Rickettsiales-like Prokaryote Research Articles

Molecular identification of a novel intracellular proteobacteria from scallop Chlamys farreri

The scallop Chlamys farreri is an important bivalve species cultured in the north of China. With the large expansion of culturing, mass mortalities have occurred persistently and caused great economic loss since 1996. An intracellular Rickettsiales-like prokaryote (RLP) was proposed as the causative agent. To identify the RLP, a partial 16S rDNA sequence of 1319 base pairs was amplified, cloned and sequenced. Phylogenetic analysis indicates that the prokaryote is a member of the α subclass of the proteobacteria. In the phylogenetic tree, it forms an independent and novel clade, which shares an evolutionary line of descent with a group of uniquely obligate intracellular organism comprised of Caedibacter caryophilus, an endosymbiont of Acanthamoeba sp., and the necrotizing hepatopancreatitis (NHP) bacterium of shrimp. Based on the obtained 16S rDNA sequence, three specific RNA targeted oligonucleotide DNA probes were designed, synthesized and labeled with digoxigenin for in situ hybridization detection. Results showed that the probes specifically bonded to the RLP inclusion in infected C. farreri tissues, indicating the amplified, cloned and sequenced 16S rDNA originated from the intracellular RLP parasitizing the C. farreri. We propose the name Sinorickettsia chlamys sp.nov. to C. farreri RLP.

An oxytetracycline bath protocol to eliminate the agent of withering syndrome, Candidatus Xenohaliotis californiensis, in captive abalone populations

Abalone (Haliotis spp.) populations are in decline due to overfishing and/or disease in all regions that historically supported important fisheries worldwide. Two species have been federally listed as Endangered in the USA. Recovery plans for these and other highly depleted species typically include outplanting of progeny from captive-reared broodstock, for which maintaining good health is essential to promote gonad development and maturation. In California, USA, the principal disease of concern for abalone is withering syndrome, caused by a gastrointestinal intracellular Rickettsiales-like prokaryote named Candidatus Xenohaliotis californiensis (Ca.Xc). We report the application of an antibiotic bath exposure method to eliminate the pathogen in captive red abalone (H. rufescens) populations. A series of eight, 24-h baths consisting of 500 ppm oxytetracycline in seawater were delivered over a period of 21 days. Testing of feces by PCR and histology indicated complete elimination of Ca.Xc by day 105 following the first bath treatment. Mean oxytetracycline levels in the digestive gland reached a peak of 670 ppm at the end of the treatment period and remained above 100 ppm for 125 days. Survival rate, body condition index, growth and a measure of foot muscle health were similar between treated and control groups, despite tetracyclines being potentially toxic or immunosuppressive in some animal systems. This method should prove useful for creating Ca.Xc specific-pathogen-free captive abalone populations for land-based production and to prevent pathogen transfer when abalone are introduced to growing regions where the pathogen is not present.

Differing responses of red abalone (Haliotis rufescens) and white abalone (H. sorenseni) to infection with phage-associated Candidatus Xenohaliotis californiensis.

The Rickettsiales-like prokaryote and causative agent of Withering Syndrome (WS)—Candidatus Xenohaliotis californiensis (Ca. Xc)—decimated black abalone populations along the Pacific coast of North America. White abalone—Haliotis sorenseni—are also susceptible to WS and have become nearly extinct in the wild due to overfishing in the 1970s. Candidatus Xenohaliotis californiensis proliferates within epithelial cells of the abalone gastrointestinal tract and causes clinical signs of starvation. In 2012, evidence of a putative bacteriophage associated with Ca. Xc in red abalone—Haliotis rufescens—was described. Recently, histologic examination of animals with Ca. Xc infection in California abalone populations universally appear to have the phage-containing inclusions. In this study, we investigated the current virulence of Ca. Xc in red abalone and white abalone at different environmental temperatures. Using a comparative experimental design, we observed differences over time between the two abalone species in mortality, body condition, and bacterial load by quantitative real time PCR (qPCR). By day 251, all white abalone exposed to the current variant of Ca. Xc held in the warm water (18.5 °C) treatment died, while red abalone exposed to the same conditions had a mortality rate of only 10%, despite a relatively heavy bacterial burden as determined by qPCR of posterior esophagus tissue and histological assessment at the termination of the experiment. These data support the current status of Ca. Xc as less virulent in red abalone, and may provide correlative evidence of a protective phage interaction. However, white abalone appear to remain highly susceptible to this disease. These findings have important implications for implementation of a white abalone recovery program, particularly with respect to the thermal regimes of locations where captively-reared individuals will be outplanted.

Hyperparasitism by the bacteriophage (Caudovirales) infecting Candidatus Xenohaliotis californiensis (Rickettsiales-like prokaryote) parasite of wild abalone Haliotis fulgens and Haliotis corrugata from the Peninsula of Baja California, Mexico

Candidatus Xenohaliotis californiensis (CXc) is a Rickettsiales-like prokaryote that is considered the causal agent of Withering Syndrome (WS), a chronic disease of abalone, from the west coast of North America and it is listed by the International Organization for Animal Health (OIE) as a reportable agent due to its pathogenicity. This bacterium in red abalone Haliotis rufescens, black abalone Haliotis cracherodii, and yellow abalone Haliotis corrugata from California, US and Baja California, Mexico has been found to be infected by a bacteriophage. To date, there is no information on the epizootiology of CXc and its bacteriophage in natural populations of abalone; furthermore, it is unknown if the bacteriophage was also present in CXc infecting blue abalone Haliotis fulgens. The objective of this study was to determine the distribution, prevalence and intensity of CXc, as well as to determine the distribution and prevalence of the bacteriophage and to study interactions between host sex and hyperparasitism in blue abalone and yellow abalone. Tissue samples were obtained from seven localities where the commercial capture of wild abalone is carried out. Samplings were conducted throughout the 2012–2013 capture seasons and a total of 182 blue abalone and 170 yellow abalone were obtained. The prevalence and intensity of CXc and the prevalence of the bacteriophage were determined by histology. The identity of CXc was confirmed by PCR, product sequence analysis and in situ hybridization while the identity of the bacteriophage was corroborated by TEM. The prevalence of CXc infected and uninfected by the bacteriophage was 80% in blue abalone and 62% in yellow abalone. Low infection intensities were found in 86% of blue abalone and 82% of yellow abalone. Infection intensity was significantly higher in undifferentiated yellow abalone. The bacteriophage in CXc showed a prevalence of 22% and 31% in blue abalone and yellow abalone respectively. These results show that CXc and its bacteriophage are widely distributed in the peninsula of Baja California and that they are well established in natural populations of blue abalone and yellow abalone. Additionally, this data constitutes the first record of a bacteriophage in blue abalone.

The hyperparasite of the rickettsiales-like prokaryote, Candidatus Xenohaliotis californiensis has morphological characteristics of a Siphoviridae (Caudovirales)

Transmission electron microscopy analysis (TEM) of the rickettsiales-like prokaryote, Candidatus Xenohaliotis californiensis (CXc), pathogen of Haliotis spp. from the West Coast of North America, were found to be infected by a bacteriophage hyperparasite previously described in red abalone from California. The hyperparasite has an icosahedrical-like capsid with a narrow long flexible tail, this morphological characteristic tentatively place this virus in the Family Siphoviridae from the order Caudovirales. TEM images also showed the bacteriophage in different stages of assembly in the cytoplasm of CXc, demonstrating its lytic cycle.

Characterization of an intracellular bacterium infecting the digestive gland of the South African abalone Haliotis midae

A Rickettsiales-like prokaryote was observed in digestive gland epithelium of farmed South African abalone Haliotis midae sampled for routine surveillance. Histological examination showed basophilic inclusions typical of rickettsial infections in shellfish (Mouton, 2000). This study aimed to identify the Rickettsiales-like prokaryote (RLP) colonising H. midae. Total genomic DNA was extracted from infected digestive gland sampled from cultured H. midae. PCR performed with “Candidatus Xenohaliotis californiensis” specific primers RA 5-1 and RA 3-6 did not yield positive results, indicating that the H. midae RLP is not the withering syndrome agent. The 16S rRNA gene sequence of the intracellular bacterium was determined following nested PCR using universal eubacterial primers fD1/Rp2 and the broad spectrum primers EHR16SD/EHR16SR which are specific for members of the Anaplasmataceae family within the Rickettsiales order. Phylogenetic and evolutionary distance analysis showed that the H. midae RLP is a unique member of the α-proteobacteria. The identity and location of the histologically observed intracellular bacterium was confirmed by in situ hybridization performed on sequential sections of infected H. midae digestive gland using a combination of three digoxigenin-labelled probes specific to various regions of the 16S rRNA gene of the RLP.Statement of relevanceThe manuscript shows that farmed abalone, showing no disease symptoms, can remain healthy despite the presence of intracellular bacteria colonising their digestive glands. The study shows that Rickettsiales-like prokaryotes detected in farmed abalone are not necessarily “Candidatus Xenohaliotis californiensis”, the causative agent of withering syndrome.

Oxytetracycline water bath treatment of juvenile blue abalone Haliotis fulgens (Philippi 1845) affected by the withering syndrom

The fatal abalone disease named withering syndrome (WS), caused by a Rickettsiales-like prokaryote classified as Xenohaliotis californiensis, has been treated by injections and oral administration of oxytetracycline (OTC) with relatively successful results. These therapeutic treatments have practical limitations for their application in aquacultural and experimental facilities due to the inability to eat of severely ill organisms, and the need to incorporate the antibiotic in pelletized food, increasing production costs. During an experiment to determine the effect of photoperiod and temperature on the growth of juvenile blue abalone Haliotis fulgens, some animals showed overt symptoms of WS. Oxytetracycline baths at 400 mg L–1 were used for one hour daily during seven days to treat diseased animals. The metabolic rate of one group of these organisms was also assessed. Recovery time (time for an organism to regain the upright position) and size were assessed on days 0, 14, and 29. By the third day of treatment, most treated animals regained a limited adhesion capacity, and began to ingest food and to produce feces. After one week, more than 50% of the treated abalone were capable of regaining the upright position within the first 10 min of being turned upside down. After 30 days all organisms showed strong adhesion to the substrate. Metabolic rate declined from 380 (initially) to 280 µL O2 ind–1 h–1 after two weeks of treatment. This study opens the door for further research on the use of OTC baths for practical use in abalone culture and management.

Response of Red Abalone Reproduction to Warm Water, Starvation, and Disease Stressors: Implications of Ocean Warming

Changes in ocean temperature can have direct and indirect effects on the population dynamics of marine invertebrates. We examined the impacts of warm water, starvation, and disease on reproduction in red abalone (Haliotis rufescens). We found that sperm production was highly sensitive to warm water and starvation, suggesting there may be a dramatic temperature threshold above which sperm production fails. Wild males from northern (72%) and southern (81%) California had sperm. In contrast, only 30% of the males exposed to warm water (18C) for 6 mo or starvation for 13 mo had sperm, with spermatogenesis dropping dramatically from 300,000 presperm cells/mm 3 (wild) to 46,000 presperm cells/mm 3 (warm water) and 84,000 presperm cells/mm 3 (starvation). In a longer warm-water experiment (12 mo), males had total reproductive failure in temperatures greater than 16C, irrespective of food treatment. Egg production was less sensitive to warm water, but was impacted more by starvation, especially food quantity relative to quality. Wild females from northern (97%) and southern (100%) California had mature oocytes averaging 3 million eggs and 21 million eggs, respectively. Females exposed to 18C water for 6 mo had diminished fecundity, averaging only 400,000 mature eggs whereas females in the starvation experiment did not produce any mature eggs. Normal sperm and egg production was found in abalone testing positive for Rickettsiales-like-prokaryote (RLP), the agent of Withering Syndrome in cool water. However, abalone with RLP also exposed to warm water developed the disease withering syndrome and did not produce any mature gametes. The temperature-mediated lethal and sublethal effects on red abalone reproduction described here, combined with temperature's known impacts on abalone growth, kelp abundance, and disease status, clearly demonstrate population-level consequences. We suggest that temperature needs to be explicitly incorporated into red abalone recovery and management planning, because California's ocean has warmed and is predicted to warm in the future.

Parasites of the Pleasure Oyster Crassostrea corteziensis Cultured in Nayarit, Mexico

The pleasure oyster Crassostrea corteziensis is collected and cultured in Nayarit on the Pacific coast of Mexico, and the improvement and promotion of its culture are seen as a possible source for the economic development of coastal populations. However, information about the parasite fauna of the pleasure oyster is almost completely lacking. A histopathological survey carried out in two estuaries, Boca del Camichín and Pozo Chino, revealed the presence of hypertrophied gametes, rickettsiales-like prokaryotes (RLPs), the protozoan Perkinsus marinus, a protozoan Nematopsis sp., Ancistrocoma-like ciliates (ALCs), Sphenophrya-like ciliates, a turbellarian Urastoma sp., and encysted crustaceans. In general, prevalence and intensity of parasites were similar in both localities except that ALCs and encysted crustaceans were more prevalent in Pozo Chino than in Boca del Camichín. Perkinsus marinus and RLPs seem to represent a more significant risk for the health of pleasure oysters than do the other parasites, and surveillance and control of these parasites are needed for the development of pleasure oyster culture.

Bacterial diversity of the digestive gland of Sydney rock oysters, Saccostrea glomerata infected with the paramyxean parasite, Marteilia sydneyi

To determine whether the infestation by the protozoan paramyxean parasite, Marteilia sydneyi, changes the bacterial community of the digestive gland of Sydney rock oysters, Saccostrea glomerata. Six 16S rDNA clone libraries were established from three M. sydneyi-infected and three un-infected oysters. Restriction enzyme analysis followed by sequencing representative clones revealed a total of 23 different operational taxonomic units (OTUs) in un-infected oysters, comprising the major phyla: Firmicutes, Proteobacteria, Cyanobacteria and Spirocheates, where the clone distribution was 44, 36, 7 and 5%, respectively. Close to half of the OTUs are not closely related to any other hitherto determined sequence. In contrast, S. glomerata infected by M. sydneyi had only one OTU present in the digestive gland. Phylogenetic analysis of the 16S rDNA sequence reveals that this dominant OTU, belonging to the alpha-Proteobacteria, is closely related to a Rickettsiales-like prokaryote (RLP). The microbiota of the digestive gland of Sydney rock oysters is changed by infection by M. sydneyi, becoming dominated by a RLP, and generally less diverse. The bacterial community of un-infected S. glomerata differs from previous studies in that we identified the dominant taxa as Firmicutes and alpha-Proteobacteria, rather than heterotrophic gamma-Proteobacteria. This is the first culture-independent study of the microbiota of the digestive glands of edible oysters to the species level. The commercial viability of the Sydney rock oyster industry in Australia is currently threatened by Queensland Unknown disease and the changes in the bacterial community of S. glomerata corresponding with infection by M. sydneyi sheds further light on the link between parasite infection and mortality in this economically damaging disease.

Health and survival of red abalone, Haliotis rufescens, under varying temperature, food supply, and exposure to the agent of withering syndrome

Withering syndrome (WS) is a disease of wild and cultured abalone caused by a Rickettsiales-like prokaryote (WS-RLP). This study compared the pathologic changes that occur during the progression of WS in red abalone to those caused by environmental stresses consisting of elevated temperature and food limitation and determined the impact of these stressors on WS prevalence and intensity. Farmed red abalone were administered a feed-based oxytetracycline therapeutic treatment to assure WS-RLP-free status prior to initiation of the experiment. Groups were then held in each of eight combinations of exposed vs. unexposed to WS-RLP, elevated vs. ambient temperature, and high vs. low food supply, for 447 days. Mortality was associated with starvation and disease but not elevated temperature alone. Elevated temperature significantly affected WS-RLP transmission: only 1.7% of WS-RLP- exposed abalone held at ambient temperature (12.3 °C) became infected compared to at least 72% of those held at elevated temperature (18.7 °C). Among exposed abalone at elevated temperature, fed animals exhibited greater infection prevalence but not greater infection intensity or digestive gland changes than starved animals, suggesting that abalone acquire infections by ingesting contaminated food. Food, temperature, WS-RLP exposure, and most of their interactions had significant effects on body condition and foot atrophy. Immunohistochemical detection of cell proliferation and apoptosis revealed no differences between normal digestive gland and that infected with WS-RLP. Body mass shrinkage, foot atrophy, elevated mortality, and decreased foot and digestive gland glycogen were observed in both WS-affected and starved, unexposed abalone, with the WS-RLP-exposed, starved group held at elevated temperature faring worst. Among exposed and unexposed animals, food supply but not temperature affected body mass and growth. These data demonstrate that the high morbidity and mortality exhibited by WS-RLP-infected abalone is a consequence of disease and not direct thermal stress. Drug residue analysis indicated oxytetracycline concentrations of up to 600 ppm in the digestive gland at 38 days post-treatment, an unusual degree of tissue retention of this therapeutant.

Characterizing the metabolic actions of natural stresses in the California red abalone, Haliotis rufescens using 1H NMR metabolomics

Withering syndrome in California red abalone (Haliotis rufescens) is caused by the Rickettsiales-like prokaryote (WS-RLP) Candidatus Xenohaliotis californiensis. WS-RLP infection is not sufficient to cause withering syndrome, and for reasons not yet well understood additional stressors such as elevated water temperature appear to influence disease development. Using nuclear magnetic resonance (NMR) based metabolomics, we have investigated the influence of food availability, temperature, and bacterial infection, both individually and in combination, on the metabolic status of the red abalone. Food limitation caused dramatic reductions in all observed classes of foot muscle metabolites, while at the same time metabolite levels within the digestive gland were preserved or increased. We also found that food limitation in combination with elevated temperature led to greater metabolic perturbations in both tissue types than those observed under food limitation alone. WS-RLP infection and food-limitation resulted in many of the same metabolic changes within the tissues studied, although the effects of infection were less severe. We observed increased levels of homarine in the digestive gland of both food-limited and WS-RLP-infected animals, yet only observed increased homarine levels in the foot muscle of infected abalone. These results further support the recently established glucose-to-homarine ratio in foot muscle as a potential marker for differentiating WS-RLP-infected animals from those of both healthy and food limited abalone. Furthermore, we found that the NMR metabolic data correlates well with histological measurements supporting the use of the metabolomics approach for characterizing both normal and pathological events in marine species, particularly during periods of environmentally relevant stress.

Ultrastructure of a haplosporidian containing Rickettsiae, associated with mortalities among cultured paua Haliotis iris.

Uninucleate and multinucleate stages of a protozoan parasite are described from cultured abalone Haliotis iris Martyn, 1784 in New Zealand. The parasite is identified as a haplosporidian by the occurrence of multinucleate plasmodia, mitochondria with tubular cristae, lipid droplets, anastomosing endoplasmic reticulum (aER), multivesicular bodies (MVBs), haplosporogenesis by the production of haplosporosome-like bodies from nuclear membrane-bound Golgi, and their maturation to haplosporosomes. Coated pits occurred in the plasma membrane and coated vesicles were scattered in the cytoplasm, particularly in association with the Golgi face away from the nucleus, and aER. It is concluded that the outward face of the Golgi may be the trans face, and that aER is the trans-Golgi network. Coated pits and bristle-coated vesicles are reported from a haplosporidian for the first time. The vesicles in the MVBs resembled the cores and inner membranes of haplosporosomes, without the outer layer. The possible inter-relationships of these features are discussed. The abalone parasite differs from previously described haplosporidians in the apparent absence of a persistent mitotic spindle, and the presence of intracytoplasmic coccoid to rod-shaped bacteria resembling Rickettsiales-like prokaryotes. Phylogenetic analysis of the 16S rRNA gene sequence of the Rickettsiales-like prokaryotes indicated that these organisms belong to the Rickettsia cluster. The prokaryotes have a high (7%) sequence divergence from known Rickettsieae, with Rickettsia sp. and R. massiliae being the closest relatives. The lack of non-molecular evidence prevents us from proposing a new rickettsial genus at this time.

Detection of 'Candidatus Xenohaliotis californiensis' (Rickettsiales-like prokaryote) inclusions in tissue squashes of abalone (Haliotis spp.) gastrointestinal epithelium using a nucleic acid fluorochrome.

Rickettsiales-like prokaryotes appear to be etiologic agents of a number of newly described diseases of fish and shellfish. 'Candidatus Xenohaliotis californiensis' is a Rickettsiales-like prokaryote responsible for withering syndrome, a fatal disease of wild and farmed Eastern Pacific abalone, Haliotis spp. The bacterium proliferates in gastrointestinal epithelial cells, forming large intracytoplasmic inclusions. We describe a method of rapidly detecting and assessing the intensity of 'Candidatus Xenohaliotis californiensis' infections in abalone gastrointestinal tissue using the nucleic acid-specific fluorochrome Hoechst 33258. In excised tissue pieces dried onto slides, rehydrated in the Hoechst stain and viewed with ultraviolet light, the large bacterial inclusions were strongly fluorescent and could be easily distinguished from smaller host cell nuclei. This provided a rapid, inexpensive alternative to paraffin section microscopy or molecular techniques, allowing detection of the pathogen within minutes of tissue excision. Comparison of the fluorochrome method with conventional histological analysis for the ability to detect inclusions in 109 samples was 90% accurate, with discrepancies due to false negative diagnosis of low-level infections. An alternative nucleic acid-specific fluorochrome, propidium iodide, showed a staining pattern identical to that of Hoechst 33258. These methods should prove useful for the rapid detection of inclusion-forming Rickettsiales-like prokaryotes in tissues from many host species.

Characterization of intracytoplasmic prokaryote infections in Dreissena sp. (Bivalvia: Dreissenidae).

This study characterizes intracytoplasmic infections with prokaryote microorganisms in Dreissena sp. (near Dreissena polymorpha) from northeastern Greece and represents the first report of such infections in freshwater bivalves. Light microscope observations of stained tissues revealed basophilic, cytoplasmic inclusion bodies in 87.5% (28/32) of the mussels sectioned. Inclusions in epithelial cells and connective tissues were noted, respectively, in 34.4 and 71.9% of the sample, with 5 mussels (15.6%) having both tissue types infected. Epithelial cell infections were observed in histological sections only in digestive gland tubules and ducts; within tubules, inclusions were present more often in secretory than digestive cells. Connective tissue infections, however, were systemic; among the 32 mussels sectioned, inclusions were found in the gills (65.6%), foot (12.5%), mantle (9.4%), labial palps (6.3%), digestive gland (6.3%), stomach (6.3%), and gonads (3.1%). Cytoplasmic inclusions (maximum dimension, 138 microm) were prominent enough in the gills to be visible in 17.0% of the 247 mussels dissected. Ultrastructurally, prokaryote cells in gill connective tissues were clearly characteristic of Chlamydiales-like organisms, with each intracytoplasmic inclusion containing a loosely packed mixture of elementary, reticulate, intermediate bodies, and blebs. Prokaryote colonies in digestive gland epithelial cells exclusively contained 1 of 4 morphological cell types and were considered Rickettsiales-like. Hexagonal, virus-like particles were present in the cytoplasm of the largest of these Rickettsiales-like prokaryotes. Although host stress was evident from localized cell necrosis and dense hemocyte infiltration, overall infection was fairly benign, with no major, adverse impact on body condition evident among sectioned or dissected mussels. A possible negative effect was partial constriction of gill water tubes, but at the infection intensity observed (typical range 1 to 7 inclusion bodies per section), significant interference with respiration and other metabolic functions of the gills was highly unlikely.

Withering Syndrome in Farmed Red Abalone Haliotis rufescens: Thermal Induction and Association with a Gastrointestinal Rickettsiales-like Prokaryote.

Withering syndrome (WS) is a chronic wasting disease responsible for mass mortality in wild populations of black abalone Haliotis cracherodii. The etiology of WS is uncertain with limited evidence for the role of a gastrointestinal Rickettsiales-like prokaryote (RLP). We documented for the first time the occurrence of animals with clinical signs of WS and associated morphological changes in another haliotid species, the red abalone H. rufescens. In this study, 60 juvenile red abalone (8 cm) were randomly selected from a farmed population raised at 14°C that was known to have low-intensity RLP infections but lacked clinical signs of WS. The abalone were held in triplicate containers receiving water of approximately 14.7°C (Control, Co) or 18.5°C (elevated temperature, ET) and were fed equally for 220 d. Survival was 100% (30/30) for the Co group and 67% (20/30) for the ET group. The ET group animals had higher RLP infection intensities and showed more clinical signs (mantle retraction, lower weight gain, lower condition index) and morphological changes (digestive gland degeneration) associated with WS. In trials conducted immediately before termination of the experiment, ET group animals fed at half the rate of Co group animals. Among ET group animals, the intensity of RLP infections in the posterior portion of the esophagus was positively correlated with WS clinical signs and morphological changes, whereas no correlations were present among Co group animals. During 1997-1998 and in conjunction with elevated seawater temperatures associated with El Niño, several abalone farms in California experienced a dramatic increase in the proportion of red abalone showing signs of WS. Examination of 66 red abalone from five commercial farms revealed that animals with more advanced RLP infections had more severe WS clinical signs and associated morphological changes. Collectively, these data demonstrate that RLP infection plays a key role in the etiology of WS in red abalone and that warm water enhances the severity of the pathogenic effects of RLP infection.

Detection of Rickettsiales-like Prokaryotes by in Situ Hybridization in Black Abalone, Haliotis cracherodii, with Withering Syndrome

Detection of Rickettsiales-like Prokaryotes by in Situ Hybridization in Black Abalone, Haliotis cracherodii, with Withering Syndrome