The fate of Lernaeocera branchialis (L.) (Crustacea; Copepoda) in Atlantic cod, Gadus morhua L.

Lernaeocera branchialis, a copepod crustacean parasite of gadoids, represents a potential threat to both wild and farmed cod, Gadus morhua. The pathological changes associated with the early stages of experimental infection have previously been reported in detail, and this article describes the lesions associated with later chronic stages of experimental infection. Chronic infection is characterised by extravascular granuloma formation and proliferation of fibrovascular tissue around intact and fragmented, degenerate parasites within both the gill arch and cardiac region. The majority of parasite granulomas are located within connective tissues of the gill arch or pericardium; however, low numbers are present within the wall of large vessels. The intraluminal parasites and thrombi of early stage infection are largely absent in these later lesions. We propose that organisation and incorporation of the parasite thrombus into the vessel wall with subsequent granuloma formation and extrusion into the surrounding connective tissue leads to the elimination of the parasite from the vascular system. Thus, rather than being a negative consequence of infection thrombosis is protective, allowing the host to survive the substantial initial vascular insult.

Infection dynamics of two renal myxozoans in hatchery reared fry and juvenile Atlantic cod Gadus morhua L.

In order to study the infection dynamics of 2 renal myxozoans, Zschokkella hildae Auerbach, 1910 and Gadimyxa atlantica Køie, Karlsbakk and Nylund, 2007 in cultured Atlantic cod, Gadus morhua L. aged 3-19 months, a specific single-round PCR assay and a double-label in situ hybridization protocol were developed. The results demonstrated that the 2 myxozoans show spatial separation of their development with regard to spore formation inside the renal tubules versus the collecting ducts and ureters, as well as temporal separation with Z. hildae proliferating and developing spores only once the G. atlantica infection decreases, despite the presence of both myxozoans in the smallest fry studied. These results strongly suggest within-host competition of the 2 myxozoans with potential suppression of Z. hildae by G. atlantica until G. morhua acquires immunity against G. atlantica. The quantification of the G. atlantica infection inside the renal tubules before and after a 29-day experimental growth performance study using fry from hatcheries with differing filtration systems showed that the intensity of infection with G. atlantica seems to be controlled if prolonged exposure to the myxozoan transmission stages takes place from hatching onwards. Surprisingly, growth rates in the trial were inversely affected suggesting that G. atlantica does not negatively influence cod fry growth performance.

Using length-frequency data to elucidate the population dynamics of Argulus foliaceus (Crustacea: Branchiura).

This study uses a novel method for discriminating cohorts and investigating the population dynamics of the parasitic crustacean, Argulus foliaceus. Analysis of parasite length-frequency data was carried out in order to elucidate the timings and drivers behind the parasite's life cycle. Up to 6 cohorts of the parasite emerge through the course of 1 year in still-water trout fisheries in England. Recruitment ceases over the winter months; however, 3 cohorts of the parasite over-winter, 2 as eggs and 1 as a hatched stage. The technique, when used in conjunction with temperature data, also allowed for the reliable prediction of growth rates and provided estimates of egg incubation times and the length of hatching periods. These data showed that growth rates increased exponentially between the observed temperatures of 4 to 22 degrees C. The method allowed for the time taken from hatching to egg laying under field conditions to be predicted and produced estimates that were validated against independent laboratory studies on the growth of the parasite.

The influence of risk factors on the abundance, egg laying habits and impact of Argulus foliaceus in stillwater trout fisheries.

Fishery managers perceive the ectoparasitic crustacean, Argulus foliaceus to be a cause of significant economic loss through reduced fish capture rates. This study investigates the influence of previously identified risk factors on the abundance, egg laying habits and impact of this parasite through a longitudinal study of five trout fisheries of varying management intensity. Low water clarity, slow stock turnover and high temperatures showed a significant association with a high abundance of A. foliaceus. High infection levels, low water clarity and low temperature were also associated with reduced rates of fish capture, suggesting abundance of A. foliaceus alone may not affect the catch rates. Depth of egg laying varied in each site throughout the study, increasing in depth as temperature and water clarity increased. Eggs were found to be most abundant in natural fish holding areas and it is hypothesized that the location of egg laying is determined by the habitat usage of host fish.

The pathology of the early stages of the crustacean parasite, Lernaeocera branchialis (L.), on Atlantic cod, Gadus morhua L.

Lernaeocera branchialis is a penellid copepod, the adults of which are commonly found on gadoids in the North Atlantic and North Sea. L. branchialis has a severe negative impact on wild gadoids and is a potentially serious pathogen of farmed cod, Gadus morhua. This paper describes an experimental infection by cohabiting the intermediate host, the flounder, Platichthys flesus, heavily infected with juvenile stages of L. branchialis, with hatchery-reared cod in order to study the pathology of the early stages of infection of the final host. Early stages of L. branchialis induce extensive local gill hyperplasia, large intravascular thrombus formation and a moderate cellular immune response in the cardiac and branchial tissues of G. morhua. The parasite appears to migrate within the lumen of the afferent branchial artery, ventral aorta and in turn, that of the bulbus arteriosus. These findings are consistent with the presence of a haematophagous, voluminous and rapidly invasive parasite.

The epidemiology of Argulus spp. (Crustacea: Branchiura) infections in stillwater trout fisheries.

Problem infections caused by species of the crustacean ectoparasite, Argulus, in UK stillwater trout fisheries appear to have increased in recent years. A cross-sectional study of 77 such fisheries was conducted to establish the perceived problem and the extent and severity of this problem, and to identify associated risk factors. An interview-based study was conducted in 2001 using a standardized questionnaire based on the management and infection status of each fishery in the previous year. Logistic regression was used to identify potential risk factors. Argulus spp. were perceived to cause economic losses in infected fisheries through a reduction in the number of anglers due to reduced aesthetic appeal and catchability of fish. Of the sites studied, 29% experienced such a problem infection in 2000. Argulus foliaceus was identified in all but one case and was found to be widely distributed throughout the UK. The remaining case was identified as Argulus coregoni. Three risk factors were associated with problem infections: the presence of an algal bloom, slow rates of stock turnover and whether water level dropped by <1 m during the summer months.

Molecular studies on the seasonal occurrence and development of five myxozoans in farmed Salmo trutta L.

Five myxozoan species, Tetracapsuloides bryosalmonae, Sphaerospora truttae, Chloromyxum schurovi, Chloromyxum truttae and a Myxobolus species were detected in farmed brown trout, Salmo trutta L. from Central Scotland. Using PCR and in situ hybridization, this study investigated the seasonal occurrence and tissue location of these species in young of the year brown trout. C. schurovi, C. truttae and Myxobolus sp. were first detected in brown trout in April, 2 months before T. bryosalmonae and S. truttae. T. bryosalmonae and S. truttae showed proliferation in the blood with intravascular stages of T. bryosalmonae accumulating in the heart. In contrast, only small amounts of PCR products of C. schurovi and C. truttae were obtained from the blood, suggesting that these species use the vascular system for transport but proliferate only in their target tissues from which large amounts of PCR product were obtained and where parasites were visible in histological sections. Large amounts of PCR product were obtained for T. bryosalmonae, S. truttae and both Chloromyxum species from the gills of brown trout, suggesting the gills as entry locus for these species. The neurotropic Myxobolus species formed plasmodia predominantly in the peripheral nerves, possibly indicating an entry route through the skin. Presporogonic stages of all other species had disappeared by September and mature spores were present from August onwards.

A molecular study of Eubothrium rugosum (Batsch, 1786) (Cestoda: Pseudophyllidea) using ITS rDNA sequences, with notes on the distribution and intraspecific sequence variation of Eubothrium crassum (Bloch, 1779).

The internal transcribed spacers (ITS-1 and ITS-2) of the ribosomal DNA (rDNA) of fish cestodes of the genus Eubothrium were sequenced. The ITS sequences of Eubothrium rugosum were determined and compared with previously analysed congeneric species, Eubothrium crassum and Eubothrium salvelini. The ITS-1 sequences of E. rugosum and E. crassum were 535 bp long, the length of E. salvelini ITS-1 was 536 bp. The ITS-2 region was found to be 403 bp in E. rugosum and E. crassum, and 401-402 bp in E. salvelini. The ITS-1 region of E. rugosum was closer to E. salvelini (identity 98.9-99.1%) than to E. crassum(97.9-98.1%), while the sequence similarity within the ITS-2 region was almost identical (97.5-98.0% for E. crassum; 97.7-98.3% for E. salvelini). Several restriction enzymes were found to be suitable for the differentiation of the three Eubothrium species by PCR-RFLP. The intraspecific sequence variation of E. crassum from different fish hosts (freshwater and marine) and European regions was very low, 0.2% for ITS-1 and 0.5% for ITS-2. Analysis of the ITS sequences of specimens from rainbow trout from three localities in Scotland revealed that both E. crassum and E. salvelini are present in this fish host.

Tracing the route of Sphaerospora truttae from the entry locus to the target organ of the host, Salmo salar L., using an optimized and specific in situ hybridization technique.

Sphaerospora truttae is an important pathogen of Atlantic salmon parr in Scottish aquaculture. To trace the early development of S. truttae and to overcome the common problem of detecting low numbers of cryptic, early myxosporean stages, a DNA-based approach was applied in this study. Specific primers were designed for S. truttae from Atlantic salmon, based on 18S rDNA sequences, obtained from isolated myxosporean spores. These were 5' biotin-labelled and used in an optimized and rapid in situ hybridization (ISH) protocol, which provided a strong and specific signal of the parasite within host tissue sections and, at the same time, minimized structural damage to tissues due to processing. This methodology provided a reliable tool enabling the detection of S. truttae stages down to single cell level. Using ISH the epithelium of the gills was identified as the predominant entry locus of the parasite. By 3 days after infection S. truttae had penetrated the vascular epithelia and thereafter proliferated in the blood for at least 10 days before exiting the vascular system through capillary walls. From day 12 post-infection onwards, the kidney, as well as the spleen and the liver, were invaded. Numbers of S. truttae invading the kidney (37.3%) differed little from numbers found in the spleen (35.3%) and the liver (27.4%). The latter organs represented a dead end in the development of S. truttae as all stages in these organs degenerated and sporogony was found to take place exclusively inside the renal tubules. Early sporogonic stages were found from day 25 post-infection but mature spores only developed after at least 15 days of proliferation within the tubules.

Does size really matter? Effects of fish surface area on the settlement and initial survival of Lepeophtheirus salmonis, an ectoparasite of Atlantic salmon Salmo salar.

The effect of the size of the surface area of a fish host on settlement and initial survival of Lepeophtheirus salmonis (Krøyer, 1837) was determined. Atlantic salmon Salmo salar L. of various sizes-small (43 +/- 4 g), medium (173 +/- 32 g) and large (644 +/- 62 g)-were initially examined to ascertain their respective body surface area, excluding gill surface. The initial examination showed a size-dependent body to fin area ratio, with the fin area of small fish representing 34% of the total body area and the fin area of medium and large fish representing 26 and 23%, respectively. Regression analysis of the body weight and standard length against total body surface area gave a good correlation and high R2 values. Two simultaneous experimental infections with approximately 7,000 copepodids of L. salmonis were carried out on fish populations of mixed sizes consisting of 30 small fish, 10 medium-sized fish and 5 large fish, with an approximate total surface area per size group of 2700 cm2. Higher numbers of parasites were found on the small size group, which also had the highest parasite density, with 0.25 and 0.45 parasites cm(-2). Comparison of samples of 5 fish per experimental group revealed that the larger fish had the highest mean numbers of parasites, but the smaller fish still retained the highest density. There was a statistically significantly higher settlement on the fins than on the remainder of the body surface in all size groups. Highest numbers of parasites were particularly found on the dorsal and pectoral fins. Examination of the surface surface area revealed that the dorsal fin had the greater parasite density, with >2 parasites cm(-2) in all fish size groups. These data provide insight into the effects of the amount of host surface area available on parasite settlement and survival, and highlight the potentially increased susceptibility of farmed salmon smolts to infection of this ectoparasite.

The life cycle of Sphaerospora truttae (Myxozoa: Myxosporea) and some features of the biology of both the actinosporean and myxosporean stages.

A previously undescribed echinactinomyxon type actinosporean was shown experimentally to be the alternate stage of Sphaerospora truttae. The echinactinomyxon type spores were found to be released from Lumbriculus variegatus and occasionally Tubifex tubifex. Overall infection prevalence of the echinactinomyxon was 0.14%. Peak release was in March-May each year. S. truttae spores developed in experimentally infected salmon 4.5 mo post-exposure to echinactinomyxon spores. Extrasporogonic stages of S. truttae first appeared in naturally infected salmon in early July and persisted for 8 to 10 wk. Correlation between peak release of echinactinomyxon spores and appearance of extrasporogonic stages of S. truttae is discussed.

Gyrodactylus sommervillae n. sp. (Monogenea) from Abramis brama (L.) and Rutilus rutilus (L.) (Cyprinidae) in Oxfordshire, UK.

Gyrodactylus sommervillae n. sp. (Monogenea) is described from the skin, fins and gills of Abramis brama (L.) and Rutilus rutilus (L.) from Blenheim Palace Lake, Oxfordshire, UK. G. sommervillae n. sp. closely resembles G. aphyae Malmberg, 1957, G. kearni Ergens, 1990 and G. lamberti Ergens, 1990 in the approximate shape of the marginal hook sickle and the anchors, but can be separated on the precise shape of the marginal hooks and the ventral bar.

An SEM study of adhesive disc skeletal structures isolated from trichodinids (Ciliophora: Peritrichida) of the genera Trichodina Ehrenberg, 1838 and Paratrichodina Lom, 1963.

Specimens of Trichodina domerguei Wallengren, 1897, T. intermedia (Lom, 1961) and Paratrichodina incissa (Lom, 1959) were sonicated to liberate skeletal components of the adhesive disc. This enabled SEM observation of the taxonomically important structures obscured in preparations of complete cells. A previously undescribed peg-like structure on the centrifugal surface of the central part of the denticles is revealed in T. domerguei. In P. incissa the ray apophysis and its supporting apophysis appear to be absent, providing an additional characteristic to discriminate it from species of the genus Trichondina Ehrenberg, 1838. From silver stained and SEM preparations of T. intermedia and P. incissa important differences in denticle blade form are apparent, underlining the value of observation of isolated skeletal structures by electron microscopy.

Ectoparasitic species of the genus Trichodina (Ciliophora: Peritrichida) parasitising British freshwater fish.

Seven species of the genus Trichodina Ehrenberg, 1838 were identified during a sampling programme of twenty freshwater fish species from approximately sixty sites in Scotland, England and Wales. Species found include: Trichodina acuta Lom, 1961 from Cyprinus carpio L., Carassius auratus L., Oncorhynchus mykiss (Walbaum), Salmo trutta L. and Phoxinus phoxinus L.; Trichodina domerguei Wallengren, 1897 from Gasterosteus aculeatus L.; Trichodina tenuidens Faure-Fremiet, 1944 from Gasterosteus aculeatus; Trichodina pediculus Ehrenberg, 1838 from Gasterosteus aculeatus; Trichodina modesta Lom, 1970 from Abramis brama L.; Trichodina nigra Lom, 1960 from Cyprinus carpio, Salmo trutta and Oncorhynchus mykiss; and Trichodina intermedia Lom, 1960 from Phoxinus. Morphological variation within and between host populations and host specificity of the Trichodina species recovered are described.

The effects of hydrogen peroxide treatment on different life-cycle stages of the salmon louse, Lepeophtheirus salmonis (Krøyer 1837).

The effects of hydrogen peroxide administered at a concentration of 1500 ppm for 20 min at 7.5 °C on different life-cycle stages of Lepeophtheirus salmonis were examined experimentally. The mobile adult and pre-adult stages of L. salmonis readily reattached to Atlantic salmon after hydrogen peroxide treatment. Adult female lice, but not adult males or pre-adults, reattached in significantly lower numbers than untreated controls. Survival of early chalimus stages (I and II) was not affected by hydrogen peroxide treatment, but their subsequent development to chalimus III and IV was delayed compared with untreated controls. Nauplii and copepodid larvae of L. salmonis were almost all dead by 1 hour post-treatment. A very few copepodids survived 24 h post-treatment. Egg strings of L. salmonis at an early stage of development failed to hatch after hydrogen peroxide treatment, but those with pigmented eggs did hatch, although in significantly reduced numbers. Treated egg strings did produce viable copepodids, although in significantly reduced numbers compared with controls.

Transmission experiments to determine the relationship between Sphaerospora sp. from Atlantic salmon, Salmo salar, and Sphaerospora truttae: a revised species description for S. truttae.

Extrasporogonic stages of Sphaerospora sp. from the kidneys of Atlantic salmon (Salmo salar L.) were successfully transmitted via intra-peritoneal injection to naive Atlantic salmon and brown trout (Salmo trutta L.). Rainbow trout (Oncorhynchus mykiss Walbaum) could not be infected in this way. Transmitted extrasporogonic stages continued their development to form sporogonic stages and mature spores in the kidney tubules. Extrasporogonic stages, sporogonic stages and mature spores of the parasite in both experimentally infected hosts were morphologically identical to the equivalent stage in naturally infected Atlantic salmon, although minor differences were seen in spore dimensions. A farm-based exposure experiment confirmed the susceptibility of brown trout to the salmon Sphaerospora. These results are consistent with the view that the salmon Sphaerospora is Sphaerospora truttae Fischer-Scherl, El-Matbouli et Hoffmann, 1986. The parasite is redescribed according to the guidelines of Lom and Arthur (1989) since details of extrasporogonic stages, the ultrastructure of extrasprogonic and sporogonic stage development, and of the parasite's epidemiology are known from Atlantic salmon but not from other reports.

Studies on the parasite fauna of juvenile Atlantic salmon, Salmo salar L., cultured in fresh water in eastern Scotland.

The parasite fauna of juvenile Atlantic salmon, Salmo salar L., at a smolt-rearing unit in eastern Scotland was monitored from two weeks after first-feeding until smolting occurred (May 1973-April 1975). A total of 625 fish was examined. Wild salmon from the river supplying the unit with unfiltered water were also examined. The cultured fish yielded eight protozoan and seven metazoan parasite species, all derived from wild fish. The parasite fauna of cultured fish became established quickly, subsequent changes involving loss of some species and increasing abundance of others. The metazoan parasite fauna was restricted to those species with free-swimming infective stages capable of entering the tanks.