Trickle and single infection with Discocotyle sagittata (Monogenea: Polyopisthocotylea): effect of exposure mode on parasite abundance and development

Experimental infection of rainbow trout Oncorhynchus mykiss (Walbaum) with the monogenean Discocotyle sagittata (Leuckart, 1842) allowed comparison between trickle and single exposure, two infection modes demonstrated to occur in the wild. Both types of infection resulted in mean larval attachment success around 50%, which was significantly dependent on dose of infective larvae used (P < 0.0001), but was not affected by mode of infection (P = 0.244). Worms recovered from fish exposed to the same number of oncomiracidia but different mode of infection differed in their rate of development. The developmental stage attained by parasites was significantly affected by number of infective larvae used (P = 0.005), and by the interaction between dose and mode of infection (P = 0.026), suggesting competition among attached larvae. Statistical analysis demonstrated that in the early stages of infestation, worm distribution over the gill arches can be explained by the relative amount of water flowing over them. One, two and three months post-infection parasite numbers were comparable (P = 0.805), but their observed distribution gradually decreased in gill arches III and IV and increased in gill arch I, suggesting that parasites migrate after initial attachment. These results reproduce phenomena observed in the field, indicating that the experimental infection system could be employed to study infection dynamics and host-parasite interactions under controlled conditions.     

Actin and major sperm protein in spermatozoa of a nematode, Graphidium strigosum (Strongylida: Trichostrongylidae).

In most amoeboid cells, the main protein involved in motility is actin. Nematode sperm are an exception, and their amoeboid motility is based on major sperm protein (MSP). We have studied the localization of actin and MSP in spermatids and spermatozoa of Graphidium strigosum (Dujardin, 1845), a species which has elongate male germ cells in which organelles are easily identified. Electrophoreses of G. strigosum sperm proteins indicate that the main protein band, about 15 kDa in molecular weight, is specifically recognized by an anti-MSP polyclonal antibody developed against MSP of Caenorhabditis elegans (Burke and Ward 1983). Actin is present in small quantities. Immunocytochemical observations reveal that actin and MSP have an identical localization in precise areas of the male germ cells. Spermatids are labelled as dots around a central unlabelled zone, and spermatozoa are labelled only at the level of the anterior cap. Observations in G. strigosum are similar to that previously obtained in Heligmosomoides polygyrus (Mansir and Justine 1996). Co-localization of actin and MSP in the anterior cap of the spermatozoon, the region associated with pseudopod production, does not demonstrate directly that actin is involved in amoeboid movements, but shows that the role of actin in the cytoskeleton of nematode sperm should be re-investigated.     

Description of Pseudorhabdosynochus seabassi sp. n. (Monogenea: Diplectanidae) from Lates calcarifer and revision of the phylogenetic position of Diplectanum grouperi (Monogenea: Diplectanidae) based on rDNA sequence data

Pseudorhabdosynochus seabassi sp. n. (Monogenea: Diplectanidae) from the gill filaments of Lates calcarifer Bloch, a marine teleost fish held in floating sea cages in Guangdong Province, China, is described based on morphological observations and molecular data. The shapes of the male copulatory organs (MCO) of Pseudorhabdosynochus spp. were the focus of this study. The typical proximal part of the MCO in most species of Pseudorhabdosynochus is reniform, heavily sclerotized, and divided into four chambers. However, the new species from L. calcarifer has a bulbous proximal region with four concentric layers of apparent muscular origin, instead of a reniform structure with four compartments. This organ is also different in Diplectanum grouperi Bu, Leong, Wong, Woo et Foo, 1999, being sclerotized, cup-shaped, wide proximally with four concentric muscular layers and tubular distally. The 3' terminal portion of the small subunit ribosomal RNA gene (ssrDNA) and the 5' terminal region (domains C1-D2) of the large subunit ribosomal RNA gene (lsrDNA) were used to reconstruct the phylogenetic relationships of P. seabassi and D. grouperi with related taxa utilizing maximum-parsimony and neighbour-joining methods. Phylogenetic analyses unequivocally placed D. grouperi amongst Pseudorhabdosynochus using either ssrDNA or lsrDNA data. All species of Pseudorhabdosynochus (including D. grouperi) used in this study clustered together, inferring monophyly. Based on molecular phylogenetic evidence, we propose that D. grouperi from Epinephelus coioides Hamilton be transferred to Pseudorhabdosynochus as P. grouperi comb. n.     

Contribution to the knowledge of Gyrodactylus gurleyi Price, 1937 (Monogenea: Gyrodactylidae)

Gyrodactylus longoacuminatus Zitnan, 1964, forma minor sensu Ergens (1985) was identified as G. gurleyi Price, 1937. Morphological and metrical evaluations of the hard parts of opisthaptor of more than 200 specimens provided data on the variability of this parasite; Cyprinus carpio haematopterus represents its new host. Since the shape of the hard parts of opisthaptor of G. gurleyi is very similar to that of G. longoacuminatus, further detailed studies of the two species will be necessary, particularly of their reaction to the changes of environmental temperature.     

The handling and fate of spermatophores in Neoentobdella diadema and N. apiocolpos (Monogenea: Capsalidae: Entobdellinae)

The elongated encased spermatophores of the capsalid (entobdelline) monogeneans Neoentobdella diadema (Monticelli, 1902) Kearn et Whittington, 2005 and N. apiocolpos (Euzet et Maillard, 1967) Kearn et Whittington, 2005 have been found attached by their proximal ends to the region of the vaginal opening, with the bulk of the spermatophore projecting from the vagina and therefore lying outside the body. In spite of previous reports, no spermatophores were found projecting from the common genital opening and if spermatophore exchange is as rapid as it is in the related entobdelline Entobdella soleae, then the chances of finding a spermatophore in this location are small. In N. diadema and N. apiocolpos it is likely that sperm enters the vagina through the open proximal end of an attached spermatophore, after which the empty spermatophore case is probably discarded. There is no evidence for a previous proposal that the whole spermatophore is engulfed by the vagina followed by digestion of the case to release the sperm. Three specimens of N. diadema were found each with two spermatophore cases projecting from the vagina and a specimen of N. apiocolpos carried three cases. Assuming that each parasite is able to donate or receive only one spermatophore at each mating, then the presence of one spermatophore does not prevent a further mating and acceptance of a fresh spermatophore. In spite of differences between the spermatophores of E. soleae and N. diadema/N. apiocolpos, the events of spermatophore exchange may be similar.     

Annulotrematoides bryconi sp. n. (Monogenea: Dactylogyridae) parasitic on Brycon cephalus (Osteichthyes: Characidae) from Brazil

Annulotrematoides bryconi sp. n. is described and illustrated from specimens collected from gills of characiform fish, Brycon cephalus (Günther, 1869), in pisciculture ponds from Pirassununga, S?o Paulo, Brazil. Diagnostic characters of the new species are the tegument of trunk showing annulations, except on the cephalic regiona, and copulatory complex comprising sclerotized male copulatory organ coiled in 1 1/2 rings. This is the first record of monogeneans parasitic on the gills of B. cephalus.     

A review of Dendromonocotyle (Monogenea: Monocotylidae) from the skin of stingrays and their control in public aquaria

Dendromonocotyle species (Monogenea: Monocotylidae) are the only monocotylids to parasitize the skin of chondrichthyan hosts. Currently 11 species are recorded from the skin of ray species in the Dasyatidae, Myliobatidae and Urolophidae. There have been increasing reports of Dendromonocotyle outbreaks on rays kept in public aquaria. This paper provides a broad review of Dendromonocotyle that should assist taxonomists and aquarists with species identification and help decisions on potential control methods for Dendromonocotyle infections. The taxonomy and host-specificity of Dendromonocotyle are discussed and a key to current species is provided. We summarise what little is known about the biology of Dendromonocotyle including egg embryonation and hatching, feeding, camouflage and reproduction. The efficacy of freshwater baths, chemical treatments and biological control measures such as the use of cleaner fish for Dendromonocotyle is also discussed. We demonstrate that effective control of Dendromonocotyle on captive rays is hampered by the lack of basic biological data on the life cycle of the parasites. A case history is provided outlining the success of a public aquarium (Underwater World, Mooloolaba, Queensland, Australia) in managing D. pipinna infections on captive Taeniura meyeni without chemical intervention simply by taking measures to reduce host stress.     

The capsalidae (Monogenea: Monopisthocotylea): a review of diversity, classification and phylogeny with a note about species complexes

The Capsalidae are monogeneans parasitizing 'skin', fins and gills of marine fishes. Some capsalids are pathogenic to cultivated fish and a few have caused epizootic events. It is a cosmopolitan family with broad host associations (elasmobranchs and teleosts, including sturgeons). Approximately 200 capsalid species are placed in nine subfamilies and 44-46 genera, some of which are well known (Benedenia, Capsala, Entobdella, Neobenedenia). Sturgeons host two capsalid species (Nitzschiinae) and 15 species in five genera are reliably reported from elasmobranchs. The combination of ancient (shark, ray, sturgeon) and modem (teleost) host fish lineages indicates that capsalid evolution is likely a blend of coevolution and host-switching, but a family phylogeny has been lacking due to deficient knowledge about homologies. The current phenetic subfamilial classification is discussed in detail using a preliminary phylogeny generated from large subunit ribosomal DNA sequence data from representatives of five subfamilies. Monophyly of the Capsalidae is supported by possession of accessory sclerites. Hypotheses are proposed for the possible radiation of capsalids. A suggestion that Neobenedenia melleni, a pathogenic species atypical due to its broad host-specificity (>100 host teleost species from >30 families in five orders), may be a complex of species is supported from genetic evidence. This may explain peculiarities in biology, taxonomy, host associations and geographic distribution of N. 'melleni' and has implications for fish health. Holistic studies using live and preserved larval and adult capsalid specimens and material for genetic analysis are emphasised to further determine identity, phylogeny and details of biology, especially for pathogenic species.     

Ultrastructure of the excretory system of Brachylaimus aequans (Trematoda: Brachylaimoidea)

The ultrastructure of the flame cell, excretory capillaries, ducts, collecting ducts, excretory bladder, and excretory pore of Brachylaimus aequans was studied 6-8 days p.i. The excretory ducts, collecting ducts and excretory bladder are provided with numerous lamellae on the luminal side. The cilia of lateral flames in the excretory ducts have a triated rootlet. The excretory pore is covered by a tegument identical with the body tegument. The syncytium of the excretory bladder is connected with the tegument of the excretory pore by means of a septate desmosome. No lipid or excretory corpuscles have been demonstrated in the excretory system.     

Ultrastructure of Archigetes sieboldi (Cestoda: Caryophyllidea): relationship between progenesis, development and evolution

Ultrastructural characteristics of progenetic and monoxenic Archigetes sieboldi Leuckart, 1878 from the oligochaete Limnodrilus hoffmeisteri Claparède are described. Our observations demonstrate that progenetic Archigetes sieboldi shares characteristics of both larval (progenetic) and adult stages. The primary larval characteristics are: the presence of a cercomer; a surface filamentous coat covering the whole worm; the presence of the penetration glands and the absence of tegumental ones; wide sarcoplasmic processes connecting the circular and longitudinal external tegumental muscles; the absence of the dense homogenous zone of the basal lamina beneath the epithelial cytoplasm of all reproductive organs and ducts; non-functional gonopores; and an orthogonal plan of nervous system with three pairs of longitudinal nerve trunks. The principle adult characteristics are: oogenesis, spermiogenesis and vitellogenesis that produce fertilized eggs; the uterine glands; a well-developed longitudinal tegumental muscle layer between tegumental cytons; and the presence of different microtriches. As a result of this progenetic development there has been a secondary reduction in the life cycle of A. sieboldi. It is postulated that a similar process of progenesis may have played a major role in the early evolution of the Caryophyllidea by first appearing in a plerocercoid stage of an ancestral strobilate cestode from fish.     

Homing of Gyrodactylus salaris and G. derjavini (Monogenea) on different hosts and response post-attachment

In natural European waters, the congeneric monogeneans Gyrodactylus derjavini Mikailov, 1975 and G. salaris Malmberg, 1957 are primarily found on brown trout Salmo trutta L. and Atlantic salmon Salmo salar L., respectively. Interestingly, rainbow trout, Oncorhynchus mykiss (Walbaum), originating from North America, is as susceptible as brown trout to G. derjavini. However, the mechanisms involved in this host specificity are poorly understood but may include behavioural, mechanical and chemical factors affecting parasite attraction, attachment, feeding, reproduction and host responses. In the present laboratory work, this question has been studied. Detached parasites (either G. derjavini or G. salaris) were offered a choice in small aquaria between fry of rainbow trout, Atlantic salmon and carp Cyprinus carpio L. Within 48 hours more than 90% of G. derjavini colonised rainbow trout and left salmon almost uninfected. Some parasites were found on carp. During the same time span, more than 60% of G. salaris attached to salmon, the rest infected rainbow trout and none were found on carp. Following attachment, the parasites need appropriate stimuli to initiate feeding and reproduction but even such a successful specific colonisation can be followed by a host response. Both humoral and cellular elements have been suggested to participate in these reactions but in the present work it was demonstrated by immunoblotting and immunocytochemistry that no antibodies in host mucus and host plasma bound to any parasite structures or epitopes.     

Histochemical demonstration of five enzymes' activities in Macrogyrodactylus clarii (Monogenea: Gyrodactylidae) from the catfish Clarias gariepinus

Histochemical techniques were applied to whole mounts, to study the distribution of the enzymes alkaline phosphatase, acid phosphatase, adenosine triphosphatase, 5'-nucleotidase and glucose-6-phosphatase in the organs and tissues of a viviparous monogenean, Macrogyrodactylus clarii Gussev, 1961, from the gills of the North African catfish Clarias gariepinus (Burchell) in Egypt. The following organs and tissues were studied: head region, anterior adhesive glands, mouth region, pharynx, intestine, testis, vesicula seminalis, male accessory gland, male accessory reservoir, copulatory organ, receptaculum seminis, egg-cell forming region, embryonic cells, excretory system, nerve cells, haptor, muscle fibres and subtegumental cell bodies (cytons). The enzymes showed marked differences in their activities among the studied organs and tissues. Alkaline phosphatase and acid phosphatase activities were detected in many organs and tissues, while the activities of adenosine triphosphatase, 5'-nucleotidase and glucose-6-phosphatase were restricted to a few organs. Although no positive reaction for any enzyme was observed in the anterior adhesive gland cells, a positive reaction for acid phosphatase was detected in the anterior adhesive areas. All enzymes showed marked activity in the digestive and excretory systems. The distribution of the enzymes in the tissues and organs of M clarii is compared with those of other monogeneans, including other gyrodactylids parasitizing the same host fish. Some possible functions of the enzymes are discussed.     

云南锦斑蛾幼虫触角和口器感器的超微结构

利用扫描电子显微镜技术,研究了云南锦斑蛾幼虫头部触角和口器上的感器.结果 表明,云南锦斑蛾幼虫头部呈椭球形.每根触角上具有1根毛形感器、1根刺形感器、5根锥形感器和1根栓锥形感器.上唇两侧各有6根刺形感器.内唇分布有6根刺形感器、4根内唇感器和2根指形感器.每个上颚表面具有2根刺形感器.下颚的感器主要分布在外颚叶和下颚...     

Gyrodactylus bohemicus sp. n. (Monogenea: Gyrodactylidae) from Oncorhynchus mykiss (Walbaum) and Salvelinus fontinalis (Mitchill) (Clupeiformes: Salmonidae) in Czechoslovakia.

Gyrodactylus bohemicus sp. n (Gyrodactylidae: Monogenea) is described from the fins, skin and gill filaments of Oncorhynchus mykiss (Walbaum) and Salvelinus fontinalis (Mitchill) (Clupeiformes: Salmonidae) from a trout farm in Czechoslovakia. G. bohemicus sp. n. is most closely related to G. thymalli Zitnan, 1960 and G. magnus Konovalov, 1967 in the shape of the anchors, the ventral and the dorsal bar, but can be distinguished from both these species by the shape of the marginal hook proper.     

Neopolystoma liewi sp. n. (Monogenea: Polystomatidae) from the eye of the Malayan box turtle (Cuora amboinensis)

Neopolystoma liewi sp. n. is described from the conjunctival cavity of the Malayan box turtle Cuora amboinensis (Daudin, 1802), in Peninsular Malaysia. This is the first record of Neopolystoma in Malaysia and the fourth polystomatid species described from C. amboinensis. Of the 27 Malayan box turtles examined, 8 were found to be infected. A maximum of 2 parasites per eye and 4 individuals per host was recorded. N. liewi sp. n. differs from all other members of the genus by possessing few and short genital spines and small marginal hooks. The oncomiracidium has 64 ciliated cells arranged symmetrically about the sagittal axis.