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.

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.

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.

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.

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.

Distribution of arsenic, silver, cadmium, lead and other trace elements in water, sediment and macrophytes in the Kenyan part of Lake Victoria: spatial, temporal and bioindicative aspects.

This is the first detailed report on the distribution of Ni, As, Sr and Ag in water, sediments and macrophytes from Lake Victoria, complemented with recent data on the heavy metals Cr, Cu, Zn, Cd and Pb. The study was conducted over an 11-month period at five sites in the Kenyan part of Lake Victoria: four sites in the Winam Gulf influenced by various anthropogenic pressures including a site near Kisumu City, and one in the main lake, perceived to have lesser direct anthropogenic influence. Compared with the main lake site, the water in Winam Gulf had significantly higher values for electrical conductivity and concentrations of dissolved components: organic carbon and bound nitrogen, as well as major and most trace elements. This contamination is also evident in surface sediments, which contained significantly higher levels of Cr, Zn, As, Ag, Cd and Pb compared with the main lake site. The mean levels of Cr, Cu, Zn, Ag and Pb exceeded probable effect levels at least at one of the gulf sites. The sediments at the Kisumu City site were classified as severely polluted with Cu (up to 259 mg/kg dw) and Pb (up to 1188 mg/kg dw). The sediment cores showed significantly higher levels of Cu, Zn, Ag, Cd and Pb in the surface (0-3 cm) versus subsurface (22-25 cm) layer at the Kisumu City site, indicating increasing pollution by these elements within the last 15 years. This is also the first report on trace elements in the emergent water plant Vossia cuspidata and submerged plant Ceratophyllum demersum from this lake. Even though the accumulation of most elements is comparable between C. demersum (whole plant) and V. cuspidata roots, the latter shows a better bioindicative potential. Contamination of the gulf with Ni, Cu, Zn, Ag, Cd and Pb is well mirrored in V. cuspidata roots. V. cuspidata strongly restricts the acropetal transport of trace elements, and hence using the shoots as fodder does not pose a risk to livestock.

Diversity of digenean trematode larvae in snails from Lake Victoria, Kenya: First reports and bioindicative aspects.

This study investigated the occurrence of digenean trematode larvae in snails from the Kenyan part of Lake Victoria. The survey included caenogastropod snails that have received less focus in parasitological studies in Africa: their trematodes are largely unknown. Out of 1145 snail specimens, 149 (13.0%) were infected with Digenea. The highest prevalence (P) was recorded in Melanoides tuberculata (64.5%), followed by Pila ovata (15.4%), Radix natalensis (9.5%), Bulinus ugandae (9.1%), Bellamya unicolor (8.9%), Biomphalaria pfeifferi (7.3%) and Biomphalaria sudanica (4.4%). Morphological and molecular analyses revealed 17 digenean species. Contrary to reports of low diversity of Digenea in caenogastropods, P. ovata harboured 8 species - at least twice as many as in each of the pulmonates. The following taxa are reported for the first time in the Lake Victoria region: Haplorchis pumilio, Thapariella prudhoei, Nudacotyle sp., Renicola sp. and Bolbophorus sp. An unknown cercaria belonging to the genus Haematoloechus is reported from P. ovata: a xiphidiocercaria possessing a long sword-shaped stylet (47-71 µm) which does not match any available literature records. From this study, H. pumilio from M. tuberculata (P = 69.4%), Fasciola gigantica from R. natalensis (P = 1.9%) and Bolbophorus sp. from Bu. ugandae (P = 4.6%) are species of veterinary or medical importance. Snails from the study site with little direct anthropogenic influence had the highest prevalence and diversity of digenean larvae, indicating that environmental conditions influence trematode occurrence.