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.     

Effects of Pseudoloma neurophilia infection on the brain transcriptome in zebrafish (Danio rerio)

Laboratory zebrafish are commonly infected with the intracellular, brain-infecting microsporidian parasite Pseudoloma neurophilia. Chronic P. neurophilia infections induce inflammation in meninges, brain and spinal cord, and have been suggested to affect neural functions since parasite clusters reside inside neurons. However, underlying neural and immunological mechanisms associated with infection have not been explored. Utilizing RNA-sequencing analysis, we found that P. neurophilia infection upregulated 175 and downregulated 45 genes in the zebrafish brain, compared to uninfected controls. Four biological pathways were enriched by the parasite, all of which were associated with immune function. In addition, 14 gene ontology (GO) terms were enriched, eight of which were associated with immune responses and five with circadian rhythm. Surprisingly, no differentially expressed genes or enriched pathways were specific for nervous system function. Upregulated immune-related genes indicate that the host generally show a pro-inflammatory immune response to infection. On the other hand, we found a general downregulation of immune response genes associated with anti-pathogen functions, suggesting an immune evasion strategy by the parasite. The results reported here provide important information on host–parasite interaction and highlight possible pathways for complex effects of parasite infections on zebrafish phenotypes.     

Epizootiology of Parvicapsulaminibicornis in Fraser River sockeye salmon, Oncorhynchusnerka (Walbaum)

Late-spawning Fraser River sockeye salmon, Oncorhynchus nerka , stocks have suffered significant prespawn mortality associated with an unusually early freshwater migration pattern and the myxosporean parasite Parvicapsula minibicornis . Surveys of migrating adult salmon from several spawning populations were conducted in 1999 and 2000 to determine the extent of infection with P. minibicornis , when and where the parasite first becomes detectable during migration, and whether early migrating stocks might be used as sentinels to assess risk of infection in late-spawning stocks. Posterior kidney, preserved in 95% ethanol, was examined for P. minibicornis in stained histological sections and using a polymerase chain reaction (PCR) test. The prevalence of this parasite in all Fraser River sockeye salmon stocks examined was high (range 47–100% infected). In contrast, P. minibicornis was not detected in the fish tested from the two sockeye salmon stocks outside the Fraser River drainage in either 1999 or 2000. The parasite was also not detected histologically or by PCR in the kidney tissue of the fish from the Fraser River that were sampled in salt water or early during their freshwater migration up the river. These findings and the progression in the prevalence and intensity of infection as the fish from three stocks (early Stuart, Weaver Creek and Cultus Lake) were monitored over time, suggest salmon acquired the parasite either in the lower Strait of Georgia or in the lower Fraser River before the confluence of the Harrison River. In both 1999 and 2000 the parasite was present in all Fraser River sockeye salmon stocks sampled, which suggests that early Stuart salmon may be valuable as a sentinel stock for the presence of the parasite in later-spawning stocks.     

The effect of foraging and ontogeny on the prevalence and intensity of the invasive parasite Anguillicola crassus in the European eel Anguilla anguilla

Infection patterns of the invasive Anguillicola crassus nematode were investigated in a population of the European eel Anguilla anguilla where parasite invasion is very recent, Loch Lomond, Scotland. Intensity levels of the parasite were associated with differences in fish ontogeny and trophic ecology. Although eels foraged on both fish and invertebrates, individuals which were smaller and fed on invertebrates (>70% contribution to diet) were found to contain a greater number of swim bladder parasites compared to larger eel with a predominance of fish (>60% contribution) in their diet. Within affected fish, a significant negative relationship was found between fish length and parasite intensity, with smaller individuals having higher parasite intensity than larger individuals. This study indicates that food intake and infection risk are linked in this recently infected host–parasite system. From a management perspective increasing our understanding of how infection intensity and repeated exposure is linked to resource use in an ecosystem is important for the future management of this endangered species in Europe.     

Infection of Cherax quadricarinatus (Decapoda: Parastacidae) by the microsporidium Thelohania sp. (Microsporida: Nosematidae)

Abstract The microsporidian parasite Thelohania sp. was present in a population of Cherax quadriearinatus in the Mitchell River, north Queensland. An infection rate of 7--8% was recorded for the population sampled. Cohabitation, passage of spores through fish, ingestion of spores and injection of blood from an infected individual failed to elicit infection in laboratory held animals. Injection of spores elicited a host responseconsisting of encapsulation and melanization of spores. Further studies are necessary to clarify the taxonomic position and mode of transmission of this parasite.     

Dynamics of challenge infections of Gyrodactylus bullatarudis Turnbull (Monogenea) on guppies, Poecilia reticulata (Peters)

Abstract. Comparison of the population dynamics of Gyrodactylus bullatarudis o fish that were reinfected at 0, 1, 2, 4, or 6 weeks post-recovery from an initial, infection were based on the following parameters: percentage establishment, recovery and death, mean peak parasite burden, mean time to peak burden, mea duration of infection, mean time to death. Of fish that recover from an initial, infection, some are immediately susceptible to a challenge infection. Other fish are able to harbour a challenge infection hut parasite numbers increase less and the infection is of shorter duration than the initial. These fish regain their fu susceptibility within 4-6 weeks post-recovery. A third group of fish become totally refractory to reinfection and only slowly regain their ability to support an infection Thus even at 6 weeks post-recovery they are only partially susceptible t reinfection. It is probable that the three groups in fact represent a continuum increasing susceptibility to challenge infection.     

Non‐invasive fast real‐time PCR assay for detection of the enteric parasite Enteromyxum scophthalmi in cultured turbot (Scophthalmus maximus L.)

Enteromyxum scophthalmi is a myxozoan parasite that causes severe parasitic diseases in cultured turbot affecting mainly the intestine of the host. It is characterized by producing acute enteritis, starvation and eventually death. Current diagnosis of E. scopthalmi use traditional techniques, based on the identification of the morphology of the parasite. These techniques take extended time to be carried out and do not favour the adoption of control measure at turbot farms and require the sacrifice of fish. This study develops a fast real‐time PCR molecular tool for the detection of E. scophthalmi in infected farmed turbot. This methodology is applicable for routine controls on the farm at every stage of the parasite infection. Results of the study demonstrate the robustness, specificity, efficiency and reliability of the technique. In addition, this study also provides a non‐invasive procedure of sampling through swaps. This allows control, prevention and diagnosis of the parasite infection at turbot farms while maintaining the welfare of the cultivated fish and avoiding sacrifice of the fish sampled.     

Immunization of channel catfish, Ictalurus punctatus Rafinesque, against Ichthyophthirius multifiliis (Fouquet): killed versus live vaccines

Abstract. Fish surviving infection with the pathogenic ciliated protozoan, Ichthyophthirius multifiliis (Fouquet, 1876), become resistant to subsequent infection by the parasite. The acquired immunity suggests that development of a vaccine against the parasite may be possible. Because of the advantages of immunoprophylaxis for treatment of the disease, an effort has been made to determine whether fish exposed to killed parasite preparations can resist subsequent lethal challenge. Both the route of administration and the effects of stage specific antigens have been examined. Channel catfish vaccinated by intraperitoneal (i.p.) injection or bath immersion with killed I multifiliis tomites show 100% mortality following a standard challenge protocol. Similarly, 100% mortality was observed in test groups injected with tomite cilia. In both cases, a consistent difference in days to death between control and test group animals was observed. Although complete mortality was seen with fish injected with tomite preparations, fish vaccinated with killed trophonts (the feeding stage of the parasite) had a much greater degree of protection with approximately 50% of fish surviving an otherwise lethal challenge. Finally, animals injected intraperitoneally with live tomites showed nearly complete immunity and were identical in their response to fish which survive natural infection. The response of fish vaccinated with live parasites indicates that animals injected intraperitoneally can develop surface immunity and that i.p. injection is a suitable route of administration for potential I. multifiliis vaccines.     

Experimental transmission of Enteromyxum leei Diamant, Lom and Dykova, 1994 in sharpsnout sea bream, Diplodus puntazzo C. and the effect on some innate immune parameters

Enteromyxum leei has been implicated in outbreaks in sharpsnout sea bream, causing a serious economic impact on Mediterranean fish farming. Information about transmission and life cycles of marine myxosporea is limited to only a few fish species. In the present study, direct transmission of the infection was achieved by cohabitation of infected and healthy Diplodus puntazzo C. for the first time. The progress of the infection was monitored by the detection of different developmental stages of the parasite in the posterior intestine. Infected fish with E. leei were firstly observed in day 5 (prevalence of 20%). The total prevalence reached 100% in day 19 and was maintained in this high level until the end of the experimental period (day 55). In addition, the initial intensity of the different parasite stages was low, but on day 26 the intensity of trophozoites was suddenly increased, followed by the increased intensity of sporoblasts and mature spores. Also, the experimental transmission of E. leei in recovered D. puntazzo previously treated for parasite elimination proved to be unsuccessful, suggesting of a possible development of protective immunity against the parasite.     

Myxobolus cerebralis (Hofer) infection risk in native cutthroat trout Oncorhynchus clarkii (Richardson) and its relationships to tributary environments in the Yellowstone Lake Basin

Conservation of native species is challenged by the introduction of non‐native pathogens and diseases into aquatic and terrestrial environments worldwide. In the Yellowstone Lake basin, Yellowstone National Park, the invasive parasite causing salmonid whirling disease Myxobolus cerebralis (Hofer) has been identified as one factor contributing to population declines of native Yellowstone cutthroat trout Oncorhynchus clarkii bouvieri (Jordan & Gilbert). In 2002 and 2003, we examined relationships between the stream environment and severity of M. cerebralis infection in native trout. Coefficients of variation of environmental features were calculated to examine variability. Ten years later, we reassessed infection levels at 22 tributaries broadly across the system. Results of principal component analysis (PCA) of physical features (2003) were negatively correlated with infection severity, mostly in lower jaw cartilage of cutthroat trout, and PCA of chemical features (and temperature) correlated with infection severity in cranial cartilage. Pelican Creek, where M. cerebralis prevalence and severity was high 2002–2003, remained high in 2012. We did not find evidence that the parasite had dispersed further within the system. Variable environmental features (physiological stress) across short spatiotemporal scales within a stream or season may possibly predispose salmonids to infection in the wild and facilitate parasite establishment.     

The effect of temperature on ability of Lepeophtheirus salmonis to infect and persist on Atlantic salmon

The salmon louse (Lepeophtheirus salmonis) is an ecologically and economically important parasite of salmonid fish. Temperature is a strong influencer of biological processes in salmon lice, with development rate increased at higher temperatures. The successful attachment of lice onto a host is also predicted to be influenced by temperature; however, the correlation of temperature with parasite survival is unknown. This study describes the effects of temperature on infection success, and survival on the host during development to the adult stage. To accurately describe infection dynamics with varying temperatures, infection success was recorded on Atlantic salmon (Salmo salar) between 2 and 10°C. Infection success ranged from 20% to 50% and was strongly correlated with temperature, with the highest success at 10°C. Parasite loss was monitored during development at eight temperatures with high loss of lice at 3 and 24°C, whilst no loss was recorded in the temperature range from 6 to 21°C. Sea temperatures thus have large effects on the outcome of salmon louse infections and should be taken into account in the management and risk assessment of this parasite. Improving understanding of the infection dynamics of salmon lice will facilitate epidemiological modelling efforts and efficiency of pest management strategies.     

The pathology caused by Pomphorhynchus laevis Müller in the alimentary tract of the stone loach, Noemacheilus barbatulus (L.)

Abstract. Changes induced in intestinal and rectal structure of the stone loach, Noemacheilus barbatulus (L.) by Pomphorhynchus laevis Müller infection are described. Within the gut, the main effect of the infection is the abrasion and flattening of the mucosa by the parasite. Where the praesoma penetrates the gut wall, there is a marked inflammatory response. This inflammatory response tissue is well vascularized and displays esterase activity. Granulocytes and phagocytic cells, which are found both within the inflammation tissue and within capillaries, are probably host leucocytes. The integument of P. laevis appears undamaged by granulocytes in its proximity and it is unlikely that the host's inflammatory response is capable of rejecting the parasite.     

The histopathology of the eye of big head carp, Aristichthys noblis (Richardson), infested with Lernaea piscinae Harding, 1950

Abstract. The histopathological changes in the eye due to the penetration and growth of the parasite, Lernaea piscinae , in the host tissue of big head carp is described. Distinct pathological changes were hyperplasia of the corneal epithelium, severe inflammatory response mainly around the anchor of the parasite, in the anterior chamber, the corneal stroma and the vitreous body and detachment of the vitreous and retinal layers. Rupture of the lens capsule and fibrous tissue response around the process of the anchor structure of the parasite were also observed. The development of an immune response to the parasite within the body of the host is thought to have led to the infection of the cornea, an avascular site.     

Bromodeoxyuridine DNA labelling reveals host and parasite proliferation in a fish‐myxozoan model

Enteromyxum leei is a myxozoan parasite responsible for enteritis in gilthead sea bream (Sparus aurata). The parasite proliferates in the paracellular space of the intestinal epithelium and induces an inflammatory reaction. To assess intestinal cell turnover and parasite proliferation, fish were infected with the parasite by anal intubation; after 17 and 64 days, the cell proliferative marker bromodeoxyuridine (BrdU) was administered; and after 24 hr, tissue samples were taken for immunohistochemical detection. Parasite exposure induced increased epithelial and immune cell proliferation in all intestinal segments at all time points, even before parasite establishment. This increased turnover was triggered early after intubation and mainly at a local level, as shown by an increased proliferating cell nuclear antigen (pcna) gene expression only at the posterior intestine after 17 days (not found in lymphohaematopoietic organs). Incorporation of BrdU in parasite secondary and tertiary daughter cells indicated that parasite endogeny is not by schizogonial division, which uses de novo synthesis pathway of pyrimidines. Altogether, BrdU immunolabelling and pcna gene expression showed the rapid proliferative response of the fish intestines upon a myxozoan infection and how this response is effectively triggered even before the parasite reaches or establishes in the site.     

Tissue reactions to microsporidian infections in fish

Abstract. Histopathological studies on natural and experimental infections of nineteen microsporidian species from fishes distinguished two types of tissue reactions.
The first type is characteristic of infections with xenoma-inducing microsporidian species and comprises three successive stages: a weakly reactive stage, a productive stage with the formation of granulomas and a stage of granuloma involution. Following the first stage, tissue reactions are directed towards the isolation of the parasite and result in its complete elimination and host tissue repair. The extent of pathological changes probably depends on the number of parasite cells which initiated the infection.
The second type is represented by Pleistophora species infecting muscles or oocytes. Host tissue reaction is surprisingly slight during the schizogony and sporogony and does not tend to isolate the invaded muscle fibres. A slight lymphocytic infiltration of myosepta indicates the first stage of tissue reaction. The tissue reaction only reaches the productive stage when mature spores completely fill the contents of the infected muscle fibre. A thick wall of fibroblasts may be formed to demarcate the parasite mass as soon as it undergoes necrotic changes. The extent of pathological changes probably depends on the ability of early developmental stages of the parasite to spread the infection within the host.
In both types of tissue reaction, the spores are destroyed by complete digestion within host phagocytic cells.     

Recruitment and effects of Discocotyle sagittata (Monogenea) infection on farmed trout

Farmed rainbow trout Oncorhynchus mykiss and brown trout Salmo trutta were monitored over 3 years for infection with the blood-feeding gill fluke Discocotyle sagittata. Parasite transmission is seasonal: new infections take place during summer/autumn, and transmission is generally negligible during winter/spring. There are 2 sources of infection for naïve fish-of-the-year: limited invasion when fish are in the raceways by riverborne larvae originating external to the farm; and internally, within the farm, when 0+ fish are transferred to ponds previously occupied by older cohorts of infected fish. Thereafter, infection levels continue to increase in rainbow trout primarily through transmission within the farm. Prevalence rose to 100% in 1+ fish by the end of their second summer. In O. mykiss, mean abundance reached 194 worms/host for 1+ fish (up to 489 worms/host) and 160 worms/host for 2+ fish. By contrast, in S. trutta, parasite prevalence never exceeded 85% and, after the first year's invasions, infection levels decreased over time: in 1+ and 2+ brown trout, parasite mean abundance was < 4 (maximum 15) worms/host. We present evidence of the detrimental effects of D. sagittata on the host: high burdens are associated with pale gills, decreased body condition and host mortality. Parasite burdens become overdispersed during the warmer part of the year, as prevalence and mean abundance increase. However, the degree of parasite overdispersion decreases over winter; we cannot distinguish whether decreased aggregation is due to parasite losses from infected fish (including immune-mediated parasite mortality) or parasite-induced host mortality.     

Tetrahymena sp. infection in guppies, Poecilia reticulata Peters: parasite characterization and pathology of infected fish

Tetrahymena sp. infection was diagnosed in guppies imported from Singapore. The parasite was isolated (Tet-NI) and optimally cultured in vitro in RM-9 medium. Cytological analyses [silver-staining and scanning electron microscopy (SEM)] revealed a pyriform-shaped, 64 × 41-μm holotrich ciliate without caudal cilium, containing a macro-nucleus (18.25 × 16.83 μm) and micro-nucleus (5.73 × 5.40 μm). Wet-mount examination and histological analyses of fish exposed to the parasite by co-habitation, immersion and infection by i.p. (intra-peritoneal) and i.m. (intra-muscular) injection revealed numerous ciliates on the skin, and in the gill and caudal fin blood vessels. Ciliates surrounded internal organs, the peri-orbital region of the eye, and were observed inside developing guppy embryos. Some muscle necrosis was associated with infection, but little or no inflammatory response. Immersion, co-habitation and i.m. injection caused relatively high infection rates and levels in the skin and tail, and lower infection in the gill blood vessels and internal organs; i.p. injection caused higher infection in the gill blood vessels and internal organs. Co-habited fish had relatively high infection levels in the hind-gut sub-mucosa. This is the first report of controlled systemic infection by Tetrahymena sp.     

Resistance of Ostrea edulis to Bonamia ostreae infection

Preliminary studies showed that oysters from a population of Ostrea edulis infected with Bonamia ostreae since at least 1963 have a substantial degree of resistance to the infection in comparison with oysters from a population with no prior exposure history to the disease. Over a 46-week period and under identical environmental conditions, 99% of non-resistant oysters exposed to the disease died while only 26% of a population of resistant oysters died. The surviving resistant oysters had lesions which indicated a chronic infection and possible degradation of the parasite within the tissues. The resistant oysters may offer a management option for the reestablishment of Ostrea edulis culture in disease enzootic areas where the parasite has virtually eliminated non-resistant populations.