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The morphological characterisation of Diplozoidae spp. is highly reliant on the details of the sclerotised components of the hooks and clamps in the haptor. Only six species of Paradiplozoon (Diplozoinae) have been described from Africa, four of which have adequate morphological and even comparative ITS2 rDNA data available. However, the descriptions of Paradiplozoon ghanense (Thomas, 1957) and Paradiplozoon aegyptense (Fischthal & Kuntz, 1963) lack essential taxonomic information, specifically the details for their haptoral sclerites. As such, all available material from museum collections for these two species were studied using light microscopy to supplement the original morphometric descriptions. The holotype and paratypes of P. aegyptense were studied, but only voucher material for P. ghanense could be sourced. However, this voucher material for P. ghanense was deposited by the species authority and bore a striking resemblance to the illustrations and collection details from the original description. They were thus identified as the type series for the taxon, with a lectotype and paralectotype designated. Both P. ghanense and P. aegyptense could be readily distinguished from other taxa based on the supplementary data generated here, supporting their distinctness. The haptoral sclerites of P. aegyptense were most similar to those of Paradiplozoon krugerense Dos Santos & Avenant-Oldewage, 2016, also described from Labeo spp., while the sclerites of P. ghanense were most similar to Paradiplozoon bingolense Civáňová, Koyun & Koubková, 2013 and Paradiplozoon iraqense Al-Nasiri & Balbuena, 2016. Additionally, a voucher of P. aegyptense collected from the alestid type host of P. ghanense was reidentified as the latter species here. This greatly simplified the known host specificity for Paradiplozoon spp. in Africa, with P. aegyptense now exclusively reported from Cypriniformes (Cyprinidae and Danionidae), and P. ghanense restricted to Characiformes (Alestidae). The occurrence of all diplozoids from non-cyprinoid hosts was also investigated and several records of diplozoids occurring on non-cyprinoid hosts were collated and scrutinised. Excluding the two instances of diplozoids described and exclusively occurring on Characiformes fishes ( P. ghanense and Paradiplozoon tetragonopterini (Sterba, 1957)), most other non-cyprinoid collections appear sporadic and unsubstantiated, but warrant further investigation supported by diligent taxonomic data. Even though the morphometric descriptions of both P. ghanense and P. aegyptense were fully reported on here, additional material will be needed to study their genetic profiles and phylogeny., Competing Interests: Annemarie Avenant-Oldewage is an Academic Editor for PeerJ., (© 2024 Dos Santos and Avenant-Oldewage.)

Females of species of Lamproglena von Nordmann, 1832 are parasitic on the gills of teleost fishes and the 38 nominal species are based on mainly morphological data. Only four of these species have been genetically characterised and no mitochondrial data are available for the genus. The present study aimed to provide representative ribosomal DNA (rDNA) data for two additional species of Lamproglena from Africa: Lamproglena clariae Fryer, 1956 and Lamproglena hoi Dippenaar, Luus-Powell et Roux, 2001, alongside mitochondrial DNA (mtDNA) for these and two other African species, Lamproglena hemprichii von Nordmann, 1832 and Lamproglena monodi Capart, 1944. The four species were collected from Clariidae, Cyprinidae, Alestidae and Cichlidae, respectively. Representative 18S rDNA and 28S rDNA data were obtained for L. clariae and L. hoi, while cox1 mtDNA was obtained for all four species. The respective haplotypes supported the distinctness of all species using all three gene regions investigated. Interestingly, species appeared to be grouped more by geographical origin than host family, with L. hoi more closely related to other African species than to Asian species also collected from cyprinid hosts. Even though the results presented here greatly add to the molecular data available for Lamproglena, there are still 32 (>80%) species for which no genetic data are available. The interpretation of the results presented here is thus preliminary and much more data are required before the phylogeny of this genus, and other members of the family, such as Lernaea Linnaeus, 1758, can be studied appropriately.

Two species of Spinitectus Fourment, 1884 have been recorded from southern Africa, namely Spinitectus polli Campana-Rouget, 1961 and Spinitectus petterae Boomker, 1993, both from the Limpopo River system. Spinitectus petterae was described from North African catfish, Clarias gariepinus (Burchell), whereas S. polli infects squeakers, Synodontis spp. During parasitological surveys in the Vaal River system (Orange River catchment), Spinitectus specimens were collected from C. gariepinus. These systems are adjacent but not connected. Therefore, this study aimed to identify the specimens collected using morphological and molecular techniques. The morphological study included light and scanning electron microscopy of whole specimens and excised spicules. Specimens were genetically characterised using 18S rDNA, 28S rDNA and cox1 mtDNA. Additionally, immature specimens of S. petterae were collected near the type locality. Morphological characteristics were most similar to S. petterae from C. gariepinus, whereas genetic data were dissimilar to all available data for the genus. Additional morphological characteristics noted for S. petterae in the present study were the details of the left and right spicule structure and the porous structures on the pseudolabia. Specimens from the Vaal River system differed from those originally described as S. petterae by additional spines posterior to the third ring, lacking caudal alae and variable total body and male oesophagus length. Based on 18S rDNA, haplotypes from the type locality varied only slightly from the study material, supporting the morphological identification. However, 28S rDNA and, more conspicuously, cox1 mtDNA displayed substantial variation between specimens from these localities, which needs further investigation. Haplotypes generated in the present study were highly dissimilar to those characterised for S. petterae from Tanzania and Egypt. Nevertheless, the nematodes collected from C. gariepinus in the Vaal River system are considered S. petterae. This study expands the geographical distribution and adds additional morphological and genetic information for S. petterae, contributing to the limited knowledge of African species of Spinitectus.

Clarias gariepinus collected from Lake Heritage, Crocodile River, were found to harbour camallanid nematodes. Previously, Boomker (1982) surveyed the Hartbeespoort Dam, downstream of the current study site, and identified a high prevalence of Procamallanus (Procamallanus) laeviconchus and Paracamallanus cyathopharynx. Since then, Procamallanus (Procamallanus) pseudolaeviconchus was described from C. gariepinus suggesting reconsideration of the identifications of Procamallanus species in historical studies from clariids . The aim of the current study was to definitively identify the nematodes collected from C. gariepinus in Lake Heritage , using morphological and molecular analyses . Morphological study consisted of light and scanning electron microscopy which confirmed the identity P. (P.) pseudolaeviconchus and P. cyathopharynx. This included descriptions of the detailed morphology of isolated buccal capsules for both species using soft tissue digestion, notably for the first time for P. (P.) pseudolaeviconchus . The morphology of isolated spiculae of both species was described for the first time using SEM. Molecular analyses included genetic characterisation of the small ribosomal subunit (18S) rDNA and cytochrome oxidase 1 (CO1) mtDNA. Genetic data supported the morphological identification of both species, however, divergence was detected in CO1 mtDNA data for P. cyathopharynx indicating two distinct lineages . Due to this variation, the morphometry of P. cyathopharynx specimens were revisited including statistical re-evaluation. No robust morphological traits were identified to support CO1 mtDNA lineages and all specimens were considered conspecific. In terms of camallanid biodiversity in the Crocodile River system, it is similar to that in Boomker (1982), despite the altered water quality from past acid mine pollution in the river., Competing Interests: The authors confirm that there is no conflict of interest., (© 2022 The Authors.)

Adult trematodes of Allocreadium Looss, 1900 (Digenea) infect the intestine of mostly freshwater fishes in Asia, Europe, Africa and the Americas. During routine parasitological surveys in the Vaal River system, adult trematodes were collected from the intestine of smallmouth yellowfish, Labeobarbus aeneus (Burchell). The trematodes were confirmed to represent a member of Allocreadium and did not match any existing taxon. Therefore, they are described as a new species, Allocreadium apokryfi sp. n. The morphology of the new species most closely resembles that of Allocreadium aswanense El-Naffar, Saoud et Hassan, 1984, but it differs from it by having a bipartite internal seminal vesicle, wider eggs, a shorter intertesticular distance, an intestinal bifurcation at the ventral sucker level, a ventral sucker that is larger than the oral sucker, and a genital pore near the intestinal bifurcation or the ventral sucker. The surface topology of the new species is notably different from that of other allocreadiids. Papillae were observed in the ventral sucker and surrounding both ventral and oral suckers, but the number and arrangement of the latter were not consistent among specimens. The protruding cirrus of A. apokryfi sp. n. was described using SEM and is the first such observation for the genus. Genetic characterisation showed that the new species was clearly distinct from other Allocreadium spp. using both 18S (nucleotide difference 1.3-9.1%) and 28S (4.7-6.5%) rDNA, forming a well-supported clade in Allocreadium. The presence of A. apokryfi sp. n. in a well-studied river is unexpected, and considering the diet of its host and the scarcity of Allocreadium in Africa, the possible biology of this species is discussed herein.

In 2016, an intense copepod infection was recorded from a reservoir in proximity to the Danube River in Hungary from visibly emaciated wels catfish, Silurus glanis . The parasite-induced pathology was described but parasite identity was not conclusive. Additional sample collections in 2017 and 2018 allowed for identification using both light and scanning electron microscopy, alongside genetic characterisation. The copepods were confirmed to be ergasilids, Sinergasilus major , distinctly different from any previous infection on silurids in Europe. This is the first record of this parasite from Hungary and the first host record from wels catfish., (© 2021 The Author(s).)

The use of molecular tools in the study of parasite taxonomy and systematics have become a substantial and crucial component of parasitology. Having genetic characterisation at the disposal of researchers has produced mostly useful, and arguably more objective conclusions. However, there are several groups for which limited genetic information is available and, coupled with the lack of standardised protocols, renders molecular study of these groups challenging. The Diplozoidae are fascinating and unique monogeneans parasitizing mainly freshwater cyprinid fishes in Europe, Asia and Africa. This group was studied from a molecular aspect since the turn of the century and as such, limitations and variability concerning the use of these techniques have not been clearly defined. In this review, all literature and molecular information, primarily from online databases such as GenBank, were compiled and scrupulously analysed for the Diplozoidae. This was done to review the information, detect possible pitfalls, and provide a "checkpoint" for future molecular studies of the family. Hindrances detected are the availability of sequence data for only a limited number of species, frequently limited to a single sequence per species, and the heavy reliance on one non-coding ribosomal marker (ITS2 rDNA) which is difficult to align objectively and displays massive divergences between taxa. Challenging species identification and limited understanding of diplozoid species diversity and plasticity are also likely restricting factors, all of which hamper the accurate taxonomic and phylogenetic study of this group. Thus, a more integrated taxonomic approach through the inclusion of additional markers, application of more rigorous morphological assessment, more structured barcoding techniques, alongside thorough capturing of species descriptions including genetypes, genophore vouchers and reference collections in open sources are encouraged. The pitfalls highlighted are not singular to the Diplozoidae, and the study of other groups may benefit from the points raised here as well.

Diplozoidae infects the gills of cyprinid fishes in Africa, Europe, and Asia. Traditionally the hardened internal structures, crucial for identification of diplozoid species, are studied using light microscopy. Recently, the sclerotised haptoral structures of an African diplozoid, Paradiplozoon vaalense, were successfully isolated and visualised using scanning electron microscopy (SEM). In this paper, the haptoral sclerites of three diplozoid species are compared using SEM for the first time. Paradiplozoon ichthyoxanthon and Paradiplozoon vaalense occur on Labeobarbus and Labeo species, respectively, in the Vaal River system, South Africa, while Diplozoon paradoxum is widely-distributed in Europe and Asia, infecting several host species. Diplozoon paradoxum is a well-studied species, as well as being the type species of the family and ideal for inclusion in an exploratory study for comparative purposes. SEM study of D. paradoxum and P. ichthyoxanthon provided valuable information regarding surface morphology of the attachment structures hitherto not observed. Elaborate morphometric study of the haptoral hooks were incorporated, adding 12 point-to-point measurements to current morphometric characteristics. The results were analysed statistically, and significant differences support absolute separation (100.00%) of the three species following discriminant analysis. These point-to-point measurements could also be used for light microscopical study in the future and aid species delimitation within the Diplozoidae., Competing Interests: The authors have declared that no competing interests exist.

A new species of Gyrodactylus von Nordmann, 1832 is described from the gills of Alcolapia grahami, a tilapian fish endemic to Lake Magadi. This alkaline soda lake in the Rift Valley in Kenya is an extreme environment with pH as high as 11, temperatures up to 42 °C, and diurnal fluctuation between hyperoxia and virtual anoxia. Nevertheless, gyrodactylid monogeneans able to survive these hostile conditions were detected from the gills the Magadi tilapia. The worms were studied using light microscopy, isolated sclerites observed using scanning electron microscopy, and molecular techniques used to genetically characterize the specimens. The gyrodactylid was described as Gyrodactylus magadiensis n. sp. and could be distinguished from other Gyrodactylus species infecting African cichlid fish based on the comparatively long and narrow hamuli, a ventral bar with small rounded anterolateral processes and a tongue-shaped posterior membrane, and marginal hooks with slender sickles which are angled forward, a trapezoid to square toe, rounded heel, a long bridge prior to reaching marginal sickle shaft, and a long lateral edge of the toe. The species is also distinct from all other Gyrodactylus taxa based on the ITS region of rDNA (ITS1-5.8s-ITS2), strongly supporting the designation of a new species. These findings represent the second record of Gyrodactylus from Kenya, with the description of G. magadiensis bringing the total number of Gyrodactylus species described from African cichlids to 18., (© Q.M. Dos Santos et al., published by EDP Sciences, 2019.)

The sclerotized structures of monogeneans have traditionally been studied by light microscopy and different staining techniques. Recently, enzymatic digestion followed by scanning electron microscopy (SEM) has enabled the examination of structural details not visible with light microscopy. In order to obtain better, and more accurate, morphological information on sclerotized structures not affected by mounting medium or cover slip pressure, the sclerites of Cichlidogyrus philander Douëllou, 1993 (Monogenea, Ancyrocephalidae), collected from Pseudocrenilabrus philander (Weber, 1897), were redescribed using SEM. Parasites were collected from Padda Dam, Gauteng, South Africa and soft tissue was digested to release the sclerotized structures. The digested tissue also provided sufficient genetic material for molecular characterization of this species. Cichlidogyrus philander is characterised by a penis with a sharp, curved, lateral termination, an accessory piece with a hook-like extremity that may appear forked terminally, and lack of a visible vagina. The transverse bars have concave and convex surfaces with ribs on the concave surface. The dorsal bar bears fenestrations at the base of the auricles and the ventral and dorsal gripi are dissimilar. Furthermore, the large first pair of uncinuli shows lateral wings on the left side of the base. On top of this wing, a ball-like structure with a small fenestration is visible. Genetic characters derived from the 28S rDNA, the COI mitochondrial DNA and ITS1 rDNA regions distinguish C. philander from all other Cichlidogyrus sequenced species., (© P.C. Igeh et al., published by EDP Sciences, 2017.)