An examination of nervous system revealed unexpected immunoreactivity of both secretory apparatus and excretory canals in plerocercoids of two broad tapeworms (Cestoda: Diphyllobothriidea)

Dibothriocephalus ditremus and Dibothriocephalus latus are diphyllobothriidean tapeworms autochthonous to Europe. Their larval stages (plerocercoids) may seriously alter health of their intermediate fish hosts (D. ditremus) or cause intestinal diphyllobothriosis of the final human host (D. latus). Despite numerous data on the internal structure of broad tapeworms, many aspects of the morphology and physiology related to host–parasite co-existence remain unclear for these 2 species. The main objective of this work was to elucidate functional morphology of the frontal part (scolex) of plerocercoids, which is crucial for their establishment in fish tissues and for an early attachment in final hosts. The whole-mount specimens were labelled with different antibodies and examined by confocal microscope to capture their complex 3-dimensional microanatomy. Both species exhibited similar general pattern of immunofluorescent signal, although some differences were observed. In the nervous system, FMRF amide-like immunoreactivity (IR) occurred in the bi-lobed brain, 2 main nerve cords and surrounding nerve plexuses. Differences between the species were found in the structure of the brain commissures and the size of the sensilla. Synapsin IR examined in D. ditremus occurred mainly around FMRF amide-like IR brain lobes and main cords. The unexpected finding was an occurrence of FMRF amide-like IR in terminal reservoirs of secretory gland ducts and excretory canals, which has not been observed previously in any tapeworm species. This may indicate that secretory/excretory products, which play a key role in host–parasite relationships, are likely to contain FMRF amide-related peptide/s

Exploring the diversity of Diplostomum (Digenea: Diplostomidae) in fishes from the River Danube using mitochondrial DNA barcodes

Abstract Background Metacercariae of Diplostomum are important fish pathogens, but reliable data on their diversity in natural fish populations are virtually lacking. This study was conducted to explore the species diversity and host-parasite association patterns of Diplostomum spp. in a large riverine system in Europe, using molecular and morphological data. Methods Twenty-eight species of fish of nine families were sampled in the River Danube at Nyergesújfalu in Hungary in 2012 and Štúrovo in Slovakia in 2015. Isolates of Diplostomum spp. were characterised morphologically and molecularly. Partial sequences of the ‘barcode’ region of the cytochrome c oxidase subunit 1 (cox1) and complete sequences of the nicotinamide adenine dinucleotide dehydrogenase subunit 3 (nad3) mitochondrial genes were amplified for 76 and 30 isolates, respectively. The partial cox1 sequences were used for molecular identification of the isolates and an assessment of haplotype diversity and possible host-associated structuring of the most prevalent parasite species. New primers were designed for amplification of the mitochondrial nad3 gene. Results Only lens-infecting Diplostomum spp. were recovered in 16 fish species of five families. Barcoding of representative isolates provided molecular identification for three species/species-level genetic lineages, D. spathaceum, D. pseudospathaceum and ‘D. mergi Lineage 2’, and three single isolates potentially representing distinct species. Molecular data helped to elucidate partially the life-cycle of ‘D. mergi Lineage 2’. Many of the haplotypes of D. spathaceum (16 in total), D. pseudospathaceum (15 in total) and ‘D. mergi Lineage 2’ (7 in total) were shared by a number of fish hosts and there was no indication of genetic structuring associated with the second intermediate host. The most frequent Diplostomum spp. exhibited a low host-specificity, predominantly infecting a wide range of cyprinid fishes, but also species of distant fish families such as the Acipenseridae, Lotidae, Percidae and Siluridae. The nad3 gene exhibited distinctly higher levels of interspecific divergence in comparison with the cox1 gene. Conclusions This first exploration of the species diversity and host ranges of Diplostomum spp., in natural fish populations in the River Danube, provided novel molecular, morphological and host-use data which will advance further ecological studies on the distribution and host ranges of these important fish parasites in Europe. Our results also indicate that the nad3 gene is a good candidate marker for multi-gene approaches to systematic estimates within the genus

Disentangling taxonomy of Biacetabulum (Cestoda, Caryophyllidea), parasites of catostomid fishes in North America: proposal of Megancestus gen. n. to accommodate B. carpiodi

A new genus, Megancestus n. gen., is proposed to accommodate the caryophyllidean tapeworm Biacetabulum carpiodi Mackiewicz, 1969 from carpsuckers and quillback (Carpiodes spp.) in North America. This species is not closely related to other species of Biacetabulum Hunter, 1927 and is transferred to a newly erected genus. This new genus is typified by the possession of a small body (total length of 3.1–7.5 mm) with a scolex that bears a pair of large acetabulum-like loculi, two pairs of shallow lateral loculi, and a slightly convex apical disc, testes arranged in one or two layers, oval, thick-walled cirrus-sac, well-developed external seminal vesicle, separate gonopores, H-shaped ovary, few median vitelline follicles, and the uterus extending by a single loop anterior to the cirrus-sac. Megancestus differs from all Nearctic caryophyllidean genera (family Capingentidae), including Biacetabulum, by vitelline follicles dorsal to the ovary that connect the preovarian and postovarian vitelline fields. The most closely related Hunterella Mackiewicz et McCrae, 1962 differs by shape of the scolex (tholate, i.e., devoid of any loculi), dumbbell-shaped ovary and the uterus not extending anterior to the cirrus-sac. Megancestus carpiodi (Mackiewicz, 1969) n. comb. is the only species of the genus and it is a stenoxenous parasite, which has been found only in the river carpsucker (Carpiodes carpio—type host), quillback (Carpiodes cyprinus) and highfin carpsucker (Carpiodes velifer) (Catostomidae: Ictiobinae) in the lower and middle Mississippi basin

Diphyllobothrium nihonkaiense Tapeworm Larvae in Salmon from North America

Diphyllobothriosis is reemerging because of global importation and increased popularity of eating raw fish. We detected Diphyllobothrium nihonkaiense plerocercoids in the musculature of wild pink salmon (Oncorhynchus gorbuscha) from Alaska, USA. Therefore, salmon from the American and Asian Pacific coasts and elsewhere pose potential dangers for persons who eat these fish raw

Archigetes Leuckart, 1878 (Cestoda, Caryophyllidea): diversity of enigmatic fish tapeworms with monoxenic life cycles

Le genre Archigetes Leuckart, 1878 (Caryophyllidea) est unique parmi les cestodes en ce sens que ses espèces peuvent atteindre la maturité chez des hôtes invertébrés (Oligochaeta), c’est-à-dire avoir un cycle de vie monoxénique (direct). Les cinq espèces ont été décrites à partir de plérocercoïdes progénétiques chez des oligochètes et deux d’entre elles aussi à partir d’adultes chez des poissons cypriniformes. Deux espèces, A. sieboldi Leuckart, 1878 et A. iowensis Calentine, 1962, ont été trouvées en Amérique du Nord chez des carpes communes ( Cyprinus carpio ), non indigènes. Une étude moléculaire des caryophyllidés du sud des États-Unis a révélé la présence de trois nouvelles espèces chez les poissons d’eau douce indigènes (Catostomidae, Ictiobinae). Archigetes loculotruncatus n. sp. d’ Ictiobus bubalus , I. niger et Carpiodes cyprinus est le plus grand représentant du genre et est caractérisé par un scolex loculotronqué. Archigetes megacephalus n. sp. d’ Ictiobus niger , I. bubalus et I. cyprinellus est caractérisé par un scolex proéminent et bothrioloculodisqué. Archigetes vadosus n. sp. d’ I. bubalus est caractérisé par un scolex globulaire avec des loges très peu profondes, et diffère d’ A. sieboldi , étroitement apparenté, par la forme du corps, avec une région du cou distincte et un scolex plus large que le reste du corps. Archigetes iowensis Calentine, 1962 devient un synonyme plus récent de Paraglaridacris limnodrili (Yamaguti, 1934). Le diagnostic générique d’ Archigetes est modifié et une clé d’identification des taxons nord-américains est fournie. Les espèces d’ Archigetes et de Paraglaridacris diffèrent de la manière la plus visible les unes des autres par la structure de l’ovaire, qui est folliculaire chez Archigetes par opposition à compact chez Paraglaridacris

A fish-parasite sentinel system in an assessment of the spatial distribution of polychlorinated biphenyls

Abstract The spatial distribution of polychlorinated biphenyls (PCBs), in the Zemplínska Šírava water reservoir and adjacent tributaries in the Bodrog River Basin were investigated using a fish-parasite sentinel system. PCB concentrations were detected in various fish matrices (dorsal and abdominal muscles, liver and intestine) of the Wels catfish (Silurus glanis) and its intestinal cestode Glanitaenia osculata. PCB concentrations in the fish from the water reservoir, located closest to the chemical plant, the primary source of the PCB pollution, were the highest. The analysis of these contaminants in catfish matrices showed the highest concentrations in the abdominal muscle, followed by the dorsal muscle, liver and intestine. Concentrations of ∑PCBs exceeding the limits for food set by European regulations were measured in the muscle tissue of catfish at all sites, even in the Bodrog River, 60 km away from the primary source of contamination, posing a significant risk to humans in the Zemplín region. For the first time, the ability of cestode G. osculata to accumulate higher amounts of PCBs compared to fish matrices has been demonstrated. Due to the enormous ability of the parasites to accumulate PCBs, we recommend this approach for alternative biomonitoring of PCBs in contaminated aquatic environments

Polyonchobothrium polypteri (Leydig, 1853) Luhe 1900

Polyonchobothrium polypteri (Leydig, 1853) Lühe, 1900 (Figs. 23–27, 33– 40) Syns: Tetrabothrium polypteri Leyding, 1853; Onchobothrium (Polyonchobothrium) septicolle Diesing, 1854; Anchistrocephalus polypteri (Leyding, 1853) Monticelli, 1890; Ptychobothrium armatum Fuhrmann, 1902; Ancistrocephalus polypteri (Leyding, 1853) Hesse, 1922; Polyonchobothrium pseudopolypteri Meggitt, 1930; Oncobothriocephalus armatum (Fuhrmann, 1902) Yamaguti, 1959; Polyoncobothrium polypteri (Leyding, 1853) Yamaguti, 1959. Type host: Polypterus bichir Lacépède (Polypteriformes: Polypteridae). Other definitive hosts: Polypterus endlicheri Heckel; Polypterus senegalus Cuvier. Type locality: Nile River in Egypt. Distribution: Congo basin – Democratic Republic of the Congo (Brazzaville); Gambia basin – Senegal; Lake Chad – Chad; Turkana basin – Kenya, Lake Turkana (Polypterus spp. occur only sporadically in saline part of the lake and P. p ol y pt e r i is therefore probably restricted to the freshwater part of the Turkana Lake, i.e. Omo River delta and upstream and adjacent northernmost part of the lake); Niger basin – Nigeria, Ivory Coast, Mali; Nile basin – Egypt, Ethiopia, the Sudan; Ogoué basin – Gabon; Zambezi basin – Malawi. Prevalence: Nile basin – the Sudan, 8–25 %, n = 21, intensity 15–50 (present study), 43 %, n = 312, 10 – 70 (Khalil 1969); Turkana Lake – Kenya, 75 %, 8, intensity up to 50 (present study); Niger basin – Nigeria, 94 %, n = 84, intensity 1–226 (mostly juveniles) (Shotter & Medaiyedu 1978). Life cycle: Not known, but plerocercoids of P. polypteri have been found in the following fish of different families, which may serve as second intermediate or paratenic hosts: Auchenoglanis occidentalis (Valenciennes) (Siluriformes: Claroteidae); Barbus bynni (Cypriniformes: Cyprinidae); Lates niloticus (Linnaeus) (Perciformes: Latidae); Mormyrops anguilloides (Linnaeus) (Mormyriformes: Mormyridae); Schilbe uranoscopus Rüppell (Siluriformes: Schilbeidae); Sarotherodon galilaeus (Linnaeus); Stigmatochromis woodi (Regan); and Oreochromis niloticus (Linnaeus) (Perciformes: Cichlidae) (see below). Type material: Not known to exist. To enable future comparison of the species with other taxa, the specimen found in P. bichir (field No. T 169 /08) from Lake Turkana – Omo River delta, Todonyang, Kenya is designated as neotype and it is deposited in IPCAS (No. C- 464). Material studied: Type material: syntype of Ptychobothrium armatum Fuhrmann, 1902 (one slide and vial with 9 scolexes and several pieces of strobila) ex Turdus parochus from Egypt (ZBM E. 2361); probably syntype of Onchobothrium septicolle Diesing, 1854 ex P. bichir from Egypt collected by Kollar in 1847 or 1852 (NMW 2612 – 3); vouchers: ex P. endlicheri from Brazzaville, Republic of the Congo (MHNG 41938 – 9; RMCA 30156); ex P. endlicheri from Mali (MNHNP C 75); ex P. senegalus from Sierra Leone (BMNH 1965.2.24.46– 53, 1977.6.28.3– 4); ex P. bichir from Kainji Dam, Nigeria (BMNH 1970.8.24.38); ex “ Silurus sp.” from Bamba, Mali (MHNG 45401); Polyonchobothrium clarias ex Chrysichthys thonneri Steindachner from Gabon (RMCA 33752); plerocercoids: Polyonchobothrium sp. ex Auchenoglanis occidentalis (Valenciennes) from unknown locality collected by McClelland (RVC C 1106 – 7); ex Barbus bynni from unknown locality collected by McClelland (RVC C 1105); ex Mormyrops anguilloides (Linnaeus) from unknown locality collected by McClelland (RVC C 1103); ex Schilbe uranoscopus from unknown locality collected by McClelland (RVC C 1109); new material (see Appendix 1 for details): around 100 worms collected from 7 / 8 (7 of 8 examined) Polypterus bichir from Kenya, Lake Turkana and the Sudan, Sennar Dam; 1 / 3 P. endlicheri from the Sudan, White Nile, Kostí; 1 / 13 P. senegalus from the Sudan. The new material is deposited in BMNH (Nos. 2012.3.120.1– 13), IPCAS (No. C- 464), MHNG (Nos. 62880, 82040–82047), USNPC (Nos. 105392–105394, 105401 – 105403) and ZMB (Nos. 7515–7516). Published records: Leydig (1853); Klaptocz (1906); Hesse (1922); Janicki (1926); Joyeux & Baer (1928); Meggitt (1930); Ukoli (1965); Khalil (1969, 1973); Shotter & Medaiyedu (1978); Troncy (1978); Jones (1980). Re-description (based on 20 complete worms from Ethiopia, Kenya and the Sudan): Bothriocephalidea, Bothriocephalidae. Strobila up to 10 cm long (up to 20 cm according to Khalil, 1969); maximum width 2.3 mm. External and internal segmentation present; segments wider than long, markedly craspedote (Fig. 33). Two pairs of longitudinal osmoregulatory canals; dorsal canals narrow (diameter up to 16); ventral canals wide (diameter up to 20), connected by transverse anastomoses. Longitudinal musculature well developed, muscle fibres diffused (Fig. 40). Surface of strobila covered with capilliform filitriches. Scolex elongate (Figs. 23, 34), 700 – 1,490 (1,065 ± 198) long by 260–430 (350 ± 44) wide (n = 19). Apical disc prominent, wider than scolex proper, usually four-lobed in apical view, 350–510 (410 ± 45) wide by 145–255 (200 ± 33) long (n = 17), armed with 27–35 (32 ± 2; n = 12) large hooks, 14–165 (106 ± 37; n = 314) long (Jones 1980 reported hooks up to 190 μm long), arranged usually in four quadrants (6–9 hooks in each quadrant). Hooks variable in size in each quadrant, smallest being on periphery and increasing to middle of quadrant, with largest hook 120–165 (152 ± 13; n = 10) in centre (Figs. 24, 35, 36). Bothria elongate, shallow, 580 – 1,020 (790 ± 37) long by 100–235 (163 ± 40) wide (n = 13) (Figs. 23, 34). Surface of scolex covered with capilliform filitriches and tumuliform globular structures (diameter around 1) (Fig. 25). Neck absent, first segments appear immediately posterior to scolex (Figs. 23, 33, 34). Immature segments 80–247 long by 270–2130 wide; length/width ratio 0.06–1.04: 1 (n = 45) (Fig. 33). Mature segments wider than long, 125–300 (178 ± 56) long by 1,400–2,300 (1,830 ± 386) wide; length/width ratio 0.06– 0.60: 1 (n = 12) (Fig. 39). Gravid segments wider than long, 170–810 (415 ± 240) long by 775 – 1,680 (1,155 ± 254) wide; length/width ratio 0.12–0.76: 1 (n = 13) (Fig. 33). Testes medullary, oval, 30–65 (48 ± 9; n = 18) in number per segment (up to 72 according to Jones 1980), 37– 81 (57 ± 14) long by 22–41 (34 ± 7) wide (n = 14), forming 2 narrow longitudinal bands (17–38 testes per band), confluent between segments, absent medially and near lateral margins (Fig. 39). Cirrus-sac large, thick-walled (thickness of sac wall 3–8), pyriform, oblique, with proximal part curved anterolaterally, 46–182 (111 ± 34) long by 51–179 (130 ± 30) wide (length/width ratio 0.70–1.06: 1) (n = 20), pre-equatorial to equatorial (at 30–51 % of length of mature segment from its anterior margin; n = 10) (Figs. 38–40). Internal seminal vesicle absent. Vas deferens forms numerous loops lateral to cirrus-sac; internal sperm duct strongly coiled; cirrus unarmed, opening into genital atrium (Figs. 26, 38–40). Numerous prostatic glands around anterior part of cirrus-sac (Figs. 38–40). Genital pore dorsal, median, near anterior margin of segment, transversely elongate (Fig. 39). Ovary slightly asymmetrical, compact, transversely elongate, 20–50 (40 ± 10) long by 100–550 (410 ± 150) wide (n = 10) (Fig. 39). Vagina a straight, thick-walled wide tube, 22–37 (28 ± 4; n = 15) in diameter, opens posterior to cirrus-sac into genital atrium; vaginal sphincter absent (Fig. 38). Vitelline follicles cortical, numerous, small, spherical, 19–41 (28 ± 5; n= 15) in diameter, form 2 wide longitudinal bands confluent between segments, separated medially to form ventral and dorsal bands (Fig. 39), rarely connected by several follicles in postovarian region. Uterine duct winding, forms numerous tightly coiled loops, filled with eggs, enlarged in gravid segments (Fig. 33). Uterus thin-walled, median, spherical, enlarged in gravid segments, occupying 23–36 % of segment surface (Fig. 33). Uterine pore thick-walled, opens anterior to midlength of uterus. Eggs oval, thin-walled, operculate, may be embryonated, 30–50 (40 ± 7) long by 20–45 (30 ± 6) wide (n = 20), fully formed oncosphere 22–37 (28 ± 4) long by 18–30 (23 ± 4) wide (n = 15) (Figs. 27, 37). Remarks: Polyonchobothrium polypteri was described as Tetrabothrium polypteri based on worms found in Polypterus bichir from the Nile River in Egypt by Leydig (1853), who described only scolex morphology. Taxonomic history of the species, which was transferred to several genera, was reviewed by Jones (1980), who redescribed the taxon on the basis of tapeworms collected by L. F. Khalil in three species of bichirs (P. bichir, P. endlicheri and P. senegalus) from the White Nile at Jebel-Awlia (south of Khartoum, the Sudan). This redescription was detailed, but it seems that tapeworms studied by Jones (1980) may have been relaxed too long in the water, because their segments were unnaturally long in relation to their width (see figs. 14 and 15 in Jones 1980). In our new material from Polypterus spp. from Kenya and the Sudan, which was observed alive, segments were invariably much wider than long and markedly craspedote. Jones (1980) provided erroneous measurements of the eggs (396–444 μm by 264–288 μm), which was apparently caused by an incorrect position of the decimal point, and reported the eggs to be unoperculate when laid. However, we observed operculate eggs in the new material from the Sudan (Figs. 27, 37). Kuchta et al. (2008 a, b) considered Polyonchobothrium to be monotypic, with P. polypteri representing its type and only species, because species previously placed in Polyonchobothrium (see Kuchta & Scholz 2007 for list of synonyms) actually belong to other genera (Kirstenella, Senga and Tetracampos). Polyonchobothrium differs from other bothriocephalidean genera in scolex morphology, especially in the possession of a prominent apical disc, usually divided into four separate lobes, each of them armed with 6–9 massive hooks up to 190 μm long. Polyonchobothrium polypteri is considered here to be a specific parasite of bichirs (Polypteridae), which represent an ancient lineage of ray-finned fish (Actinopterygii) endemic to Africa. Polypterids are phylogenetically distant from teleosts and other freshwater fish, such as paddlefish, sturgeons, gars and bowfins (Suzuki et al. 2010). Adult worms were found in other hosts, such as Chrysichthys thanneri (present study; RMCA 33752) or “ Silurus sp.” (present study, Joyeux & Baer 1928; MHNG 45401), but they may represent just atypical hosts. Fuhrmann (1902) described Ptychobothrium armatum from a trush reported under the name Turdus parochus from Egypt. However, trush of this name has never been described, which casts doubts upon the actual host of these specimens. Examination of the type material (ZBM E. 2361) has shown that Polyonchobothrium polypteri was misidentified. The most probable explanation of this unusual finding of a specific parasite of bichir in a passeriform bird is mislabelling of samples. Bichi & Yelwa (2010) reported P. polypteri in clariid catfish (Clarias gariepinus) in Nigeria. This finding may represent misidentification of Tetracampos ciliotheca, which is a typical parasite frequently infecting that host (see below). Larvae (plerocercoids), juvenile or immature specimens of P. polypteri have been found by the present authors in the intestine of several unrelated fish, such as claroteid catfish, mormyrids, barbels, Nile perch and tilapias, which may serve as paratenic or postcyclic hosts (Appendix 1). Seasonality in the occurrence and maturation of P. polypteri has been indicated by previous authors, because immature specimens were found in spring (March and April by Jones, 1980) in the Sudan and from July to September in Nigeria (Ukoli 1965), whereas fully mature specimens were present only in autumn (October) in the Sudan (Jones 1980). Even though immature worms dominated in all newly collected samples, material from Kenya and the Sudan was represented by mixture of immature, mature and gravid worms in March 2006 (the Sudan), September 2008 and 2009 (Kenya). In November 2008 (the Sudan) only immature worms were found. Shotter & Medaiyedu (1977) reported the highest prevalence and intensity of infection P. polypteri in bichirs from Nigeria in fish of the total length of 300–350 mm. Based on a high proportion of immature worms to mature ones (3,879 versus 866 specimens, i.e. ratio 4.5: 1), these authors supposed that many juvenile worms did not reach maturity, possibly due to effective immune reaction of fish hosts.Published as part of Kuchta, Roman, Burianová, Alena, Jirkú, Miloslav, Chambrier, Alain, Oros, Mikuláš, Brabec, Jan & Scholz, Tomáš, 2012, Bothriocephalidean tapeworms (Cestoda) of freshwater fish in Africa, including erection of Kirstenella n. gen. and description of Tetracampos martinae n. sp., pp. 1-35 in Zootaxa 3309 on pages 12-16, DOI: 10.11646/zootaxa.3309.1.1, http://zenodo.org/record/28099

Tetracampos ciliotheca Wedl 1861

Tetracampos ciliotheca Wedl, 1861 (Figs. 28–32, 41– 52) Syns: Clestobothrium clarias Woodland, 1925; Polyonchobothrium cylindraceum forma major Janicki, 1926; P. cylindraceum forma minor Janicki, 1926; Polyonchobothrium fulgidum Meggitt, 1930; Polyonchobothrium clarias (Woodland, 1925) Meggitt, 1930; Polyonchobothrium ciliotheca (Wedl, 1861) Dollfus, 1934; Polyoncobothrium ciliotheca (Wedl, 1861) Yamaguti, 1959; Polyoncobothrium clarias (Woodland, 1925) Yamaguti, 1959. Type host: Clarias anguillaris (Linnaeus) (Siluriformes: Clariidae). Other definitive hosts: Clarias gariepinus (Burchell), Clarias liocephalus Boulenger, Clarias werneri Boulenger. Life cycle: Khalil & Thurston (1973) observed hatching of eggs in 10 minutes after their transfer to tap water. Liberated coracidia had embryophore 36–42 μm long by 30–35 μm wide, cilia 18 μm long and embryonic hooks 1 μm in length (Diab 2007). Freshwater copepods serve as the first intermediate hosts, in which procercoids developed within 20–26 days. Developed procercoids (252–610 μm long) were infective for small fish, such as tilapias (Oreochromis niloticus). Experimentally infected tilapias were exposed to C. gariepinus, in which adult worms were found (Diab 2007; Ramadan 2007). Small fish that harbour immature cestodes in natural conditions, such as schilbeid and mochokid catfish (Schilbe uranoscopus, Synodontis membranacea and S. zambezensis) and tilapias (Oreochromis niloticus, Sarotherodon galilaeus) (Douellou 1992; Owolabi 2008; Eissa et al. 2011 a, b), may play a role of paratenic hosts. Type locality: Egypt, Nile River. Distribution: Lower Guinea – Gabon; Gambia basin – Senegal; Turkana basin – Kenya (all parts of the Lake Turkana); Limpopo basin – South Africa; Upper Guinea – Sierra Leone (Moa River); Niger basin – Mali, Nigeria; Nile basin – Egypt, Ethiopia, the Sudan, Tanzania, Uganda; Volta basin – Ghana; Zambezi basin – Zimbabwe, Malawi. Besides Africa, T. ciliotheca has been reported also from Asia – Israel and Turkey, probably as a consequence of introduction with host – see Remarks (Paperna 1964; Soylu & Emre 2005; present study). Prevalence and intensity of infection: Usually high, with values between 52 % and 100 % in most studies from Egypt, Nigeria and South Africa (Aderounmu & Adeniyi 1972; Shotter 1980; Faisal et al. 1989; Anosike et al. 1992; Barson & Avenant-Oldewage 2006). In the present study the overall prevalence was 5–17 % in the Sudan, 26 % in Ethiopia and 33 % in Kenya (Appendix 1). Type material: Not known to exist. To enable taxonomic comparative studies in the future, the specimen from Clarias sp. from Blue Nile, Sennar Dam, the Sudan (field No. Sud 438) is designated as neotype and is deposited in IPCAS (No. C- 466). Material studied: Type material: Clestobothrium clarias Woodland, 1925 ex C. anguillaris (BMNH 1965.2.24.29– 35); Polyonchobothrium fulgidum Meggitt, 1930 ex C. anguillaris (BMNH 1932.5.31.801– 806); Polyonchobothrium interruptus – nomen nudum (USNPC 74291 – 2); vouchers: P. cylindraceum ex C. anguillaris from Mali, Diafarabe (MNHNP C 79); P. clarias ex C. anguillaris from Senegal, Guerina (RMCA 34773) and Ghana (BMNH 1976.4.12.155– 161); ex C. gariepinus from Nigeria, Lekki Lagoon and Kainji Dam (BMNH 2004.2.18.38, 1970.8.24.37); from Tanzania, Lake Victoria, Mwanza Gulf (MHNG 33983), Zimbabwe, Save-Runde River Floodplain (BMNH 2006.9.1.6) and Sierra Leone (BMNH 1965.2.24.59– 6); ex Heterobranchus bidorsalis from Senegal, Guerina (RMCA 34723); ex Schilbe uranoscopus from unknown locality, collected by McClelland (RVC C 1108); T. ciliotheca ex Clarias sp. from Egypt, Luxor, collected by A. de Chambrier (MHNG 31547; 17.iv. 2001); ex C. gariepinus from South Africa, Rietvlei Dam, collected by M. Barson and from Turkey, Antalya (IPCAS C– 466); new material: tens of T. ciliotheca ex 2 / 18 C. anguillaris from the Sudan, Kostí and Sennar Dam; 12 worms ex 3 / 23 C. anguillaris from Senegal, Niokolo-Koba National Park, Gambia River collected by B. Koubková (2004; Sen 52, 53, 121); 84 / 322 C. gariepinus from Ethiopia, Lake Tana and Great Rift Lakes (Awasa, Langano and Ziway), 14 / 43 C. gariepinus from Kenya, Lake Turkana and 5 / 30 C. gariepinus from the Sudan, Al Kawa, Khartoum, Er Roseires Dam, Sennar Dam; 4 / 88 Clarias sp. from the Sudan, Khartoum, Lake Nubia (Asuan Dam), Sennar Dam; one C. gariepinus from Lake Malawi, collected by S. Hendrix (SSH 96 -09- M-1). The new material is deposited in BMNH (Nos. 2012.3.20.16– 25), IPCAS (No. C- 466), MHNG (Nos. 55309, 55337, 55338, 62879, 62904, 63006–63328), USNPC (Nos. 105395–105400, 105404 – 105408) and ZMB (Nos. 7517–7523). Published records: Wedl (1861); Woodland (1925); Janicki (1926); Meggitt (1930); Tadros (1968); Khalil (1969, 1973); Aderounmu & Adeniyi (1972); Khalil & Thurston (1973); Amin (1978); Tadros et al. (1979); Shotter (1980); Wabuke-Bunoti (1980); Onwuliri & Mgbemena (1987); Faisal et al. (1989); Mashego & Saayman (1989); Imam & El-Askalany (1990); Imam et al. (1991 a, b); Anosike et al. (1992); Douellou (1992); Al-Bassel (2003); El-Garhy (2003); Rizkalla et al. (2003); Hamanda & Abdrabouh (2004); Oniye et al. (2004); Akinsanya & Otubanjo (2006); Barson & Avenant-Oldewage (2006); Olofintoye (2006); Diab (2007); Ayanda (2008, 2009a, b); Barson et al. (2008); Mwita & Nkwengulila (2008); Moyo et al. (2009); Bichi & Yelwa (2010); Madanire-Moyo & Barson (2010); Madanire-Moyo et al. (2010); Eissa et al. (2011 a, b). Re-description (based on 25 complete worms from Ethiopia, Kenya and the Sudan): Bothriocephalidea, Bothriocephalidae. Strobila small, oval or almost spherical in cross section, up to 30 mm long; maximum width 475. External and internal segmentation present; segments wider than long, acraspedote (Figs. 31, 41, 44). Two pairs of osmoregulatory canals; dorsal canals narrow; ventral canals wide, connected by transverse anastomoses. Inner longitudinal musculature well developed, muscle fibres diffused (Fig. 46). Surface of strobila covered with capilliform filitriches. Scolex elongate to ovoid, 285–510 (396 ± 62) long by 115–245 (165 ± 42) wide (n = 20) (Figs. 28, 42). Apical disc weakly developed, 104–290 (156 ± 63) wide and 35–120 (97 ± 24) high (n = 20), armed with 25–35 (29 ± 2; n = 18) small hooks (Amin 1978 reported as many as 41 hooks) 12–51 (37 ± 7; n = 537) long, arranged in two lateral semicircles separated from each other on dorsal and ventral side. Hooks variable in size in each semicircle, with largest hook 40–51 (46 ± 3; n = 20) in each corner of apical dic (Figs. 29, 43, 45). Bothria elongate, shallow, 200– 410 (308 ± 56) long by 57–120 (79 ± 22) wide (n = 20) (Figs. 28, 42). Surface of scolex covered with capilliform filitriches and numerous tumuliform globular structures (diameter around 1) (Fig. 30). Neck absent, first segments appearing immediately posterior to scolex (Fig. 41). Immature segments 80–235 (144 ± 39) long by 84–261 (167 ± 57) wide; length/width ratio 0.41–2.58: 1 (n = 38) (Fig. 41). Mature segments wider than long by, 90–400 (182 ± 68) long by 135–480 (255 ± 96) wide; length/ width ratio 0.3–1.0: 1 (n = 41) (Fig. 41). Gravid segments wider than long, 178–488 (198 ± 69) long by 180–455 (316 ± 71) wide; length/width ratio 0.5–1.2: 1 (n = 35) (Figs. 31, 41, 44, 48). Testes medullary, spherical, 5–15 (10 ± 3; n = 21) in number per segment, 21–48 (33 ± 7; n = 60) in diameter, forming 2 narrow longitudinal bands (4–9 testes per band), confluent between segments, absent medially and near lateral margins (Fig. 44). Cirrus-sac large, thin-walled (thickness of sac wall up to 4), oval, 32–66 (48 ± 9) long by 28–68 (45 ± 10) wide (length/width ratio 0.77–1.73: 1) (n = 15), equatorial (39–59 % of length of mature segment; n = 10) (Fig. 46). Internal seminal vesicle absent; cirrus unarmed, opening into genital atrium. Vas deferens forms numerous loops lateral to cirrus-sac; internal sperm ducts strongly coiled. Genital pore dorsal, median, pre-equatorial. Ovary symmetrical, forming two spherical lobes, 31–91 (59 ± 18) long by 76–183 (113 ± 29) wide (n = 14) (Fig. 44). Vagina a straight, thin-walled tube, 6–16 (11 ± 4; n = 9) in diameter, opens posterior to cirrus-sac into genital atrium; vaginal sphincter absent. Vitelline follicles few, small, spherical, 12–40 (19 ± 8; n = 28) in diameter, medullary, distributed among testes, visible only in some mature and gravid proglottides (Fig. 44). Uterine duct winding, short, filled with eggs (Fig. 41). Uterus thin-walled, median, spherical, enlarged in gravid segments, occupying 57–80 % of segment surface (Figs. 41, 48). Uterine pore thick-walled, opens in centre of uterus. Eggs widely oval to spherical, 28–72 (46 ± 9) long by 27–51 (40 ± 6) wide (n = 46), with external hyaline membrane and internal granular layer surrounding fully formed oncospheres, 17–45 (27 ± 8) long by 17–31 (23 ± 4) wide (n = 41) in terminal segments; eggs enlarging during their development in uterus (Figs. 32, 47). Remarks: Taxonomic history of bothriocephalideans parasitic in clariid catfish in Africa is complicated because apparently conspecific tapeworms were reported under different species names and were placed in several genera. Most commonly, they were identified as Polyonchobothrium clarias (Woodland, 1925), but this species is a junior synonym of Tetracampos ciliotheca (see Kuchta et al. 2008 b). Wedl (1861) described T. ciliotheca from cestodes parasitic in Heterobranchus anguillaris (= Clarias anguillaris) from Egypt. Since the original description was incomplete, most subsequent authors considered T. ciliotheca as a nomen nudum or placed it in the order Proteocephalidea or even Tetraphyllidea, because its eggs possess a transparent, hyaline external envelope (Southwell 1925; Janicki 1926). Kuchta et al. (2008 a, b) resurrected the genus with T. ciliotheca as its type and only species because it differs from other bothriocephalideans in egg morphology, the possession of an unflattened strobila, almost round in cross section, and medullary position of vitelline follicles. The latter characteristic is also present in two other bothriocephalidean cestodes, Ptychobothrium Lönnberg, 1889 and Taphrobothrium Lühe, 1899, but they parasitize marine teleosts and their morphology is otherwise markedly different (see Kuchta et al. 2008 b). Tetracampos ciliotheca is a common parasite of clariid catfish and it is widely distributed throughout Africa, with most published reports from Egypt, Nigeria, South Africa and the Sudan (see above). The cestode has also been reported from Israel (Paperna 1964 – as P. clarias) and Turkey [Soylu & Emre 2005 – as Polyonchobothrium magnum (Zmeev, 1936); present study], apparently as a consequence of import of African species of Clarias to these countries. Records of T. ciliotheca in other catfish, such as Heterobranchus bidorsalis from Senegal (present study; Khalil 1973; RMCA 34723), Bagrus bayad from Egypt (Imam et al. 1991 a) and Chrysichthys auratus from the Sudan (present study), may represent incidental infections or these fish may serve as postcyclic or accidental hosts. Omar M. Amin deposited tapeworms found in C. anguillaris from Egypt under the name Polyonchobothrium interruptus (USNPC 74291 – 2), but that species has never been formally described and thus represents nomen nudum. In 1978 Amin himself identified these tapeworms as Polyonchobothrium clarias (= T. ciliotheca). Host-parasite relationships of T. ciliotheca and its fish host have been studied by several authors (most of them referred to this species as Polyonchobothrium clarias – see above). The tapeworms penetrates deeply into the mucosa of the intestinal wall and may cause mechanical injury by the attachment of the apical crown of hooks on the scolex (Tadros 1979; Akinsanya & Otubanjo 2006; present study Figs. 49–52). Adults of T. ciliotheca were also found in the gall bladder (Amin 1978; Shotter 1980; Faisal et al. 1989; Barson et al. 2008), where tapeworms may cause formation of nodular outgrowths in the mucosa (Wabuke-Bunoti 1980). Faisal et al. (1989) reported complete penetration of T. ciliotheca tapeworms through the intestinal wall, with their attachment in the liver, spleen and ovary. In fish with perforated intestine, the intestinal contents filled the peritoneal cavity (Wabuke-Bunoti 1980). Despite the high number of examined hosts and observed cestodes, we never found T. ciliotheca tapeworms in extraintestinal localization.Published as part of Kuchta, Roman, Burianová, Alena, Jirkú, Miloslav, Chambrier, Alain, Oros, Mikuláš, Brabec, Jan & Scholz, Tomáš, 2012, Bothriocephalidean tapeworms (Cestoda) of freshwater fish in Africa, including erection of Kirstenella n. gen. and description of Tetracampos martinae n. sp., pp. 1-35 in Zootaxa 3309 on pages 16-20, DOI: 10.11646/zootaxa.3309.1.1, http://zenodo.org/record/28099

Tetracampos martinae Kuchta, n. sp.

Tetracampos martinae Kuchta n. sp. (Figs 53–60) Type host: Bagrus meridionalis (Linnaeus) (Siluriformes: Bagridae). Type locality: Deep waters of the southeast arm of Lake Malawi, Malawi (14 °06ʹS, 35 °03ʹE). Distribution: Zambezi basin – Lake Malawi. Type material: Holotype and paratype (IPCAS C- 608) (field No. SSH 96 -09-K- 1). Material studied: Two type specimens (IPCAS C- 608). Etymology: The new species is named in honour of Martina Borovková, Institute of Parasitology, Academy of Sciences of the Czech Republic, for her extraordinary technical support. Description (based on 2 specimens, 1 without scolex): Bothriocephalidea, Bothriocephalidae. Strobila up to 19 cm long, oval in cross section; maximum width 1 mm. External and internal segmentation present; segments wider than long, slightly craspedote (Fig. 53). Two pairs of longitudinal osmoregulatory canals; dorsal canals narrow (diameter up to 10); ventral canals wide (diameter up to 25), connected by transverse anastomoses. Inner longitudinal musculature well developed, muscle fibres diffused (Fig. 60). Scolex elongate, with maximum width near posterior margin of scolex (Fig. 54), 620 long by 160 wide. Apical disc weakly developed, 100 wide and 28 high, armed with 39 small hooks 44–100 (75 ± 5; n = 15) long, arranged in two lateral semicircles separated from each other on dorsal and ventral side (Fig 56). Hooks variable in size in each semicircle, with largest hook in each corner of apical disc (Figs 55, 56). Bothria elongate, shallow, 409 long by 80 wide. Neck present (Fig. 53). Immature segments 78–135 long by 191–560 wide (n = 10) (Fig. 53). Mature segments wider than long, 103– 211 long by 690–724 wide; segment length/width ratio 0.15–0.29: 1 (n = 10). Gravid segments wider than long, 220–380 long by 1015–1059 wide; segment length/width ratio 0.21–0.37: 1 (n = 10) (Figs 53, 57, 59). Testes medullary, oval, 15–19 (n = 10) in number per segment, 36–72 (51 ± 10; 10) in diameter, forming 2 narrow longitudinal bands (7–10 testes per band), confluent between segments, absent medially and near lateral margins (Fig. 57). Cirrus-sac small, thin-walled (thickness of sac wall up to 6), pyriform, 46–59 long by 59–75 wide (length/width ratio 0.61–0.93: 1) (n = 10), pre-equatorial (at 13–44 % of length of mature segment from anterior margin; n = 10) (Figs. 57, 60). Internal seminal vesicle absent; cirrus unarmed (Fig. 60). Vas deferens forms numerous loops posterolateral to cirrus-sac; internal sperm ducts strongly coiled. Genital pore dorsal, median, near anterior margin of segment, transversely elongate (Fig. 60). Ovary symmetrical, circular biscuit-shaped, lobulate, 84–107 long by 199–242 wide (n = 10) (Fig. 57). Vagina a straight, thin-walled tube, 5–11 in diameter, opens posterior to cirrus-sac into genital atrium; vaginal sphincter absent. Vitelline follicles numerous, small, spherical, 24–40 in diameter (n = 25), medullary, form 2 wide longitudinal bands confluent between segments, separated medially, rarely connected by several follicles in postovarian region (Fig. 57). Uterine duct winding, short, filled with eggs. Uterus thin-walled, median, spherical, enlarged in gravid segments, up to 277 long and 450 wide (Fig. 59). Uterine pore thick-walled, opens in centre of uterus. Eggs oval to spherical, 40–64 long by 41–53 wide (n = 10), with external hyaline membrane and internal granular layer surrounding fully formed oncospheres 28–45 long by 30–36 wide (n = 10) in terminal segments; eggs enlarging during their development in uterus (Figs 53, 58). Remarks: Tapeworms from Bagrus meridionalis from Lake Malawi are placed in Tetracampos based on the medullary position of vitelline follicles and morphology of eggs, which are unique characters of the genus, missing in all other bothriocephalideans possessing the scolex armed with hooks on the apical disc, i.e. species of Kirstenella, Oncodiscus Yamaguti, 1934, Polyonchobothrium and Senga (see Kuchta et al. 2008 b). The new species differs from congeneric T. ciliotheca in the following characteristics: (i) much longer and dorsoventrally flattened strobila (19 cm vs. oval, much smaller strobila less than 3 cm); (ii) larger hooks (up to 100 µm long vs. shorter than 52 µm); and (iii) mature and gravid segments much wider than long vs. almost quadrate in T. ciliotheca. The definitive host of T. martinae is Bagrus meridionalis, which is endemic to Lake Malawi, whereas T. ciliotheca is specific to Clarias spp. Tetracampos martinae is the second species of Tetracampos, the diagnosis of which is emended below to reflect morphological differences between both species of the genus.Published as part of Kuchta, Roman, Burianová, Alena, Jirkú, Miloslav, Chambrier, Alain, Oros, Mikuláš, Brabec, Jan & Scholz, Tomáš, 2012, Bothriocephalidean tapeworms (Cestoda) of freshwater fish in Africa, including erection of Kirstenella n. gen. and description of Tetracampos martinae n. sp., pp. 1-35 in Zootaxa 3309 on page 22, DOI: 10.11646/zootaxa.3309.1.1, http://zenodo.org/record/28099