Underreported and taxonomically problematic: characterization of sanguinicolid larvae from freshwater limpets (Burnupiidae), with comments on the phylogeny and intermediate hosts of sanguinicolids.

Blood flukes of freshwater fish are understudied worldwide. Consequently, genetic information and data on their intramolluscan stages are scarce. In the current study, freshwater limpets of the genus Burnupia (Burnupiidae) from South Africa were examined for digeneans. Of 1645 specimens, 3.10% were infected by Sanguinicolidae larvae. Four sanguinicolids were distinguished by body size, number of penetration glands, tegumental spines' patterns and relative sizes of the finfolds on the body and furcae. Analyses of 28S, 18S and ITS rDNA sequences showed that the morphotypes were distinct from each other and from sanguinicolids whose genetic data are available. The present study is the first genetic characterization of sanguinicolids from Africa. Phylogenetic reconstruction revealed that the present species clustered with a sanguinicolid from Poland and were sister to Sanguinicola and Pseudosanguinicola from Russia and USA, respectively. The results indicate that the current species represent an unknown genus. What is more, blood fluke sequences from East Africa (presumed to be sanguinicolids), were distant from Sanguinicolidae and showed a closer relationship with acipensericolids from the USA. Freshwater fish blood flukes seem to be more diverse than previously recorded and use species of at least 13 gastropod families as intermediate hosts.

Pathology caused by introduced Neoergasilus japonicus (Copepoda: Ergasilidae) to the skin of indigenous Tilapia sparrmanii in South Africa and scanning electron microscopy study of wound-inflicting structures.

Neoergasilus japonicus, a fish ectoparasite indigenous to eastern and southern Asia, has been introduced to various regions, including Europe and North and South America, where it spreads at an alarming rate. The parasite is not host-specific. It was also collected from the Vaal-Orange and Limpopo river systems in South Africa. This report describes the morphology of the wound-inflecting structures in N. japonicus and relates their morphology to the pathological effect on Tilapia sparrmanii. Host tissue with parasites attached was imbedded in resin for histological sectioning and stained with haematoxylin and eosin. Additional specimens were prepared for scanning electron microscopy. Adult female parasites attach by inserting the hooked tips of the antennae into the epidermis. The maxillulae are minute but have sharp tips. The maxillae have sharp tips on the endopod and brush-like setae on the exopod. The mandibles are armed with sharp tips. These structures are used to remove the epidermis of the host and brush it into the buccal cavity. Host tissue in the intestine confirms that adult females consume host tissue leaving the dermis exposed to opportunistic diseases. The microscopic size of the parasite prevents casual observation, and its occurrence in Africa is, therefore, probably underreported.

Additional Taxonomic Information for Lamproglena hemprichii (Copepoda: Lernaeidae) Based on Scanning Electron Microscopy and Genetic Characterization, Alongside Some Aspects of Its Ecology.

Additional taxonomic and ecological data for the lernaeid copepod Lamproglena hemprichii Nordmann, 1832 infecting the African tigerfish, Hydrocynus vittatus Castelnau, 1861, are presented with scanning electron micrographs, molecular characterization, and selected ecological parameters. Eighty fish were collected from Lake Kariba between October 2014 and July 2015. Scanning electron microscopy provided additional data for the morphology, including structures on the antennulae, antennae, and legs. The 18S and 28S rDNA fragments of this species were distinct from those of other Lamproglena taxa but placed this species within this genus. Phylogenetically, L. hemprichii appears closest to L. monodi Capart, 1944, the only other African species for which molecular data is available. The anterior part of the second gill filament was the preferred attachment site. There was a positive correlation (Pearson correlation coefficient r2 = 0.64; P = 0.77) between the length of the parasite and the length of the fish. A positive correlation (Pearson correlation coefficient r2 = 0.61; P = 0.03) between the length of the parasite and the length of the gill filament was also observed. Although the specimens studied here are morphologically highly similar to previous reports of L. hemprichii, some morphological variation was observed, and a revision (morphometric and genetic) of the taxon is suggested. This study provides the first detailed genetic characterization and phylogenetic information for the species.

Light and scanning electron microscopy of the effects of Macrogyrodactylus congolensis (Prudhoe, 1957) on the skin of the African sharptooth catfish Clarias gariepinus (Burchell, 1822).

Clarias gariepinus (Burchell, 1822) is one of the two most actively cultured freshwater fish in Africa and therefore, economically important. Specimens of this species were purchased from a fish farm near Hartbeespoort Dam (North West, South Africa) and introduced into the tanks of the research aquarium in the Department of Zoology at the University of Johannesburg. However, the skin of these fish was infected with Macrogyrodactylus congolensis (Prudhoe, 1957), which proliferated profusely in the favourable conditions of the aquarium, posing a potential threat to its host. The current study was aimed at examining the pathology caused by M. congolensis on the skin of C. gariepinus. Species identification of the parasite was confirmed using light microscopy (LM), scanning electron microscopy (SEM) and DNA barcoding of the internal transcribed spacer region. Examination of the pathology was studied using LM of haematoxylin and eosin-stained sections (epoxy embedded) and SEM of parasites attached to the hosts' skin. Infected skin exhibited excessive mucus production, corroborated by an increased number of mucus cells alongside proliferated and abnormally enlarged club cells, resulting in varying thickness of the epidermal layer. At the site of attachment, the basement membrane detached from the dermis. Hamulus points and marginal hooks of the parasite pierce through the hosts' skin resulting in tearing. Epidermal cells and melanin granules were observed in the intestinal lumen of the parasite. Melanin granules were absorbed by the parasite's intestinal epithelium confirming that the parasite feeds on host skin tissue.

Aspects of the pathology induced by Spinitectus petterae Boomker, 1993 in the stomach of Clarias gariepinus (Burchell, 1822) using light and scanning electron microscopy.

Spinitectus spp. (Rhabdochonidae) are enteric nematodes characterized by annular spines. At the anterior end, these spines assist attachment and aid penetration into the host tissue. During parasitological surveys of the Vaal River system from three localities, below the Vaal River Barrage in the Vaal Dam surrounding UJ island and below the Grootdraai Dam, Spinitectus specimens were collected from the stomach lining of the sharptooth catfish, Clarias gariepinus (Burchell, 1822). Histopathological effects induced by Spinitectus petterae Boomker, 1993 on C. gariepinus has not been studied. Thus, the aim of this study was to describe the histopathology induced by S. petterae. For light microscopic examination, tissue samples with attached S. petterae were sectioned and stained with haematoxylin and eosin. Additionally, attached nematodes were also studied using scanning electron microscopy. Leukocytes were counted with the Disector principle. Standard infection parameters (prevalence, mean intensity and abundance) were calculated and compared to host parameters. Prevalence varied greatly (11.77% to 100%) between localities. Histopathology induced by S. petterae to C. gariepinus stomach (cardiac region) consisted of significant leukocyte infiltration, acute ulcerations and chronic granuloma formation. This was similar to the pathology of other Spinitectus occurring in host anterior intestine and stomach, but granuloma formation had not been previously reported and this suggests chronic infection in wild caught fish.

First isolation and scanning electron microscopy of haptoral sclerites of Macrogyrodactylus (Monogenea).

Macrogyrodactylus congolensis (Prudhoe, 1957) is one of six species of Macrogyrodactylus, all of which are endemic to Africa. This monogenean is a host-specific ectoparasite of the African sharptooth catfish, Clarias gariepinus (Burchell, 1822). It attaches to the host with a posterior haptor armed with sclerites. The specific morphology of sclerites is taxonomically significant and usually studied using light microscopy. The aim of the present study was to confirm the identification of macrogyrodactylid parasites using classic morphology (light microscopy of glycerine ammonium picrate mounted specimens) and molecular techniques (18S rDNA, ITS rDNA and cytochrome oxidase subunit 1 (COI) mtDNA). Additionally, the sclerites were accurately described with a technique not previously used for the genus, whereby haptoral sclerites were isolated by removing the encapsulating soft tissue with a digestion buffer and studied with scanning electron microscopy (SEM). Morphology and morphometry of studied specimens corresponded to available data for M. congolensis, confirming the identity of the parasite. All previous descriptions were summarized in a table and discrepancies discussed. Molecular analysis also confirmed the specimens to be M. congolensis, but ITS rDNA and COI mtDNA was more reliable than 18S rDNA in this regard. The isolation of haptoral sclerites and their study using SEM was successful, resolving the morphology of all sclerites. This study provided the first reconstruction of the haptor of a Macrogyrodactylus species following SEM analysis, as well as the first mtDNA for M. congolensis. Further study of isolated haptoral sclerites of other macrogyrodactylids is required to determine the full benefits of studying their isolated sclerites.

Parasite diversity of introduced fish Lates niloticus, Oreochromis niloticus and endemic Haplochromis spp. of Lake Victoria, Kenya.

This study reports on the status of metazoan fish parasites in Lake Victoria following the establishment of introduced Lates niloticus (Latidae) and Oreochromis niloticus (Cichlidae) and changes in environmental quality. For this study, 412 fish specimens were examined for parasites: 103 L. niloticus, 165 O. niloticus, 82 Haplochromis piceatus and 62 H. humilior (endemic cichlids). In total, 25 parasite taxa were identified: Lates niloticus (6), O. niloticus (19) and Haplochromis spp. (13). The myxosporean Henneguya ghaffari, prevalence (P) = 79% and the monogenean Diplectanum lacustris (P = 34%), were the dominant parasites on L. niloticus. Myxobolus sp. (P = 44%) was dominant on O. niloticus while for the haplochromines, metacercariae of 'Neascus' sp. (Diplostomidae) was dominant (P = 37%). Contrary to reports of high diversity of monogeneans on endemic species, the haplochromines harboured only Cichlidogyrus gillardinae (P = 6.9%). Oreochromis niloticus harboured seven monogenean species: Cichlidogyrus sclerosus, C. halli, C. tilapiae, C. quaestio, Scutogyrus longicornis, Gyrodactylus cichlidarum and G. malalai-they appear to have been co-introduced with the fish. Cichlidogyrus gillardinae, C. quaestio, G. malalai and the acanthocephalan Acanthogyrus (Acathosentis) tilapiae are recorded for the first time in Lake Victoria, representing new geographical records. Presence of A. (A) tilapiae in Haplochromis spp. indicates possible spillover from O. niloticus. Low prevalence and diversity of monogeneans, crustaceans and glochidia on the fish corresponded with increased pollution in the lake. Overall, changing environmental conditions and introductions of fish species have contributed to parasite community changes in Lake Victoria.

Pathological effects of Cichlidogyrus philander Douëllou, 1993 (Monogenea, Ancyrocephalidae) on the gills of Pseudocrenilabrus philander (Weber, 1897) (Cichlidae).

Histopathological changes of Cichlidogyrus philander Douëllou, 1993 on the gills of Pseudocrenilabrus philander (Weber, 1897) were studied using light and scanning electron microscopy. Observations revealed that C. philander attaches to its host by alternating the prohaptor (for temporary attachment or feeding) or haptor (using haptoral parts for firm and secured attachment). The sharp terminal ends of the anchors are inserted basally into the gill lamella, between two adjacent secondary gill lamellae and the marginal hooklets assist by superficially penetrating, holding and lifting epithelial tissue in the proximal region of the secondary gill lamella. The attachment of C. philander resulted in compression, rupturing of the interlamellar epithelium, change in the organization of epithelial cells in both primary and secondary gill lamella, displacement of the extracellular cartilaginous matrix, occasional rupturing of blood vessels and erythrocytes and some cells becoming ill-defined. At the site of attachment, the host response comprises of hyperplasia, increase in the number of mucous cells and infiltration with neutrophils. It was concluded that the effect of C. philander is mild in natural conditions.

You are how you eat: differences in trophic position of two parasite species infecting a single host according to stable isotopes.

Parasitism is commonly recognised as a consumer strategy, although, the interaction of parasites in communities and ecosystems are generally poorly understood. As parasites are integral parts of food webs, analysis of the trophic interactions between parasites and hosts was assessed through comparison of stable isotope ratios of carbon (13C/12C) and nitrogen (15N/14N). Largemouth yellowfish (Labeobarbus kimberleyensis) infected with the Asian tapeworm (Schyzocotyle acheilognathi) were collected from the Vaal Dam. Signatures of δ13C and δ15N were assessed in host muscle and liver tissue, and cestodes using an elemental analyser coupled with an isotope ratio-mass spectrometer (EA-IRMS). Hosts were enriched by 4.1‰ in the heavy nitrogen isotope with respect to the S. acheilognathi and therefore occupy a higher trophic position than the parasite. Comparison of δ13C indicates that dietary sources of carbon in cestodes are derived from the host liver. Comparison of stable isotope signatures between Paradiplozoon ichthyoxanthon (another common parasite of the Largemouth yellowfish in the Vaal River) and S. acheilognathi showed that the monogenean was enriched by 5.3‰ in 15N which accounts for a difference of almost two trophic positions. Isotope differences in the host-parasite system considered indicate that differences can be related to the mode of nutrient acquisition employed by host and parasites. Cestodes, being depleted in both 13C and 15N relative to the host and monogenean (P. ichthyoxanthon), indicate that S. acheilognathi assimilates nutrients derived from the host metabolism which are released from the liver.

Trace Elements in Crustaceans, Mollusks and Fish in the Kenyan Part of Lake Victoria: Bioaccumulation, Bioindication and Health Risk Analysis.

This is the first comprehensive report on the accumulation of Cr, Ni, As, and Ag in the fish species Nile tilapia Oreochromis niloticus and Nile perch Lates niloticus from Lake Victoria, complemented with recent data on Cu, Zn, Cd, and Pb. This also is the first report on Cr, Ni, As, and Ag levels in invertebrates: the shrimp Caridina nilotica, gastropod Pila ovata, and bivalve Mutela bourguignati. The study was conducted at five sites in the Kenyan part of Lake Victoria: four sites in Winam Gulf influenced by various anthropogenic pressures, including a site near Kisumu City, and one in the main lake, with lesser direct anthropogenic influence. Apart from Cu and Ag, which were highest in O. niloticus liver, the invertebrates had higher levels of trace elements than fish. Contamination of the gulf with trace elements was best mirrored by the invertebrates, whose mobility is limited; they accumulated Cr, Cd, Ag, and Pb corresponding to the levels in the surface sediment. The accumulation of trace elements in fish species and their bioindicative potential corresponded to their habitats and feeding behaviour. The tissue contents of most trace elements were higher in the inshore-dwelling, omnivorous O. niloticus compared to the pelagic, piscivorous L. niloticus. Cu (465 ± 689 mg/kg dw) and Ag (3.45 ± 1.49 mg/kg dw) in the liver of O. niloticus were up to 10 and 119 times higher than in L. niloticus, respectively. Oreochromis niloticus therefore has bioindicative potential for Cu and Ag contamination. Both the invertebrates and fish showed positive correlations between Cu and Ag concentrations, indicating similar source and/or uptake route. The target hazard quotients (THQ) show that there is no human health risk associated with the consumption of these fish. However, the levels of Zn, Cd, and Pb in P. ovata surpassed maximum food safety limits and are hence potentially unsafe for human consumption.

Histopathological changes induced by the digenean intestinal parasite Masenia nkomatiensis Dumbo, Dos Santos, & Avenant-Oldewage, 2019 of the catfish Clarias gariepinus (Burchell) from Incomati Basin, Mozambique.

The intestines of 154 Clarias gariepinus were examined of which 29 were naturally infected with Masenia nkomatiensis, and of these, seven (intensity ranging from 8 to 231) were examined for pathology. Destruction of the epithelium covering the villi, detachment of epithelial cells and parts of villi were observed. Excessive mucus secretion occurred in the vicinity of the worm and catarrh was observed, indicative of an inflammatory response. The number of mucous and mast cells was higher at the attachment site than at an area 5,000 µm away and in uninfected individuals, suggesting that the parasite triggered a localized innate immune response. The number of neutrophils, basophils and lymphocytes in infected tissue was not significantly different from uninfected tissue confirming that no acquired immune response was produced against the maseniid. The caecae in the anterior part of the parasites' intestine consisted of convoluted epithelium forming invaginations or "crypts." Contraction of the thick layer of circular muscle fibres of the caeca facilitates the movement of digested material. Observation of digested host cells and cell debris within the caecae provides further evidence that M. nkomatiensis is consuming host cells.

Masenia nkomatiensis n. sp. (Digenea: Cephalogonimidae) from Clarias gariepinus (Burchell) (Clariidae) in Incomati Basin, Mozambique.

A new species of Masenia Chatterji, 1933 is described based on material from the intestine of Clarias gariepinus (Burchell) in the Incomati River, Mozambique. The combination of morphological characteristics and analysis of 18S and 28S rDNA sequences delineated the specimens found in the present study as a distinct species. The new form is distinguished from other Masenia spp. in having a large reniform seminal receptacle, a cirrus-sac ending anterior to the ventral sucker, intestinal caeca extending into the hindbody to the level of the posterior testis, and the vitelline fields extending anteriorly to the ventral sucker and posteriorly to the middle of the ovary. Notably, the new form is the only record of African species having a sac-shaped excretory vesicle. Analysis of 28S rDNA sequence data supported its placement in the Cephalogonimidae Looss, 1899. 18S analyses also supported its placement in this family but showed it was not closely related to Masenia bangweulensis (Beverley-Burton, 1962), the sole other African species for which genetic data is currently accessible. The total pairwise differences for 18S and 28S sequences showed the new form differing from other cephalogonimids. Thus, both morphological and genetic characteristics indicate that the present form represents a distinct species, here described as Masenia nkomatiensis n. sp. An updated key to African Masenia spp., now five, is provided.

Trace Elements in Water, Sediments and the Elongate Tigerfish Hydrocynus forskahlii (Cuvier 1819) from Lake Turkana, Kenya Including a Comprehensive Health Risk Analysis.

This study presents the distribution of 17 major and trace elements in surface water, sediments and fish tissues from Lake Turkana, Kenya. Eight sediment and ten water samples from the west bank of the lake, as well as 34 specimens of the elongate tigerfish Hydrocynus forskahlii caught in that region were examined. It is the first report for Li, Rb, Sr, Mo from the lake and the first report on most of the trace elements for this fish species. The concentrations of elements in the water and sediments showed no sign of pollution. In fish muscle, Li, Zn and Cd showed relatively high abundances, with mean concentrations of 206, 427 and 0.56 mg/kg dw, respectively. The calculated target hazard quotient values for Li, Zn, Sr and Cd were 138.7, 1.9, 4.1 and 0.76, respectively; therefore the consumption of these fish poses a health risk to humans in the area.

Additional morphological information on Dipteropeltis hirundo Calman, 1912, and a description of Dipteropeltis campanaformis n. sp. (Crustacea: Branchiura) from two characiform benthopelagic fish hosts from two Northern rivers of the Brazilian Amazon.

Dipteropeltis is a monotypic genus of the Branchiura and endemic to South America. Twelve specimens of Dipteropeltis sp. and micrographs of a thirteenth specimen were obtained from four institutions. Measurements and light micrographs were taken of all the specimens. The specimens were compared to all known descriptions of D. hirundo and sorted into two groups, those that conformed to the description of D. hirundo; and those that differed morphologically. Two specimens, one from each group¸ were stained with lignin pink and subsequently prepared for Scanning Electron Microscopy. The differences observed in the specimens indicated that a new species exists. Thus, Dipteropeltis campanaformis n. sp. is described from seven specimens collected from two characiform benthopelagic fish species and compared to D. hirundo. Dipteropeltis campanaformis n. sp. differs from D. hirundo in the head shape, the length and shape of the carapace lobes, the membrane composition of the maxillules, the shape of the maxillae, the shape of the mouth, and the shape of the natatory lobes.

Gastropod invasions in anthropogenically impacted impoundments in South Africa: Tracing their origins and exploring field evidence of parasite spillback and amplification.

Invasive snails are associated with ecological problems in freshwater bodies worldwide. However, their impact on the transmission of digenean infections remain underreported. In the present study, 1708 specimens representing four snail species were sampled from four impoundments in the Limpopo River system in South Africa. Gyraulus chinensis (Planorbidae), Physella acuta (Physidae) and Pseudosuccinea columella (Lymnaeidae), which are invasive, were found in all the sampling sites. In contrast, the native lymnaeid Radix natalensis occurred at only one study site. Digeneans were observed only from R. natalensis (prevalence = 49%) and Ps. columella (prevalence = 23%). Morphological and genetic analyses revealed four digeneans: Fasciola nyanzae, Orientocreadium sp., Petasiger sp. and Patagifer vioscai. Pseudosuccinea columella was infected by the four digeneans while R. natalensis harboured only Orientocreadium sp. and Petasiger sp. Partial sequences of Orientocreadium sp. from the current study differed from congeners whose DNA data are available on GenBank, by p-distances of at least 1.84 and 2.2% for 28S and the internal transcribed spacer (ITS) rDNA, respectively. Phylogenetic analyses demonstrated that the present species is sister to Orientocreadium batrachoides. Genetic and phylogenetic data based on 28S and ITS rDNA suggested that Petasiger sp. from the present study and isolates of three unidentified Petasiger spp. from Kenya, Hungary and Australia, were representatives of the same species. This is the first known report of Orientocreadium, Petasiger and Patagifer from Ps. columella. The occurrence of F. nyanzae in Ps. columella indicates spillback from R. natalensis. These findings echo the concerns raised in previous studies about the potential role of Ps. columella in the amplification of digenean diseases in its introduced range. Phylogenetic analyses of partial sequences of the cytochrome c oxidase subunit 1 mitochondrial gene (cox1) showed multiple lineages of Ps. columella in North and South America. Pseudosuccinea columella specimens from the present study belong to an invasive genotype that has spread globally and has been reported from Zimbabwe, Egypt, Portugal, Australia, Argentina, Colombia and New Mexico (USA). Physella acuta from the current study had a stronger genetic relationship with isolates from Canada and Iceland, than with isolates from other parts of Africa, suggesting several invasion routes into Africa. This is the first known DNA characterisation of G. chinensis from Africa. Phylogenetic reconstruction indicated multiple exit events of G. chinensis from Asia into Europe and Africa. South African isolates clustered in a recent branch containing isolates from the Czech Republic and Hong Kong, China. Considering the presence of invasive snails in all the sampling sites in the present study, it is necessary to investigate the factors that enhance their establishment and to monitor their effects on the native snail populations.

Effects of water quality on fish parasite biodiversity and physiological responses in the host fish Clarias gariepinus from a eutrophic lake subjected to acid mine drainage in South Africa.

Elevated concentrations of contaminants have negative impacts on aquatic organisms and their parasites. Changes in parasite infections have been proposed as a technique for monitoring the health of aquatic ecosystems. Furthermore, alterations in physiological responses (biomarkers) of organisms have also been used to delineate ecosystem quality. Lake Heritage is situated along the Crocodile River in Muldersdrift, Gauteng, South Africa, and is subject to contamination by acid mine drainage. Clarias gariepinus is a well-studied bioindicator species and host to numerous endoparasites and ectoparasites. The aims of this study were to delineate the health status of Lake Heritage through a multifaceted approach by comparing the water quality, biomarker responses, and parasite biodiversity of C. gariepinus, compared to unexposed laboratory-reared fish. Physical and chemical water quality parameters were determined using a hand-held probe, test kits, and element analysis with inductively coupled plasma-mass spectrometry. Biomarker responses in the gill, liver, and muscle tissues from C. gariepinus were assessed for total protein, metallothioneins, superoxide dismutase (SOD), and reduced glutathione (GSH) concentrations and activities of acetylcholinesterase and catalase. Results for water quality variables showed higher pH, nitrate, hardness, and copper levels compared with the South African Target Water Quality Guidelines. Catalase activity and concentrations of SOD and reduced GSH showed a response in C. gariepinus to the water quality. Ectoparasites had lower prevalence and mean intensity than endoparasites. However, there were no differences in the physiological responses between infected and uninfected hosts. The study shows that the eutrophic conditions in Lake Heritage cause biomarker responses in the host when compared to host fish in laboratory conditions. Integr Environ Assess Manag 2024;20:1539-1553. © 2023 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

Revisiting the type material of two African Diplozoinae (Diplozoidae: Monogenea), with remarks on morphology, systematics and diplozoid specificity.

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 Civánová, 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.

Integrated characterisation of Daubaylia burnupiae n. sp. (Nematoda: Daubayliidae) from a freshwater gastropod in South Africa, with comments on the biology of Daubaylia spp.

Gastropod-nematode associations are underreported worldwide. In the present study, juvenile and adult nematodes were found in the freshwater gastropod Burnupia stenochorias (Melvill & Ponsonby, 1903), from the Vaal River, South Africa. The nematodes were confirmed to belong to the genus Daubaylia chitwood & chitwood, 1934 (Daubayliidae). This is the first report of Daubaylia from a snail belonging to the family Burnupiidae, and the first report of this nematode taxon in southern Africa. Like D. pearsoni and D. malayanum from Australia and Malaysia respectively, adult females of the current species possess multiple well-developed eggs in the uteri, with larvae developing in utero. Morphological and molecular characteristics showed that the nematodes are distinct from all the described species of Daubaylia. Thus, they are considered a new species, Daubaylia burnupiae n. sp. The species differs from its congeners based on spicule shape, the short tail of the male, an anal cuticular knob-like protrusion on the female, and oesophagi with short isthmi and short glandular basal bulbs in both sexes. Three club-shaped pharyngeal lobes, extending slightly above the surface of the cephalic lips in both sexes and a pre-cloacal median papilla on the male were described using scanning electron microscopy, the first of such observations for the daubayliids. Genetic analyses showed that partial sequences of D. burnupiae n. sp. differed from species for which genetic data are available, by at least 26 and 9 base pair differences for 28S and 18S rDNA, respectively. Our results show that low prevalence and abundance of nematodes in the snails, corresponded with increased pollution in the river. We suspect that exposure to pollutants reduces the viability of the infective gravid female nematode during transmission. Therefore, the nematode is a potential bioindicator for aquatic pollution.

Additional data on Spinitectus petterae (Nematoda: Rhabditida) from Clarias gariepinus (Siluriformes: Clariidae) in the Vaal River system: conserved morphology or high intraspecific genetic variability?

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.

Biomineralisation and metal sequestration in a crustacean ectoparasite infecting the gills of a freshwater fish.

It has been suggested that parasites are effective bioindicators as they are sensitive to environmental changes and, in some cases, accumulate trace elements in higher concentrations than their hosts. Accumulated elements sequester in different organs. In monogenean and crustacean ectoparasites, sclerotised structures and egg yolk appear to be the preferred site for element sequestration. In this study, the sequestration of trace elements; Mg, Al, Ca, Fe, Cu, and Zn in Lamproglena clariae was studied from two rivers. Adult L. clariae were collected from the gills of Clarias gariepinus from Lake Heritage in the Crocodile River and in the Vaal River below the Vaal Dam, South Africa. Collected parasites were flash frozen in liquid nitrogen and sectioned with a cryomicrotome. Sections were treated with Phen-Green to observe fluorescent signals. Trace elements in the parasite were analysed using a scanning electron microscope with an energy-dispersive spectroscope (SEM-EDS). Results showed more intense fluorescence signals in the exoskeleton compared to tissues, and in the egg yolk. Analysis by SEM-EDS confirmed the presence of elements in the parasite from both sites. Levels of Al were higher in L. clariae from the Vaal River than those from Lake Heritage, and Fe was higher in L. clariae from Lake Heritage. Element distribution patterns in the parasite matched those in the water from the sites. Unlike other crustaceans, regulation of metals in adult females of L. clariae does not occur through moulting, but high levels occurred in the yolk.