNA sequences, showing the position of phylogentic relationship of parasitic Tetrahymena species. Numbers at nodes represent the bootstrap values of Maximum likelihood out of 1,000 replicates and the posterior probability of Bayesian Inference. Fully supported (100%/1.00) branches are marked with solid circles. The scale bar corresponds to one substitutions per 100 nucleotide positions. HA refers ‘Harbin population’.","description":"","filename":"Fig.4.jpg","url":"https://assets-eu.researchsquare.com/files/rs-15239/v1/Fig.4.jpg"},{"id":13491983,"identity":"724e74db-4f09-41ab-a1d4-77a630399118","added_by":"auto","created_at":"2021-09-16 22:29:54","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1022907,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-15239/v1/b49e1b09-cd48-4991-9691-e9eb5b110195.pdf"},{"id":595050,"identity":"c315f6e5-7a29-4c36-8bb0-4091d30535a5","added_by":"auto","created_at":"2020-03-04 17:11:50","extension":"jpg","order_by":0,"title":"","display":"","copyAsset":false,"role":"supplement","size":6930587,"visible":true,"origin":"","legend":"","description":"","filename":"GAJFP.jpg","url":"https://assets-eu.researchsquare.com/files/rs-15239/v1/GA-JFP.jpg"}],"financialInterests":"","formattedTitle":"Taxonomy, diversity and diagnosis of Tetrahymenosis, and its recent identification measures in aquaculture inferred from infections in Northeastern China","fulltext":[{"header":"Background","content":" \u003cp\u003e \u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003eTetrahymena\u003c/span\u003e, assigned to class Oligohymenophorea and family Tetrahymenidae, is a pyriform-shaped ciliate, with buccal cavity equipped with one paroral membrane and three serially arranged membranelles [\u003cspan additionalcitationids=\"CR2 CR3 CR4 CR5 CR6 CR7 CR8 CR9 CR10 CR11 CR12 CR13 CR14\" citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. Occasionally, this microorganism can become parasitic and affect a wide range of hosts, including ornamental fish, edible fish, beetle, dragonfly, salmon, slug, midge larvae and freshwater mussel [\u003cspan additionalcitationids=\"CR17 CR18 CR19 CR20\" citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. Pathogenic characteristics of infected fishes by \u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003eTetrahymena\u003c/span\u003e spp. are similar: possessing typical whitish lesions on the body surface, and affected organs include the skin, musculature, viscera, eye socket and spinal cord; masses of ciliates can be detected in copious amount of mucus and between spaces in the damaged tissues [\u003cspan additionalcitationids=\"CR17\" citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e, \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. Some of them are facultative parasites while others might be obligate parasites [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. The most pathogenic species of this genus, according to published literature, is \u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003eT. corlissi\u003c/span\u003e, which often leads to a systemic infection and well-known as \u0026ldquo;guppy killer disease\u0026rdquo; [\u003cspan additionalcitationids=\"CR24\" citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. They are responsible for serious economic losses in commercial fish farms around the world.\u003c/p\u003e \u003cp\u003eWhile morphological method is the routine way to identify \u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003eTetrahymena\u003c/span\u003e spp., it has the limitations as follows: 1) the commonly used silver staining methods for reliable identification need considerable experience and staining failure is often the case, making species identification difficult, 2) parasitic \u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003eTetrahymena\u003c/span\u003e species are hard to control when massive infection occurs. Thus, early detection, quantification, and control of these infectious pathogens are important in aquaculture [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. FISH with specific fluorochrome-labeled oligonucleotide probes is a staining technique that allows molecular identification of targeted organisms in mixed assemblages by means of a fluorescence microscopy, this method can overcome the drawbacks of the morphological method and yield prompt detection, and has been applied successfully to pathogenic ciliates (e. g. \u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003ePseudocohnilembus persalinus, Boveria labialis\u003c/span\u003e and \u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003eB. subcylindrica\u003c/span\u003e) [\u003cspan additionalcitationids=\"CR28 CR29\" citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. However, to date, there are no general fluorochrome-labeled oligonucleotide probes for parasitic \u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003eTetrahymena\u003c/span\u003e spp.\u003c/p\u003e "},{"header":"Methods","content":" \u003cp\u003e \u003cspan type=\"Bold\" class=\"Bold\" name=\"Emphasis\"\u003eCiliates isolation, cultivation and morphological identification\u003c/span\u003e.\u003c/p\u003e \u003cp\u003e \u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003eTetrahymena pyriformis, T. vorax, T. chironomi\u003c/span\u003e and \u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003eT. bergeri\u003c/span\u003e were collected from commercially farmed fishes e. g. \u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003eMalapterurus electricus\u003c/span\u003e, \u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003eTetraodon palembangensis\u003c/span\u003e, \u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003ePoecilia latipinna\u003c/span\u003e, \u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003eChanna aurantimaculata\u003c/span\u003e, \u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003eCarassius auratus\u003c/span\u003e, zebrafish, \u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003ePhoxinus lagowskii, Pelteobagrus fulvidrac\u003c/span\u003e, from fish farms or markets in Harbin, northeastern China between September 2017 and November 2019. Mucus or tissue were spread on glass slides using glass slides and the fresh mounts examined under a light microscope (20X and 40X objective) to confirm infection. Slides containing ciliates were selected for silver staining and morphological description. Cells were cultured at 25\u0026nbsp;\u0026deg;C and maintained in a culture medium (sterilized distilled water boiled with fish meat, stored at -20 \u0026deg; C). Live observations were made under a microscope with phase-contrast illumination. Silver carbonate [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e] and protargol [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e] staining methods were used to reveal the infraciliature. All measurements of stained specimens were made under oil immersion (1250 x).\u003c/p\u003e \u003cp\u003e \u003cspan type=\"Bold\" class=\"Bold\" name=\"Emphasis\"\u003eDesign and test of the specific oligonucleotide probe and FISH staining\u003c/span\u003e.\u003c/p\u003e \u003cp\u003eProbe (nucleotides, 19; GC contents, 47.37%; nucleotide-nucleotide Tm, 53.1 \u003csup\u003eo\u003c/sup\u003eC) was designed using the probe design tool as implemented in the ARB software package for the SSU-rDNA sequence of the present \u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003eTetrahymena pyriformis, T. vorax, T. chironomi\u003c/span\u003e and \u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003eT. bergeri\u003c/span\u003e, and other \u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003eTetrahymena\u003c/span\u003e species from GenBank [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. Generated probes were checked against the GenBank sequence collection by a standard nucleotide-nucleotide BLAST search [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]. FISH was used to visualize \u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003eTetrahymena\u003c/span\u003e spp. above both in field samples and a mixture of species as well as \u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003ePseudocohnilembus persalinus\u003c/span\u003e and \u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003eUronema marinum\u003c/span\u003e that frequently occurred in the same hosts as the negative controls. Cells were fixed with 50% Bouin\u0026rsquo;s solution and filtered onto a 2-\u0026micro;m-pore-size cellulose nitrate membrane (25\u0026nbsp;mm in diameter) using low under pressure. The membrane was then washed five times with 2\u0026nbsp;ml of filtered sterile water. The basic hybridization followed the protocol of Stoeck et al. (2003) and Zhan et al. (2014).\u003c/p\u003e \u003cp\u003e \u003cspan type=\"Bold\" class=\"Bold\" name=\"Emphasis\"\u003eHoechst33342 staining methods\u003c/span\u003e.\u003c/p\u003e \u003cp\u003eCells of parasitic \u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003eTetrahymena\u003c/span\u003e species were stained and fixed in 4% PFA for 20\u0026nbsp;min at room temperature, then were washed with PBS for 5\u0026nbsp;min at room temperature. Cells were then incubated with Hoechst 33342 (100\u0026nbsp;ng/mL in PBS) in the dark for 15\u0026nbsp;min at room temperature, then cells were washed with PBS for 5\u0026nbsp;min at room temperature. At last, cells were viewed using a fluorescence microscope with emission 350\u0026nbsp;nm and a blue/cyan emission filter (460\u0026nbsp;nm) (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e3\u003c/span\u003e) [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cspan type=\"Bold\" class=\"Bold\" name=\"Emphasis\"\u003eGene sequencing and Phylogenetic analyses\u003c/span\u003e.\u003c/p\u003e \u003cp\u003eSingle cells of \u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003eTetrahymena pyriformis, T. vorax, T. chironomi\u003c/span\u003e and \u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003eT. bergeri\u003c/span\u003e were washed individually with distilled water. Genomic DNA was extracted from five cells using a DNeasy Blood \u0026amp; Tissue Kit (Qiagen, Hilden, Germany), following the manufacturer\u0026rsquo;s instructions. The SSU rRNA gene was amplified with the primers 82F- (5\u0026rsquo;-GAA ACT GCG AAT GGC TC-3\u0026rsquo;) and 18\u0026nbsp;s-R (5\u0026rsquo;-TGA TCC TTC TGC AGG TTC ACC TAC-3\u0026rsquo;) [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]. Bidirectional sequencing was performed by the Shanghai Sunny Biotechnology Company (Shanghai, China). The SSU rRNA gene sequences of \u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003eT. pyriformis, T. vorax, T. chironomi\u003c/span\u003e and \u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003eT. bergeri\u003c/span\u003e were aligned with sequences of other related taxa on the European Bioinformatics Institute web server (URL: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://www.ebi.ac.uk/Tools/msa/muscle/\u003c/span\u003e\u003c/span\u003e) using the MUSCLE package. Resulting alignments were refined by trimming both ends using BioEdit 7.0.5.2 [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e]. Bayesian inference (BI) analysis was carried out with MrBayes on XSEDE v3.2.6 (Ronquist and Huelsenbeck 2003) on CIPRES Science Gateway [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e] using the GTR\u0026thinsp;+\u0026thinsp;I\u0026thinsp;+\u0026thinsp;G evolutionary model as the best-fit model selected by MrModeltest v.2 [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e] according to the Akaike Information Criterion (AIC). A Maximum Likelihood (ML) tree was constructed using RAxML-HPC2 v. 8.2.10 [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e] on the CIPRES Science Gateway with the optimal model GTR\u0026thinsp;+\u0026thinsp;I\u0026thinsp;+\u0026thinsp;G evolutionary model as the best model according to the AIC criterion selected by the program Modeltest v.3.4 [\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e]. Node support came from 1000 bootstrap replicates. TreeView v.1.6.6 [\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e] and MEGA v5 were used to visualize tree topologies.\u003c/p\u003e "},{"header":"Results","content":"\u003cp\u003e\u003cstrong\u003eTaxonomy\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe term Tetrahymenosis covers diseases affecting a number of fishes, crustaceans, mollus, beetle, dragonfly, salmon, slug, midge larvae and freshwater mussel species that caused by ciliates of the genus \u003cem\u003eTetrahymena\u003c/em\u003e which constitutes an abundant group inhabit various aquaculture and natural habitats (Table 1). \u003cem\u003eTetrahymena \u003c/em\u003ebelongs to the phylum Ciliophora, Class Oligohymenophorea, order Tetrahymenida and family Tetrahymenidae [4]. Currently, the genus \u003cem\u003eTetrahymena\u003c/em\u003e Furgason, 1940 (Ciliophora, Hymenostomatida) is comprised of over 40 species [2\u0026ndash;5, 13, 19]. \u003cem\u003eTetrahymena\u003c/em\u003e is generally pyriform-shaped, uniformly covered with meridional kineties, the buccal cavity is equipped with one curved undulating membrane with paired basal bodies organized in zigzag pattern (i.e. stichodyad), located on right edge of the buccal cavity, and three parallel membranelles. Within species, size variations depend on culture medium, physiological state and life stage (Corliss, 1973). The life cycle of \u003cem\u003eTetrahymena\u003c/em\u003e species is typified by the following developmental stages: theront, trophont, tomont and tomite.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDiversity of parasitic \u003cem\u003eTetrahymena\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eTetrahymena\u003c/em\u003e species commonly exhibits a whole range of existence from the completely free-living state through stages of facultative parasitism to obligate endoparasitism [6]. Corliss (1971a) classified \u003cem\u003eTetrahymena\u003c/em\u003e parasitism into three types: facultatively parasitic forms (generally found free-living in nature but for which a parasitic existence is not uncommon), facultatively free-living forms (typically found in association with a host but capable in nature as well as experimentally) and parasitic forms (as endoparasites associated with specific hosts). To date, about ten \u003cem\u003eTetrahymena \u003c/em\u003especies (\u003cem\u003eT. corlissi, T. pyriformis, \u003c/em\u003e\u003cem\u003eT. stegomyiae\u003c/em\u003e\u003cem\u003e, T. chironomi, T. vorax, T. rostrata, \u003c/em\u003e\u003cem\u003eT. limacis, T. rotunda,\u0026nbsp;T. glochidiophila \u003c/em\u003eand \u003cem\u003eT. papula\u003c/em\u003e; Table 1) have been generally reported as pathogenicity to various invertebrates and vertebrates [16\u0026ndash;18, 22].\u003c/p\u003e\n\u003cp\u003eIn the current work, \u003cem\u003eT.\u003c/em\u003e\u003cem\u003e pyriformis, T. vorax, \u003c/em\u003e\u003cem\u003eT. \u003c/em\u003e\u003cem\u003echironomi\u003c/em\u003e and\u003cem\u003e T. bergeri \u003c/em\u003eare the four species isolated from fish farms or markets in Harbin. The cells commonly showed a high densities and vitalities when just isolated from the various fishes (Figs 1, 2). The detailed morphology of the four species are reported before, and the Harbin populations of the four species are similar with those previous descriptions (Figure 2) [1, 3, 7, 42]. The topologies of BI trees are similar to that of ML trees, so only the ML trees are shown in Figure 4 with branch support values for both analyses. \u0026shy;\u0026shy;\u0026shy;\u0026shy;In the consensus topology, all the parasitic \u003cem\u003eTetrahymena \u003c/em\u003espp. (Table 1) having sequence data are selected to constructed trees. As is shown, all the parasitic forms cluster in the \u0026lsquo;borealis\u0026rsquo; group. The clustering pattern indicates that \u0026lsquo;borealis\u0026rdquo; \u003cem\u003eTetrahymena\u003c/em\u003e spp. might have a greater probability to become parasitism [13, 43]. In addition, Harbin population of \u003cem\u003eT.\u003c/em\u003e\u003cem\u003e pyriformis, T. vorax,\u003c/em\u003e and\u003cem\u003e T. bergeri\u003c/em\u003e cluster with their previous populations, respectively, which also supports the morphological identifications.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDiagnostic features: \u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePathogenicity/Histopathology.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAs pathogenic parasites, histopathology of parasitic \u003cem\u003eTetrahymena\u003c/em\u003e spp. have been studied in detail and reported many times [16, 18, 22, 44\u0026ndash;48]: possessing typical whitish lesions on the body surface, and affected organs include the skin, musculature, viscera, eye socket and spinal cord; masses of ciliates can be detected in copious amount of mucus and between spaces in the damaged tissues. In fishes, extensive studies reporting the organ-specific pathological changes when infected with Tetrahymenosis are available, and typical gross lesions of \u003cem\u003eTetrahymena \u003c/em\u003einfection are characterized by protrusion of scales, swelling, ulcerative wounds on the skin, blindness, and protrusion of eyes [47, 48, 22, 18]. In our recent study, four \u003cem\u003eTetrahymena\u003c/em\u003e species were isolated \u003cem\u003eMalapterurus electricus\u003c/em\u003e, \u003cem\u003eTetraodon palembangensis\u003c/em\u003e, \u003cem\u003ePoecilia latipinna\u003c/em\u003e, \u003cem\u003eChanna aurantimaculata\u003c/em\u003e, \u003cem\u003eCarassius auratus\u003c/em\u003e, zebrafish, \u003cem\u003ePhoxinus lagowskii, Pelteobagrus fulvidrac\u003c/em\u003e from farms in Harbin\u003cem\u003e. \u003c/em\u003eAt arrival, fish did not have any apparent gross signs of \u003cem\u003eTetrahymena\u003c/em\u003e infection. Nonetheless, the parasite was observed in fresh-mount preparations on the skin, in the gills and in internal organs (Figure 2). Noticeable, many cells of \u003cem\u003eTetrahymena bergeri\u003c/em\u003e were observed in the skin mucus of \u003cem\u003ePhoxinus lagowskii\u003c/em\u003e, individuals of \u003cem\u003eP. lagowskii\u003c/em\u003e exhibited symptoms that no feeding, staying upside down, gasping attack edges and secreting a lot of mucus (Figure 3G, H, arrowhead). \u003cem\u003eTetrahymena \u003c/em\u003e\u003cem\u003epyriformis \u003c/em\u003ewere observed from guts of diseased zebrafish (Figure 3F, arrowhead). \u003cem\u003ePelteobagrus fulvidraco\u003c/em\u003e had many large, round deep skin lesions all over the body with hyperaemic margins exposing the skeletal muscle, and scrapings revealed the presence of \u003cem\u003eT. vorax \u003c/em\u003e(Figure 2G, arrowheads). Gills of all diseased fish were shriveling, dark and anaemic, gills of several fish had dark-red ulceration (Figure 2I; arrowhead). Some cells of \u003cem\u003eT. vorax \u003c/em\u003ewere also detected in gaps among the gill filaments.\u003c/p\u003e\n\u003cp\u003eSkin lesions or mucus of fishes were directly stained in vivo by Hoechst33342 staining method, and Figure 3A\u0026ndash;C show the process of facultatively parasitic \u003cem\u003eTetrahymena \u003c/em\u003esp. eating bacteria around the cell (Figure 3B), and \u003cem\u003eTetrahymena \u003c/em\u003esp. eating tissues (skin or gill) as well as bacteria of host (Figure 3C). Cytoplasm of \u003cem\u003eTetrahymena \u003c/em\u003esp. always contain many (approximately 2\u0026ndash;5 \u0026mu;m in diameter) bacteria-or fish tissue- filled food vacuoles and variable-sized granules, distributed randomly (Figure 3C, arrowhead). The result indicates that facultatively parasitic \u003cem\u003eTetrahymena \u003c/em\u003esp. can physically devour tissue, not only by producing enzyme [49].\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDetection and identification using FISH\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe probe Tetr2020 evaluated with the probe match tool of the ARB software package and the GenBank BLAST tool showed that they are specific to \u003cem\u003eTetrahymena \u003c/em\u003espp. The probe Tetr2020 (5\u003csup\u003e\u0026rsquo;\u003c/sup\u003e- TGTAGTAGCCGTTTCTCAG-3\u003csup\u003e\u0026rsquo;\u003c/sup\u003e) had at least two mismatches to other closely related species like \u003cem\u003eUronema \u003c/em\u003espp., \u003cem\u003ePseudocohnilembus\u003c/em\u003e spp. and \u003cem\u003eCyclidium \u003c/em\u003espp. FISH with the probes Tetr2020 resulted in the presence of a red fluorescence signal each for \u003cem\u003eT.\u003c/em\u003e\u003cem\u003e pyriformis, T. vorax, \u003c/em\u003e\u003cem\u003eT. \u003c/em\u003e\u003cem\u003echironomi\u003c/em\u003e and \u003cem\u003eT. bergeri\u003c/em\u003e (Figure 3D, E), clearly distinguishable from the faint autofluorescence signal achieved with negative-control hybridizations using the Tetr2020 probe to hybridize the untargeted ciliates \u003cem\u003ePseudocohnilembus persalinus\u003c/em\u003e, \u003cem\u003eUronema marinum\u003c/em\u003e and isolated from the same hosts (Figure 3F). Thus, the probes Tetr2020 is also a general probe suitable for widely and rapidly detecting Tetrahymenosis.\u0026nbsp;\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eCorliss (1971a) classified \u003cem\u003eTetrahymena\u003c/em\u003e parasitism into facultatively parasitic forms (\u003cem\u003eT. pyriformis\u003c/em\u003e and \u003cem\u003eT. rostrata\u003c/em\u003e), facultatively free-living forms (\u003cem\u003eT. chironomi\u003c/em\u003e and \u003cem\u003eT. corlissi\u003c/em\u003e) and parasitic forms (\u003cem\u003eT. stegomyiae\u003c/em\u003e and \u003cem\u003eT. limacis\u003c/em\u003e). However, based on later and the present works (Tables 1, 2), supplementary classification is as follows: facultatively parasitic forms (\u003cem\u003eT. pyriformis\u003c/em\u003e, \u003cem\u003eT. rostrate\u003c/em\u003e, \u003cem\u003eT. bergeri \u003c/em\u003eand \u003cem\u003eT. vorax\u003c/em\u003e), facultatively free-living forms (\u003cem\u003eT. chironomi\u003c/em\u003e, \u003cem\u003eT. corlissi, \u003c/em\u003e\u003cem\u003eT. rotunda,\u0026nbsp;T. glochidiophila \u003c/em\u003eand \u003cem\u003eT. papula\u003c/em\u003e) and parasitic forms (\u003cem\u003eT. stegomyiae\u003c/em\u003e and \u003cem\u003eT. limacis\u003c/em\u003e). and the morphology of pathogens and the hosts and data sources are compared in Tables 1 and 2. \u003cem\u003eTetrahymena\u003c/em\u003e\u003cem\u003e bergeri \u003c/em\u003eis reported in the present work as facultatively parasitic forms in \u003cem\u003eChanna aurantimaculata\u003c/em\u003e for the first time (free-living reported before), the reason possibly is that high latitude cold areas may have multiple life-styles when cold weather, due to the slow degradations of microorganisms and insufficiency of food [50]. Except for the ten species showed above, many unidentified \u003cem\u003eTetrahymena \u003c/em\u003esp. conducted infections were also recorded, e. g. Stolk (1960) isolated \u003cem\u003eTetrahymena \u003c/em\u003esp. from the central nervous system of larvae of \u003cem\u003eCyprinous carpio\u003c/em\u003e, \u003cem\u003eAbramis brama \u003c/em\u003eand \u003cem\u003eAmeiurus \u003c/em\u003esp., Kim et al., (2002) examined the fish imported from Southeast Asian countries into Korea and reported of infection from the consignment of ornamental fish, Laoprasert et al., (2001) reported of tetrahymenosis in guppies from local fish farms, appearing as a white ulcerative disease. Thilakaratne et al., (2003) reported of \u003cem\u003eTetrahymena \u003c/em\u003esp. infection in guppies from commercial ornamental fish farms in Sri Lanka, along with other parasites in different ornamental fish species. Pimenta-Leibowitz et al., (2005) reported that poor environmental and adverse physiological conditions pre-dispose to infections with \u003cem\u003eTetrahymena \u003c/em\u003esp.\u003c/p\u003e\n\u003cp\u003eAs a histophagous parasite, \u003cem\u003eTetrahymena \u003c/em\u003edisintegrates host tissue and feeds on cell debris. Infection is showed by whitish patches due to the masses of ciliates in copious amount of mucus, patches usually located on the dorso-lateral skin around the abdomen (Johnson, 1978). Typical gross lesions of \u003cem\u003eTetrahymena \u003c/em\u003einfection are characterized by protrusion of scales, swelling, ulcerative wounds on the skin, blindness, and protrusion of eyes (Johnson, 1978). Numerous \u003cem\u003eTetrahymena \u003c/em\u003espp. may form a rim around the eye orbit (spectacled eye). The parasites route of entry into the body has not been reported. In histopathological analyses, the ciliates were found to be distributed between spaces in the damaged tissues, and extensive necrotic changes occur in the muscle and subdermal tissue. Necrosis of the epithelial cells, extending to the musculature, and disfigured dermal and subcutaneous tissue with edema and hemorrhage was also reported [24, 51].\u003c/p\u003e\n\u003cp\u003eCompared with Scuticociliatosis, Tetrahymenosis can be found in a more wide-ranging hosts (e. g. slugs, hick embryos, dragonfly, helgramite, roach, cockroach and caterpillar), and Tetrahymenosis is easier to discovered in freshwater fish (vs. mostly in marine habitat fishes for Scuticociliatosis) (24, 46, 48, 52). However, their major clinico-pathological manifestations are similar: anemia, weight loss, dark coloration, enteritis, excessive body mucus, yellowish intestinal mucus, loss of scales, hemorrhagic and/or bleached spots on the skin, and dermal necrotic lesions that finally destroy tissues lead to high mortalities [8, 16, 50, 52].\u003c/p\u003e\n\u003cp\u003eAs a test case using probes for the identification of pathogenic ciliates in aquaculture, the present study utilized the probe Tetr2020 to unambiguously detect Tetrahymenosis (Figure 3, D). The general identification by strong fluorescence signals from the oligonucleotide probe can be used for quick and early detection of Tetrahymenosis infections on both invertebrates and vertebrates, especially fish. The FISH probe designed here has the potential to be used for confirming the geographical distribution of parasitic \u003cem\u003eTetrahymena\u003c/em\u003e spp. and detecting the possible dispersal of this facultative pathogen [27\u0026ndash;30]. The FISH approach also provided some morphological information such as body shape, macro and micronuclei shape and number of micronuclei of parasitic \u003cem\u003eTetrahymena\u003c/em\u003e spp.\u003c/p\u003e"},{"header":"Conclusions","content":" \u003cp\u003eThe manifestations of histopathology in fish typically include lesions on the body surface, and affected organs include the skin, musculature, viscera, eye socket and spinal cord; masses of ciliates can be detected in copious amount of mucus and between spaces in the damaged tissues. Improved classification of parasitic \u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003eTetrahymena\u003c/span\u003e is provided: facultatively parasitic forms (\u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003eT. pyriformis\u003c/span\u003e, \u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003eT. rostrate\u003c/span\u003e, \u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003eT. bergeri\u003c/span\u003e and \u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003eT. vorax\u003c/span\u003e), facultatively free-living forms (\u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003eT. chironomi\u003c/span\u003e, \u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003eT. corlissi, T. rotunda, T. glochidiophila\u003c/span\u003e and \u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003eT. papula\u003c/span\u003e) and parasitic forms (\u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003eT. stegomyiae\u003c/span\u003e and \u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003eT. limacis\u003c/span\u003e).The phylogenic results indicate that \u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003eTetrahymena\u003c/span\u003e spp. belonging to \u0026lsquo;borealis\u0026rsquo; group have a greater probability to become parasitism. the method of which can be used for quick and early detection of Tetrahymenosis.\u003c/p\u003e "},{"header":"List of Abbreviations","content":" \u003cdiv class=\"DefinitionList\"\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eFluorescence \u003cspan type=\"Italic\" class=\"Italic\" name=\"Emphasis\"\u003ein situ\u003c/span\u003e hybridization\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eFISH\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003c/div\u003e "},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate \u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication Availability of data and materials \u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe datasets supporting the conclusions of this article are included within the article. Representative sequences are submitted to the GenBank database (accession numbers are provided in Figure\u0026nbsp;4).\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests \u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003ePXM and CY work at Harbin Normal University, Harbin, China. The study was conducted as part of a research program to investigate Tetrahymenosis in aquaculture in Northeastern China.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding \u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNatural Science Foundation of China (project numbers: 31970498), Heilongjiang Province Science Foundation for Youths (project number: QC2017017).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors' contributions \u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe study design, protocol and were prepared by ZSX, PMM, LWY, SM, WCN, LJL, WX and ZL, and report of the study and reviewed by PXM and CY. PXM and his team at Harbin Normal University were responsible for the animal phase and data collection. PXM were responsible for compiling the first draft of the manuscript. All authors revised and approved the final version.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements \u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis work was supported by the Natural Science Foundation of China (project numbers: 31970498), Heilongjiang Province Science Foundation for Youths (project number: QC2017017).\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eCorliss JO. \u003cem\u003eTetrahymena chironomi\u003c/em\u003e nov., a ciliate from midge larvae, and the current status of facultative parasitism in the genus \u003cem\u003eTetrahymena\u003c/em\u003e.\u0026nbsp;Parasitology.\u0026nbsp;1960;50:111\u0026ndash;153.\u003c/li\u003e\n\u003cli\u003eCorliss JO. Facultative parasitism in Protozoa and its possible evolutionary significance. (Abstr.) 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A disease of freshwater fishes caused by \u003cem\u003eTetrahymena corliss \u003c/em\u003eThompson, 1955, and a key for identification of holotrich ciliates of freshwater fishes. J Parasitol. 1975;6:217\u0026ndash;\u003c/li\u003e\n\u003cli\u003eJohnson SK. Tet diseases of tropical fishes and an evaluation of correction techniques. Fish Disease Diagnostic Laboratory F12, Texas A \u0026amp; M University, Austin. 1978.\u003c/li\u003e\n\u003cli\u003eKazubski SL. \u003cem\u003eTrichia lubomirskii \u003c/em\u003e (Helicidae), a new host of \u003cem\u003eTetrahymena limacis \u003c/em\u003e(Warren, 1932) Kozloff, 1946 \u003cem\u003e(Ciliata) \u003c/em\u003eand \u003cem\u003eZonitoides nitidus \u003c/em\u003eMull. (Zonitidae), a new host of \u003cem\u003eT. rostrata \u003c/em\u003e(Kahl, 1926) Corliss, 1952, in Poland. Bull Acad Polonaise Sci. 1958;6:247\u0026ndash;52.\u003c/li\u003e\n\u003cli\u003eKim JH, Hayward CJ, Joh SJ, Heo GJ. Parasitic infections in live freshwater tropical fishes imported to Korea. Dis Aquat Organ. 2002;52:169\u0026ndash;\u003c/li\u003e\n\u003cli\u003eKozloff EN. The morphology and systematic position of a holotrichous ciliate parasitizing \u003cem\u003eDeroceras agreste \u003c/em\u003e(L.). J. Morph. 1946;79:445\u0026ndash;65\u003c/li\u003e\n\u003c/ol\u003e\n\u003col start=\"59\"\u003e\n\u003cli\u003eKozloff EN. A comparison of the parasitic phase of \u003cem\u003eTetrahymena limacis \u003c/em\u003e(Warren) with clones in culture, with particular reference to variability in the number of primary ciliary meridians. J Protozool. 1956a;3:20\u0026ndash;\u003c/li\u003e\n\u003cli\u003eKozloff EN. Tetrahymena limacis from the pulmonate gastropods \u003cem\u003eMonadenia fidelis \u003c/em\u003eand \u003cem\u003eProphysaon andersoni. \u003c/em\u003eJ Protozool. 1956b;3:204\u0026ndash;\u003c/li\u003e\n\u003cli\u003eKozloff EN. A species of \u003cem\u003eTetrahymena \u003c/em\u003eparasitic in the renal organ of the slug \u003cem\u003eDerocercas reticidatum.\u003c/em\u003e J Protozool. 1957;4:75\u0026ndash;\u003c/li\u003e\n\u003cli\u003eLaird M. Parasites of Singapore mosquitoes, with particular reference to the significance of larval epibionts as an index of habitat pollution. Ecology. 1959;40:206\u0026ndash;\u003c/li\u003e\n\u003cli\u003eLaoprasert T, Khaisri C, Chinabut S. Study on white ulcerative disease in guppy (\u003cem\u003ePoecilia reticulata\u003c/em\u003e). Proceeding of the Seminar on Fisheries 2001. The Aquatic Animal Health Research Institute (AAHRI) newsletter, abstracts from Volume 10 No.2. Bangkok, Thailand. 2001.\u003c/li\u003e\n\u003cli\u003eLynn DH, Molloy D, LeBrun R. \u003cem\u003eTetrahymena rotunda\u003c/em\u003e sp. (Hymenostomatida: Tetrahymenidae), a ciliate parasite of the hemolymph of Simulium (Diptera: Simuliidae).\u0026nbsp;Trans Am Microsc Soc. 1981;134\u0026ndash;141.\u003c/li\u003e\n\u003cli\u003eMichelson EH. Distribution and pathogenicity of \u003cem\u003eTetrahymena limacis\u003c/em\u003e in the slug \u003cem\u003eDeroceras reticulatum\u003c/em\u003e.\u0026nbsp;Parasitology.\u0026nbsp;1971;62:125\u0026ndash;\u003c/li\u003e\n\u003cli\u003eRonquist F, Huelsenbeck J. MrBayes 3: Bayesian phylogenetic inference under mixed models. Bioinformatics. 2003;19:1572\u0026ndash;1574.\u003c/li\u003e\n\u003cli\u003eSeaman GR, Tosney T, Berglund R, Goldberg G. Infectivity and recovery of \u003cem\u003eTetrahymena pyriformis\u003c/em\u003e strain S from adult female cockroaches (\u003cem\u003ePeriplaneta americana\u003c/em\u003e).\u0026nbsp;J Protozool.\u0026nbsp;1972;19:644\u0026ndash;\u003c/li\u003e\n\u003cli\u003eStout JD. The ecology, life history and parasitism of \u003cem\u003eTetrahymena\u003c/em\u003e [\u003cem\u003eParaglaucoma\u003c/em\u003e] rostrata (Kahl) Corliss.\u0026nbsp;J Protozool.1954;1:211\u0026ndash;\u003c/li\u003e\n\u003cli\u003eWakita K, Imai S, Hatai K. \"Histopathological study on \u003cem\u003eTetrahymena\u003c/em\u003e infection in Dwarf Gourami (Colisa lalia).\"\u0026nbsp;5th Symposium on Diseases in Asian Aquaculture. 2002.\u003c/li\u003e\n\u003cli\u003eWang Q, Luo S, Ghonimy A, Chen Y, Guo Z, Liu H, Zhang D. Effect of dietary l-carnitine on growth performance and antioxidant response in Amur minnow (\u003cem\u003ePhoxinus lagowskii Dybowskii\u003c/em\u003e). Aquacult Nutr. 2019;25:749\u0026ndash;760.\u003c/li\u003e\n\u003cli\u003eWilliams NE. The polymorphic life history of \u003cem\u003eTetrahymena patula\u003c/em\u003e. J Protozool. 1960; 7:10\u0026ndash;\u003c/li\u003e\n\u003cli\u003eWilson MJ, Coyne C, Glen DM. Low temperatures suppress growth of the ciliate parasite, \u003cem\u003eTetrahymena rostrata\u003c/em\u003e, and pathogenicity to field slugs, \u003cem\u003eDeroceras reticulatum\u003c/em\u003e.\u0026nbsp;Biocontrol. Sci. Techn.\u0026nbsp;1998;8:181\u0026ndash;\u003c/li\u003e\n\u003cli\u003eWright JF. \u003cem\u003eTetrahymena pyriformis \u003c/em\u003e(Ehrenberg) and \u003cem\u003e corlissi \u003c/em\u003eThompson parasitic in stream dwelling triclads (Platyhelminthes: Turbellaria). J Parasitol. 1981;67:131\u0026ndash;133.\u003c/li\u003e\n\u003cli\u003eXue Z, Zhang Y, Lin M, Sun S, Gao W, Wang W. Effects of habitat fragmentation on the population genetic diversity of the Amur minnow (\u003cem\u003ePhoxinus lagowskii\u003c/em\u003e), Mitochondrial DNA Part B 2017;2:331\u0026ndash;336.\u003c/li\u003e\n\u003cli\u003eLwoff, A. Infection experimentale a\u003cem\u003e Glaucoma piriformis\u003c/em\u003e chez \u003cem\u003eQalleria mellonella \u003c/em\u003e(Lepidoptere). C.R. Acad Sci Paris. 1924;178:1106\u0026ndash;\u003c/li\u003e\n\u003cli\u003eWindsor DA. \u003cem\u003eColpoda steinii \u003c/em\u003eand \u003cem\u003eTetrahymena limacis \u003c/em\u003efrom terrestrial pulmonated gastropods collected in Illinois. (Abstr.) J Protozool. 1959;6:33.\u003c/li\u003e\n\u003cli\u003eJanda V, J\u0026Iacute;RovEC O. Uber kunstlieh hervorgerufenen Parasitismus eines freilebenden Ciliaten \u003cem\u003eGlaucoma piriformis \u003c/em\u003eund Infektionsversuch mit \u003cem\u003eEuglena gracilis \u003c/em\u003eand \u003cem\u003eSpirochaeta biftexa.\u003c/em\u003e Vestnik Ceskoslov. Zool. spolec. Praze. (Mem. Soc. zool. tchicosl.) 1937;5:34\u0026ndash;\u003c/li\u003e\n\u003cli\u003eCorliss JO. 1951. Morphology and systematic status of pure culture strains in the \u003cem\u003eColpidium-Glaucoma-Leucophrys-Tetrahymena \u003c/em\u003e (Abstr.) Proc Amer Soc Protozool. 2:11.\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Tables","content":"\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cstrong\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eTable 1.\u003c/span\u003e\u003c/strong\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e Morphological identification characteristics of parasitic \u003cem\u003eTetrahymena\u003c/em\u003e spp.\u003c/span\u003e\u003c/p\u003e\n\u003ctable style=\"border-collapse: collapse; border: none;\" width=\"699\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 91.9pt; border: solid windowtext 1.0pt; border-top: solid windowtext 2.25pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eSpecies\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 78.25pt; border-top: solid windowtext 2.25pt; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"104\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eSize in vivo\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 91.85pt; border-top: solid windowtext 2.25pt; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"122\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eBody shape \u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49.9pt; border-top: solid windowtext 2.25pt; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"67\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eCaudal cilia\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 42.25pt; border-top: solid windowtext 2.25pt; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"56\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eSK No.\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 35.45pt; border-top: solid windowtext 2.25pt; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"47\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eMi No.\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49.85pt; border-top: solid windowtext 2.25pt; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"66\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eCVP No. \u0026amp; position\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 85.05pt; border-top: solid windowtext 2.25pt; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"113\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eReferences\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 91.9pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. corlissi\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 78.25pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"104\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003ean average 52 \u0026times; 32 \u0026mu;m\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 91.85pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"122\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eelongate, pointed anteriorly and rounded posteriorly\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49.9pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"67\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003epresent\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 42.25pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"56\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e25-31\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 35.45pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"47\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e1\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49.85pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"66\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 10.0pt; font-family: 'Times New Roman',serif;\"\u003e2 or 3; SK6-10\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 85.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"113\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e16\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 91.9pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. pyriformis\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 78.25pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"104\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eca. 50 \u0026times; 30 \u0026mu;m \u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 91.85pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"122\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003epear-shaped\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49.9pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"67\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eabsent\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 42.25pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"56\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e15-25\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 35.45pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"47\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eN/A\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49.85pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"66\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 10.0pt; font-family: 'Times New Roman',serif;\"\u003e2; SK5, 6\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 85.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"113\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e5\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 91.9pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. stegomyiae\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 78.25pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"104\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e60-100 \u0026mu;m\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 91.85pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"122\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003epear-shaped\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49.9pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"67\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eabsent\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 42.25pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"56\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e25-30\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 35.45pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"47\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e1\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49.85pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"66\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 10.0pt; font-family: 'Times New Roman',serif;\"\u003e-\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 85.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"113\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e1\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 91.9pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. chironomi\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 78.25pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"104\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eca. 40 \u0026times; 23 \u0026mu;m\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 91.85pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"122\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eelongate\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49.9pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"67\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eabsent\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 42.25pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"56\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e23-28\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 35.45pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"47\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e1\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49.85pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"66\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 10.0pt; font-family: 'Times New Roman',serif;\"\u003e3; SK6-9 \u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 85.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"113\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e1\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 91.9pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. vorax\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 78.25pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"104\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e\u0026lt; 150 \u0026mu;m \u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 91.85pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"122\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eposterior end is drawn out into a tail\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49.9pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"67\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eabsent\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 42.25pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"56\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e18-23\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 35.45pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"47\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e0\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49.85pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"66\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 10.0pt; font-family: 'Times New Roman',serif;\"\u003e2-6; SK5-10\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 85.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"113\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e7\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 91.9pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. rostrata\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 78.25pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"104\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e50-60 \u0026times; 30-40 \u0026mu;m\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 91.85pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"122\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eVary\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49.9pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"67\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003epresent\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 42.25pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"56\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e36-58\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 35.45pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"47\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e1\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49.85pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"66\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 10.0pt; font-family: 'Times New Roman',serif;\"\u003e3 or 4; SK6-12\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 85.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"113\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e5,7\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 91.9pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. limacis\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 78.25pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"104\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e40-55 \u0026times; 30-40 \u0026mu;m\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 91.85pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"122\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eCucumber-shaped with strongly apiculate anterior end\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49.9pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"67\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eabsent\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 42.25pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"56\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e26-46\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 35.45pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"47\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e1\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49.85pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"66\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 10.0pt; font-family: 'Times New Roman',serif;\"\u003e2 or 3; 5-8\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 85.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"113\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e58\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 91.9pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. rotunda\u0026nbsp;\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 78.25pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"104\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e40-90 \u0026mu;m (after staining)\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 91.85pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"122\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003ebluntly ovoid to globose\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49.9pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"67\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003epresent\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 42.25pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"56\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e50-66;\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 35.45pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"47\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e--\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49.85pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"66\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 10.0pt; font-family: 'Times New Roman',serif;\"\u003enot observed\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 85.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"113\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e64\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 91.9pt; border-top: none; border-left: solid windowtext 1.0pt; border-bottom: solid windowtext 2.25pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. papula\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 78.25pt; border-top: none; border-left: none; border-bottom: solid windowtext 2.25pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"104\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e80-100 \u0026mu;m\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 91.85pt; border-top: none; border-left: none; border-bottom: solid windowtext 2.25pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"122\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003epiriform, bag-shaped\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49.9pt; border-top: none; border-left: none; border-bottom: solid windowtext 2.25pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"67\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eabsent\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 42.25pt; border-top: none; border-left: none; border-bottom: solid windowtext 2.25pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"56\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e33-45\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 35.45pt; border-top: none; border-left: none; border-bottom: solid windowtext 2.25pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"47\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e1\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49.85pt; border-top: none; border-left: none; border-bottom: solid windowtext 2.25pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"66\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 10.0pt; font-family: 'Times New Roman',serif;\"\u003e2-6; SK9-16\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 85.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 2.25pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"113\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e78\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eTable 2. Reports of parasitic \u003cem\u003eTetrahymena \u003c/em\u003especies and their hosts\u003cem\u003e.\u003c/em\u003e\u003c/span\u003e\u003c/p\u003e\n\u003ctable style=\"border-collapse: collapse; border: none;\" width=\"614\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border-top: solid windowtext 2.25pt; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eParasite\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: solid windowtext 2.25pt; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eHost\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: solid windowtext 2.25pt; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eData source\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. bergeri\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eChanna aurantimaculata\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eFrom the present work\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. chironomi\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eFourth-instar larva (\u003cem\u003eChironomus plumosus\u003c/em\u003e)\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif; color: #222222; background: white;\"\u003e1\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. chironomi\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003ePhoxinus lagowskii\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eFrom the present work\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. corlissi\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eBeetle, Dragonfly, Guppies, Helgramite, Tadpoles, Tent caterpillar\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e16\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. corlissi\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eGuppies (\u003cem\u003ePoecilia reticulatus\u003c/em\u003e)\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e54\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. corlissi\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eFreshwater triclads\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif; color: #211d1e;\"\u003e73\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. corlissi\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eAtlantic Salmon \u003cem\u003eSalmon salar\u003c/em\u003e\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e23\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. corlissi\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eFishes\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e46\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. corlissi\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eFresh fish\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e17\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. corlissi\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eGuppies \u003cspan style=\"color: #211d1e;\"\u003e(\u003cem\u003ePoecilia reticulata\u003c/em\u003e) \u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e24\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. corlissi\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eGuppies\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e25\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. corlissi\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eGuppies \u003cspan style=\"color: #211d1e;\"\u003e(\u003cem\u003ePoecilia reticulata\u003c/em\u003e)\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e22\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. corlissi\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif; color: #221e1f;\"\u003eDwarf Gourami (\u003cem\u003eColisa lalia\u003c/em\u003e)\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif; color: #211d1e;\"\u003e69\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. corlissi\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif; color: #222222; background: white;\"\u003eFreshwater ornamental fish\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif; color: #222222; background: white;\"\u003e48\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. corlissi\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eGuppies \u003cspan style=\"color: #211d1e;\"\u003e(\u003cem\u003ePoecilia reticulata\u003c/em\u003e)\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e18\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. corlissi\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif; color: #222222; background: white;\"\u003eGolden perch (\u003cem\u003eMacquaria ambiqua\u003c/em\u003e)\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e50\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. glochidiophila\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eFreshwater mussel\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e21\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. limacis\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eEuropean slug (\u003cem\u003eDeroceras retioulatum)\u003c/em\u003e\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e58\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. limacis\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eEuropean slug (\u003cem\u003eDeroceras reticulatum\u003c/em\u003e)\u003cem\u003e, \u003c/em\u003enative slug (\u003cem\u003eProphysaon andersoni\u003c/em\u003e)\u003cem\u003e, \u003c/em\u003enative snail (\u003cem\u003eMonadenia fidelis\u003c/em\u003e)\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e59, 60\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. limacis\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eFresh-water clams\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e53\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. limacis\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eLand snail (\u003cem\u003eTrichia lubomirskii\u003c/em\u003e)\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e56\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. limacis\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eEuropean slug (\u003cem\u003eDeroceras reticulatum\u003c/em\u003e)\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e76\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. limacis\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif; color: #222222; background: white;\"\u003eSlug (\u003cem\u003eDeroceras reticulatum\u003c/em\u003e)\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif; color: #222222; background: white;\"\u003e65\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. papula\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eTent caterpillar (\u003cem\u003eMalacosoma americana \u003c/em\u003elarva)\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e16\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. pyriformis\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eInsects\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e75\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. pyriformis\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eInsects\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e77 \u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. pyriformis\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eClams\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e53\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. pyriformis\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eBeetle, Chick embryos, Dragonfly, Helgramite, Roach, cockroach, Tent caterpillar\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e16\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. pyriformis\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eOligochaete, \u003cem\u003eAllolobophora caliginosa \u003c/em\u003evar. \u003cem\u003etrapezoides\u003c/em\u003e\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e17\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. pyriformis\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eMosquitoes\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e62\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. pyriformis\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif; color: #222222; background: white;\"\u003eAdult female cockroaches (\u003cem\u003ePeriplaneta americana\u003c/em\u003e)\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif; color: #222222; background: white;\"\u003e67\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. pyriformis\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eChannel catfish (\u003cem\u003eIctalurus punctatus\u003c/em\u003e)\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e44\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. pyriformis\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eFreshwater triclads\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; color: #211d1e;\"\u003e73\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. pyriformis\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eFishes\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif; color: #211d1e;\"\u003e45\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. pyriformis\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left; text-autospace: none;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif; color: #211d1e;\"\u003eFreshwater crayfish (\u003cem\u003eCherax quadricarinatus\u003c/em\u003e) \u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif; color: #211d1e;\"\u003e47\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. pyriformis\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif; color: #211d1e;\"\u003eOrnamental fishes \u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e51\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. pyriformis\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif; color: #222222; background: white;\"\u003eFreshwater ornamental fish\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif; color: #222222; background: white;\"\u003e48\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. pyriformis\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003ePhenacogrammus interruptus\u0026nbsp; \u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eFrom the present work\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. rostrata\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eEuropean slug (\u003cem\u003eDeroceras reticulatum\u003c/em\u003e)\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e61\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. rostrata\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eLand snail (\u003cem\u003eZonitoides nitidua\u003c/em\u003e)\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e56\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. rostrata\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eEuropean slug (\u003cem\u003eDeroceras reticulatum\u003c/em\u003e)\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e16\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. rostrata\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif; color: #222222; background: white;\"\u003eField slugs (\u003cem\u003eDeroceras reticulatum\u003c/em\u003e)\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif; color: #222222; background: white;\"\u003e72\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. rotunda\u0026nbsp;\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eSimulium\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e9\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: solid windowtext 1.0pt; border-top: none; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. stegomyiae\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eMosquito\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border-top: none; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e1\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border-top: none; border-left: solid windowtext 1.0pt; border-bottom: none; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. vorax\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border: none; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eChick embryos, Cockroach, Dragonfly, Helgramite, Tent caterpillar\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border: none; border-right: solid windowtext 1.0pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e16\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 92.15pt; border: none; border-bottom: solid windowtext 2.25pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"123\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eT. vorax\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 248.05pt; border: none; border-bottom: solid windowtext 2.25pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"331\"\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cem\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003ePelteobagrus fulvidraco\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 120.5pt; border: none; border-bottom: solid windowtext 2.25pt; padding: 0in 5.4pt 0in 5.4pt;\" width=\"161\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003eFrom the present work\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cstrong\u003e\u003cspan style=\"font-size: 11.0pt; font-family: 'Times New Roman',serif;\"\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Tetrahymenosis, facultative parasitic ciliates, Simultaneous fluorescence in situ hybridization, diversity, histopathology","lastPublishedDoi":"10.21203/rs.3.rs-15239/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-15239/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eBackground Tetrahymenosis caused by about ten parasitic Tetrahymena species belonging to the Phylum Ciliophora has been recognized as an emerging problem inflicting significant economic loss in aquaculture industry in the world. Increasing knowledge and identification of Tetrahymenosis are important. \u003c/p\u003e\u003cp\u003eMethods Four parasitic Tetrahymena species were collected from eight commercially farmed fishes in Harbin, northeastern China. Specific oligonucleotide probe and fluorescence in situ hybridization staining was designed and tested. Hoechst33342 staining methods, gene sequencing and phylogenetic analyses were also conducted. \u003c/p\u003e\u003cp\u003eResults Tetrahymena pyriformis, T. vorax, T. chironomi and T. bergeri are the four species responsible for Tetrahymenosis in various fishes in Harbin. Taxonomy, diagnosis, diversity, pathogenicity, and histopathology of Tetrahymenosis were supplemented, analyzed and summaried, based on the present and previous work. The term Tetrahymenosis is diagnosised as diseases affecting a number of fishes, crustaceans, mollus, beetle, dragonfly, salmon, slug, midge larvae and freshwater mussel species that caused by ciliates of the genus Tetrahymena which constitutes an abundant group inhabit various aquaculture and natural habitats. Improved classification of parasitic Tetrahymena was provided. Phylogeny of parasitic Tetrahymena species was studied, and an SSU-rDNA targeted oligonucleotide probe labeled with a fluorochrome was designed, and the FISH protocol was optimized for identification of Tetrahymena species, \u003c/p\u003e\u003cp\u003eConclusions The manifestations of histopathology in fish typically include lesions on the body surface, and affected organs include the skin, musculature, viscera, eye socket and spinal cord; masses of ciliates can be detected in copious amount of mucus and between spaces in the damaged tissues. Improved classification of parasitic Tetrahymena is provided: facultatively parasitic forms ( T. pyriformis , T. rostrate , T. bergeri and T. vorax ), facultatively free-living forms ( T. chironomi , T. corlissi, T. rotunda, T. glochidiophila and T. papula ) and parasitic forms ( T. stegomyiae and T. limacis ).The phylogenic results indicate that Tetrahymena spp. belonging to ‘borealis’ group have a greater probability to become parasitism. the method of which can be used for quick and early detection of Tetrahymenosis.\u003c/p\u003e","manuscriptTitle":"Taxonomy, diversity and diagnosis of Tetrahymenosis, and its recent identification measures in aquaculture inferred from infections in Northeastern China","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2020-03-04 17:11:50","doi":"10.21203/rs.3.rs-15239/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"a235f783-58a5-41d2-8dc6-490967b03079","owner":[],"postedDate":"March 4th, 2020","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":66075,"name":"Parasitology"},{"id":66076,"name":"Aquaculture and Mariculture"}],"tags":[],"updatedAt":"","versionOfRecord":[],"versionCreatedAt":"2020-03-04 17:11:50","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-15239","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-15239","identity":"rs-15239","version":["v1"]},"buildId":"-HB7Z8yhvgn0wM9Nzuekk","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}