Gyrodactylus sprostonae Ling, 1962 infects an indigenous cyprinid in southern Africa: An expanded description.

Gyrodactylus sprostonae Ling, 1962 is a highly invasive parasite reported across freshwater environments of the northern hemisphere. The taxon was originally described from Carassius auratus (Linnaeus, 1758) and Cyprinus carpio Linnaeus, 1758 in China. This parasite has never been reported in Africa or the southern hemisphere. Recently, this taxon was collected from an indigenous yellowfish, Labeobarbus aeneus (Burchell, 1822), in the Vaal River, South Africa. The present study includes the conclusive identification of the gyrodactylid parasites collected from L. aeneus, including additional taxonomic data, using microscopy and molecular techniques. Microscopy included light microscopy (LM) of whole worms and scanning electron microscopy (SEM) of isolated haptoral sclerites. Additionally, morphometric data were obtained from SEM and compared to that generated using LM. For molecular analysis, the internal transcribed spacer (ITS) region of rDNA was amplified and phylogenetic topologies constructed. The specimens were morphometrically and genetically highly similar to other data for G. sprostonae. Additional point-to-point measurements and ITS rDNA sequences were generated for the taxon, contributing to the morphometric and molecular data for G. sprostonae. The study also includes the first study of the isolated haptoral sclerites of the taxon using SEM, with similar morphometric results to LM. This is the first record of G. sprostonae in the southern hemisphere and from a new, indigenous African host, L. aeneus, indicating host switching to smallmouth yellowfish. Furthermore, these results expand on the knowledge of the distribution of invasive parasites in South Africa, as well as Gyrodactylus species diversity in Africa.

An improved method for isolating camallanid (Nematoda) spicules for scanning electron microscopy.

In nematodes, the structure of male copulatory organs is a significant taxonomic distinguisher and includes the morphometry of the spicules. The description of these structures mainly relies on the study of whole mounts using light microscopy. In rare instances, protruding spicules have been described with scanning electron microscopy. Even fewer studies have described the ultrastructure of isolated spicules following their isolation. In the present study, two different methods of spicule isolation were performed on two parasitic camallanid nematodes, Procamallanus (Procamallanus) pseudolaeviconchus Moravec & van As, 2015 and Paracamallanus cyathopharynx (Baylis, 1923), from African sharptooth catfish to determine the practicality and efficiency of the methodologies. The first method involved using sharpened tungsten needles and microdissection of the spicule pouch to free the spicules, followed by soft tissue digestion if necessary. Alternatively, the spicules were isolated through mechanical release instead of dissection in a method developed in the current study. This involved freeing the spicules from surrounding soft tissue by placing live specimens between a coverslip and a glass slide in a drop of water and exerting pressure with small rotational movements. Both methods yielded favourable results, but Method 2 is recommended for future studies due to the many advantages.

The monogenean Paradiplozoon ichthyoxanthon behaves like a micropredator on two of its hosts, as indicated by stable isotopes.

The analysis of stable isotopes of carbon and nitrogen has been used as a fingerprint for understanding the trophic interactions of organisms. Most of these studies have been applied to free-living organisms, while parasites have largely been neglected. Studies dealing with parasites so far have assessed the carbon and nitrogen signatures in endoparasites or ectoparasites of different hosts, without showing general trends concerning the nutritional relationships within host-parasite associations. Moreover, in most cases such systems involved a single host and parasite species. The present study is therefore the first to detail the trophic interactions of a freshwater monogenean-host model using δ13C and δ15N, where a single monogenean species infects two distinctly different hosts. Host fishes, Labeobarbus aeneus and Labeobarbus kimberleyensis from the Vaal Dam, South Africa, were assessed for the monogenean parasite Paradiplozoon ichthyoxanthon, individuals of which were removed from the gills of the hosts. The parasites and host muscle samples were analysed for signatures of δ13C and δ15N using an elemental analyser connected to an isotope ratio mass spectrometer. Host fish appear to use partly different food sources, with L. aeneus having slightly elevated δ13C signatures compared to L. kimberleyensis, and showed only small differences with regard to their nitrogen signatures, suggesting that both species range on the same trophic level. Carbon and nitrogen signatures in P. ichthyoxanthon showed that the parasites mirrored the small differences in dietary carbon sources of the host but, according to δ15N signatures, the parasite ranged on a higher trophic level than the hosts. This relationship resembles predator-prey relationships and therefore suggests that P. ichthyoxanthon might act as a micropredator, similar to blood-sucking arthropods such as mites and fleas.

Histopathological changes on the gills of asp (Aspius aspius) and European catfish (Silurus glanis) caused by Lamproglena pulchella and a Lamproglena sp. (Copepoda: Lernaeidae), respectively.

In a parasitology survey of Hungarian fishes, heavy infections of parasitic copepods Lamproglena pulchella and a Lamproglena sp. were found in the gills of the asp and the European catfish, respectively. Individuals of both fish species were emaciated and infected with hundreds of Lamproglena. Copepods located close to the tip of gill filaments and formed a depression at the attachment sites. In histological sections, cell degenerations and local haemorrhages were present adjacent to the maxillipeds and where the maxillary claws pierced the gill tissue. Around maxillae and in the midgut of the Lamproglena, damaged piscine blood cells and remains of the gill tissue were observed. Host reaction was expressed by proliferation of epithelioid cells, increase in both number and size of goblet and mast cells and formation of giant cells.

Soft tissue digestion of Paradiplozoon vaalense for SEM of sclerites and simultaneous molecular analysis.

Classification of most monogeneans is primarily based on size, shape, and arrangement of haptoral sclerites. These structures are often obscured or misinterpreted when studied using light microscopy, leading to confusion regarding defining characters. Scanning electron microscopy (SEM) has predominantly been used to study haptoral sclerites in smaller monogeneans, focusing on hooks and anchors. In the Diplozoidae, SEM has not been used to study haptoral sclerites. Using new and modified techniques, the sclerites of diplozoids collected in South Africa were successfully studied using SEM. The digestion buffer from a DNA extraction kit was used to digest the surrounding tissue, and Poly-L-lysine-coated and concavity slides were employed to limit the movement and loss of sclerites, with the latter being more user-friendly. In addition to the success of visualizing the sclerites using SEM, the digested tissue from as little as half of the haptor provided viable genetic material for molecular characterization. From the results presented here, the study of the sclerites of larger monogeneans using SEM, including those bearing clamps, is a viable possibility for future research. Also, this method may be beneficial for the study of other, non-haptoral sclerites, such as cirri in other families of monogeneans. During this study, Labeo capensis was noted as a valid host of Paradiplozoon vaalense in a region of the Vaal River where the type host, Labeo umbratus, appears to be absent.

Paradiplozoon vaalense n. sp. (Monogenea: Diplozoidae) from the gills of moggel, Labeo umbratus (Smith, 1841), in the Vaal River System, South Africa.

An unidentified monogenean diplozoid species was collected from the gills of moggel in the Vaal River and Vaal Dam, South Africa. Specimens were removed from gills of the hosts and observed using light and electron microscopy to compare these diplozoids with known species. The second internal transcribed spacer (ITS2) of the ribosomal gene was amplified, sequenced and compared to that of other diplozoid taxa. Morphological species delimitation was used to determine the identity of these diplozoids, but they did not match the description of any diplozoid taxa. This species is recognized by the specific size of the hooks, number of plicae in posterior and trapezoid anterior projection of the median sclerite connecting to the clamp jaws via a single sclerite, occasionally with two small additional sclerites. Genetic characteristics based on sequence data from the ITS2 region also distinguish this taxon from all other diplozoid taxa. This South African diplozoid grouped in the same clade as Paradiplozoon ichthyoxanthon Avenant-Oldewage, 2013. Data clearly indicate that diplozoids collected from moggel represent a new, distinct taxon of Paradiplozoon Akhmerov, 1974 and are described here as Paradiplozoon vaalense n. sp.

Paradiplozoon ichthyoxanthon n. sp. (Monogenea: Diplozoidae) from Labeobarbus aeneus (Cyprinidae) in the Vaal River, South Africa.

A new species, Paradiplozoon ichthyoxanthon (Monogenea: Diplozoidae) is described from the gills of Labeobarbus aeneus (Cyprinidae) in the Vaal Dam and Vaal River Barrage in the Vaal River, South Africa. This species is the first in this genus from southern Africa; Paradiplozoon aegtyptense and Paradiplozoon ghanense have been described from northern Africa. The new species differs from all other Paradiplozoon species on the basis of the combination of morphological characteristics, namely disk-like haptor, eggs without filaments, clamp and central hook morphology and size of specimens. The species description is accompanied by a description of the reproductive organs. The internal transcribed spacer of the ribosomal gene (ITS2) was studied and the South African individuals formed a distinct clade, supporting the establishment of a new species.

The effect of starvation on the ultrastructure of the digestive cells of Dolops ranarum (Stuhlmann, 1891) (Crustacea: Branchiura).

Transmission electron microscopy was conducted on the digestive epithelium of the crustacean ectoparasite Dolops ranarum to elucidate its ultrastructure for the first time, both in a nourished and starved condition. Specimens were collected from the Limpopo Drainage System in South Africa, and the specimens were killed and dissected in Todd's fixative. The anterior midgut is composed mostly of absorptive cells or R cells, while the diverticula are composed of R cells and of F cells, which are moderately abundant in rough endoplasmic reticulum. They are probably responsible for producing digestive enzymes. The posterior midgut is composed of papilliform B cells with large apically located vesicles and R' cells devoid of cell inclusions. Under starvation, specimens survive for a maximum of 12 days; R cells show the most conspicuous changes in ultrastructural characteristics. It is concluded that D. ranarum has adapted to short-term survival only without a host.

The ultrastructure of the digestive cells of Argulus japonicus, Thiele 1900 (Crustacea: Branchiura).

The ultrastructure of the cells of the digestive system of Argulus japonicus is described with the use of transmission electron microscopy. Specimens of Argulus japonicus were collected from the Vaal Dam in South Africa and fixed in Todd's fixative. The samples were post fixed in osmium tetroxide and embedded in resin. The anterior midgut is composed mostly of R cells while the enteral diverticula are composed mainly of R cells in the proximal diverticules and of F cells in the distal diverticula. The posterior midgut is composed of very large papilliform B cells and of R cells. The R cells in the anterior midgut probably absorb nutrients including lipids. The F cells are filled mostly with rough endoplasmic reticulum, suggesting enzyme synthesis, while the B cells portrayed endocytotic vesicles, indicating intracellular digestion of predigested food. The R cells of the posterior midgut are less active than cells present in the anterior midgut. E cells and peritrophic membrane were not observed.

Ecological parameters of Lamproglena hoi (Copepoda: Lernaeidae) infection on the Bushveld smallscale yellowfish, Labeobarbus polylepis (Boulenger, 1907).

This study describes the distribution and aspects of the ecology of Lamproglena hoi. Bushveld small-scale yellowfish, Labeobarbus polylepis (Boulenger, 1907) were collected during June 2006 from the Phongolo and Assegaai rivers, March 2005 and October 2006 from the Elands River, and January 2007 and June 2008 from the Komati River in Mpumalanga, South Africa and examined for the presence of parasites. Lamproglena hoi specimens were collected from the gill filaments of the host. Specimens were fixed with warm AFA (alcohol-formaldehyde-acetic acid) and preserved in 70% ethanol. The identification of parasites took place in the laboratories of the University of Johannesburg. Twenty-five copepods (prevalence 21%, mean intensity = 4.17, abundance = 0.86) were collected on 29 fish in the Phongolo River and 46 copepods (prevalence 40%, mean intensity = 3.83, abundance = 1.53) were collected on 30 fish in the Assegaai River. One hundred and sixty eight copepods (prevalence 52%, mean intensity = 12.92, abundance = 6.72) were collected on 25 fish in 2005, and 527 copepods (prevalence 95%, mean intensity = 27.74, abundance = 26.35) were collected on 20 fish in the Elands River. One hundred and sixteen copepods (prevalence 75%, mean intensity = 7.73, abundance = 5.80) were collected on 20 fish in 2007, and 273 copepods (prevalence 63%, mean intensity = 16.06, abundance = 10.11) were collected on 27 fish in 2008 in the Komati River. Labeobarbus polylepis from these four rivers was found to have a relatively high L. hoi infection. Inseminated L. hoi females (immature) attach to the host in winter and their ovaries become conspicuous (mature). In spring fertilized eggs are stored in egg sacs hanging from the body (gravid), indicating that fertilized eggs start to hatch in spring and continued hatching into summer. Parasites prefer the median part of the second gill arch for attachment. No correlation exists between the number of parasites recorded on the gills and the sizes (total lengths) of yellowfish sampled.

First maxillae suction discs in Branchiura (Crustacea): development and evolution in light of the first molecular phylogeny of Branchiura, Pentastomida, and other "Maxillopoda".

The fish ectoparasites Branchiura (Crustacea) display two different ways of attachment to the fish surface as adults: the first maxillae are either hooks (Dolops) or suction discs (Argulus, Chonopeltis, and Dipteropeltis). In larval Argulus foliaceus the first maxillae are hooks. With the first molecular phylogeny of the Branchiura as a background, the present paper discusses the evolutionary scenarios leading to hooks versus suction discs. Specific homologies exist between larval Argulus foliaceus hooks and adult Dolops ranarum hooks. These include the presence of a comparable number of segments/portions and a distal segment terminating in a double structure: a distal two-part hook (in Argulus) or one hook and an associate spine-like structure (in Dolops). In the phylogenetic reconstruction based on three molecular markers (mitochondrial 16S rRNA, nuclear 18S and 28S rRNA), Dolops ranarum is found to be in a sister group position to all other Branchiura, which in this analysis include six Argulus and one Chonopeltis sequences. Based on the molecular phylogeny a likely evolutionary scenario is that the ancestral branchiuran used hooks (on the first maxilla) for attachment, as seen in Dolops, of which the proximal part was subsequently modified into suction discs in Argulus and Chonopeltis (and Dipteropeltis). The sister group relationship of the Branchiura and Pentastomida is confirmed based on the most comprehensive taxon sampling until now. No evidence was found for a branchiuran in-group position of the Pentastomida.

Nematode parasites of Clarias gariepinus (Burchell, 1822) from the Rietvlei Dam, South Africa.

Catfish, Clarias gariepinus, from the Rietvlei Dam near Pretoria, South Africa were examined for nematode parasites. Two species, Procamallanus laeviconchus in the stomach and Contracaecum spp. larvae in the abdominal cavity, were found. The morphology of these species, based on light and scanning electron microscopy, and how they compare with previously described specimens are discussed. Infection rates were mild compared to previous surveys although Contracaecum spp. had a high prevalence of 86%.

On cestode and digenean parasites of Clarias gariepinus (Burchell, 1822) from the Rietvlei Dam, South Africa.

Sharptooth catfish, Clarias gariepinus, from the Rietvlei Dam near Pretoria, South Africa were examined for internal platyhelminth parasites. Two adult cestodes, Polyonchobothrium clarias (stomach) (prevalence 71%, mean intensity = 5, n = 7) and Proteocephalus glanduliger (anterior intestine) (prevalence 14%, mean intensity = 2, n = 7), were found in the gut while metacercariae of one larval digenean, Ornithodiplostomum sp. (prevalence 14%, mean intensity = 140, n = 7), were found encysted in the muscles. The morphology of these species, based on light and scanning electron microscopy as well as histological analysis, and how they differ from previously described specimens, are discussed. Ornithodiplostomum is a new record in southern Africa. Infection levels of the host fish were mild compared to records from previous surveys.

The fluorescent detection of Paradiplozoon sp. (Monogenea: Diplozoidae) attachment clamps' sclerites and integumental proteins.

In the genus Paradiplozoon, few hard structures are present therefore most of the taxonomic studies have focused on the unique ultrastructure of the sclerites. Alcohol-fixed specimens were transferred to BSA-saline for 5 min. before staining indefinitely with 5 microl WGA-TRX, 5 microl SYTO 9 and 5 microl of Calcofluor White M2R. Rhodamine, Fluorescein and DAPI bandpass filters on the microscope enabled selective light wavelength illumination of the three flourochromes by a mercury light source. This method provided an easy and rapid methodology to show the internal sclerites of attachment clamps. It is suitable for alcohol preserved specimens and may have additional applications in other helminth organisms.

Aspects of the ecology of the Asian tapeworm, Bothriocephalus acheilognathi Yamaguti, 1934 in yellowfish in the Vaal Dam, South Africa.

Seasonal surveys were conducted at the Vaal Dam between April 2000 and January 2001. Twenty smallmouth yellowfish (Labeobarbus aeneus) and 20 largemouth yellowfish (Labeobarbus kimberleyensis) were collected with the aid of gill nets. Surface water quality variables were included. The cestodes were identified as either Bothriocephalus acheilognathi Yamaguti, 1934 or "other cestode spp.". The majority (99.8%) of the cestodes found in both yellowfish species were identified as B. acheilognathi (Asian tapeworm). The prevalence, mean intensity and abundance of B. acheilognathi in both yellowfish species were calculated. Ecological parameters including species specificity, seasonality, gender specificity and relationships between fish size and the Asian tapeworm prevalence were also included. In this study, B. acheilognathi preferred L. kimberleyensis over L. aeneus although a low intensity was observed in smallmouth yellowfish. Furthermore, the infection (in terms of prevalence, abundance and mean intensity) in largemouth yellowfish was markedly higher. Seasonal patterns observed in the Asian tapeworm's infection of smallmouth yellowfish are attributed to breeding and subsequent feeding patterns of this fish species with relatively high infections recorded in winter and spring. For L. kimberleyensis no explanation can be given regarding the seasonal patterns observed for the mean intensity and abundance of B. acheilognathi. The maximum and minimum mean intensity and abundance values in largemouth yellowfish were recorded in autumn and spring, respectively. In addition, the prevalence of B. acheilognathi was consistently high in all four seasons.