Working towards a conservation plan for fish parasites: Cyprinid parasites from the south African cape fold freshwater ecoregion as a case study.

The preservation of the world's biodiversity for future generations has been a global objective for many years, with the establishment of the International Union for Conservation of Nature (IUCN) Red List of Threatened Species in 1964. However, the conservation of parasites is a more recent development and, due to the difficulty in obtaining data and studying some of the parasitic species, comes its own series of challenges. Using parasites of cyprinid hosts (one critically endangered, one endangered and three near threatened) collected from South Africa's Cape Fold freshwater ecoregion (CF) as a case study, this paper discusses the challenges and possible solutions for implementing a fish parasite conservation project. Novel data on the fish parasites (1819 metazoan parasite individuals, representing the Acanthocephala, Cestoda, Copepoda, Digenea, Monogenea and Nematoda) of the CF are provided from the five endemic hosts, Cheilobarbus serra (Peters, 1864), Labeobarbus seeberi (Gilchrist et Thompson, 1913), Pseudobarbus phlegethon (Barnard, 1938), Sedercypris calidus (Barnard, 1938), and Sedercypris erubescens (Skelton, 1974). Conservation statuses for selected parasite taxa are also proposed based on the conservation statuses of the fish hosts, according to the Conservation Assessment Methodology for Animal Parasites (CAMAP).

The use of fish parasitic isopods as element accumulation indicators in marine pollution monitoring.

Marine ecosystems are continuously under threat due to pollutants, which endanger marine biodiversity. The present study determines the potential use of the parasitic isopod, Cinusa tetrodontis Schjödte et Meinert, 1884, together with its fish host, Amblyrhynchotes honckenii (Bloch) for marine bioaccumulation monitoring. The concentrations of As, Cd, Cu, Mn, Ni, Pb, Se, and Zn were determined in muscle and liver tissues of infested and uninfested fish, and male and female parasites on the South African temperate south coast. The concentrations of Cu and Ni in C. tetrodontis differed significantly between two sampling sites, a near-pristine (Breede River Estuary, Witsand) and a more polluted site (harbour area in Mossel Bay). Mossel Bay isopods had higher concentrations of Ni, while Witsand isopods had higher concentrations of Cu. In contrast to fish hosts, parasitic isopods accumulated significantly higher levels of all elements except Cd. Most significant relationships between elements accumulated by C. tetrodontis and an increase of elements in fish tissues were seen in liver, rather than muscle tissue samples. Specimens of C. tetrodontis can be defined as good bioindicators for elements such as As, Cu, Mn, Ni, Pb, Se, and Zn, as they possess high bioaccumulation capabilities. This study addresses one of several future directions needed within environmental parasitology and highlights the importance of studying and utilising this host-ectoparasite model system.

Bioaccumulation and trophic transfer of total mercury through the aquatic food webs of an African sub-tropical wetland system.

Anthropogenic activities, including combustion of fossil fuels, coal, and gold mining, are significant sources of mercury (Hg) emissions into aquatic ecosystems. South Africa is a major contributor to global Hg emissions (46.4 tons Hg in 2018), with coal-fired power stations as the main source. Atmospheric transport of Hg emissions is the dominant cause of contamination, especially on the east coast of southern Africa where the Phongolo River Floodplain (PRF) is located. The PRF is the largest floodplain system in South Africa, with unique wetlands and high biodiversity, and provides essential ecosystem services to local communities who rely on fish as a protein source. We assessed the bioaccumulation of Hg in various biota, the trophic positions and food webs, as well as the biomagnification of Hg through the food webs in the PRF. Elevated Hg concentrations were found in sediments, macroinvertebrates and fish from the main rivers and associated floodplains in the PRF. Mercury biomagnification was observed through the food webs, with the apex predator tigerfish, Hydrocynus vittatus, having the highest Hg concentration. Our study shows that Hg in the PRF is bioavailable, accumulates in biota and biomagnifies in food webs.

Total mercury concentrations in sharks, skates and rays along the South African coast.

Global declines in elasmobranch populations resulting from several stressors raises conservation concern. Additionally, apex predators bioaccumulate high concentrations of total mercury (THg), due to biomagnification. Although South Africa is considered one of the top ten contributors of Hg emissions globally, information on Hg concentrations in elasmobranchs is limited. The aim of this study was to evaluate the THg concentrations in 22 species of elasmobranchs along the South African coastline. Concentrations ranged between 0.22 and 5.8 mg/kg in Haploblepharus pictus (dark shysharks) and Rostroraja alba (white skates) on the south coast, respectively. Along the east coast it ranged between 0.21 and 17.8 mg/kg in Mobula kuhlii (shortfin devil rays) and Sphyrna lewini (scalloped hammerheads), respectively. Mercury concentrations on the east coast were in the same range or higher compared to the same species sampled between 2005-10 from the same region, with generally higher concentrations compared to the same species sampled globally.

Human health risks associated with consumption of fish contaminated with trace elements from intensive mining activities in a peri-urban region.

Worldwide, numerous rural communities and low-income groups depend on fish harvested by subsistence fishers from local rivers and its impoundments as a source of protein. The aim of the present study was to determine the trace element bioaccumulation (As, Cd, Cr, Cu, Ni, Pb, Pt, Zn) in three edible fish species (Cyprinus carpio, Clarias gariepinus, Oreochromis mossambicus) from two impoundments in the Hex River system, South Africa, as well as the chronic health risk these trace elements pose to regular fish consumers. Trace element concentrations in the Hex River are naturally high (geogenic source), however, increased anthropogenic activities, such as intensive platinum mining activities, elevate the already high background concentrations. Concentrations of As, Cr, and Pt in C. carpio and C. gariepinus, as well as Ni and Zn in O. mossambicus were significantly higher in the impacted impoundment as compared to the reference impoundment. Concentrations of Cr and Cu were at both sampling sites the highest in O. mossambicus. From the human health risk assessment, As poses non-carcinogenic (HQ = 2-7) and carcinogenic risks (33-93 out of 10,000 people), while Cr (3-10 out of 10,000 people) and Ni (2-6 out of 10,000 people) pose only carcinogenic risks for the regular consumption of all three fish species from both impoundments, indicating a high probability of adverse human health effects. For As, Cr and Ni, also the sediment concentrations exceeded the levels of concern within the consensus based sediment quality guideline (CBSQG), while Cd, Cu, Ni and Zn exceeded the water quality guideline values. Thus, the CBSQG approach could be a promising tool for predicting human health risk associated with fish consumption. Since the present study only focused on the individual trace element risks, mixed toxicity of these trace elements and possible other pollutants within these fish species may pose an even greater risk to people who consume these fish regularly.

High parasite diversity in a neglected host: larval trematodes of Bithynia tentaculata in Central Europe.

Bithynids snails are a widespread group of molluscs in European freshwater systems. However, not much information is available on trematode communities from molluscs of this family. Here, we investigate the trematode diversity of Bithynia tentaculata, based on molecular and morphological data. A total of 682 snails from the rivers Lippe and Rhine in North Rhine-Westphalia, Germany, and 121 B. tentaculata from Curonian Lagoon, Lithuania were screened for infections with digeneans. In total, B. tentaculata showed a trematode prevalence of 12.9% and 14%, respectively. The phylogenetic analyses based on 55 novel sequences for 36 isolates demonstrated a high diversity of digeneans. Analyses of the molecular and morphological data revealed a species-rich trematode fauna, comprising 20 species, belonging to ten families. Interestingly, the larval trematode community of B. tentaculata shows little overlap with the well-studied trematode fauna of lymnaeids and planorbids, and some of the detected species (Echinochasmus beleocephalus and E. coaxatus) constitute first records for B. tentaculata in Central Europe. Our study revealed an abundant, diverse and distinct trematode fauna in B. tentaculata, which highlights the need for further research on this so far understudied host-parasite system. Therefore, we might currently be underestimating the ecological roles of several parasite communities of non-pulmonate snail host families in European fresh waters.

Metal accumulation in riverine macroinvertebrates from a platinum mining region.

South Africa is the world's main supplier of Pt. The Bushveld Igneous Complex in South Africa contains 75% of the world's Pt resources. Mining of this precious metal requires large volumes of water for production and removal of waste products. Most of this wastewater is discharged into river systems. Although the source of contamination with Pt in aquatic systems due to mining activities is known, little to no information is available about the impact of Pt on aquatic organisms. Additionally, other metals are released as byproducts of Pt mining, which might also be discharged into the environment. Therefore, concentrations of Cr, Ni, Cu, Zn, Cd, Pt and Pb were determined in water, sediment and macroinvertebrate samples from a reference site (Site 1), a highly impacted site (Site 2) and a moderately impacted site (Site 3) along the Hex River, South Africa. Aquatic invertebrate families representing different functional feeding groups i.e. scraper-grazers (Lymnaeidae), collector-gatherers (Potamonautidae, Hydropsychidae, Tubificidae and Chironomidae), shredders (Baetidae) and predators (Coenagrionidae and Libellulidae) were studied. In the sediments, the concentrations of Cr and Pt were significantly higher at Site 2 than at Sites 1 and 3, respectively, whereas concentrations of Ni, Cu, Cd, and Pb showed no significant differences between the sites. Depending on the metal, the aquatic invertebrate families showed different grades of bioaccumulation. The results from especially Lymnaeidae, Baetidae, Tubificidae and Chironomidae showed great promise for the use of these taxa for biomonitoring of metal contaminations. The macroinvertebrates accumulated metals associated with Pt mining, with epi-benthic dwelling taxa (Tubificidae) accumulating higher concentrations of Pt and Cr than other families (e.g. Potamonautidae, Coenagrionidae and Lymnaeidae). These results provide valuable information on the behavior of metals related to Pt mining in aquatic ecosystems and therefore can contribute to the risk assessment of these intensive mining activities.

Epizootic ulcerative syndrome - First report of evidence from South Africa's largest and premier conservation area, the Kruger National Park.

This study reports on the first evidence of genomic material of the causative agent for epizootic ulcerative syndrome (EUS), Aphanomyces invadans, from fish in the Limpopo River system and the Kruger National Park, South Africa. Fourteen fish species were collected from various depressions in the floodplains of the Limpopo and Luvuvhu Rivers in the Makuleke Wetlands during 2015 and 2017. A single individual of Clarias gariepinus was found to have a suspected epizootic ulcerative syndrome (EUS) lesion. Samples were collected and evidence of A. invadans DNA in the samples was found through PCR and amplicon sequencing. The spread of EUS into this premier conservation area is of concern as it could potentially spread across borders and into other naïve river systems with important conservation statuses.

Molecular and morphological characterisation of four diplostomid metacercariae infecting Tilapia sparrmanii (Perciformes: Cichlidae) in the North West Province, South Africa.

Despite their pathogenic effects on fish, the diversity of trematodes from the family Diplostomidae remains vastly unexplored in Africa and specifically South Africa. To date, only six species of diplostomids have been reported from freshwater fishes in this country, with only two species being molecularly characterised. In this study, combined morphological and molecular analyses were used to identify and describe metacercariae of the Diplostomidae (Digenea) parasitising banded tilapia Tilapia sparrmanii (Perciformes: Cichlidae) collected within the North West Province, South Africa. Metacercariae found on the body surface and muscles of the fish were separated into four groups based on the infection site, the colour of the cysts and the morphology of excysted specimens. Isolates from each group were further identified through molecular analyses. Comparative analyses of the newly generated 28S rDNA, ITS1-5.8S-ITS2 and cox1 sequences revealed the presence of four species of which three were identified as Bolbophorus sp. 3 (28S rDNA, ITS1-5.8S-ITS2 and cox1), Posthodiplostomum sp. 9 (28S rDNA and ITS1-5.8S-ITS2) and Uvulifer sp. 4 (28S rDNA, ITS1-5.8S-ITS2 and cox1), respectively, and the fourth species belonging to the Diplostomidae gen. sp. (28S rDNA, ITS1-5.8S-ITS2 and cox1). Morphology of metacercariae of Posthodiplostomum sp. was compared with metacercariae of this genus previously reported in fishes in Africa. This study presents the first molecular data for species of Bolbophorus Dubois, 1935, Posthodiplostomum Dubois, 1936 and Uvulifer Yamaguti, 1934 from Africa, and it highlights the need for future research on the diversity of diplostomid parasites in South Africa and in Africa as whole.

Biomarkers in tigerfish (Hydrocynus vittatus) as indicators of metal and organic pollution in ecologically sensitive subtropical rivers.

Studies have shown high levels of contamination of both metals and organochlorine pesticides (OCPs) in aquatic systems of the world renowned Kruger National Park, South Africa. With effects evident in top predators, including, unexplained Crocodylus niloticus deaths and organ level and histological changes in Hydrocynus vittatus. A suite of biomarkers reflecting exposure and were selected to evaluate biological responses of H. vittatus to anthropogenic stressors as well as to evaluate whether the chosen suite of biomarkers could successfully distinguish between the different pollution profiles present in the selected rivers. During this study a clear relationship was found between exposure to environmental contaminants and the concomitant responses of H. vittatus to these stressors. The ensuing biomarker responses indicated that there is a physiological attempt to deal with, and mitigate the deleterious effects that metals and OCPs may induce. In the Luvuvhu River there is a clear indication in H. vittatus of the stimulation of anti-oxidant protective mechanisms in response to internal OCP exposure. This is reflected by the increasing cytochrome P-450, superoxide dismutase, and more specifically reduced glutathione, which resulted in decreased lipid and protein breakdown (reflected in decreased lipid peroxidation and protein carbonyl levels). Consequently H. vittatus populations of the Luvuvhu River are under greater cumulative stress and this is reflected in the lower energy budgets. Our results further show the integrated application value of the current suite of biomarkers in assessing responses of subtropical fish to metal and OCP exposure as the entire suite of biomarkers when used in conjunction were able to explain 100% of the variation in the data.

Human Health Risk from Consumption of Marine Fish Contaminated with DDT and Its Metabolites in Maputo Bay, Mozambique.

Many countries with incidence of malaria, including those surrounding Maputo Bay, use dichloro-diphenyl-trichloroethane (DDT) to reduce mosquitoes. This study is the first to estimate the human health risk associated with consumption of marine fish from Maputo Bay contaminated with DDTs. The median for ∑DDTs was 3.8 ng/g ww (maximum 280.9 ng/g ww). The overall hazard ratio for samples was 1.5 at the 75th percentile concentration and 28.2 at the 95th percentile. These calculations show increased potential cancer risks due to contamination by DDTs, data which will help policy makers perform a risk-benefit analysis of DDT use in malaria control programs in the region.

Concentrations and human health risk assessment of DDT and its metabolites in free-range and commercial chicken products from KwaZulu-Natal, South Africa.

Organochlorine pesticides such as dichlorodiphenyltrichloroethane (DDT) have been used in agriculture and for disease control purposes over many decades. Reports suggest that DDT exposure may result in a number of adverse effects in humans. In the KwaZulu-Natal Province of South Africa, DDT is sprayed annually in homes (indoor residual spraying) to control the mosquito vector of malaria. In the northern part of the Province, samples of free-range chicken meat (n = 48) and eggs (n = 13), and commercially produced chicken meat (n = 6) and eggs (n = 11), were collected and analysed. Of the free-range chicken meat samples, 94% (45/48) contained DDTs (ΣDDTs median 6.1 ng/g wet weight [ww], maximum 79.1 ng/g ww). Chicken egg contents were also contaminated (ΣDDTs in free-range eggs median 9544 ng/g ww, maximum 96.666 ng/g ww; and in commercial eggs median 1.3 ng/g ww, maximum 4.6 ng/g ww). The predominant DDT congener detected was p,p'-DDE in both free-range meat (>63%) and eggs (>66%), followed by p,p'-DDT and then p,p'-DDD. Based on estimated daily intake values, calculated human risk ratio (carcinogenic) values were >1 for DDTs detected in both free-range chicken products. Consumption of free-range eggs poses a particularly high health risk.

First observation of African tigerfish Hydrocynus vittatus predating on barn swallows Hirundo rustica in flight.

A population of African tigerfish Hydrocynus vittatus from the Schroda Dam, actively prey on barn swallows Hirundo rustica in flight. This behaviour was discovered during a radio telemetry study and documented using a motion picture video camera. These results show that an avivorous diet is a part of the feeding biology of H. vittatus, and may occur in other populations.

Comparative histomorphological assessment of the testes of two Clarias species from the Okavango Delta Panhandle, Botswana.

The testes of two Clarias species from the Okavango Delta Panhandle were histomorphologically assessed for differences in structure mainly because the testes varied externally in colour, with Clarias ngamensis having black testes while those of Clarias gariepinus were off-white. Although a detailed histological description of normal testes in C. gariepinus has been compiled based on laboratory experimentation, there is limited histomorphological field data available on both species which could be used as reference material in the histology and histomorphology assessment of gonads both in the laboratory and field experiments. In August 2006 and 2007 fifteen fish per species of C. ngamensis and C. gariepinus were collected from the Shakawe Panhandle in the Okavango Delta. Testicular samples excised from the fish were fixed in 10% neutral buffered formalin and later transported to the University of Johannesburg where laboratory analyses using standard histological procedures were conducted. Microscopic assessments were used to describe the testicular morphology and the reproductive developmental stages. Haematoxylin and Eosin (H&E), Periodic Acid Schiff (PAS), Perl's Prussian Blue and Gordon and Sweets Silver Stain Solution were used to enhance finer cellular detail and the morphology of various structures present in the catfish testes. Results emanating from this study (a) form a histomorphological baseline set of data from an undisturbed wetland system for comparison between normal morphology and anomalies within the catfish species, (b) contribute to the limited database of the normal histomorphology of gonads in Southern African freshwater fish species and (c) expand the academic knowledge and skills required for the conservation of wetland ecosystems in Southern Africa.

Hemoproteids (Apicomplexa: Haemoproteidae) from South African tortoises (Cryptodira: Testudinidae).

Terrestrial tortoises, Chersina angulata (Schweigger, 1812), Kinixys belliana belliana Gray, 1831, K. lobatsiana Power, 1927, K. natalensis Hewitt, 1935, and Stigmochelys pardalis (Bell, 1828), both wild (n  =  84) and captive (n  =  70), were examined for hematozoans from the provinces of Gauteng, Kwazulu-Natal, North West, and Western Cape, South Africa. In Giemsa-stained blood films, 2 species of hemoproteids were identified, 1 previously described, that is, Haemoproteus testudinalis ( Laveran, 1905 ) Wenyon, 1915, the other unknown. Haemoproteus testudinalis occurred in 2/27 wild K. lobatsiana and 1/41 S. pardalis and 2/2 captive K. lobatsiana from the North West and Gauteng, respectively, whereas Haemoproteus sp. A was recorded in 1 of 3 captive K. natalensis from Kwazulu-Natal. Haemoproteus testudinalis had pale blue-stained, often vacuolated, halteridial macrogametocytes, pale pink-stained microhalterdial to halteridial microgametocytes, and dark, scattered, often bacillus-like hemozoin granules. Haemoproteus natalensis n. sp. had ameboid growth stages, blue-stained halteridial to circumnuclear macrogametocytes, purple-stained circumnuclear microgametocytes, and brownish, often clustered, pigment granules. It is recommended that Haemoproteus balazuci Dias, 1953, become a junior synonym of H. testudinalis.