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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.

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.

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.

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.

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.

The effect of increased aluminium concentration on the hatchability and survival of Lamproglena clariae is unknown. During October 2019 and October 2020, infected C. gariepinus were collected from the Vaal River and transferred to a controlled aquarium room. Parasite infection was established on acclimated C. gariepinus. Adult female parasites (F2 and F3 generation) on live fish were inspected for egg strings. Viable egg strings were removed, leaving females attached to produce more eggs. Bioassays were performed in varying concentrations of Al (control, 5 µg/L, 30 µg/L, 60 µg/L, and 120 µg/L). Egg development was monitored. In situ physical and chemical water quality parameters were measured, and water samples were collected every 24 h for metal analysis using inductively coupled mass spectrometry. The experiment terminated when all juveniles perished. Five percent of exposed eggs did not hatch in the control solution, compared to 26% in 120 µg/L Al. Hatchability and survival of L. clariae were negatively affected by increased Al concentrations. By removing this ectoparasite from living fish, the need to euthanise the host organism is eliminated, emphasising the usefulness of L. clariae as a bioindicator for metal pollution.

Abstract 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.

Parasites and environmental features could synergistically act as stressors to the health of their hosts. The objectives of this study were to evaluate the effect of: (i) water quality, host sex, size and body condition on adult digenean parasite infections; (ii) digenean infections and host sex and size on the oxidative stress biomarkers and body condition of hosts; and (iii) water quality on the oxidative stress biomarkers and body condition in Clarias gariepinus. Water quality variables were measured and C. gariepinus were collected each month for a year for examination of two intestinal digeneans, Masenia nkomatiensis and Glossidium pedatum, and determination of body condition and measurement of biomarkers in the host. The results indicated that the intensity of M. nkomatiensis was positively correlated with electrical conductivity and total dissolved solids. Prevalence of G. pedatum was negatively correlated with electrical conductivity, salinity and total dissolved solids. High summer water temperature was strongly associated with high digenean infections. There was no host body condition, sex or size bias for any of the parasite infection variables. Differences in the biomarker levels and body condition between uninfected fish and those infected with M. nkomatiensis or G. pedatum were insignificant indicating a low effect of the digenean parasites on oxidative stress biomarkers and body condition in the fish. However, total protein levels were positively associated with host size, and lipid peroxidation was negatively related to host body condition; total protein levels were also positively correlated with temperature and negatively correlated with dissolved oxygen. Host body condition was only negatively correlated with dissolved oxygen. Overall the trends observed in the data showed that the parasites have a negligible effect on oxidative stress in host fish and the trends observed for all variables (water quality, stress biomarkers, body condition and parasite infections) showed a strong seasonal pattern.

Stable isotope analysis offers a unique tool for comparing trophic interactions and food web architecture in ecosystems based on analysis of stable isotope ratios of carbon (13C/12C) and nitrogen (15N/14N) in organisms. Clarias gariepinus were collected from six sites along the Vaal River, South Africa and were assessed for ectoparasites and endoparasites. Lamproglena clariae (Copepoda), Tetracampos ciliotheca and Proteocephalus glanduligerus (Cestoda), and larval Contracaecum sp. (Nematoda) were collected from the gills, intestine and mesenteries, respectively. Signatures of δ13C and δ15N were analysed in host muscle tissue and parasites using bulk stable isotope analysis. Variable stable isotope enrichment between parasites and host were observed; L. clariae and the host shared similar δ15N signatures and endoparasites being depleted in δ13C and δ15N relative to the host. Differences in stable isotope enrichment between parasites could be related to the feeding strategy of each parasite species collected. Geographic and spatial differences in enrichment of stable isotopes observed in hosts were mirrored by parasites. As parasites rely on a single host for meeting their nutritional demands, stable isotope variability in parasites relates to the dietary differences of host organisms and therefore variations in baseline stable isotope signatures of food items consumed by hosts.

Monogenea is a Class of ecologically and economically significant platyhelminth parasites of freshwater and marine hosts. Contamination of the environment has been shown to variably affect parasites resulting in alterations at the population and organismal levels. The aim of this study was to perform a meta-analysis of published data to assess similarities among responses of monogeneans to pollution and estimate effect sizes of contaminant exposure on these parasites. Effect size was calculated according to Cohen’s d based on reported mean abundance for monogeneans from individual studies. A total of 57 studies were used in the present study. Each Monogenea–contaminant record represented a single data point in the meta-analysis, and this accounted for a total of 217 useable records. Most studies were concerned with species belong to Gyrodactylidae, Diplozoidae, Ancyrocephalidae and Dactylogyridae. Assessment of the effects of specific pollutants on monogeneans have mostly been restricted to exposure to eutrophication, pulp-mill effluent and metals. Comparison of effect sizes showed strong negative effects for metals and strong positive effects for eutrophication on monogeneans. Other contaminant types had weak effect sizes on monogeneans. Comparison of effect sizes among monogenean families showed that none of the effect sizes calculated were significant but metals and disturbance contaminants showed strong negative and positive effects respectively for Gyrodactylidae. The current comparisons were based on changes in the population structure and infections of monogeneans, however, organ level effects associated with exposure to poor water quality have also been noted. In this instance, exposure has been suggested to result in asymmetry of the attachment organs of some monogeneans. Despite numerous records of impacts of pollution exposure at the population level, understanding of the physiological effects and mechanisms employed by Monogenea to survive and adapt to changes in the environment are yet to be explored. Study of these aspects would enable better understanding of the variation in the trends observed at a population level for monogeneans in relation to the quality of the macroenvironment.

The upper reaches of the Crocodile River (West) system are located in the western basin of the Witwatersrand mountain chain, an area heavily influenced by gold mining. After the spill of highly acidic and contaminated mining-influenced water into the river system in 2002, chemical water treatment was implemented to compensate for the consequences of acid mine drainage. Some studies thereafter have shown metal concentrations of concern in the system, but never considered longer periods of time. For this study, we investigated water and sediments over a period of 13 months from 7 sampling sites in the system. In addition to physico-chemical parameters such as temperature, pH, and electrical conductivity, a multi-element analysis was conducted. We used various versions of atomic-absorption-spectroscopy, total reflection x-ray fluorescence spectroscopy, and inductively coupled plasma mass spectrometry in filtered and acidified water samples as well as sediment leachates. Concentrations of Ni, Zn, As, Pb, and U in the sediment were clearly elevated at the site closest to the mine as well as further downstream, some of them far exceeding quality guidelines. Moreover, dissolved Mn, Fe, Ni, Zn and U occurred irregularly in concentrations of concern at the site of mining-influenced water inflow. Our findings clearly indicate a risk of further and chronic mobilization of toxic elements from this site and a possible threat to the connected river system.

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Lake Kariba is a tropical lake with slight variations in seasonal temperature. Temperature is an important physical variable in the biology of both fish and their parasites. Currently, there is no information on the seasonal occurrence of fish parasites in Lake Kariba. The objective of this study was to investigate the seasonal occurrence of metazoan parasites in Hydrocynus vittatus in Lake Kariba, Zimbabwe. Twenty fish specimens were collected by seine netting per season between October 2014 and July 2015 in the Sanyati Basin, Lake Kariba, and examined for metazoan parasites. Mean water temperatures ranged from 24.1 °C to 31.2 °C with slight variations between the seasons. Metazoan parasites consisting of Monogenea (Annulotrema pikei, Annulotrema pseudonili, Annulotrema bracteatum), Nematoda (Contracaecum larvae), Copepoda (Lamproglena hemprichii), Cestoda (larval cestodes, Ichthybothrium sp.) and Pentastomida (pentastomid larvae) were recorded. Larval cestodes were recorded in autumn and spring, while pentastome larvae were recorded in summer and spring. The Ichthybothrium sp. was recorded once in winter. Annulotrema pikei and A. pseudonili were observed on the gills and A. bracteatum on both the gills and the skin. Contracaecum larvae, L. hemprichii and A. bracteatum (from the skin) were recorded in all the seasons, with slight variations in prevalence, mean abundance and mean intensity. However, these variations were not statistically significant (analysis of variance or ANOVA, p > 0.05). The slight variations in occurrence of the parasites were probably because of the thermal stability of the lake where variation in temperature was small between seasons. Both A. bracteatum and Contracaecum larvae were aggregated on the fish host, whereas L. hemprichii exhibited a random distribution. Parasite diversity was at its highest during winter.

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.