Rickettsia-like organism associated with tremor disease and mortality of the Chinese mitten crab Eriocheir sinensis

We report the discovery of a rickettsia-like organism (RLO) in cultured freshwater Chinese mitten crab Eriocheir sinensis. The RLO caused tremor disease and was apparently responsible for a mass mortality (30 to 90%) in 2 provinces in southeast China. Moribund crabs were investigated from different districts during outbreaks in 1999 and 2000. With electron microscopy, 3 different pathogens were detected in moribund crabs: a rickettsia-like organism (RLO), virus-like particles (VLP) and a microsporidian-like protozoan (MLP). Based on the high prevalence, infection intensity and cytopathological signs, the RLO was considered to be the probable cause of the high mortality. Both VLP and MLP occurred at low prevalences and were considered secondary infections. The RLO was 0.22 to 0.35 microm in diameter, granular or clavate, bounded by a cell wall and membrane, and possessed no nucleus but a nucleoid was present. When dividing, RLOs occurred in irregular shapes, such as dumbbells, awls, and crescents. The RLOs exhibited a predilection for muscle and connective tissues and were probably transported to various tissues and organs by hemocytes.     

Histology, ultrastructure, and morphogenesis of a rickettsia-like organism causing disease in the oyster, Crassostrea ariakensis Gould

Moribund specimens of the oyster, Crassostrea ariakensis Gould, aged 2-3 years were collected from Hailing Bay in Yangxi County of Guangdong Province from February to May and November to December in the years 2001, 2002, and 2003. A massive infection by an obligate intracellular prokaryote, specifically a rickettsia-like organism (RLO), was found. Here we report investigations of this RLO in the tissues of the oyster C. ariakensis Gould and describe the histology, ultrastructure, and morphogenesis of this pathogen in C. ariakensis Gould. Light microscopic observations of stained tissues revealed cytoplasmic inclusion bodies typical of prokaryote infection in about 87% (26/30) of the oysters. Most inclusions were observed in epithelial cells and connective tissues of the gill, mantle, and digestive gland of most of the infected oysters. The shape, size, and color of inclusions from different tissues were polymorphic. Electron microscopic examination of digestive gland, gill, and mantle tissues showed that the RLOs were intracytoplasmic. RLOs were often round, dumb-bell-shaped (undergoing binary fission), or occasionally rod-shaped and ranged from approximately 0.58 to 1.20microm in size. The organisms exhibited an ultrastructure characteristic of prokaryotic bacteria-like cells, including a trilaminar cell wall, electron-dense periplasmic ribosome zone, and a DNA nucleoid. Reproductive stages, including transverse binary fission, were observed by TEM. These stages were frequently observed within membrane-bound cytoplasmic vacuoles. Hexagonal phage-like particles in the cytoplasm of RLOs were also observed.     

类立克次体侵染中华绒螯蟹神经组织的光镜和电镜观察

对患颤抖病的中华绒螯解（Eriocheir sinensis）神经和肌肉组织进行光镜和电镜观察,结果显示类立克次体广泛存在于胸神经节中的神经胶质细胞、结缔组织以及细胞间隙中。在神经细胞与步足肌肉细胞连接处,即运动终板部位有大量类立克次体侵染,并且侵染方式与在胸神经节中一样,均以聚集成团的形式存在。胸神经节是支配步足的神经丛,而颤抖病是以步足颤抖为病症的疾病。类立克次体大量侵染胸神经节和运动终板部位,表明该微生物与病蟹步足颤抖的病症有密切关系,进一步证明类立克次体是颤抖病的致病微生物。     

Purification and antigenic characteristics of a rickettsia-like organism from the oyster Crassostrea ariakensis

A rickettsia-like organism (RLO) has been suggested to be the etiological agent responsible for heavy losses of the oyster Crassostrea ariakensis Gould in China. Because of the lack of molluscan cell lines for in vitro culture of intracellular prokaryotes, antigenic analysis of RLOs has been limited by the inherent difficulties of their purification. In this report, we describe the use of differential speed centrifugation and renografin density gradient centrifugation to purify the RLO directly from infected oyster tissues. The purity and integrity of purified prokaryotes were validated by transmission electron microscopy. Thirteen major constituent proteins, with molecular weights ranging between 17 and 99 kDa, were electrophoretically identified by silver staining, and 8 major proteins were identified with Coomassie blue R staining. Specific mouse polyclonal antiserum was prepared for serological characterization of the RLO and was used in an immunoblot assay, and 3 major antigen groups were identified. The present results advance our knowledge of RLO protein antigens, and several proteins have been identified that could potentially be useful for diagnostic assays or for production of experimental immunostimulants.     

Mycobacteriosis in wild rabbitfish Siganus rivulatus associated with cage farming in the Gulf of Eilat, Red Sea

Infection patterns of Mycobacterium marinum were studied over a period of 3 yr in wild rabbitfish Siganus nivulatus populations associated with commercial mariculture cages and inhabiting various sites along the Israeli Red Sea coastline. Mycobacteriosis was first recorded from the Red Sea in 1990 in farmed sea bass Dicentrarchus labrax and is absent from records of studies on parasites and diseases of wild rabbitfish carried out in the 1970s and 1980s. A sharp increase in the prevalence of the disease in cultured and wild fish in the region has occurred since. A total of 1142 rabbitfish were examined over a 3 yr period from inside mariculture net cages, from the cage surroundings and from several sites along the coast. Histological sections of spleens were examined for presence of granulomatous lesions. Overall prevalence levels of 50% were recorded in the rabbitfish sampled inside the net cages and 39% at the cages' close surroundings, 21% at a sandy beach site 1.2 km westwards, 35% at Eilat harbour 3 km to the south and 42% at a coral reef site about 10 km south of the cages. In addition, 147 fish belonging to 18 native Red Sea species were sampled from 2 sites, the net cage farm perimeter and the coral reef area, and examined for similar lesions. None of those from the coral reef were infected with Mycobacterium; however, 9 of 14 species collected from the cage surroundings were infected. An increase in prevalence of mycobacteriosis in the mariculture farm area was noted from 1995 to 1997. At the same time, a significant increase in prevalence was also apparent at the coral reef sampling site. Two M. marinum isolates from rabbitfish captured at Eilat harbour and the coral reef site were shown by 16S rDNA sequencing analysis to be identical to isolates from rabbitfish trapped inside the mariculture cages as well as isolates from locally cultured sea bass D. labrax. The implications of spreading of M. marinum infection in wild fish populations in the Gulf of Eilat are discussed.     

Effects of dietary protein and lipid levels on growth and feed utilization of wild‐caught striped sea bream,Lithognathus mormyrus

A feeding trial was carried out to determine the effects of dietary protein and lipid levels on the growth performance and feed utilization of wild‐caught striped sea bream (Lithognathus mormyrus). The experimental fish were collected from a local lagoon (Çardak Lagoon, Çanakkale, Turkey), transferred to the Marine Net Cage Unit and fed by hand to apparent satiation with a commercial sea bream feed (Biomar; 42% crude protein, 16% crude lipid). Approximately 4 weeks were needed to acclimate the fish to farming conditions. No pathological signs were observed and no fish losses occurred during the adaptation period. For the test trials four test diets with different levels of protein and lipid were formulated [low protein and low lipid (LP:LL), low protein and high lipid (LP:HL), high protein and low lipid (HP:LL), and high protein and high lipid (HP:HL)] and fed to L. mormyrus (mean weight 85.0 ± 4.6 g SEM) in the net cages (Ø 2 m, depth 2.5 m) for 60 days. During the experiment water temperature varied between 21.1 and 26.4°C; dissolved oxygen 8.4–9.6 mg L^?1; pH 7.2–8.6; and salinity 23.3–25.6‰. Growth performances of fish fed high protein diets were higher compared to fish fed low protein diets, irrespective of the dietary lipid level (P < 0.05). Feed conversion ratio (FCR) and protein efficiency ratio (PER) were not influenced by dietary protein or lipid levels (P > 0.05). Preliminary results indicate that striped sea bream can be easily adapted to farming conditions in net cages, and that a diet containing 50% crude protein and 15% crude lipid (HP:LL) levels with 23.0 g protein MJ^?1 gross energy of protein/energy ratio would be suitable for striped sea bream growth.     

The pathology and seawater performance of farmed Atlantic salmon infected with glochidia of Margaritifera margaritifera

The pathology of glochidial infection of the freshwater mussel Margaritifera margaritifera was examined in farmed Atlantic salmon Salmo salar in fresh water and for 150 days after transfer of salmon to sea water. Prevalence of infection in fresh water was 95%, mean abundance 134 per fish and mean infection intensity 140. Prevalence in sea water was 80–94% in the first 7 weeks after transfer but glochidia were absent, apart from remains, after 50 days in sea water. Glochidia on salmon in fresh water were associated with localized hyperplasia and fusion of secondary gill lamellae. Focally extensive epithelial hyperplasia and fusion of secondary lamellae were present 4–10 days after transfer to sea water. Twenty-three days after transfer, small nodules with a more discrete appearance were present suggesting partial resolution of tissue response; hyperplastic responses associated with glochidia were much reduced after 50 days. Plasma chloride in infected fish 10 days after transfer was 153 mmol. 1⁻¹, significantly higher than fish without infection, suggesting poorer adaptation to sea water. No mortalities due to glochidial infection in sea water were recorded and there was no significant difference in mean weight between infected and control fish.     

Lactococcus garvieae and Streptococcus iniae infections in rainbow trout Oncorhynchus mykiss: similar, but different diseases.

Clinical and macroscopic findings (anorexia, lethargy, loss of orientation and exophthalmia) indicate that Streptococcus iniae and Lactococcus garvieae infections of trout share some common features, but histopathology reveals notable differences between the 2 diseases. Meningitis and panophthalmitis are the main lesions among S. iniae infected trout, whereas L. garvieae infection results in a hyperacute systemic disease. Differences in the LD50s of the 2 pathogens and the sudden onset of signs and death correlate with the histopathological findings, indicating the severity of L. garvieae infection of trout.     

Carriage of potentially fish-pathogenic bacteria in Sparus aurata cultured in Mediterranean fish farms

A bacteriological survey of gilthead sea bream Sparus aurata from different fish farms and culture systems on the Spanish Mediterranean coast was conducted. Three different studies were performed. Study A included hatchery-reared larvae; Study B, periodic examination of randomly sampled growing fish; and Study C, growing fish sampled only during mortality/morbidity events. In Studies B and C, sea cages, earth ponds and indoor tanks were surveyed, and in both cases diseased (showing clinical signs) and non-diseased fish were included. In Study A, a shift from Vibrio spp. (30 d after hatching) to oxidative species (60 d after hatching) was detected, and no mortality events were registered. The percentage of fish yielding bacterial growth were similar in Studies B and C, reaching 57.4 and 61.3%, respectively. A statistically significant relationship between the bacterial carriage and the type of facility was only found in Study B, showing that fish from sea cages had a higher bacterial occurrence than fish from other facilities. A statistically significant relationship between bacterial carriage and signs of disease was found, although the pattern differed in each study. Thus, in Study B only 36.2% of fish yielding abundant bacterial growth were diseased, versus 68.0% in Study C. In total, 25.0% of the fish examined were diseased. Bacterial species composition was similar in asymptomatic and diseased fish, except for a group of V. ichthyoenteri-like isolates that occurred almost exclusively in asymptomatic fish. Dominant bacterial species were V. harveyi and V. splendidus, followed by V. ichthyoenteri-like isolates, Photobacterium damselae ssp. damselae and V. fisheri. Non-fermenters were less frequent but, among them, unidentified halophilic Cytophaga-Flavobacterium isolates and Pseudoalteromonas haloplanktis were the most abundant. An association of individual species with disease was not clear, which suggests the involvement of mixed infections.     

Histopathology of cultured sea bream Sparus aurata infected with sanguinicolid trematodes

The present study is the first report of a sanguinicolid infection affecting sea bream Sparus aurata cultured in net cages in the NE of Spain. The disease was associated with trickling mortalities during the cold season (1999 and 2000). Examination of gill wet mounts of the affected population revealed that sanguinicolid infection was present in 82.6 and 100% of the fish sampled in 1999 and 2000, respectively. Adult flukes, which were located in the kidney, were tentatively identified as members of the family Sanguinicolidae, subfamily Cardicolinae. Eggs and miracidia were found in the gill vascular structures. The inflammatory response triggered by the parasites was moderate and the lesions caused by either eggs and miracidia in the gills or adult flukes in the kidney were not extremely severe, possibly because of the moderate intensity of the parasitosis. Histological observations of sanguinicolid infected sea bream presented here are compared with those reported in other fish species. The role played on sea bream morbility and mortality by other factors (occurrence of a simultaneous moderate monogenean infection, immunological impairement related to low water temperatures) is discussed.     

Differences in proximate lipid composition,heavy metals,and mineral contents in bogue (Boops boops,Linnaueus, 1758) captured far away and directly at sea bass and sea bream cage farms

The objective of this study was to investigate the differences in proximate lipid composition, heavy metals, and mineral contents between bogue (Boops boops, Linnaeus, 1758) captured directly around and near a cage fish farm and those captured far away from it (“wild”). Wild fish were obtained from the fishery of the Aegean Sea and Northeastern Mediterranean. Fish were also caught at a distance of at least 100 meters away from the net cages using a commercial trammel set net. The fish captured near cages were taken at farms cultivating mainly sea bass and sea bream. Samples were taken in March 2016 to determine crude lipid content, moisture, and ash amounts. There were some differences between wild fish and near farm captured fish for both genders. Crude lipid contents of female wild fish and near farm captured fish were found to be higher than those in males (p < .05). The moisture contents of the males near the farms were higher than those in females. Results showed that the predominant macro minerals were potassium in females and males for both capture locations (near and far away from farms: range of 1070–1205 mg/kg). The near farm captured fish had a higher heavy metal content than those captured further away. This may be the result of environmental influences caused by the farm operations.     

Evaluating the effect of clustering when monitoring the abundance of sea lice populations on farmed Atlantic salmon

Using cluster random sampling theory and empirical estimates of the intra‐class correlations for sea lice Lepeophtheirus salmonis abundances, methods on how best to sample Atlantic salmon Salmo salar from cages on farms were derived. Estimates of intra‐class correlations for the abundance of the chalimus and mobile sea lice stages on Atlantic salmon in Scottish farms are given. These correlations were higher for mobile stages than for chalimus, and they had a substantive effect on increasing the number of cages and fish to be sampled for all sea lice stages. An important finding is that it is better to have a procedure that randomly samples a large number of cages using a small number of fish from each. This finding and the cluster random sampling approach have relevance to the monitoring of all marine species being farmed in cages or tanks.     

Important disease conditions of newly cultured species in intensive freshwater farms in Greece: first incidence of nodavirus infection in Acipenser sp

We describe here the main pathological conditions of freshwater fish recently introduced for intensive rearing (open ponds and recirculating freshwater systems) in Greece. Sturgeon were susceptible to skeletal abnormalities of the spine (scoliosis and lordosis) of unknown aetiology. Horizontal transmission of nodavirus from infected sea bass to sturgeon was detected for the first time. This caused serious pathology and clinical signs, such as lethargy and imbalance, leading to secondary infections with Aeromonas hydrophila and Trichodina sp. and chronic, but steady, mortality. Sea bass were very susceptible to nodavirus infection, monogenean infections and gas bubble disease. Mullet reared under recirculated and open-flow conditions were very sensitive to Chilodonella sp. infection, whereas catfish were susceptible to infection with Ichthyophthirius sp. leading to secondary infections with A. hydrophila, Saprolegnia sp. and Myxobacteria spp. Tilapia were very susceptible to gas bubble disease and A. hydrophila. This bacterium was associated with management manipulations for all species and fully responsive to corrective hygiene methods.     

Epidemiology of Cryptosporidium molnari in Spanish Gilthead Sea Bream (Sparus aurata L.) and European Sea Bass (Dicentrarchus labrax L.) Cultures: from Hatchery to Market Size

A long-term epidemiological study of Cryptosporidium molnari in aquacultured European sea bass (ESB) and gilthead sea bream (GSB) was performed in different types of facilities on the Atlantic, Cantabric, and Mediterranean coasts. Four types of studies were carried out. In study A, fish raised from juveniles to marketable size (ongrowing stage) were periodically sampled in three different types of cultures. Studies B and C focused on hatchery and nursery facilities. In study D, occasional samplings were performed during mortality or morbidity outbreaks. As a general trend, C. molnari was more prevalent in GSB than in ESB. Data on the distribution pattern of C. molnari in total sampled GSB (studies A, B, and D) had a variance higher than the mean (overdispersion). In GSB (study A), the type of ongrowing system (sea cages, earth ponds, or indoor tanks) was found to have no significant effect. There was a significant relationship between the presence of the parasite and both fish weight and season. The highest infection values were recorded in spring. Prevalence and intensity had convex weight profiles, with a peak in 30- to 100-g fish. In study D, the prevalence of infection was higher in fish recently introduced in sea cages and in preongrowing systems. In studies B and C, fish were almost never infected before entering the postlarval and nursery facilities. The parasite seems to enter the host mainly through the water in production steps with less stringent water treatment. Recirculation systems and fish cannibalism could contribute to oocyst concentration and dispersion in aquaculture facilities.     

Cymothoid parasite Ceratothoa parallela inflicts great losses on cultured gilthead sea bream sparus aurata in Greece

For the first time Ceratothoa parallela (Otto, 1828), a cymothoid isopod, is reported parasitizing cage-cultured gilthead sea bream Sparus aurata, in Greece. The specimens observed are larvae (Pullus secundus). They were found in the branchial and buccal cavity of young gilthead sea bream of 2 g mean body weight, upon introduction in the cages in an intensive cage farm facility. The species was previously known from several species of wild fish, particularly Sparidae (chiefly Boops boops) in the Mediterranean Sea, Adriatic Sea and Atlantic Ocean. However, this is the first documentation of this parasite in cage-cultured gilthead sea bream. Serious lesions were macroscopically visible and typical of a crustacean infection. The cumulative mortality over a 2 mo period was over 50%. The parasitic problem was not successfully dealt with due to high stocking densities and the non removal of the dead fishes, resulting in constant reinfection.     

Does reduced feeding prior to release improve the marine migration of hatchery brown trout Salmo trutta smolts?

The aim of this study was to test the hypothesis that hatchery brown trout Salmo trutta smolts, with 50% reduced or no feeding over the last 5 months before release, were more likely to migrate to the sea than individuals with standard feeding ratios. The juvenile fish were divided into three groups 176 days before release: (A) with no feeding, (B) with 50% and (C) with 100% feeding. To study their seaward migration, 40 fish from each feeding group were tagged with acoustic transmitters and tracked by automatic listening stations in the River Nidelva, Trondheim, Norway, its estuary and in the nearest marine environment. At the time of release, mean condition factor was significantly lower in group A and the fish from groups A and B had higher levels of Na+, K+‐ATPase. Significantly more fish from group A migrated to the sea, but the rate of downstream progression from release to the estuary did not differ between the three groups. In conclusion, the S. trutta smolts with no access to food in the last 176 day before release were more likely to migrate to the sea. Fish from all three feeding groups, however, appeared to smoltify and had the same rate of downstream progression to the estuary. This indicates that differences in migratory behaviour between individuals from the three feeding groups begin from the time when the fish reach saline waters. It is suggested that feeding in hatcheries has to be greatly reduced (by 50% or more) over several months to have a pronounced effect on the migratory behaviour in S. trutta.     

Experimental infections of Atlantic salmon Salmo salar with Spironucleus barkhanus

Atlantic salmon Salmo salar L. (Salmonidae) were experimentally infected with Spironucleus barkhanus (Diplomonadida: Hexamitidae). Parasites were found in the blood 1 to 8 wk after infection, after which they disappeared from the blood and were found mainly in the internal organs (e.g. spleen and liver), eye socket or muscles. Mortality (38 out of 40 infected fish) occurred when fish had lesions in internal organs and/or on the body surface. Uninfected fish cohabiting with infected fish became infected after 4 wk, indicating direct transmission. There was no difference in susceptibility to spironucleosis between 3 different families of Atlantic salmon. All families developed the disease with a similar pattern of parasitaemia in the blood, similar clinical signs and gross pathology, and with very high mortality (29 out of 30). Clinical signs of systemic spironucleosis may include anemia, skin blisters, muscle ulcerations or unilateral exophthalmia. Gross pathologies include hemorrhaging of internal organs, splenomegaly or deformed (globulated) spleen, or granulomatous lesions in the spleen and liver.     

Serological evidence of infectious salmon anaemia virus (ISAV) infection in farmed fishes,using an indirect enzyme-linked immunosorbent assay (ELISA)

Antibody detection tests are rarely used for diagnostic purposes in fish diseases. Infectious salmon anaemia (ISA) caused by ISA virus (ISAV) is an emerging disease of Atlantic salmon Salmo salar L. The virus has also been isolated from diseased coho salmon Oncorhynchus kisutch in Chile. An indirect enzyme-linked immunosorbent assay (ELISA) that should facilitate serodiagnosis of ISAV infection, the study of epidemiology, and the control of ISA in farmed fishes has been developed using purified ISAV as the coating antigen, and monoclonal antibodies that detect fish immunoglobulins bound to the antigen on the plate. Application of the test to a random sample of farmed Atlantic salmon from the Bay of Fundy, New Brunswick, Canada, positively identified 5 of the 7 ISAV RT-PCR-positive fish, and all 10 RT-PCR-negative fish were also negative in the ELISA. Some RT-PCR-negative fish had an elevated non-specific antibody reactivity suggestive of chronic infection or resistance to ISAV. This test was also able to detect 11 of the 14 coho salmon pooled serum samples from a clinically affected farm in Chile that were positive by the virus neutralization (VN) test, and 2 of the 4 VN-negative samples. We conclude that this ELISA would be suitable as a routine test for ISAV infection or for assessing ISAV vaccine efficacy before placing smolts in sea cages, and for testing fishes in sea cages to detect level of resistance to ISA. The assay enables vaccination in combination with depopulation control methods.     

Epidemiology of Cryptosporidium molnari in Spanish gilthead sea bream (Sparus aurata L.) and European sea bass (Dicentrarchus labrax L.) cultures: from hatchery to market size

A long-term epidemiological study of Cryptosporidium molnari in aquacultured European sea bass (ESB) and gilthead sea bream (GSB) was performed in different types of facilities on the Atlantic, Cantabric, and Mediterranean coasts. Four types of studies were carried out. In study A, fish raised from juveniles to marketable size (ongrowing stage) were periodically sampled in three different types of cultures. Studies B and C focused on hatchery and nursery facilities. In study D, occasional samplings were performed during mortality or morbidity outbreaks. As a general trend, C. molnari was more prevalent in GSB than in ESB. Data on the distribution pattern of C. molnari in total sampled GSB (studies A, B, and D) had a variance higher than the mean (overdispersion). In GSB (study A), the type of ongrowing system (sea cages, earth ponds, or indoor tanks) was found to have no significant effect. There was a significant relationship between the presence of the parasite and both fish weight and season. The highest infection values were recorded in spring. Prevalence and intensity had convex weight profiles, with a peak in 30- to 100-g fish. In study D, the prevalence of infection was higher in fish recently introduced in sea cages and in preongrowing systems. In studies B and C, fish were almost never infected before entering the postlarval and nursery facilities. The parasite seems to enter the host mainly through the water in production steps with less stringent water treatment. Recirculation systems and fish cannibalism could contribute to oocyst concentration and dispersion in aquaculture facilities.     

Putative Phage Hyperparasite in the Rickettsial Pathogen of Abalone, “Candidatus Xenohaliotis californiensis”

Studies on the ecology of microbial parasites and their hosts are predicated on understanding the assemblage of and relationship among the species present. Changes in organismal morphology and physiology can have profound effects on host–parasite interactions and associated microbial community structure. The marine rickettsial organism, “Candidatus Xenohaliotis californiensis” (WS-RLO), that causes withering syndrome of abalones has had a consistent morphology based on light and electron microscopy. However, a morphological variant of the WS-RLO has recently been observed infecting red abalone from California. We used light and electron microscopy, in situ hybridization and16S rDNA sequence analysis to compare the WS-RLO and the morphologically distinct RLO variant (RLOv). The WS-RLO forms oblong inclusions within the abalone posterior esophagus (PE) and digestive gland (DG) tissues that contain small rod-shaped bacteria; individual bacteria within the light purple inclusions upon hematoxylin and eosin staining cannot be discerned by light microscopy. Like the WS-RLO, the RLOv forms oblong inclusions in the PE and DG but contain large, pleomorphic bacteria that stain dark navy blue with hematoxylin and eosin. Transmission electron microscopy (TEM) examination revealed that the large pleomorphic bacteria within RLOv inclusions were infected with a spherical to icosahedral-shaped putative phage hyperparasite. TEM also revealed the presence of rod-shaped bacteria along the periphery of the RLOv inclusions that were morphologically indistinguishable from the WS-RLO. Binding of the WS-RLO-specific in situ hybridization probe to the RLOv inclusions demonstrated sequence similarity between these RLOs. In addition, sequence analysis revealed 98.9–99.4 % similarity between 16S rDNA sequences of the WS-RLO and RLOv. Collectively, these data suggest that both of these RLOs infecting California abalone are “Candidatus Xenohaliotis californiensis,” and that the novel variant is infected by a putative phage hyperparasite that induced morphological variation of its RLO host.