Genetic variation of fish parasite populations in historically connected habitats: undetected habitat fragmentation effect on populations of the nematode Procamallanus fulvidraconis in the catfish Pelteobagrus fulvidraco

Habitat fragmentation may have some significant effects on population genetic structure because geographic distance and physical barriers may impede gene flow between populations. In this study, we investigated whether recent habitat fragmentation affected genetic structure and diversity of populations of the nematode Procamallanus fulvidraconis in the yellowhead catfish, Pelteobagrus fin't4draco. The nematode was collected from 12 localities in 7 floodplain lakes of the Yangtze River. Using I I intersirnple sequence repeat markers, analysis of molecular variance showed that genetic diversity occurred mainly within populations (70.26%). Expected heterozygosity (He) of P. fulvidraconis was barely different between connected (0.2105) and unconnected lakes (0.2083). Population subdivision (Fst) between connected lakes (0.2177) was higher than in unconnected lakes (0. 1676). However, the connected and unconnected lakes did not Cluster into 2 clades. A Mantel test revealed significant positive correlation between genetic and geographic distances (R = 0.5335, P < 0.01). These results suggest that habitat fragmentation did not cause genetic differentiation among populations or a reduction of diversity in isolated populations of P. fulvidraconis. At least 2 factors may increase the dispersal range of the nematode, i.e., flash flooding in summer and other species of fish that may serve as the definitive hosts. Moreover, lake fragmentation is probably a recent process; population size of the nematode in these lakes is large enough to maintain Population structure.

Population dynamics and maturation cycle of Camallanus cotti (Nematoda : Camallanidae) in the Chinese hooksnout carp Opsariichthys bidens (Osteichthyes : Cyprinidae) from a reservoir in China

The seasonal population dynamics and maturation cycle of the nematode Camallanus cotti in the posterior intestine of Chinese hooksnout carp Opsariichthys bidens have been studied in the Danjiangkou Reservoir of the Hubei Province in central China from September 2004 to November, 2005. The overall prevalence, mean abundance and intensity of C cotti among fish sampled (n = 700 fish) were 47%, 2.29 +/- 12.38 ( +/- S.D.) and 1-307 (average 4.89 +/- 17.74), respectively. The overall sexual ratio of female to male nematodes (excluding L3 and L4 juveniles) was 1.17:1. Statistical results showed weakly positive correlations betweerl fish length and the number of nematodes per host. The dynamics of infection of the nematode exhibited significant seasonal pattern in changes in mean abundance. A similar pattern was found for changes in nematode prevalence, although this was not statistically significant. Higher levels of infection were observed among fish sampled in summer months and the lower in the winter. Neither the prevalence nor the abundance of the parasite was significantly different between male and female hosts. The pattern of frequency distribution of the parasite in the host was found to be over-dispersed throughout the sampling period. In addition, studies on the development and maturation of the parasite in O. bidens revealed that development (maturation), recruitment of the next generation, and reproduction may be continuous year-round, although reproduction may peak during the winter. (c) 2007 Elsevier B.V. All rights reserved.

Occurrence of Camallanus cotti in greatly diverse fish species from Danjiangkou Reservoir in central China

Two thousand four hundred fifty-eight fish comprised of 53 species were captured in the Danjiangkou Reservoir, in the northwestern part of Hubei Province, central China during 2004, to examine Camallanus cotti infections. We found that 19 cypriniform, 3 siluriforme, and 4 perciforme fishes were infected by the nematode. Our study revealed the species, Hemiculter bleekeri bleekeri, Culter oxycephaloide, Pseudolaubuca sinensis, Acanthobrama simony, Mylopharyngodon piceus, Ctenopharyngodon idella, Gnathopogon imberbis, G. argentatus, Saurogobio dabryi, S. dumerili, Gobiobotia ichangensis, Liobagrus marginatoides, and Ctenogobius shennongensis as new hosts of the worm. The number and range of fish host species found in this survey were much greater than any of the previous investigations. The mean prevalence, prevalence, mean abundance, and intensity of infection varied in different fish species, indicating a possible host preference. Moreover, we suggest that this nematode is a native parasite of cypriniform fishes in China, perhaps initially in the reaches of the Yangtze River.

Evidence of host specificity and congruence between phylogenies of bitterling and freshwater mussels

Evidence of host specificity and congruence between phylogenies of bitterling and freshwater mussels. Zoological Studies 45(3): 428-434. Bitterling (Cyprinidae: Acheilognathinae) are freshwater fishes with a unique spawning relationship with freshwater mussels on whose gills they lay their eggs. During the breeding season of bitterling fishes, we collected 843 mussels belonging to 16 species from Lake Qinglan, central China and examined their gill chambers for the presence of bitterling larvae. Three species of bitterling larvae were identified; Acheilognathus tonkinensis, Ach. cf. meridianus, and Ach. barbatulus, in 3 species of mussel: Unio douglasiae, Lamprotula caveata, and L. tortuosa, suggesting host specialization. Using our own and other published data, we compared the respective phylogenies of bitterling and mussels, but failed to show clear congruence. However, broad specializations are evident, with Acheilognathus and Tanakia showing preferences for mussels with a relatively simple gill structure (Ableminae), and Rhodeus spp. showing preferences for mussels of the Anodontinae and Unioninae, which have more-complex gill structures.

First description of the male and redescription of the female of Philometra clavaeceps Dogiel and Akhmerov, 1959 (Nematoda : Philometridae), a parasite of east Asian cyprinid fish

The first description of the male and a redescription of the female of the nematode Philometra clavaeceps Dogiel and Akhmerov, 1959, a parasite of east Asian cyprinids, are presented on the basis of specimens collected from Culter erythropterus Basilewsky and Culler dabryi Bleeker from Liangzi Lake (the Yangtze River basin), Hubei Province, central China. Gravid females from the fish abdominal cavity, penetrating often into ovaries, occurred in May-June, whereas conspecific males and young mature females on the swimbladder were recorded in January. Philometra clavaeceps seems to have a pronounced annual maturation cycle in the locality. The finding of P. clavaeceps in C. dabryi represents a new host record.

Redescription of Camallanus hypophthalmichthys Dogel and Akhmerov, 1959 (Nematoda : Camallanidae) and its first record from fishes in China

The nematode Camallanus hypophthalmichthys Dogel and Akhmerov, 1959 is redescribed from specimens collected from the intestine of the bighead carp Aristichthys nobilis, from Liangzihu Lake (Yangtze River basin), Hubei Province, central China. The light and scanning electron microscopical examination made it possible to study in detail the morphology of this so far little-known species and to confirm its validity. The main specific features of C. hypophthalmichthys distinguishing it from the most similar Camallanus spp. is the presence of 3 small caudal processes on the male tail tip, 13-16 longitudinal ridges on the inner surface of the valve of the buccal capsule, and the arrangement of preanal and postanal genital papillae in the male. This finding represents a new host record, the first record of this parasite in the Yangtze River basin, and the first documented record of C. hypophthalmichthys from China. Camallanus hypophthalmichthys is considered a specific intestinal parasite of fishes of the cyprinid Hypophthalmichthyinae.

Broad host range plasmid-based gene transfer system in the cyanobacterium Gloeobacter violaceus which lacks thylakoids

Gloeobacter violaceus, a cyanobacterium lack of thylakoids, is refractory to genetic manipulations because its cells are enveloped by a thick gelatinous sheath and in colonial form. In this study, a large number of single cells were obtained by repeated pumping with a syringe with the gelatinous sheath removed. And an exogenous broad host range plasmid pKT210 was conjugatively transferred into G. violaceus. Analyses with dot-blot hybridization and restriction mapping showed that the exogenous plasmid pKT210 had been introduced into G. violaceus and stably maintained with no alteration in its structure. pKT210 extracted from G. violaceus exconjugants could be transformed into the mcr - mrr - E. coli strain DH10B but not the mcr(+) mrr(+) strain DH5alpha, which suggests that a methylase system may be present in G. violaceus.

Scanning electron microscopy of Aspidogaster ijimai Kawamura, 1913 and A-conchicola Baer, 1827 (Aspidogastrea, Aspidogastridae) with reference to their fish definitive-host specificity

Two species of aspidogastreans, namely Aspidogaster ijimai and A. conchicola, were studied by scanning electron microscopy. In nine lakes and an old river course, the Tian'ezhou oxbow, investigated in the flood plain of the Yangtze River, A. ijimai was obtained from the common carp (Cyprinus carpio) in three lakes, and A. conchicola from the black carp Mylopharyngodon piceus in three lakes and the oxbow. In none of the localities, however, were the two species found together. It is suggested that A. ijimai may be considered as a specialist parasite for the common carp, at least in the flood-plain lakes of the Yangtze River. The two parasites were similar in many aspects of their morphology. Their bodies can both be separated into a dorsal part and a ventral disc, with the body surface of the dorsal part elevated by transverse folds, and the disc subdivided into alveoli by transverse and longitudinal septa, although the number of alveoli was different in the two species. The depression on the ventral surface of the neck region was prominent for both species, and their ventral disc was covered densely with non-ciliated bulbous papillae. The position of mouth, osmo-regulatory pore and marginal organ was also similar for A. ijimai and A. conchicola. However, microridges in the trough of the folds in the neck region and numerous small pits on the upper part of the septa were found exclusively in A. ijimai, but uniciliated sensory papillae in A. conchicola.

Some nematodes of fishes from central China, with the redescription of Procamallanus (Spirocamallanus)fulvidraconis (Camallanidae)

The present paper comprises a systematic survey of nematodes based on helminthological examinations of 176 specimens of freshwater fishes, belonging to 22 species, from central China (mostly lakes in Hubei Province) collected during the autumn of 2001. The following six species were recorded: Procamallanus (Spirocamallanus) fulvidraconis Li, 1935, Camallanus cotti Fujita, 1927, Dentiphilometra monopteri Moravec et Wang, 2002, Pingus sinensis Hsu, 1933, Proleptinae gen. sp. larv., and Eustrongylides sp. larv. Data on their morphology, morphological variability, host range, prevalence, intensity and distribution are provided. SEM studies of P. fulvidraconis and larval Physalopterinae, used for the first time in these species, revealed some additional morphological details and made it possible to redescribe the former. In contrast to the existing description of P. fulvidraconis, this species was found to possess two spicules and a V-shaped gubernaculum with unequal arms (originally mistaken for the left spicule), as well as deirids, whose location can be considered an important taxonomic feature. Larvae of the Physalopterinae have not previously been reported from fishes in China. The finding of larval Eustrongylides in Paramisgurnus dabryanus represents a new host record. All but one nematode species from this zoogeographically interesting region are briefly described and illustrated.

Immunization of rainbow trout fry with Ichthyophthirius multifiliis sonicate: protection of host and immunological changes

Rainbow trout fry (10 weeks post hatch) were immunized (injection or immersion) with sonicated formalin-killed trophonts of the fish parasitic ciliate Ichthyophthirius multifiliis. Challenge infections 22 days after immunization showed a relative protection represented by significantly fewer established parasites and lower prevalence in the immunized groups compared to the controls. Associations between the obtained protection and changes in differential leukocyte counts, haematocrit values, anti Ichthyophthirius multifiliis antibodies, mucous cell density and some epidermal cell markers were investigated. No changes in antibody titers, haematocrit values and mucous cell counts were associated with the response; however, a minor change in peripheral blood neutrophils and epidermal cell markers were found.

Molecular variation of Bothriocephalus acheilognathi Yamaguti, 1934 (Cestoda : Pseudophyllidea) in different fish host species based on ITS rDNA sequences

The molecular variation in Bothriocephalus acheilognathi Yamaguti, 1934 from 11 species of freshwater fish collected in Australia, China, the Czech Republic, England and Hawaii was investigated by determining the nucleotide sequences of the internal transcribed spacer region. The length of the first and second internal transcribed spacer sequences of multiple individuals ranged from 553 to 571 bp and 553 to 615 bp, and the G + C content from 53.1 to 53.5%. The percentage sequence divergence varied between 0 and 0.9% in the ITS1 and 0 and 6.6% in the ITS2, respectively, indicating the occurrence of intraspecific variation. It is demonstrated that the fragment length variation resulted primarily from microsatellite polymorphisms present in the ITS region, especially in the ITS2 region. Phylogenetic analyses revealed that B. acheilognathi examined in this study consisted of three closely related genotypes with certain degrees of host-specificity, and the genotype representing isolates from Cyprinus carpio L. was the most common and diverse form within the species B. acheilognathi.

Manipulation of host density affects infestation of a peritrich ciliate (Epistylis lacustris) on a calanoid copepod (Leptodiaptomus minutus) in Crystal Lake, Wisconsin, USA

Specimens of the calanoid copepod, Leptodiaptomus minutus, collected in June 1994 in oligotrophic: north temperate Crystal Lake, were infested with the stalked ciliate Epistylis lacustris. E. lacustris was highly specific to L. minutus and no other coexisting zooplankters were infested. Excluding nauplii, nearly 70% of copepods carried 1-20 ciliates, although the maximum load was as high as 250 ciliates. A lower percentage of nauplii were infested by the ciliate; those that were infested had a lower ciliate load than other copepod stages. Infestation by ciliates had no significant influence on the average egg number of female copepods. In a field experiment, higher copepod densities in enclosures resulted in a significantly higher infestation rate, but the ciliate load per individual copepod did not differ significantly among treatments.

Purification and characterization of White Spot Syndrome Virus (WSSV) produced in an alternate host: crayfish, Cambarus clarkii

Penaeid shrimp is the natural host of White Spot Syndrome Virus (WSSV) that can cause high mortality in the infected hosts. Attempts to obtain sufficient amounts of purified intact WSSV for characterization have been unsuccessful. Using crayfish, Cambarus clarkii as a proliferation system, a large amount of infectious WSSV was reproduced and intact WSSV viral particles were purified with a new isolation medium by ultra-centrifugation. Purified WSSV particles were very sensitive to organic solvents and the detergent, Triton X-100. The size of the rod-shape, somewhat elliptical, intact WSSV was 110-130 x 260-350 mm with a long, tail-like envelope extension. The naked viral nucleocapsid was about 80 x 350 nm, and it possessed 15 spiral and cylindrical helices composed of 14 globular capsomers along its long axis, and a 'ring' structure at one terminus. Distinct WSSV genome DNA patterns were obtained when the purified genomic dsDNA of WSSV was digested with five different restriction enzymes (HindIII, XhoI, B(BamHI, SalI, and SacI). In addition, at least 13 major and distinct protein bands could be observed when purified intact WSSV viruses were separated by SDS-PAGE followed by Coomassie Brilliant R-250 staining. The estimated molecular weights of these proteins were 190, 84, 75, 69, 68, 58, 52, 44, 28, 27.5, 23, 19, and 16 kD, respectively. Both the 44 and 190 kD proteins were easily removed if the hemolymph from the: WSSV infected crayfish was transiently treated with 1%, Triton X-100 before it was subjected to gradient centrifugation, indicating that both of them are located on the surface of the viral envelope. These characteristics are consistent with WSSV isolated from the penaeid shrimp. (C) 2001 Elsevier Science B.V. All rights reserved.

The freeliving nematode genus Parodontophora Timm 1963 (Nematoda : Axonolaimidae) is not exclusively marine: Parodontophora limnophila sp nov from freshwater in China

Parodontophora limnophila sp. nov. is described from Poyang Lake, the largest freshwater lake of China. It is characterized by having an amphid with its posterior end close to the base of the stoma, relatively short cephalic setae, opisthocephalic setae arranged as two subdorsal groups of three longitudinally arranged setae and two single subventral setae, excretory pore at the level of the anterior part of the stoma and renette gland 34-47% of the oesophageal length. To date, the new species is the only Parodontophora species found in freshwater habitats.

Evidence of host blood feeding by the monogenean, Ancyrocephalus mogurndae (Monogenea : Ancyrocephalidae) from the gills of the mandarin fish, Siniperca chuatsi

By examining iron contents, it is demonstrated that the monogenean Ancyrocephalus mogurndae (Yamaguti, 1940) feeds on the blood of its host, the mandarin fish Siniperca chuatsi (Basilewsky). The iron content and then the quantity of blood necessary to produce this amount of iron are found different in young and fully-matured worms. Young worms contain higher levels of iron and estimated amount of blood. It is suggested that A. mogurndae may start to feed on host blood as attached on gills, and the amount of blood ingested by young worms may vary from 0.01 to 1.00 mu l before reproduction. The difference between young and fully-matured worms may be accounted for by the elimination of haematin and change of food composition in matured worms and may also be affected by reproduction. Experimental infections of the monogenean may provide supportive information for explaining the difference, and further studies should also examine the effect of immune components in host blood ol mucus on the intestines of the parasite.