Xenoma Formation Research Articles

More about the biodiversity of parasites in Vietnam: a first report of microsporidia (opistosporidia, microsporidia) in coral fish

This is the first report of microsporidia infection in marine fish from Vietnam. Microsporidia (Opistosporidia, Microsporidia) were identified in 4 species of coral fish Cheilinus chlorourus (Labridae), Abudefduf bengalensis (Pomacentridae), Pomacentrus brachialis (Pomacentridae), Atule mate (Carangidae) from Nha Trang Bay (East Sea), Khanh Hoa Province, Vietnam, in 2018 and 2019. Dry smears of blood, intestine and liver were stained using the Giemsa method. Gut and liver were fixed in Bouin’s fluid, processed for routine paraffin infiltration, and sections were stained with Hematoxylin and Eosin. Strong microsporidia infections were the same for all three species revealed in 1 of every 6 fish samples (prevalence 16.7%) of C. chlorourus, A. bengalensis, and P. brachialis. In C. chlorourus there were meronts, sporogonial plasmodia, and sporoblasts found in all layers of intestine (enterocytes, myocytes, mesotheliocytes), pancreas and liver. In A. bengalensis parasites were found in enterocytes of the small intestine and liver. In P. brachialis meronts, sporogonial plasmodia and sporoblasts were in enterocytes of the small intestine. Intracellular parasites are located in parasitophorous vacuoles covered with additional shell, which appeared to be in the process of forming xenoma. Microsporidia infection was revealed in 1 of 31 fish samples of A. mate (prevalence 3 %). Up to 30 of round and oval white xenomas were detected in the gills of fresh samples by microscopy. We tentatively assign the microsporidia to Glugeidae due to host affiliation (fish), localization (digestive system) and xenoma formation. They differ from Pleistophoridae by localization in non-muscular cells.

Description of hyperparasitism of microsporidia in nematode (Ascaridia galli) parasitic in fowl, Gallus gallus domesticus

Microsporidia are intracellular, obligate parasites. They have wide infection range from diverse phyla of invertebrates to vertebrates and also these eukaryotic pathogens parasitize those hosts which themselves are parasites, thus revealing hyperparasitism. The present study describe intracellular pathogen microsporidia which is isolated from a nematode parasite, Ascaridia sp. which itself is a parasite of fowl, Gallus gallus domesticus. The initial detection by optical microscopy represented numerous refractive oval spores in the homogenate samples of nematodes with mean spore length and width of 4.32±0.32μm×1.57±0.26μm (N=10). Giemsa staining exhibited demarcating stages of spore development. Spores developing in sporophorous vesicle, mature spores of considerable size and shape, dividing sporonts and sporoblasts, were clearly noticeable. However, the phases of proliferation (merogony) were indistinguishable. The ultrastructural study by Scanning electron microscopy illustrated intracellular development of microspor-idian spore inside nematode tissues. Xenoma formation as well as spore adherence to the epithelial wall most likely indicated the degradation and auto-infection by the microsporidian spores in the host tissue. A spore infecting host tissue by ejecting its polar tubule was also captured. Thus, the overall study focuses on developing microsporidia as hyperparasite in nematodes, although further classification for determining the genus has been left to be explored.

Pseudokabatana alburnus n. gen. n. sp., (Microsporidia) from the liver of topmouth culter Culter alburnus (Actinopterygii, Cyprinidae) from China.

We describe the type species of a novel genus of microsporidian parasite, Pseudokabatana alburnus n. gen. n. sp., infecting the liver of topmouth culter, Culter alburnus Basilewsky, 1855, from Lake Poyang off Xingzi county, Jiangxi Province, China. The parasite elicits formation of spherical xenomas of up to 1.2mm in diameter containing all observed life stages from early merogonal plasmodia to mature spores contained within the cytoplasm of host hepatocytes. Merogonal plasmodia existed in direct contact with the host cytoplasm and contained up to 20 visible nuclei. Plasmotomy of the multinucleate plasmodium led to formation of uninucleate cells in which the nucleus underwent further division to form bi-nucleate presporonts, sporonts (defined by cells with a thickened endospore) and eventually sporoblasts (containing pre-cursors of the spore extrusion apparatus). Mature spores were pyriform and monokaryotic, measuring 2.3 ± 0.19μm long and 1.3 ± 0.10μm wide. Spores possessed a bipartite polaroplast and 5-6 coils of a polar filament, in a single rank. The obtained partial SSU rRNA gene sequence, 1383bp in length, did not match any of microsporidia available in GenBank. SSU rDNA-based phylogenetic analysis indicated a new taxon branching with Kabatana rondoni, a parasite infecting the skeletal muscle of Gymnorhamphichthys rondoni from the Amazon River. Due to different host and tissue tropism, the novel taxon did not fit the diagnostic criteria for the genus Kabatana. Further, based on SSU rDNA-inferred phylogenetic analyses, different ultrastructural features of developmental stages, and ecological considerations, a new genus Pseudokabatana and type species Pseudokabatana alburnus n. sp. was erected for the parasite in topmouth culter.

Occurrence of microsporidians Glugea hertwigi and Pleistophora ladogensis, in smelt Osmerus eperlanus from two German rivers, North Sea coast.

Monthly samples of smelt Osmerus eperlanus (Linnaeus, 1758) were collected from July 1985 to May 1986, in the river Elbe (Germany), and examined for infections with microsporidians. Two microsporidians were found: Glugea hertwigi Weissenberg, 1911, infecting the digestive tract and Pleistophora ladogenis Voronin, 1978, infecting the skeletal musculature. G. hertwigi infection led to the formation of xenomas, whereas P. ladogensis was characterized by diffuse infections, with the production of macroscopic visible thread-like or oval-shaped infection foci. Development of G. hertwigi in the host cells showed characteristics typical of the genus Glugea. The ultrastructural development of P. ladogensis showed features typical of the genus Pleistophora, without evidence of the production of 2 types of spores. Host reaction consisted of inflammatory tissue surrounding some of the infection foci as well as phagocytosis of spores. G. hertwigi was only found in juvenile smelt (<10 cm in length), whereas P. ladogensis infected smelts from 6 to 26 cm in length. Prevalence increased with fish length to a maximum value of 9.6%. Seasonal fluctuations in prevalence of infection were also found, with the lowest value in the winter months (2.5% in January 1986) and the highest in summer (11.8% in July 1985). The differences in prevalence of infection with fish length and date of sampling were significant. Additionally, samples of smelt caught in April 1986 from the rivers Eider and Ems revealed infections with P. ladogensis in the first river system only.

Unusually abundant and large ciliate xenomas in oysters, Crassostrea virginica, from Great Bay, New Hampshire, USA

During routine histological examination of oysters (Crassostrea virginica) from Great Bay, New Hampshire, USA, a high prevalence and intensity of ciliate xenomas has been noted since sampling began in 1997. Xenomas are hypertrophic lesions on the gills of bivalve molluscs caused by intracellular ciliates, likely Sphenophrya sp. Although not known to cause mortality in oysters, xenomas have not previously been reported at this high abundance. The objectives of this study were to characterize the xenomas, describe the ciliates, and gather baseline epizootiological data with correlations to environmental and biological parameters.Upon gross examination, xenomas appeared as white nodules, up to 3mm in diameter, located in the gill tissue and occasionally fusing into large masses along the gill filaments. Light microscopy of histological sections revealed xenomas located in the gill water tubes, which they often completely blocked. Higher magnification revealed dual nuclei, eight kineties, and conjugation of the ciliates. Transmission electron microscopy revealed dual nuclei that varied in density, a maximum of twenty cilia in each kinety radiating from the oral apparatus to the posterior, and a 9+2 axoneme structure within the cilia. These traits place the ciliates into the Order Rhynchodida, but insufficient molecular data exist to confirm classification of this ciliate to the Genus Sphenophrya. Since 1997, xenoma prevalence has fluctuated with peaks in 2000, 2004, and 2011. Infected oysters generally contained <30 xenomas, but 2.1% contained >100, sharply contrasting the rare prevalence and low intensity reported elsewhere. Prevalence increased with oyster size, leveling off near 50% in oysters >60mm. Infection intensity peaked in 70–90mm oysters and declined in larger oysters. Individual oyster condition was not associated with xenoma intensity, but sites with oysters in higher condition generally had a greater prevalence and intensity of xenoma infections. Seasonal data indicated an infection cycle increasing from summer to fall, peaking at 55–65% in November and dropping to <10% by spring. The oyster population in Great Bay, NH warrants further examination to understand the mechanisms and conditions controlling xenoma formation.

Distribution, prevalence, and pathology of a microsporidian infecting freshwater sculpins.

Microsporidian infections are common in many fish species, yet detailed studies of these parasites in ecologically important wild populations are rare. Phylogenetic analysis using rDNA sequence data and parasite morphology indicate that mottled sculpin Cottus bairdii and slimy sculpin C. cognatus are hosts for Glugea sp. microsporidia in the northern USA. Glugea sp. is common in the Michigan populations sampled for this study, and prevalence was ≥ 70% in 4 of 6 infected populations (range -4 to 80%). Glugea sp. infection causes the formation of xenomas associated with the body wall, fat body, gonads, and kidneys. Infections range from mild to very heavy, with variable xenoma numbers and sizes. Female sculpin experience heavier infections and more frequent infection of the gonads relative to males. Glugea sp. is transmitted horizontally between hosts through ingestion of spores. Vertical transmission may also be possible, either by spores infecting eggs directly or by spores contaminating the surface of eggs in the ovary or in the nest. The frequency and route of vertical transmission requires further study, but if it occurs, it may partly explain the high prevalence of infection. Our study combined with previous research suggests that additional molecular data and cross-infection experiments should be conducted to clarify species designations in the genus Glugea.

Effects of Loma morhua (Microsporidia) infection on the cardiorespiratory performance of Atlantic cod Gadus morhua (L).

The microsporidian Loma morhua infects Atlantic cod (Gadus morhua) in the wild and in culture and results in the formation of xenomas within the gill filaments, heart and spleen. Given the importance of the two former organs to metabolic capacity and thermal tolerance, the cardiorespiratory performance of cod with a naturally acquired infection of Loma was measured during an acute temperature increase (2 °C h(-1)) from 10 °C to the fish's critical thermal maximum (CT(Max)). In addition, oxygen consumption and swimming performance were measured during two successive critical swimming speed (U(crit)) tests at 10 °C. While Loma infection had a negative impact on cod cardiac function at warm temperatures, and on metabolic capacity in both the CT(Max) and U(crit) tests (i.e. a reduction of 30-40%), it appears that the Atlantic cod can largely compensate for these Loma-induced cardiorespiratory limitations. For example, (i) CT(Max) (21.0 ± 0.3 °C) and U(crit) (~1.75 BL s(-1)) were very comparable to those reported in previous studies using uninfected fish from the same founder population; and (ii) our data suggest that tissue oxygen extraction, and potentially the capacity for anaerobic metabolism, is enhanced in fish infected with this microsporidian.

Immunohistochemical localization of inflammatory cells and cell cycle proteins in the gills of Loma salmonae infected rainbow trout (Oncorhynchus mykiss)

Microsporidial gill diseases particularly those caused by Loma salmonae incur significant economic losses to the salmonid aquaculture industry. The gill responses to infection include the formation of xenomas and the acute hyperplastic inflammatory responses once the xenomas rupture releasing infective spores. The aim of this work was to characterize the inflammatory responses of the gill to both the presence of the xenomas as well as the hyperplasia associated with L. salmonae infection in the rainbow trout gill following an experimental infection using immunohistochemistry. Hyperplastic lesions demonstrated numerous cells expressing PCNA as well as an apparent increased expression of caspase-3 and number of apoptotic cells (TUNEL positive cells). There was an expression of TNFα in individual cells within the gill and increased expression of a myeloid cell line antigen indicating the presence of granulocyte infiltration of both the hyperplastic lesions as well as the xenomas. Similar immune-reactivity was seen in gill EGCs. Hyperplastic gill lesions showed a marked infiltration of CD8+ cells and expression of MHC class I antigens. These findings suggest that L. salmonae xenomas may be subject to infiltration by the host immune cells as well as the mounting or a marked cellular cytotoxic immunoreaction in the resultant hyperplasia following xenoma rupture and spore release.

Efficacy of Intraperitoneally and Orally Administered ProVale, a Yeast β‐(1,3)/(1,6)‐D‐glucan Product, in Inhibiting Xenoma Formation by the Microsporidian Loma salmonae on Rainbow Trout Gills

AbstractThe objectives of the research were to compare the efficacy of ProVale yeast beta‐glucan (β‐glucan) with that of a previously tested research‐grade yeast β‐glucan preparation when administered as an intraperitoneal (IP) injection and to also pilot test the effectiveness of ProVale yeast β‐glucan as a feed additive for reducing Loma salmonae xenoma formation on the gills of rainbow trout Oncorhynchus mykiss. Rainbow trout received IP injections of ProVale (4, 10, or 20 mg of ProVale/kg of fish) or Sigma β‐glucan (4 mg/kg). Oral challenge with L. salmonae occurred 1 week after IP injection with the β‐glucan products. For the second objective, 400 rainbow trout were separated into tanks and duplicate groups were treated with 0, 50, 100, and 200 g of ProVale/1,000 kg of feed. Starting at 3 weeks prior to challenge and continuing 2 weeks after challenge, rainbow trout were fed the various ProVale doses daily at a feeding rate of 1% of the fish biomass. Commencing at 4 weeks postchallenge, the fish in each trial were evaluated for the presence of xenomas on the first left gill arch. The most protective IP dose of commercial ProVale was 10 mg/kg when compared with the laboratory‐grade IP dose of Sigma β‐glucan (4 mg/kg). Both of these intraperitoneally administered β‐glucan products were effective in reducing the mean xenoma count. ProVale used as a feed coating (200 g/1,000 kg) was able to reduce the mean xenoma count by 50%.

First record of a Kabatana sp. microsporidium infecting fish in the Atlantic Ocean

Two-spotted goby Gobiusculus flavescens from the Swedish Gullmarsfjord regularly present subcutaneous creamy-white patches in the body musculature, associated with Kabatana sp. infection. Analysis of the 16S rRNA gene of the microsporidium showed 98.54% homology with Kabatana newberryi infecting a marine goby from California, indicating that the Swedish microsporidium is either a different strain of K. newberryi or a closely related species. This represents the first record of a Kabatana species in the Atlantic Ocean. The genetic similarity of the 2 microsporidia was paralleled by close infection phenotypes. Infected muscle fibres were swollen compared to adjacent non-infected fibres, and mature spore masses were found throughout the skeletal musculature. No xenoma formation was detected. Since G. flavescens is an established model species in behavioural ecology, the host-parasite system is ideally suited for testing how microsporidian infections affect host behaviour and fitness.

First report of microsporidiosis in fairy shrimp Branchinella thailandensis (Sanoamuang, Saengphan and Murugan, 2002)

The diagnosis of microsporidia infestation in the muscle of the fairy shrimp ( Branchinella thailandensis [Sanoamuang, L., Saengphan, N., Murugan, G., 2002. First record of the family Thamnocephalidae, (Crustacea, Anostraca) from Southeast Asia and description of a new species of Branchinella. Hydrobiologia 486, 63–69.]) culture was examined by optical microscopy and transmission electron microscopy. The clinical sign of microsporidiosis in fairy shrimp was white tubular masses in the muscle tissue. There was no apparent host encapsulation or xenoma formation within the muscle fibers. Spores of this parasite were uninucleate. The length of the spores was about 2.829–3.269 (3.086) µm and the width was about 1.686–2.857 (1.850) µm. The polar filament had 12–13 coils, arranged in a single row at the posterior end of the spore. Based on the gross pathology, morphology, histopathology and ultrastructure of the spore, we suggest that this parasite is a species of Pleistophora sp. Although this disease was not acute, the fairy shrimp showed continuous mortality. Secondary bacterial infections occurred frequently in infected fairy shrimp. Fairy shrimp died within one to five days of contracting a bacterial infection. Finally, microsporidiosis affected the productivity of the fairy shrimp culture. Prophylaxis was performed by treating underground water that might be contaminated with this microspore before use in culture.

Whole body net ion fluxes, plasma electrolyte concentrations and haematology during a Loma salmonae infection in juvenile rainbow trout, Oncorhynchus mykiss (Walbaum)

Loma salmonae infections of salmonids culminate in the development of branchial xenomas and subsequent focal hyperplasia of the lamellar or filament epithelium following xenoma rupture and spore release. The effects of this acute branchial disruption upon net ionic flux rates and plasma electrolyte concentrations were determined in juvenile rainbow trout given an experimental oral exposure to L. salmonae. Mean numbers of branchial xenomas peaked at week 5 post-exposure (PE), which coincided with a reduction in the specific growth rate, although there were no significant differences in mass, length or condition of Loma-exposed fish compared with unexposed controls. Following exposure, negative net whole body Na(+) and K(+) fluxes decreased, whereas net Cl(-) fluxes remained unchanged compared with non-exposed control fish. At week 3 PE during the initial branchial xenoma formation stage, there was a significant negative whole body net K(+) flux in Loma-exposed trout compared with other points during the exposure and subsequent infection. Additionally, Loma-exposed fish had marginally elevated plasma Na(+) and Cl(-) concentrations, whilst K(+) levels remained unchanged, compared with control fish. Although there was a progressive decrease in leucocrit, haematocrit remained unchanged over the course of the Loma exposure and subsequent infection. These results suggest that ionic compensation can occur at the gills during the development of xenomas during exposure to L. salmonae and the resultant infection, therefore allowing defence of plasma electrolyte concentrations, unlike the acute ionic disturbances seen with some other parasitic diseases.

Microsporidian xenomas in fish seen in wider perspective

The history of understanding xenoparasitic complexes or xenomas provoked in the host cell by various protists and especially by microsporidia is outlined. Microsporidia have been known to produce xenomas in oligochaetes (e.g., genera Bacillidium, Burkea, Hrabyeia, Jirovecia, species of the collective group Microsporidium), crustaceans (e.g., Abelspora, Mrazekia), insects (e.g., Polydispyrenia, Thelohania) and poikilothermic vertebrates, mostly fish (Alloglugea, Amazonspora, Glugea, Ichthyosporidium, Loma, Microfilum, Microgemma, Neonosemoides, Pseudoloma, Spraguea, Tetramicra). An overview of characters of xenomas caused by species of these genera is presented. The study of microsporidia causing xenomas in fish offers an insight into cell pathology and is of interest since many of these species are important agents of diseases in commercial fish. Xenomas produced from a few types of target cell display a complete change of organisation of the host cell and differ, according to the agent, in their structure. Recent data show that proliferation of the parasite may have already started in the cells transporting the parasites to the final site of xenoma formation. However, these are preliminary revelations and most of the facets of the life cycle are still to be clarified. Curiously, xenoma-forming microsporidia do not seem to be strictly host specific. The salient features of fish microsporidian xenomas are discussed, such as role of the xenoma, whether its features are host- or microsporidium-dependent, development and demise of the xenoma in the course of time, and host reaction phenomena. The need of further research is emphasised.

Review of the sequential development of Loma salmonae (Microsporidia) based on experimental infections of rainbow trout (Oncorhynchus mykiss) and Chinook salmon (O. tshawytscha)

Loma salmonae is a common gill parasite of salmonids, and essentially all species in the genus Oncorhynchus are susceptible. Infections occur in both fresh and salt water. Loma salmonae is directly transmissible by ingestion of spores or infected tissue. The parasite infects the wall of blood vessels of various organs, but the gill is the primary site of infection. Initial infection occurs in the intestine, and xenomas are easily detected in the gills by standard histology at 4-6 wk post-exposure. A few presporogonic stages of the parasite are found in the heart endothelium prior to xenoma formation in the gills. Ultrastructure studies of early infections demonstrated that wandering blood cells transport the meronts to the gills, and that merogony occurs in pillar cells and other cells underlying the gill endothelium. Xenomas develop in these cells, resulting in hypertrophied host cells filled with spores. Xenomas ultimately rupture, and are associated with severe inflammation in which free spores are found in macrophages. The parasites are most pathogenic during this phase of the infection, resulting in severe vasculitis and clinical disease. Both rainbow trout (Oncorhynchus mykiss) and Chinook salmon (Oncorhynchus ishawytscha) recover from infections, but free spores persist in kidney and spleen phagocytes for many months after xenomas are absent in Chinook salmon. Fish that have recovered from the infection show strong immunity against the parasite, lasting up to 1 year. Fish are susceptible to infection by other routes of exposure by spores; co-habitation, anal gavage, and intramuscular, intraperitoneal and intravascular injection. Autoinfection probably occurs following release of spores in blood vessels after xenomas rupture. The optimal temperature for L. salmonae infections is 15-17 degrees C, with a permissive range of 11-20 degrees C.

Microgemma vivaresi n. sp. (Microsporidia, Tetramicridae), Infecting Liver and Skeletal Muscle of Sea Scorpions, Taurulus bubalis (Euphrasen 1786) (Osteichthyes, Cottidae), an Inshore, Littoral Fish

The ultrastructure of a new microsporidian species Microgemma vivaresi n. sp. causing liver cell xenoma formation in sea scorpions, Taurulus bubalis, is described. Stages of merogony, sporogony, and sporogenesis are mixed in the central cytoplasm of developing xenomas. All stages have unpaired nuclei. Uninucleate and multinucleate meronts lie within vacuoles formed from host endoplasmic reticulum and divide by binary or multiple fission. Sporonts, no longer in vacuoles, deposit plaques of surface coat on the plasma membrane that cause the surface to pucker. Division occurs at the puckered stage into sporoblast mother cells, on which plaques join up to complete the surface coat. A final binary fission gives rise to sporoblasts. A dense globule, thought to be involved in polar tube synthesis, is gradually dispersed during spore maturation. Spores are broadly ovoid, have a large posterior vacuole, and measure 3.6 microm x 2.1 microm (fresh). The polar tube has a short wide anterior section that constricts abruptly, then runs posteriad to coil about eight times around the posterior vacuole with granular contents. The polaroplast has up to 40 membranes arranged in pairs mostly attached to the wide region of the polar tube and directed posteriorly around a cytoplasm of a coarsely granular appearance. The species is placed alongside the type species Microgemma hepaticusRalphs and Matthews 1986 within the family Tetramicridae, which is transferred from the class Dihaplophasea to the class Haplophasea, as there is no evidence for the occurrence of a diplokaryotic phase.

Resolution of a taxonomic conundrum: an ultrastructural and molecular description of the life cycle of Pleistophora mulleri (Pfeiffer 1895; Georgevitch 1929).

The classification of a microsporidian parasite observed in the abdominal muscles of amphipod hosts has been repeatedly revised but still remains inconclusive. This parasite has variable spore numbers within a sporophorous vesicle and has been assigned to the genera Glugea, Pleistophora, Stempellia, and Thelohania. We used electron microscopy and molecular evidence to resolve the previous taxonomic confusion and confirm its identification as Pleistophora mulleri. The life cycle of P. mulleri is described from the freshwater amphipod host Gammarus duebeni celticus. Infection appeared as white tubular masses within the abdominal muscle of the host. Light and transmission electron microscope examination revealed the presence of an active microsporidian infection that was diffuse within the muscle block with no evidence of xenoma formation. Paucinucleate merogonial plasmodia were surrounded by an amorphous coat immediately external to the plasmalemma. The amorphous coat developed into a merontogenetic sporophorous vesicle that was present throughout sporulation. Sporogony was polysporous resulting in uninucleate spores, with a bipartite polaroplast, an anisofilar polar filament and a large posterior vacuole. SSU rDNA analysis supported the ultrastructural evidence clearly placing this parasite within the genus Pleistophora. This paper indicates that Pleistophora species are not restricted to vertebrate hosts.

Effects of monensin dose and treatment time on xenoma reduction in microsporidial gill disease in rainbow trout, Oncorhynchus mykiss (Walbaum)

AbstractThe objectives of the study were (1) to determine an acceptable dose of dietary monensin against microsporidial gill disease (MGD) of salmon caused by Loma salmonae and (2) to determine the life stage of L. salmonae at which the action of monensin was most effective at reducing clinical disease. Rainbow trout (RBT) were fed monensin diets at concentrations of 0, 10, 100, 1000 and 5000 ppm starting 1 week before per os exposure to L. salmonae infected gill material. Lethal sampling occurred at weeks 6, 7, 8 and 9 post‐exposure (PE). The acceptable dose of dietary monensin was determined to be 1000 ppm because it was the lowest concentration that produced a significant and constant reduction in xenoma formation. RBT were fed monensin at 1000 ppm beginning at 0, 1, 2 and 3 weeks before exposure and at 1, 2 and 3 weeks after exposure and continuing until week 9 PE. Fish in which treatment was initiated at the time of exposure or 1 week before, showed the greatest reduction in xenoma production when compared with the untreated RBT.

Xenoma formation during microsporidial gill disease of salmonids caused by Loma salmonae is affected by host species (Oncorhynchus tshawytscha, O. kisutch, O. mykiss) but not by salinity.

Host species and salinity often affect the development of disease in aquatic species. Eighty chinook salmon Oncorhynchus tshawytscha, 80 coho salmon O. kisutch and 80 rainbow trout O. mykiss were infected with Loma salmonae. Forty of each species were reared in seawater and 40 in freshwater. The mean number of xenomas per gill filament was 8 to 33 times greater in chinook salmon than in rainbow trout (RBT). Coho salmon had a mean xenoma intensity intermediate to that of chinook salmon and RBT. In contrast to the differences between species, salinity had no significant effect on xenoma intensity in any of these host species. The onset of xenoma formation occurred at Week 5 postexposure (PE) for chinook salmon and RBT, and at Week 6 PE for coho salmon. RBT had cleared all visible branchial xenomas by Week 9 PE, whereas xenomas persisted in coho and chinook salmon at Week 9 PE. Histologically, xenomas were visible in the filament arteries of the branchial arch in chinook and coho salmon gills but were absent from RBT gills. Fewer xenomas were seen in the central venous sinusoids of RBT than in chinook and coho salmon. The lower xenoma intensity, shorter duration of infection and pathological characteristics, common to microsporidial gill disease in RBT, suggest a degree of resistance to clinical disease that is not seen in coho and chinook salmon.

Evaluating protection against Loma salmonae generated from primary exposure of rainbow trout, Oncorhynchus mykiss (Walbaum), outside of the xenoma‐expression temperature boundaries

A series of challenge and re‐challenge studies was conducted in which juvenile rainbow trout were exposed to the pathogen Loma salmonae, a microsporidian which typically causes xenoma formation during sporogony and inflammation in the gills as the xenomas undergo dissolution. The specific goal was to determine if a primary exposure, conducted at a water temperature outside of the range which permits the parasite to undergo sporogony and form branchial xenomas, would stimulate a protective response in the fish to a later challenge conducted under temperature conditions optimal for the parasite. Primary challenge of fish to L. salmonae at 7 °C or 21 °C blocked or limited xenoma formation, as discussed in a previous study. However, these fish had a relative percentage protection (RPP) against a second optimized exposure which matched, or was not significantly less than, the degree of protection (100%) that developed in other groups of fish that received a primary exposure throughout the range of water temperatures which permits xenoma formation. When the primary exposure was conducted at 5 °C, the RPP against the second exposure was adversely affected and declined to 61%. These findings have application to the control of L. salmonae within aquaculture, in that it may be possible to expose hatchery stocks of susceptible salmonid species to spores of L. salmonae when hatchery water temperature is at 7 °C. At this temperature, the risks of disease stemming from this primary exposure appear minimal, since xenomas fail to form. However, the degree of protection appears promising, and may be sufficient to protect fish from spore exposure occurring at netpen marine sites where the parasite may be endemic.

A Preliminary Investigation of Alternatives to Fumagillin for the Treatment of Loma salmonae Infection in Rainbow Trout

The effects of the following six treatments against Loma salmonae, a microÍsporidian gill pathogen, were examined in rainbow trout: fumagillin (high dose), pyrimethamine + sulphaquinoxaline, albendazole, amprolium, fumagillin (low dose), and metronidazole. The fish were infected by mouth and the treatments were administered at intervals for a period of several weeks. The results were assessed on the basis of (1) delay in the formation of xenomas, and (2) the number of xenomas per gill arch. The first five treatments, in descending order of efficacy, delayed the formation of xenomas (P<0·01), but metronidazole had no such effect. Fumagillin (high or low dose) and albendazole both reduced the number of xenomas present 10 weeks after infection (P<0·01), but the other three treatments did not do so. From these results, both fumagillin and albendazole appeared to be of potential value in controlling L. salmonae infection in trout.