Six species of nonmazocraeid monogeneans including two new species from clupeiform fishes of Visakhapatnam coast, Bay of Bengal.

During a survey on monogenean parasites of clupeiform fishes of Visakhapatnam coast, Bay of Bengal, apart from typical mazocraeids, 6 species of nonmazocraeids, including two new species were encountered. These belonged to 5 genera distributed over 4 families: Family Gastrocotylidae: Engraulicola thrissocles from Thryssa mystax, T. setirostris, T. malabarica; Engraulicola longisomum n.sp. from Thryssa mystax, T. setirostris, T. malabarica; Pellonicola elongatus from Ilisha filigera; family Megamicrocotylidae: Megamicrocotyle chirocentrus from Chirocentrus dorab; family Microcotylidae: Microcotyle clupei n.sp. from Thryssa mystax and T. setirostris; family Diclidophoridae: Keralina opisthopterus from Opisthopterus tardoore. Engraulicola longisomum n. sp. differs from E. thrissocles in the larger body size, in the relatively smaller size of haptor and also in the number of clamps and alignment of reproductive organs. Microcotyle clupei n.sp. the first species of the genus to be reported from clupeiform fishes differs from other species of the genus, mainly in the presence of a circlet of spines on the oral sucker. Descriptions are provided for all the species collected. Megamicrocotyle johnstoni is synonymized with M. chirocentrus.

Assessment of Salinomycin's Potential to Treat Microcotyle sebastis in Korean Rockfish (Sebastes schlegelii).

Aquaculture, a crucial sector of the global food industry, faces a myriad of issues due to parasitic invasions. One such parasite, Microcotyle sebastis, which afflicts Korean rockfish in South Korea, has a significant economic impact. The impending danger of resistance to traditional anthelmintics necessitates the exploration of new antiparasitic candidates. Although the efficacy of salinomycin against aquatic parasites such as ciliates and sporozoans is known, its influence on monogeneans has yet to be studied. Therefore, this study investigated the efficacy and safety of salinomycin for the treatment of M. sebastis infections, presenting the first exploration of salinomycin's therapeutic potential against monogeneans. In vitro examinations revealed a minimum effective concentration of salinomycin of 5 mg/kg, which led to necrosis of the haptor upon dislodging from the gill filaments. The one-time oral administration of the drug at concentrations of 5 mg/kg and 10 mg/kg showed a significant dose-dependent reduction in parasite counts, with no apparent behavioral side effects in Korean rockfish. Biochemical analyses monitored the liver, heart, and kidney enzymes, specifically aspartate transaminase (AST), alanine transaminase (ALT), blood urea nitrogen (BUN), and creatine kinase-myocardial band (CK-MB). At both 20 °C and 13 °C, no significant differences were observed in the levels of AST and ALT. However, at 20 °C, alterations in BUN levels were evident on Day 14, a deviation not observed at 13 °C. The CK-MB analysis revealed elevated enzyme levels at both temperatures when compared to the control group, reflecting the similar changes observed in terrestrial animals administered salinomycin. The biochemical data suggest that the oral administration of salinomycin is potentially more favorable at 13 °C than at 20 °C. Although our findings warrant further comprehensive studies, including on the long-term and potential effects on nontarget species and water quality, they also suggest that salinomycin could be considered as an alternative or adjunctive treatment if resistance to the currently used praziquantel against M. sebastis is confirmed.

Microcotyle justinei n. sp. (Monogenea: Microcotylidae) from the Gills of the Cardinal Fish Apogon imberbis (Teleostei: Apogonidae) off the Algerian Coast of the Western Mediterranean.

PURPOSE: A new monogenean Microcotyle justinei n. sp. (Monogenea: Microcotylidae) is described based on specimens found on the gill filaments of the cardinal fish Apogon imberbis (Apogonidae) off the Algerian coast of the Western Mediterranean. METHODS: Monogeneans were examined, measured and drawn for a comparative morphological study with other species of Microcotyle and characterised molecularly using a partial fragment of the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene. The identification of fish was confirmed by molecular barcoding using the cox1 gene. RESULTS: The new species is distinguished from all other species of the genus by a combination of features, such as the number and size of the clamps, the shape and size of the genital atrium and the number of testes. The molecular analysis of the cox1 gene sequences showed that interspecific differences between Microcotyle justinei n. sp. and published sequences of Microcotyle spp. was greater than 8.8%, strongly suggesting that the new species is distinct from other congeners with sequences available on GenBank. CONCLUSION: The morphological and molecular analyses support the status of M. justinei as a new species. The present finding extends the list of Microcotyle spp. to 72.

Exploring the Efficacy and Safety of Levamisole Hydrochloride against Microcotyle sebastis in Korean Rockfish (Sebastes schlegelii): An In Vitro and In Vivo Approach.

Parasitic infections pose significant challenges in aquaculture, and the increasing resistance to conventional anthelmintics necessitates the exploration of alternative treatments. Levamisole hydrochloride (HCl) has demonstrated efficacy against monogenean infections in various fish species; however, research focused on Microcotyle sebastis infections in Korean rockfish (Sebastes schlegelii) remains limited. Therefore, this study aimed to evaluate the efficacy of levamisole HCl against M. sebastis infections in Korean rockfish with the goal of optimizing anthelmintic usage in aquaculture. In this study, we first assessed the susceptibility of M. sebastis to levamisole HCl in vitro. Subsequently, in vivo evaluations were conducted to assess the drug's efficacy, safety, and to identify optimal administration methods. In vitro experiments revealed concentration-dependent sensitivity of M. sebastis to levamisole HCl, with a minimum effective concentration (MEC) of 100 mg/L. In vivo experiments employed oral administration, intraperitoneal injection, and immersion treatments based on the MEC. Oral administration proved to be a safe method, yielding efficacy rates of 27.3% and 41.6% for 100 mg/kg and 200 mg/kg doses, respectively, in contrast to the immersion and injection methods, which induced symptoms of abnormal swimming, vomiting, and death. Biochemical analyses conducted to assess the safety of levamisole HCl revealed a transient, statistically significant elevation in the levels of glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT) on day three post-administration at 20 °C. Following this, no substantial differences were observed. However, at 13 °C, the enzyme levels remained relatively consistent, emphasizing the role of water temperature conditions in influencing the action of levamisole HCl. Our research findings substantiate the efficacy of levamisole HCl against M. sebastis in Korean rockfish, underscoring its potential for safe oral administration. These results provide valuable insights for developing parasite control strategies involving levamisole HCl in Korean rockfish populations while minimizing adverse impacts on fish health and the environment. However, this study bears limitations due to its controlled setting and narrow focus. Future research should expand on these findings by testing levamisole HCl in diverse environments, exploring different administration protocols, and examining wider temperature ranges.

Correlation of endemicity between monogenean trematode, Microcotyle sebastis, and parasitic copepods in Korean rockfish, Sebastes schlegelii, from a fish farm in Tongyeong-si, Gyeongsangnam-do, Korea.

Infestations of parasites, particularly those caused by copepods and monogeneans, are a major hindrance to aquaculture and have a big negative economic impact. Sebastes schlegelii, a farmed Korean rockfish, is particularly prone to copepods and monogeneans. This study comprehended how parasitic copepods and rockfish from a farm in Tongyeong-si, Gyeongsangnam-do, Korea, are related to the monogenean trematode Microcotyle sebastis. Our research revealed that monogeneans predominated, with a rate of 98.4% and an average infection intensity of 7 per infected fish. With an average infection intensity of 4 per sick fish, the infection rate for parasitic copepods was equally considerable, coming in at 91.7%. The high co-infection rate of both parasite families, at 90.8%, is noteworthy. Furthermore, Lepeophtheirus elegans and Peniculus truncatus, 2 parasitic copepod species, were discovered to have M. sebastis eggs attached to their bodies. This demonstrates a connection between parasitic copepod infestations in farmed Korean rockfish and the monogenean trematode M. sebastis. These discoveries highlight the necessity for creating more potent parasite control methods for the aquaculture sector and shed light on the intricate relationships between various parasite species.

The genus Microcotyle in Mediterranean scorpaenoids (Teleostei), including the description of Microcotyle merche n. sp. from Helicolenus dactylopterus (Delaroche, 1809).

More than 65 species of the genus Microcotyle Van Beneden & Hesse, 1863, have been described to date, most of them infecting Perciformes. Among the scorpaenoids (Perciformes, formerly Scorpaeniformes) the species of the genus Microcotyle parasitize sebastids and scorpaenids worldwide. In this study, we provide new morphological and molecular data for Microcotyle spp. in sebastids and scorpaenids from the Western Mediterranean and north-east Atlantic. Specimens of Helicolenus dactylopterus (Delaroche, 1809) (n = 107) and Scorpaena spp. (n = 107) were examined and their microcotylid specimens morphologically and molecularly characterized. Microcotyle merche n. sp. ex H. dactylopterus and specimens of Microcotyle algeriensis Ayadi, Gey, Justine & Tazerouti, 2016 from a new host and locality (Scorpaena scrofa from the north-east Atlantic) are herein described. Both species are phylogenetically close, but their morphology is markedly different mostly because the anterior lobe of the haptor present in other Microcotyle species is almost absent in M. algeriensis. Findings of M. merche n. sp. in the Mediterranean also excludes the presence of Microcotyle sebastis in this sea, encouraging the review of the exceptionally large host range and geographical distribution of this species.

Development of a PCR-RFLP method for differential identification of Microcotyle sebastis and Microcotyle caudata isolated from cultured rockfish in Korea.

Microcotylid monogeneans can cause considerable health problems in cultured fish, and several Microcotyle species are reported from scorpaenid fish, an economically important aquaculture target species in Korea. We developed a PCR-RFLP assay targeting the mitochondrial cox1 gene, for discriminating Microcotyle sebastis and M. caudata from cultured Korean rockfish Sebastes schlegelii and dark-banded rockfish S. inermis. AseI enzyme treatment of the PCR products showed that M. sebastis sequence was cleaved while M. caudata was not. A total of 95.2% (118/124) of monogeneans from S. schlegelii were identified as M. sebastis, and 96.2% (126/131) of monogeneans from S. inermis were identified as M. caudata by PCR-RFLP. However, the remaining parasites from each host showed the opposite digestion pattern. Additional analyses of these specimens by targeting the ITS region by PCR-RFLP showed the same results, suggesting that cross-species infection by the parasites may have occurred. In Korea, S. inermis net cages are commonly located nearby S. schlegelii net cages, and this encaged proximity might have provided the opportunity for cross-infection to occur. Further examination of wild host populations and experimental cross-infection will be necessary to explain this phenomenon. The PCR-RFLP method in this study will help investigate the epidemiology and infection dynamics of Microcotyle species in S. inermis.

Occurrence and Molecular Identification of Microcotyle sebastis Isolated from Fish Farms of the Korean Rockfish, Sebastes schlegelii.

Microcotyle sebastis is a gill monogenean ectoparasite that causes serious problems in the mariculture of the Korean rockfish, Sebastes schlegelii. In this study, we isolated the parasite from fish farms along the coasts of Tongyeong, South Korea in 2016, and characterized its infection, morphology and molecular phylogeny. The prevalence of M. sebastis infection during the study period ranged from 46.7% to 96.7%, and the mean intensity was 2.3 to 31.4 ind./fish, indicating that the fish was constantly exposed to parasitic infections throughout the year. Morphological observations under light and scanning electron microscopes of the M. sebastis isolates in this study showed the typical characteristics of the anterior prohaptor and posterior opisthaptor of monogenean parasites. In phylogenetic trees reconstructed using the nuclear 28S ribosomal RNA gene and the mitochondrial cytochrome c oxidase I gene (cox1), they consistently clustered together with their congeneric species, and showed the closest phylogenetic relationships to M. caudata and M. kasago in the cox1 tree.

Considerations on the taxonomy and morphology of Microcotyle spp.: redescription of M. erythrini van Beneden & Hesse, 1863 (sensu stricto) (Monogenea: Microcotylidae) and the description of a new species from Dentex dentex (L.) (Teleostei: Sparidae).

BACKGROUND: Microcotyle erythrini van Beneden & Hesse, 1863 (Platyhelminthes: Monogenea) and other closely related species of the genus are often considered as cryptic. Records in hosts other than the type-host with no species confirmation by molecular analyses have contributed to this situation. METHODS: Gill parasites of five sparid fishes, Boops boops (L.), Pagellus erythrinus (L.), P. acarne (Risso), Dentex dentex (L.) and Pagrus pagrus (L.), from the Western Mediterranean off Spain were collected. Specimens of Microcotyle spp. were characterised both molecularly and morphologically. Partial fragments (domains D1-D3) of the 28S rRNA gene and the cytochrome c oxidase subunit 1 (cox1) gene were amplified and used for molecular identification and phylogenetic reconstruction. Principal components analysis was used to look for patterns of morphological separation. RESULTS: Molecular analyses confirmed the identity of three species: M. erythrini ex P. erythrinus and Pa. pagrus; M. isyebi Bouguerche, Gey, Justine & Tazerouti, 2019 ex B. boops; and a species new to science described herein, M. whittingtoni n. sp. ex D. dentex. The specific morphological traits and confirmed hosts (P. erythrinus and Pa. pagrus) are delimited here in order to avoid misidentifications of M. erythrini (sensu stricto). Microcotyle erythrini (s.s.) is mostly differentiated by the shape of its haptor, which is also longer than in the other congeners. New morphological and molecular data are provided for M. isyebi from the Spanish Mediterranean enlarging the data on its geographical range. Microcotyle whittingtoni n. sp. is described from D. dentex and distinguished from the remaining currently recognised species of the genus by the number and robustness of the clamps. CONCLUSIONS: New diagnostic morphological traits useful to differentiate Microcotyle spp. are suggested: (i) haptor dimensions including lobes; (ii) the thickness of the clamps; (iii) the size and shape of spines of the genital atrium; (iv) the extension of the posterior extremities of vitelline fields; and (v) the shape of egg filaments. The use of new morphological approaches may allow considering these species of Microcotyle as being pseudocryptic. The use of representative undamaged specimens that have been genetically confirmed as conspecific is considered crucial to avoid abnormally wide morphological ranges that prevent species differentiation.

Morphological and molecular (28S rRNA) data of monogeneans (Platyhelminthes) infecting the gill lamellae of marine fishes in the Campeche Bank, southwest Gulf of Mexico.

During the examination of 913 fish specimens belonging to four families in the Campeche Bank (Gulf of Mexico), 23 gill ectoparasitic monogenean species were found, which belong to three families: Dactylogyridae, Microcotylidae and Diclidophoridae. The species Euryhaliotremaamydrum, E.carbuncularium, E.dunlapae, E.fajeravilae, E.fastigatum, E.longibaculum, E.paracanthi, E.tubocirrus, Haliotrematoidescornigerum, H.gracilihamus, H.heteracantha, H.longihamus, H.magnigastrohamus, H.striatohamus, Hamatopedunculariabagre, Neotetraonchusbravohollisae, and N.felis (all Dactylogyridae) were found on the hosts Lutjanussynagris, L.griseus, Ariopsisfelis, Bagremarinus, Archosargusrhomboidalis, and Haemulonplumieri. Additionally, Microcotylearchosargi, Microcotyle sp., and Microcotyloidesincisa (all Microcotylidae) were found on L.griseus and A.rhomboidalis; finally, Choricotyle sp. 1, Choricotyle sp. 2, and Choricotyle sp. 3 (all Diclidophoridae) were found on H.plumieri. The prevalence, abundance, mean intensity of infection, and supplementary taxonomic revisions for all monogeneans found are provided. Partial sequences of the 28S rRNA gene were also obtained for monogeneans of ariid, sparid, and haemulid host fishes to explore their systematic position within the Monogenea. New locality and host records for some previously described species of Euryhaliotrema, Hamatopeduncularia, Microcotyle, and Choricotyle from lutjanid, ariid, sparid, and haemulid hosts were reported. The present study adds evidence supporting the interoceanic occurrence of the same monogenean species (on lutjanids) on the west-east Atlantic and Pacific Oceans (= amphiamerican species). As previously suggested, there are at least, two possibilities to explain that parasite distribution: differentiation of morphological features in these monogeneans have resulted in only slight to insignificant morphological changes developing over the extended period of 3.2 mya (when the Isthmus of Panama was closing) and/or speciation is only evident at molecular level.