Rickettsia-like Organisms Research Articles

Abalone farm discharges the withering syndrome pathogen into the wild

An intracellular bacterium Candidatus Xenohaliotis californiensis, also called Withering-Syndrome Rickettsia-Like Organism (WS-RLO), is the cause of mass mortalities that are the chief reason for endangerment of black abalone (Haliotis cracherodii). Using a real-time PCR assay, we found that a shore-based abalone farm (AF) in Santa Barbara, CA, USA discharged WS-RLO DNA into the ocean. Several other shore-based AFs discharge effluent into critical habitat for black abalone in California and this might affect the recovery of wild black abalone. Existing regulatory frameworks exist that could help protect wild species from pathogens released from shore-based aquaculture.

Parasites of economically important bivalves from the southern coast of Bahia State, Brazil

This study investigated the parasites of three commercially important bivalve species (Crassostrea rhizophorae, Mytella guyanensis and Lucina pectinata) from the southern coast of Bahia, Brazil. A total of 540 specimens were collected in August 2009 and February 2010, at three localities. The bivalve specimens were measured on their longest axis, opened, and macroscopically examined for the presence of parasites or signs of disease. They were then fixed in Davidson' solution and subjected to routine histological processing, with paraffin embedding and H&E staining; next, the specimens were examined under a light microscope. No parasites were observed associated with L. pectinata. Rickettsia-like organisms (RLOs), Sphenophrya sp. (Ciliophora), Nematopsis sp. (Apicomplexa), Urastoma sp. (Turbellaria) and Bucephalus sp. (Digenea) were observed in both C. rhizophorae and M. guyanensis, as well as Ancistrocoma sp. (Ciliophora) and Tylocephalum sp. (Cestoda) in the former. A high prevalence of Nematopsis sp. was seen, but caused no apparent damage to the host. Bucephalus sp. caused the destruction of tissues, with castration, but showed low prevalence. The other parasites occurred in low prevalence and intensity, without causing significant damage.

HMGB in Mollusk Crassostrea ariakensis Gould: Structure, Pro-Inflammatory Cytokine Function Characterization and Anti-Infection Role of Its Antibody

Background Crassostrea ariakensis Gould is a representative bivalve species and an economically important oyster in China, but suffers severe mortalities in recent years that are caused by rickettsia-like organism (RLO). Prevention and control of this disease is a priority for the development of oyster aquaculture. It has been proven that mammalian HMGB (high mobility group box) can be released extracellularly and acts as an important pro-inflammatory cytokine and late mediator of inflammatory reactions. In vertebrates, HMGB’s antibody (anti-HMGB) has been shown to confer significant protection against certain local and systemic inflammatory diseases. Therefore, we investigated the functions of Ca-HMGB (oyster HMGB) and anti-CaHMGB (Ca-HMGB’s antibody) in oyster RLO/LPS (RLO or LPS)-induced disease or inflammation.Methodology/Principal FindingsSequencing analysis revealed Ca-HMGB shares conserved structures with mammalians. Tissue-specific expression indicates that Ca-HMGB has higher relative expression in hemocytes. Significant continuous up-regulation of Ca-HMGB was detected when the hemocytes were stimulated with RLO/LPS. Recombinant Ca-HMGB protein significantly up-regulated the expression levels of some cytokines. Indirect immunofluorescence study revealed that Ca-HMGB localized both in the hemocyte nucleus and cytoplasm before RLO challenge, but mainly in the cytoplasm 12 h after challenge. Western blot analysis demonstrated Ca-HMGB was released extracellularly 4–12 h after RLO challenge. Anti-CaHMGB was added to the RLO/LPS-challenged hemocyte monolayer and real-time RT-PCR showed that administration of anti-CaHMGB dramatically reduced the rate of RLO/LPS-induced up-regulation of LITAF at 4–12 h after treatment. Flow cytometry analysis indicated that administration of anti-CaHMGB reduced RLO/LPS-induced hemocyte apoptosis and necrosis rates.Conclusions/SignificanceCa-HMGB can be released extracellularly and its subcellular localization varies when stimulated with RLO. Ca-HMGB is involved in oyster immune reactions and functions as a pro-inflammatory cytokine. Anti-CaHMGB can significantly suppress RLO/LPS-induced inflammatory responses and hemocyte necrosis and apoptosis, suggesting that Ca-HMGB is a potential target to prevent and control RLO/LPS-induced disease or inflammation.

Tick cell culture isolation and growth of Rickettsia raoultii from Dutch Dermacentor reticulatus ticks

Tick cell lines play an important role in research on ticks and tick-borne pathogenic and symbiotic microorganisms. In an attempt to derive continuous Dermacentor reticulatus cell lines, embryo-derived primary cell cultures were set up from eggs laid by field ticks originally collected as unfed adults in The Netherlands and maintained for up to 16 months. After several months, it became evident that cells in the primary cultures were infected with a Rickettsia-like intracellular organism. Supernatant medium containing some D. reticulatus cells was inoculated into cultures of 2 Rhipicephalus (Boophilus) microplus cell lines, BME/CTVM2 and BME/CTVM23, where abundant growth of the bacteria occurred intracellularly on transfer to both cell lines. Bacterial growth was monitored by light (live, inverted microscope, Giemsa-stained cytocentrifuge smears) and transmission electron microscopy revealing heavy infection with typical intracytoplasmic Rickettsia-like bacteria, not present in uninfected cultures. DNA was extracted from bacteria-infected and uninfected control cultures, and primers specific for Rickettsia 16S rRNA, ompB, and sca4 genes were used to generate PCR products that were subsequently sequenced. D. reticulatus primary cultures and both infected tick cell lines were positive for all 3 Rickettsia genes. Sequencing of PCR products revealed 99–100% identity with published Rickettsia raoultii sequences. The R. raoultii also grew abundantly in the D. nitens cell line ANE58, poorly in the D. albipictus cell line DALBE3, and not at all in the D. andersoni cell line DAE15. In conclusion, primary tick cell cultures and cell lines are useful systems for isolation and propagation of fastidious tick-borne microorganisms. In vitro isolation of R. raoultii from Dutch D. reticulatus confirms previous PCR-based detection in field ticks, and presence of the bacteria in the tick eggs used to initiate the primary cultures confirms that transovarial transmission of this Rickettsia occurs.

Phage particles infecting branchial Rickettsiales-like organisms in banded carpet shell Polititapes virgineus (Bivalvia) from Galicia (NW Spain)

Basophilic intracellular prokaryotic-like colonies were observed in the gills of banded carpet shell Polititapes virgineus (= Tapes rhomboides) (Linnaeus, 1767) from a natural bed in Galicia (NW Spain). Light microscope observations suggested the presence of 2 types of colonies, but transmission electron microscopy revealed that these were the same Rickettsiales-like colonies, one infected and the other uninfected by phage particles. This is the first report of the presence of phage particles in Rickettsiales-like organisms in the gills of P. virgineus.

Report of pathogens and parasites in Perumytilus purpuratus from San Jorge Bay, Antofagasta, Chile

Perumytilus purpuratus is a mussel commonly found on the entire Chilean coastline. The knowledge of pathogens and parasites of this mollusc, as well as for many other species, inhabiting the Chilean coast remains largely unknown. This study aims to survey parasites and pathogens in P. purpuratus from San Jorge Bay, Antofagasta, Chile. From September to October 2009, 160 specimens of this bivalve were collected. Ciliates, virus-like, rickettsia-like organisms (RLO) and digeneans were recorded. The most prevalent parasites were metacercariae of digeneans in gonads (50.83%), gill ciliates (37.5%), and digestive gland viruses-like (37.5%). Only the prevalence of digeneans increased with host size, but any relationship between these variables was found in the other pathogens.

Parasites of the mangrove mussel Mytella guyanensis (Bivalvia: Mytilidae) in Camamu Bay, Bahia, Brazil

This contribution reports the parasites found in the mangrove mussel Mytella guyanensis in Camamu Bay, Bahia, Brazil. Samples were collected monthly from September 2006 through October 2007. A total of 460 individuals were collected, fixed in Davidson's solution, and processed by standard histological techniques, and the sections were stained with Harris hematoxylin and eosin (H&E). The water temperature ranged from 23.5 to 31.6 ºC, and the salinity from 25 to 37‰. Microscopic analysis showed Rickettsia-like organisms (RLOs), Nematopsis sp. (Apicomplexa), and Platyhelminthes, including a turbellarian, sporocysts of Bucephalus sp., metacercariae, and metacestodes of Tylocephalum sp. Parasites were observed mainly in the gills, mantle, and digestive gland. The prevalence of Nematopsis sp. was 100%, and in heavily infected mussels the tissues of the labial palps were damaged. RLOs occurred in high prevalence and intensity of infection in some periods. The digenean sporocysts showed moderate prevalence but high intensity of infection, and caused parasitic castration. In general, there was no significant spatial or temporal variation (p > 0.05) of the parasites, which is probably attributable to the small variations of temperature and salinity in the region.

Artificial Infection of the Bed Bug WithRickettsia parkeri

Although a variety of disease agents have been reported from bed bugs, the mechanical and biological disease transmission potential of bed bugs remains unelucidated. In this study we assayed survivability of the mildly pathogenic spotted fever group rickettsia, Rickettsia parkeri, in bed bugs after feeding on R. parkeri-infected chicken blood. Two groups of 15 adult bed bugs each were fed on infected or noninfected blood, and two groups of fourth-instar bed bugs also were fed on either infected or noninfected blood. One group of 15 adult bed bugs received no bloodmeal and was included as an additional control. Two weeks postfeeding, two pools of five live bed bugs from each group were surface sterilized, macerated, and placed in Vero cell cultures in an attempt to grow live organism. The remaining five individual bed bugs from each group were dissected, their salivary glands were removed for immunofluorescence assay (IFA) staining, and the remaining body parts were processed for polymerase chain reaction (PCR) analysis. Results indicated that no immature (now molted to fifth instar) bed bugs were positive for R. parkeri by IFA or PCR, indicating that organisms did not survive the molting process. After 4 wk of cell culture, no organisms were seen in cultures from any of the treatment or control groups, nor were any cultures PCR positive. However, two of the adult bed bugs were IFA positive for rickettsia-like organisms, and these two specimens were also PCR positive using R. parkeri-specific primers. These IFA and PCR results indicate that remnants of Rickettsia parkeri (possibly whole organisms) survived in the bugs for 2 wk, but the viability of the organisms in these two specimens could not be determined.

Identification and function of LPS induced tumor necrosis factor-alpha (LITAF) gene from Crassostrea ariakensis stimulated by Rickettsia-like organism

LPS induced tumor necrosis factor-alpha (LITAF) plays a role in regulating the expression of TNF-α. The present study was undertaken to identify a gene, Ca-LITAF, from oyster Crassostrea ariakensis challenged with rickettsia-likea organism (RLO). The full-length cDNA consists of 750 bp with an ORF encoding 115 amino acid residues. Amino acid sequence analysis revealed that Ca-LITAF shared conserved signature motifs with other LITAF proteins. The expression of Ca-LITAF was found in all examined tissues with a higher expression level in gills. The recombinant Ca-LITAF was successfully expressed in Escherichia coli BL21 cells and purified for the preparation of polyclonal antibody. The obvious expression of Ca-LITAF induced by RLO in hemocytes monolayers was revealed by western blotting using antibody against Ca-LITAF. These results suggest that Ca-LITAF is a LITAF homologue and may be involved in host immune responses against RLO.

Physiological performance of juvenile Haliotis rufescens and Haliotis discus hannai abalone exposed to the withering syndrome agent

Withering syndrome (WS) is a serious chronic disease caused by infection with the bacterium Candidatus Xenohaliotis californiensis, a Rickettsiales-like organism (WS-RLO) that affects multiple abalone species in both natural and farmed populations. However, there is no available information regarding the effects of this disease on the physiological performance of infected abalone. We studied the effect of different levels of infection on components of energy balance and physiological indices (rates of absorption and assimilation, O/N ratio, and scope for growth) in the abalone species Haliotis rufescens and Haliotis discus hannai. Juveniles were exposed to C. X. californiensis transmission for 130days, during which time the presence/absence of WS-RLOs was evaluated by PCR (following DNA sequencing-based confirmation of 100% identity with the sequence of C. X. californiensis from California), and the prevalence and intensity of infection were evaluated via histological analysis. Among H. rufescens juveniles exposed to the bacterium, 92% became infected (positive by histology), and the intensity of infection ranged from low (degree 1) to moderate (degree 2). In contrast, no H. discus hannai juveniles were positive for WS-RLO by histology, although 23% were positive by PCR, possibly indicating incipient WS-RLO infection that did not develop during the experimental period or to mere presence of WS-RLO DNA in the sample. Infection of H. rufescens juveniles by WS-RLOs negatively affected all components of the energy balance and physiological indices, such as scope for growth and the O/N ratio, in direct relation to the degree of infection. The most strongly affected functions were the rate of ingestion, standard metabolism, and production of feces, which were reduced by 60–80% in the most highly infected individuals. The reduced energy intake in the organisms produced a strong energy imbalance such that the energy available for growth was reduced by 49% in infected organisms. In contrast, juveniles of H. discus hannai carrying the bacterium developed no infection and showed no alterations of physiological function. Our results indicate that the level of early infection by WS-RLOs may exert a negative effect on physiological activity in H. rufescens, even when the disease is not evident.

Seasonal dynamics of zebra mussel parasite populations

AB Aquatic Biology Contact the journal Twitter RSS Mailing List Subscribe to our mailing list via Mailchimp HomeLatest VolumeAbout the JournalEditorsTheme Sections AB 15:145-151 (2012) - DOI: https://doi.org/10.3354/ab00418 Seasonal dynamics of zebra mussel parasite populations Laëtitia Minguez, Laure Giambérini* Laboratoire des Interactions, Ecotoxicologie, Biodiversité, Ecosystèmes (LIEBE), CNRS UMR 7146, Université de Lorraine, Campus Bridoux, Rue du Général Delestraint, 57070 Metz, France *Email: giamb@univ-metz.fr ABSTRACT: The seasonal dynamics of 8 parasite taxa in zebra mussels Dreissena polymorpha were studied in the Meuse River (NE France) over a 3 yr period. In total, 69.5% of sampled organisms were infected. Among observed parasite species, Ophryoglena spp. and to a lesser extent Rickettsiales-like organisms (RLOs) displayed seasonal dynamics, with lower prevalence during warmer months negatively correlated with gonadal development in the host, water temperature and phytoplankton concentration. Furthermore, this study highlights a positive correlation between the prevalence rates of Ophryoglena spp. and RLOs. No temporal variations were observed for the ciliates Sphenophrya dreissenae and Conchophthirus acuminatus. Concerning trematode infection, no defined seasonal pattern was observed. Thus, for more reliability in future biomonitoring studies involving zebra mussels, it will be important to include the influence of parasites and their temporal dynamics. KEY WORDS: Parasitism · Season · Dreissena polymorpha · Biomonitoring Full text in pdf format PreviousNextCite this article as: Minguez L, Giambérini L (2012) Seasonal dynamics of zebra mussel parasite populations. Aquat Biol 15:145-151. https://doi.org/10.3354/ab00418 Export citation RSS - Facebook - Tweet - linkedIn Cited by Published in AB Vol. 15, No. 2. Online publication date: May 15, 2012 Print ISSN: 1864-7782; Online ISSN: 1864-7790 Copyright © 2012 Inter-Research.

Effects of the toxic dinoflagellate Alexandrium catenella on histopathogical and escape responses of the Northern scallop Argopecten purpuratus

Juvenile Northern scallops Argopecten purpuratus were exposed to cultures of the paralytic shellfish toxin (PST) producing dinoflagellate, Alexandrium catenella, or a non-toxic microalga as a control, T-iso. After 3 and 6 days of exposure to either A. catenella or T-iso, scallops were stimulated to elicit an escape response by exposing them to the predatory sea star Meyenaster gelatinosus. We monitored the escape response of the scallops in terms of reaction time after first contact with the sea star, number of claps (burst of rapid valve closures) until exhaustion, clapping time, clapping rate, the time scallops spent closed when exhausted, and recovery from the initial number of claps, clapping time and clapping rate. Additionally, histopathological and stress responses (through heat-shock protein [hsp70] induction), as well as accumulation of Paralytic Shellfish Poisoning (PSP) toxins, were monitored on scallops after 3 and 6 days of exposure to A. catenella. After 6 days of exposure, scallops exposed to A. catenella accumulated PSTs and reacted more rapidly with a higher clapping rate, however the duration of their escape response was shorter than controls, when exposed to M. gelatinosus. Additionally, scallops exposed to A. catenella showed histopathological features, especially after 6 days of exposure, including increased melanization of the tissues and myopathy, with high levels of degeneration of the muscle fibers. A six-day exposure to A. catenella also caused an increase in prevalence of rickettsiales-like organisms within scallop tissues. This study suggests that PST accumulation can affect the interaction between the Northern scallop and both pathogens and predators, potentially increasing their susceptibility to either of them.

Identification of expressed genes in cDNA library of hemocytes from the RLO-challenged oyster, Crassostrea ariakensis Gould with special functional implication of three complement-related fragments (CaC1q1, CaC1q2 and CaC3)

A SMARTer™ cDNA library of hemocyte from Rickettsia-like organism (RLO) challenged oyster, Crassostrea ariakensis Gould was constructed. Random clones (400) were selected and single-pass sequenced, resulted in 200 unique sequences containing 96 known genes and 104 unknown genes. The 96 known genes were categorized into 11 groups based on their biological process. Furthermore, we identified and characterized three complement-related fragments (CaC1q1, CaC1q2 and CaC3). Tissue distribution analysis revealed that all of three fragments were ubiquitously expressed in all tissues studied including hemocyte, gills, mantle, digestive glands, gonads and adductor muscle, while the highest level was seen in the hemocyte. Temporal expression profile in the hemocyte monolayers reveled that the mRNA expression levels of three fragments presented huge increase after the RLO incubation at 3 h and 6 h in post-challenge, respectively. And the maximal expression levels at 3 h in post-challenge are about 256, 104 and 64 times higher than the values detected in the control of CaC1q1, CaC1q2 and CaC3, respectively.

Rickettsia-like organism infection associated with mass mortalities of blood clam, Tegillarca granosa, in the Yueqing Bay in China

A series of mass mortalities of the cultured blood clam, Tegillarca granosa, occurred in the Yueqing Bay of China from 2005 to 2009. An obligate intracellular prokaryote, designated as rickettsia-like organism (RLO), was frequently found in the moribund or dead blood clam sample during ultrastructural examination. These organisms were usually round, ellipsoid or occasionally dumbbell-shaped, ranged from approximately 0.28 to 0.71 µm in size and had a trilaminar cell wall. Two reproductive modes of organisms, transverse binary fission and budding, were observed. The organisms were able to form eosinophilic inclusions. Most inclusions were found within epithelial and connective tissues of the mantle, gills and digestive tube. The biological and morphological characteristics indicate that these organisms may belong to the family Rickettsiaceae. RLOs exhibited significant pathogenicity. Cytopathological examinations revealed extensive necrosis and destruction in the infected cell. The degree of tissue destruction was positively related to the number of RLO inclusions in the tissues, and the cytopathological effects were positively related to the number of intracellular RLO. RLOs and their inclusions were discovered throughout different disease areas and in different time periods. The infection intensity of the RLOs was positively correlated with the mortality rate of clams. Therefore, RLO infection might be associated with mass mortalities of cultured blood clams in the Yueqing Bay.

Is there a link between shell morphology and parasites of zebra mussels?

The shell morphology of zebra mussels, Dreissena polymorpha, was analyzed to determine if alterations in shell shape and asymmetry between valves were related to its infection status, i.e. infected or not by microparasites like ciliates Ophryoglena spp. or intracellular bacteria Rickettsiales-like organisms (RLOs), and by macroparasites like trematodes Phyllodistomum folium and Bucephalus polymorphus. For microparasites, two groups of mussels were observed depending on shell measurements. Mussels with the more concave shells were the most parasitized by ciliates. This could be more a consequence than a cause and we hypothesized that a modification of the water flow through the mantle cavity could promote the infection with a ciliate. There were more RLOs present in the most symmetrical individuals. A potential explanation involved a canalization of the left–right asymmetry as a by-product of the parasite infection. Trematode infections were associated with different responses in valve width. Females infected by P. folium displayed significantly higher symmetry in valve width compared with non-infected congeners, whereas the infection involved an opposite pattern in males. B. polymorphus was also linked to a decrease in valve width asymmetry. This study suggested that a relationship exists between parasitism and shell morphology through the physiological condition of host zebra mussels.

Cross-effects of nickel contamination and parasitism on zebra mussel physiology

Aquatic organisms are exposed to pollution which may make them more susceptible to infections and diseases. The present investigation evaluated effects of nickel contamination and parasitism (ciliates Ophryoglena spp. and intracellular bacteria Rickettsiales-like organisms), alone and in combination, on biological responses of the zebra mussel Dreissena polymorpha, and also the infestation abilities of parasites, under laboratory controlled conditions. Results showed that after 48 h, more organisms were infected in nickel-exposed groups, which could be related to weakening of their immune system. Acting separately, nickel contamination and infections were already stressful conditions; however, their combined action caused stronger biological responses in zebra mussels. Our data, therefore, confirm that the parasitism in D. polymorpha represents a potential confounding factor in ecotoxicological studies that involve this bivalve.

Quantitative PCR demonstrates a positive correlation between a Rickettsia‐like organism and severity of strawberry disease lesions in rainbow trout, Oncorhynchus mykiss (Walbaum)

Strawberry disease (SD) is an inflammatory skin disorder in rainbow trout, Oncorhynchus mykiss (Walbaum). The aetiology of SD is unknown although the 16S rDNA sequence of a Rickettsia-like organism (RLO) has been associated with SD lesions using a nested PCR assay. In this study, we developed a Taqman quantitative PCR assay (qPCR) that targeted the RLO 16S rDNA sequence to examine the distribution of RLO relative to lesion status. We compared 18 lesion samples from 13 fish representing high or low lesion severity as judged by gross examination. QPCR results showed that there was a higher number of RLO sequences in high severity lesions (mean of 12,068 copies) compared with fewer copies of RLO sequence in low severity lesions (mean of 3287 copies, P = 0.012). Grossly normal skin samples (n = 13) from SD-affected fish were all negative by qPCR except two samples (121 and 139 copies). The qPCR assay described herein is a useful tool to investigate the role of RLO in SD in the absence of a culture system for RLO. Our results demonstrate a positive correlation between copy number and lesion severity consistent with the hypothesis that the RLO is the aetiologic agent of SD.

Pathologies in commercial bivalve species from Santa Catarina State, southern Brazil

The State of Santa Catarina in southern Brazil is the most important Brazilian producer region. The Pacific oysters Crassostrea gigas and the brown mussels Perna perna are the main species currently produced. The occurrence and impact of parasites on bivalves from natural or culture populations from Brazil are still poorly studied. This work describes several diseases, parasites and commensals from four edible bivalve species, from cultured and exploited stocks, from five sites at Santa Catarina State. Unspecific histopathological symptoms of stress, parasites and commensals were found, including viral gametocytic hypertrophy and rickettsia-like organisms with very low prevalences; the protozoan Nematopsis sp. with high prevalence and, ovarian parasites with low prevalences; and, diverse metazoans, such as turbellarian, sporocysts and metacercariae of trematodes, metacestoda larvae and spionid polychaete. Ray's fluid thioglycollate medium assays for detection of Perkinsus spp. did not detect any infected bivalves. Polymerase chain reaction assays for Marteiliodies chungmuensis in oysters with ovarian lesions produced negative results.

A new blood parasite within the relict endemic New Zealand gecko Hoplodactylus duvaucelii

Duvaucel geckos ( Hoplodactylus duvaucelii) are large endemic lizards that have been extirpated from the New Zealand mainland due to introduced mammalian predators. This species has subsequently become the subject of species translocation conservation management in an endeavour to increase the number of populations and as a part of island ecological restoration. Blood sampling of a captive adult male after tail autotomy led to the discovery of a Rickettsia-like organism within this gecko's erythrocytes. We conclude that this infection was acquired at the gecko's source location, a remote island closed to the general public and lacking potential invasive parasite reservoir reptiles. The likely vector is the native mite Geckobia naultina. This finding represents valuable new baseline information about the health parameters of this threatened species. Particularly in the light of the paucity of reported blood parasitism in New Zealand reptiles, the conservation management of this species through relocation and captive-breeding, and the on-going concerns regarding the introduction of novel parasites to New Zealand.

Association of red‐mark syndrome with a Rickettsia‐like organism and its connection with strawberry disease in the USA

Red-mark syndrome (RMS), a disease seen mostly in rainbow trout, Oncorhynchus mykiss, is of unknown aetiology. The research presented here indicates the presence of an intracellular bacterium in RMS-affected fish. A positive reaction was observed using immunohistochemistry (IHC) with skin lesions, liver, kidney and spleen of affected fish sampled from several locations within the United Kingdom using two different polyclonal antisera raised against Piscirickettsia salmonis. The same reaction was also seen with a number of different anti-P.salmonis monoclonal antibodies (MAbs). A disease with similar clinical signs to RMS, referred to as strawberry disease (SD), has been reported in the USA. A Rickettsia-like organism (RLO) has recently been associated with SD based on analysis of 16S rDNA sequences. Using the same panel of anti-P.salmonis antibodies used to screen the RMS samples, similar staining was obtained in tissue of SD-affected fish by IHC. A polymerase chain reaction (PCR) using RLO-specific primers was also performed on RMS-affected fish from the United Kingdom, and the samples were positive for the RLO 16S rRNA sequence. These findings suggest that the same aetiological agent may be responsible for RMS in the United Kingdom and SD in the USA.