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Cystodiscus elachistocleis sp. nov. urn:lsid:zoobank.org:act: E2AE8907-E225-4804-A2F9-E3920AD6298D Figs 1–3 Type host Elachistocleis cesarii (Miranda-Ribeiro, 1920). Site of infection Gallbladder. Etymology The species epithet is derived from the genus Elachistocleis of the host. Material examined Hapantotypes BRAZIL • 10+ myxospores; Mato Grosso State, Araguaiana municipality; 14°35′5.29″ S, 51°41′19.15″ W; glycerogelatin slide; GenBank MZ645740 - MZ645741; INPA79. Description Numerous pseudoplasmodia were found free in the bile. Pseudoplasmodia (Fig. 1C) of approximately 2 mm were rounded in shape, appeared to be formed by a ʻgelatinousʼ substance, and were floating in the host’s bile containing several myxospores in its interior. The morphology of myxospores found in the gallbaldder of E. cesarii corresponded to the genus Cystodiscus. The myxospores (Figs 1A–B, 2) were ellipsoid to ovoid. The two myxospore valves were joined by a raised ridge that appeared straight or slightly curved (S-shaped in sutural view) along the medial axis of the myxospore. Transverse depressions on the surface of the myxospores appeared connected to the sutural ridge depression. Their measurements presented as mean ± SD (range) were: myxospore length 10.6 ± 0.3 (9.8–11.2) μm, myxospore width 6.2 ± 0.5 (5.6–6.6) μm. The polar capsules were pyriform and equal in size, situated each one in a myxospore extremity and measured 3.6 ± 0.4 (2.8–4.3) μm in length and 2.6 ± 0.3 (2.2– 3.1) μm in width. The polar filament present within the polar capsule had 2–4 coils. The myxospores showed 3–5 transverse ridges and a binucleated sporoplasm. Filiform polar appendages were not observed. Remarks Cystodiscus elachistocleis sp. nov. was morphometrically compared to all Cystodiscus spp. described worldwide (Table 2). The species that most resembled C. elachistocleis sp. nov. was C. haldari (Sarkar, 1982), which showed similarity in all measurements available for comparison. However, the geographical distance (Brazil vs India) and the different type host (E. cesarii vs Hyla arborea (Linnaeus, 1758)) allow us to separate the two species. Furthermore, C. haldari does not present any comparative values such as the number of turns of the polar filament and the number of transverse ridges, which could differentiate the two species. Cystodiscus thyponius Gray (1993) also presented measures of length and width of myxospores similar to those found in C. elachistocleis sp. nov. However, differences were observed in the polar capsule width (2.6 ± 0.3 (2.2–3.1) vs 3.6 (3.3–5.2)) and the number of transverse ridges (3–5 vs 9–11). Regarding the Cystodiscus spp. described from Brazil, C. elachistocleis sp. nov. was morphometrically compared with two species parasitizing the gallbladder of Rhinella marina (Linnaeus, 1758). Cystodiscus immersus, Kudo & Sprague (1940) presented a longer body length than that found in C. elachistocleis sp. nov. (11.8–13.3 vs 10.6 ± 0.3 (9.8–11.2)), in addition to a greater number of transverse ridges (7–9 vs 3–5). Cystodiscus lyndoyense Carini, 1932 showed the length of the body (11.0–12.0 vs 10.6 ± 0.3 (9.8– 11.2)) and the body width (7.5–8.0 vs 6.2 ± 0.5 (5.6–6.6)) longer than that found for C. elachistocleis sp. nov. The other Cystodiscus spp. already described presented a body length longer than that observed in C. elachistocleis sp. nov. Molecular analyses Fragments of 1730-bp and 1916-bp of the SSU rDNA gene were generated. The fragments showed 100% similarity when aligned. The BLAST search of the sequences did not reveal a direct match with myxozoan sequences available in GenBank. The genetically closest species was C. immersus, which exhibited a similarity of 97.3%, and a difference of 24 out of 885 nucleotides (Table 3). A well-supported phylogenetic tree divided into three main groups was obtained (Fig. 3).A monophyletic group composed of Sphaeromyxa Thélohan, 1892 spp. that parasitizes fish, a polyphyletic group composed of Zschokkella Auerbach, 1909 and Myxidium Bütschli, 1882 spp. that infect fish, and finally a monophyletic group composed of Cystodiscus spp. that parasitize amphibians. Still, there is a small monophyletic group composed of Myxidium ssp. found in reptiles, fish, and birds. Cystodiscus elachistocleis sp. nov. appears as a sister species of Cystodiscus cf. immersus 1, in a subclade formed by species that parasitize the amphibians gallbladder., Published as part of Vieira, Diego Henrique Mirandola Dias, Úngari, Letícia Pereira, Alcantara, Edna Paulino De, Emmerich, Enzo, Santos, André Luiz Quagliatto, O'Dwyer, Lucia Helena & Silva, Reinaldo José Da, 2021, Description of Cystodiscus elachistocleis sp. nov. (Cnidaria: Myxosporea) parasitizing the gallbladder of Elachistocleis cesarii from Brazil, based on morphological and molecular analyses, pp. 107-118 in European Journal of Taxonomy 775 on pages 110-113, DOI: 10.5852/ejt.2021.775.1549, http://zenodo.org/record/5589675, {"references":["Lutz A. 1889. Uber ein Myxosporidium aus der Gallenblase brasilianischer Batrachier. Centralblatt fur Bakteriologie und Parasitenkunde 5: 84 - 88.","Hartigan A., Fiala I., Dykova I., Rose K., Phalen D. N. & Slapeta J. 2012 a. New species of Myxosporea from frogs and resurrection of the genus Cystodiscus Lutz, 1889 for species with myxospores in gallbladders of amphibians. Parasitology 139 (4): 478 - 496. https: // doi. org / 10.1017 / S 0031182011002149","Hartigan A., Dhand N. K., Rose K., Slapeta J. & Phalen D. N. 2012 c. Comparative pathology and ecological implications of two myxosporean parasites in native Australian frogs and the invasive cane toad. PLoS One 7 (10): e 43780. https: // doi. org / 10.1371 / journal. pone. 0043780"]}

Numerous pseudoplasmodia containing myxospores belonging to the genus Cystodiscus were found in the gallbladder of Elachistocleis cesarii from Mato Grosso State, Brazil. Herein, we describe Cystodiscus elachistocleis sp. nov., using morphological and small subunit ribosomal DNA sequences. The mature myxospores were ellipsoid to ovoid, measuring in average 10.6 (9.8–11.2) μm in length and 6.2 (5.6–6.6) μm in width. Polar capsules were pyriform and equal in size measuring in average 3.6 (2.8–4.3) μm in length and 2.6 (2.2–3.1) μm in width. Polar filaments had 2–4 coils. The myxospores had 3–5 transverse ridges. The phylogenetic analysis showed Cystodiscus elachistocleis sp. nov. as a sister species of Cystodiscus cf. immersus 1, in a subclade formed by species that parasitize the amphibians gallbladder. This is the first species of Cystodiscus described parasitizing a species of Elachistocleis and the third species of Cystodiscus described in Brazil.

Submitted by DIEGO HENRIQUE MIRÂNDOLA DIAS VIEIRA (diego.vieira@unesp.br) on 2020-03-10T14:34:32Z No. of bitstreams: 1 Tese.pdf: 6116410 bytes, checksum: 17190f6280f0f79572baa9edbb5a6356 (MD5) Approved for entry into archive by ROSANGELA APARECIDA LOBO null (rosangelalobo@btu.unesp.br) on 2020-03-11T17:55:53Z (GMT) No. of bitstreams: 1 vieira_dhmd_dr_bot_par.pdf: 1043292 bytes, checksum: b66d75a8f798a48c43f6edc784921248 (MD5) Made available in DSpace on 2020-03-11T17:55:53Z (GMT). No. of bitstreams: 1 vieira_dhmd_dr_bot_par.pdf: 1043292 bytes, checksum: b66d75a8f798a48c43f6edc784921248 (MD5) Previous issue date: 2020-02-18 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Parasitos de organismos aquáticos apresentam considerável relevância para estudo devido ao seu impacto nos sistemas de aquicultura e na pesca em todo o mundo. Mixozoários são microparasitos que tem um ciclo de vida complexo, alternando entre um hospedeiro vertebrado e um invertebrado, podendo ser responsáveis por causar doenças principalmente em peixes, com altas taxas de mortalidade. Muitos mixozoários permanecem desconhecidos pela comunidade científica e seu potencial em causar doenças em peixes segue indeterminado em alguns casos. Desta forma, o presente trabalho teve como objetivo analisar de forma morfológica e molecular mixozoários parasitos de peixes de rios interiores do estado de São Paulo, assim como caracterizar as relações filogenéticas desses parasitos e seu potencial patológico. Durante os anos de 2016 a 2019 foram coletados espécimes de cinco espécies de peixes das ordens Characiformes e Siluriformes. Foram utilizadas técnicas de biologia molecular, histologia e análise ultraestrutural na descrição taxonômica das espécies. Myxobolus batalhensis n. sp. encontrado parasitando Salminus hilarii (Bryconidae), Myxobolus imparfinis n. sp. encontrando parasitando Imparfinis mirini (Heptapteridae), Henneguya sp. 1 encontrado parasitando Prochilodus lineatus (Prochilodontidae) e Henneguya sp. 2 encontrado parasitando Astyanax lacustris (Characidae), tiveram a caracterização morfológica e molecular realizadas. Foi feito um estudo suplementar sobre Henneguya guanduensis, encontrado parasitando Hoplosternum littorale (Callichthyidae), com análise molecular, ultraestrutural e filogenética. Foi realizada ainda a análise filogenética das espécies de mixozoários da família Myxobolidae que parasitam peixes do Brasil. A diversidade de mixozoários encontrados nas espécies de peixes alvos deste estudo evidencia a necessidade de se intensificar o estudo da diversidade e do potencial patogênico destes parasitos. Parasites of aquatic organisms present considerable relevance to study because of their impact on aquaculture and fisheries worldwide. Myxozoans are microparasites that have a complex life cycle, alternating between a vertebrate and an invertebrate host, being able to be responsible for causing diseases mainly in fish, with high mortality rates. Many myxozoans remain unknown to the scientific community and their potential to cause disease in fish remains undetermined in some cases. Thus, the present study aimed to analyze morphologically and molecularly myxozoans parasites of fish from inland rivers of the state of São Paulo, as well as to characterize the phylogenetic relationships of these parasites and their pathological potential. From 2016 to 2019 specimens from five species from Characiformes and Siluriformes orders were collected. Molecular biology, histology, and ultrastructural analysis techniques were used taxonomic description of species. Myxobolus batalhensis n. sp. found parasitizing Salminus hilarii (Bryconidae), Myxobolus imparfinis n. sp. found parasitizing Imparfinis mirini (Heptapteridae), Henneguya sp. 1 found parasitizing Prochilodus lineatus (Prochilodontidae) and Henneguya sp. 2 found parasitizing Astyanax lacustris (Characidae), had the morphological and molecular characterization performed. A supplementary study on Henneguya guanduensis found parasitizing Hoplosternum littorale (Callichthyidae) was done, with molecular, ultrastructural and phylogenetic analysis. A phylogenetic analysis of the myxozoans species of Myxobolidae family parasitizing fish from Brazil was performed. The diversity of myxozoans found in the target fish species of this study highlights the need to intensify the study of the diversity and pathogenic potential of these parasites. FAPESP: 2015/24901-2

Submitted by DIEGO HENRIQUE MIRANDOLA DIAS VIEIRA null (diegovieira.50@gmail.com) on 2016-04-12T17:02:12Z No. of bitstreams: 1 Dissertação de mestrado_Diego_Vieira.pdf: 4207286 bytes, checksum: 09374ce24480ed9aa883dcaaecc5ce63 (MD5) Rejected by Ana Paula Grisoto (grisotoana@reitoria.unesp.br), reason: Solicitamos que realize uma nova submissão seguindo as orientações abaixo: No campo “Versão a ser disponibilizada online imediatamente” foi informado que seria disponibilizado o texto completo porém no campo “Data para a disponibilização do texto completo” foi informado que o texto completo deverá ser disponibilizado apenas 18 meses após a defesa. Caso opte pela disponibilização do texto completo apenas 18 meses após a defesa selecione no campo “Versão a ser disponibilizada online imediatamente” a opção “Texto parcial”. 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Agradecemos a compreensão. on 2016-04-13T17:53:40Z (GMT) Submitted by DIEGO HENRIQUE MIRANDOLA DIAS VIEIRA null (diegovieira.50@gmail.com) on 2016-04-13T18:07:34Z No. of bitstreams: 1 Dissertação de mestrado_Diego_Vieira.pdf: 4207286 bytes, checksum: 09374ce24480ed9aa883dcaaecc5ce63 (MD5) Approved for entry into archive by Ana Paula Grisoto (grisotoana@reitoria.unesp.br) on 2016-04-13T19:39:28Z (GMT) No. of bitstreams: 1 vieira_dhmd_me_bot_par.pdf: 568974 bytes, checksum: 8a52b1cedc517c38ae34f618834c31b4 (MD5) Made available in DSpace on 2016-04-13T19:39:28Z (GMT). No. of bitstreams: 1 vieira_dhmd_me_bot_par.pdf: 568974 bytes, checksum: 8a52b1cedc517c38ae34f618834c31b4 (MD5) Previous issue date: 2016-02-22 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Mixozoários são metazoários parasitos obrigatórios principalmente de peixes, sendo alguns dos mais importantes patógenos desses hospedeiros. Existem cerca de 2200 espécies descritas para a classe Myxosporea, porém no Brasil os estudos ainda continuam escassos. Nesse estudo foram coletados peixes de diferentes espécies entre março de 2014 e agosto de 2015, para ser verificado a presença de mixozoários parasitos. Foram encontrados mixozoários do gênero Henneguya e Myxobolus parasitando peixes das ordens Characiformes e Siluriformes. Os parasitos coletados foram processados visando uma análise morfológica através de microscopia de luz, histopatologia e análise ultraestrutural, observando-se ainda a relação parasito-hospedeiro. Henneguya sp. 1 foi encontrado parasitando as brânquias de Astyanax altiparanae e Henneguya sp. 2 parasitando as brânquias de Leporinus friderici. Myxobolus prochilodus, Henneguya sp. 3 e Myxobolus sp. 1 foram encontrados parasitando as brânquias de Prochilodus lineatus causando deformações nas lamelas primárias dos peixes. Henneguya sp. 4 foi encontrada parasitando as brânquias de Cyphocharax modestus, causando uma junção das lamelas secundárias e diminuindo a área de respiração útil do peixe. A análise ultraestrutural permitiu a compreensão da interface parasito-hospedeiro. Também foram observados Henneguya guanduensis parasitando o intestino de Hoplosternum litoralle e Myxobolus sp. 2 parasitando o intestino de Rhamdia quelen. Myxobolus aureus foi encontrado parasitando as gônadas e o fígado de Salminus hilarii. A análise histopatológica indicou a infertilidade dos ovócitos afetados. Foram comparadas as medidas morfológicas dos parasitos encontrados e aqui nomeados como sp., com as já registradas na literatura e não foram encontradas espécies que se assemelhavam no tamanho. Concluiu-se que se tratam de espécies novas com a necessidade de análises moleculares para a confirmação do estudo. Myxozoans are metazoan parasites mainly on fish, with some of the most important pathogens of these hosts. There are about 2,200 described species for Myxosporea class, but in Brazil the studies are still scarce. In this study fish of different species were collected between March 2014 and August 2015 to be verified the presence of myxozoans parasites. The genera of myxozoans found were Henneguya and Myxobolus parasitizing fish of Characiformes and Siluriformes orders. The parasites collected were processed aiming a morphological analysis by light microscopy, histopathology and ultrastructural analysis, observing the host-parasite relationship. Henneguya sp. 1 was found parasitizing the gills of Astyanax altiparanae and Henneguya sp. 2 parasitizing the gills of Leporinus friderici. Myxobolus lomi (Myxobolus prochilodus), Henneguya sp. 3 and Myxobolus sp. 1 were found parasitizing the gills of Prochilodus lineatus causing deformities in fish primary lamellae. Henneguya sp. 4 was found parasitizing the gills of Cyphocharax Modestus, causing a junction of secondary lamellae and decreasing the useful breathing area of the fish. The ultrastructural analysis allowed the understanding of parasite-host interface. Also were observed Henneguya guanduensis parasitizing the intestine of Hoplosternum litoralle and Myxobolus sp. 2 parasitizing the intestine of Rhamdia quelen. Myxobolus aureus was found parasitizing the gonads and liver of Salminus hilarii. Histopathological analysis indicated the infertility of the affected oocytes. Morphological measures of parasites found were compared with those reported in the literature and did not found species that were similar in size. It was concluded that these are new species with the necessity of molecular analysis to confirm the study. FAPESP: 2013/25623-0