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Pergamon

Annual Review of Fish Diseases, Vol. 6, pp. 3-13, 1996 Copyright 0 1997 Published by Elsevier Science Ltd Printed in Great Britain. All rights reserved 0959-8030/97 $32.00 + 0.00

PII:SO959-8030(96)00001-4

RICKETTSIAL INFECTIONS OF FISH*
J. L. Fryer? and C. N. Lannan
Department of Microbiology, Oregon State University, Nash Hall 220, Corvallis, OR 97331-3804, USA

Absrract. Pisciricketfsia salmonis is the first of the previously unrecognized rickettsial pathogens of fish to be isolated, characterized, and demonstrated to be the etiologic agent of an epizootic disease. However, since the isolation of P salmonis in 1989. the scope and impact of these pathogens has become increasingly apparent. This growing awareness of the role of the rickettsiae in fish health has
led to documentation of rickettsial diseases in diverse species of fish and in disparate geographic locations and aquatic environments. Considerable work remains in order to establish the source, reservoir, and normal mode(s) of transmission of these agents and to devise appropriate methods of disease prevention and control. 0 1997 Published by Elsevier Science Ltd

Krywords: Rickettsia, Fish disease, Piscirickertsia salmonis, Piscirickettsiosis

INTRODUCTION

Aquatic animals are host to a wide variety of intracellular rickettsia-like and chlamydia-like Gram negative bacteria (1). Of these, only the diverse group of chlamydia-like organisms responsible for epitheliocystis, although never cultured or characterized, have traditionally been considered pathogens of fish. However, in recent times, this limited view of the role of intracellular bacteria in fish disease has dramatically changed, and rickettsial agents have assumed an important place among those bacterial pathogens with significant impact on fish health.

HISTORY

The role of the rickettsiae in fish disease became apparent in 1989 (2), when a previously unrecognized rickettsia [Piscirickettsia salmonis gen. nov., sp. nov., (3)] was isolated in cultures of the chinook salmon embryo cell line, CHSE-214 (4), and demonstrated to be the cause of devastating epizootics in marine-netpen-reared coho salmon Oncorhynchus kisutch in southern Chile (5). It was originally thought that the infections were specific for coho salmon, but subsequently, piscirickettsiosis was also observed in the other salmonid species cultured in the area [chinook salmon Oncorhynchus tshawyfscha, Atlantic salmon Salmo salar, rainbow trout Oncorhynchus mykiss , and masu salmon Oncorhynchus masou; (6)]. Piscirickettsia salmonis was the first rickettsia of aquatic poikilotherms to be isolated, characterized, and demonstrated to produce disease in fish. On two earlier occasions, rickettsialike agents were observed in non-salmonid fish species at diverse locations and in differing aquatic environments around the world (7,8) and one such agent from rainbow trout was isolated in cell culture (9), but none of these organisms was identified or characterized and their role in the fish disease process remains unknown. Since 1989, a growing number of reports have documented rickettsial diseases in fish (Table 1). Currently, there are at least 50 publications that detail one or more aspects of the infection of
*Oregon Agricultural Experiment Station Technical Paper 10,846

tTo whom correspondence should be addressed.

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Of these. This disease was caused by an organism morphologically identical to F! salmonis. Following the identification of I! sulmonis in Chilean salmonids (2) similar infections were identified in salmonid fish at three locations in the northern hemisphere. These researchers also reported that an analogous disease occurred. We believe the trend toward formal documentation of the role of the rickettsiae in fish health will and should increase. However. fewer than half appear in the peer-reviewed literature (Table 2). d I = isolated. c 0 = observed. it produced only low level mortality in the infected fish. Rickettsial and rickettsia-like organisms observed in or isolated from fish Host species Tetrodon fahaka Geographic Region Egypt Salt/Fresh Water F Isolated/Observed 0’ Reference (7) Rainbow trout (Oncorhynchus mykiss) Europe Wales Chile Chile F Sb S s Id 0 I* o’ * (9) 03) (2) (5) Dragonet (Callionymus lyra) Coho salmon (Oncorhynchus kisutch) Rainbow trout Chinook salmon (Oncorhynchus rshawyrscha) Atlantic salmon (Salmo salar) Chinook salmon Atlantic salmon Coho salmon Pink salmon (Oncorhynchus gorbuscha) Pacific Coast Canada S O- (29) Atlantic salmon Atlantic salmon Sakura salmon (Oncorhynchus masou) Ireland Norway Chile Taiwan Colombia Chile Chile Japan France S S S S&F F F F F S 0* oI* 0 1 0 1* I 0 0 (12) (11) (6) (13) (14) (16) (17) (30) (15) Tilapia spp. in pink salmon Oncorhynchus gorbuscha. but unlike the P safmonis infections in Chile. at least 20 peer-reviewed papers on this topic can be found. held in seawater tanks in British Columbia. Rickettsiae morphologically similar to those implicated in the diseases in Chile and in Canada also caused low level mortality in Atlantic salmon held in .N. Blue-eyed plecostomus (Panaque surtoni) Coho salmon Rainbow trout Atlantic salmon Tilapia nilotica x Oreochromis nilotica Sea bass (Dicenrrarchus labrax) a F = fresh water. as early as 1970. Brocklebank er al. and at present. an increasing number of scientific papers concerning rickettsial infections in fish are published in journals subject to peer review. Lannan Table I. e = subsequently isolated. b S = salt water. (10) found a disease in maricultured Atlantic salmon that resulted in pathology indistinguishable from that observed in the salmonids in Chile.L. In the western Canadian province of British Columbia. fish by rickettsial or rickettsia-like organisms (Tables 2 and 3).4 J. and in coho and chinook salmon seafarmed there in the 1980’ s. Fryer and C. * = identified as Piscirickettsia salmonis.

M.L. Fryer. all documented rickettsial diseases of fish occurred in salmonids cultured in sea water. L. H.. E. P. Hokkaido University Press.M. Dennis. L. (1994). Fish. C..A. Larenas. J. E. Bactetiol.H.. nov. Fryer.J. J.L.L. Rickettsia-like organisms in the blue-eyed plecostomus. (1992).A.H.like agent in farmed Atlantic salmon. C. Extracellular survival of Piscirickettsia salmonis.-F. J. 29(2):61-71. Garcbs. (ed. Tung..N. nov.] Fryer. A. aquat. 17:115-121. C. Drinan.. C. Rodger.M.. Bakt.N. and crustaceans. G.D. Games A. Schwartz-Pfitzner.-P. Khoo. Fish Dis.. salmonis .J.L. Larenas H.L. Lannan. 1. Smith.. Oreochromis niloticus. H.N. (1994). Lannan. Dis. Lannan. J.J.. Like the ongoing epizootics in Chilean salmonid fish.-C. A. Games. Lewbart. Abt Orig. L. (1993). Branson. P. In: Kimura. Until 1994. Development of polymerase chain reaction assays for detection. J. (1994) Rickettsial and chlamydial infections of freshwater and marine fishes.N.. A. An immunohistochemical diagnostic test for rickettsial disease. (In Spanish with English abstract) Avarices en Ciencias Veterinatias 10(1):53-58. C..-C.L. J. 14:147-156. S&to solar L. Lannan. Ozel. T. Fish Dis. Fish Dis. J. M. J. Oreochronuis niloticus (L. Description of a new disease condition occurring in farmed coho salmon. Dis. (1992). Zbl. R. 18:157-164. (1994).. J..D.L. Vet. (1993). Fryer. C... Fryer.H. Wood. All of these agents were identified as F! by fluorescent antibody tests using a polyclonal antiserum. Fish Dis.Egtved-Virus (VHS) der Regenbogenforelle (Salmo gairdneri) und Rickettsienahnliche Organismen. Wapia zillii.. 17:545-548.. S. Tsai.) Salmonid Diseases. Giovannoni. J. C.N. Fryer. [Translated into Japanese by Professor Tokuo Sano and appears in Fish.. Fryer.. the causative agent of an epizootic disease in salmonid fishes.E..-S.. Evelyn T. (1991). Org.).B. J. Cvitanich. and Tilapia hornorum) in both freshwater and seawater ponds in Taiwan. C. 230:1-14. and rainbow trout. Rodger. Giovannoni. Wang. (1995) First isolation of Piscirickettsiu salmonis from coho salmon. Sandino. J. Latenas. (1990) Isolation of a rickettsiales-like organism from diseased coho salmon Oncorhynchus kisutch in Chile... l1:93-97. S. (1991).. C. systemic granulomas caused by a rickettsia-like organism in Nile tilapia. Aquat. Armstrong. aquat. J. However. J.D. Reports on rickettsial and rickettsia-like pathogens of fish: The peer-reviewed literature Alday-Sanz. J.D. identification. Org. Smith.L. 16:361-369. Observation of a rickettsia-like organism in Atlantic salmon. Lannan. D. Fish Dis.-C.S. Smith S. Hakodate. (1991).N. C. H. J. Richards. Chen. Septicemia suspected to be caused by a rickettsia.. Fryer.. Chen. pp. Oncorhynchus kisutch (Walbaum).. Lin. (1994). M... Garate N. Adams.. Int. Res. Oncorhynchus mykiss (Walbaum).N.J. J. D. bivalves. in South America.. and differentiation of Piscirickettsia salmonis. Zool. Smith. 42(1):120--126. N. l4:12l-145.. R... during the freshwater stage of their life cycle. (1975). from southern Taiwan. Gaggero.. Lannan. The isolation of a rickettsia-like organism causing disease and mortality in Chilean salmonids and its confirmation by Koch’ s postulate. Mauel. seawater cages in Norway (11) and in Ireland (12). 17591-599. J. sp. J... Outbreaks of a disease caused by a rickettsia-like organism in cultured tilapias in Taiwan. J... J. Lannan. H. (1995) 14(3):54--65..A.. Isolation of a rickettsia from diseased salmonids. (1991). Speare. Can. J.. (1995). (1996). Sandino. 1. 26: 189-195.... 25(2):107-l 14.J.. Adams. Fryer.Rickettsial infections of fish 5 Table 2. Oreochromis aureus. Nieto Diaz-Munoz. J. 17:189-191. Syst. Pannque suttoni (Eigenmann & Eigenmann). J. Chern and Chao (13) found an epizootic rickettsial disease in several species of tilapia (Oreochromis mossambicus. J. S.N. Res. Hidalgo V. Lannan.R. Turnbull. Fish Pathol. Studies 33(2):95-107..-C. Chem.. J.. Chen. 151-157. J. Ewing.. S. 33:407-408. Piscirickettsia salmonis gen. Vergleichende elektronenmikroskopische Untersuchungen an Rhabdoviren pflanzlicher und tierischer Herkunft: 111. l&277-279.. Infectivity of a rickettsia isolated from coho salmon (Oncorhynchus kisutch).J. Chao.. Hyg..L.. 0. A. (1992). Animal Hlth 3:229-234.A. P. V. A fluorescent antibody test for detection of the rickettsia causing disease in Chilean salmonids. Oncorhynchus kisutch (Walbaum). Fish Dis. S.. S. Fryer.. J. Brocklebank. S. T.L. Fish Dis. Fish Pathol. P. M. Castro. R. Piscirickettsiosis: Lesiones en salm6n del Atlarttico (S&no s&r) infectados naturahnente con Piscirickettsia salmonis. P. R. it is now apparent that these pathogens affect fish over a broad host and geographic range and in both freshwater and marine environments.. Fish Dis. L. (1995). Piscirickettsio sohnonis. a major pathogen of salmonid fish in Chile. in Ireland.

. M.L. Quart. Animal Health..N. Dubuque. ‘ Diseases of Fish and Shellfish”.15. 214. B. Plassiart.. 1994. Aquat. E. ID. Pamdes. In: Short Communications of the 1991 International Marine Biotechnology Conference (IMBC ‘ 91). (Abstract) American Society for Rickettsiology and Rickettsial Diseases 12th Sesqui-Annual Meeting. Evidence of genetic diversity within Piscirickertsia salmonis..J. C. Smith. (1995)... Brocklebank. WA. Palma de Mallorca. Spec. Sot. V. Silva P. Pacific Grove. A.. Fish Pathol. FHS/AFS Newsletter 18(1):1-2. Assoc. S. M. S. Alberdi. Pacific Grove.. Animal Health. First report of Piscitickeftsia salmonis in freshwater.D. Pathol.. (1992).. Symp..E. Evensen. June 6-9. 1996. J. C. (1995). Bull. 1995. 116. L... C. J.L. (1986). C.P.N. Gaggero.. Fryer. Reports on tickettsial and rickettsia-like pathogens of fish: The non-peer-reviewed Literature Anonymous (1991). Serv. Sept. 1995. pp. 7th Intemat. Isolation of a new rickettsia-like organism from Atlantic salmon in Chile. Pacific Northwest Fish Health Protection Committee Informational Report 6.L. C. Conf. Smith. Larenas. Alberdi. Actual situation of the disease produced by Piscirickerrsia salmonis in Chile. Union Microbial. Bull.. J. Tenth Region” (Chile). Lannan Table 3. (Abstract) Eur. C.. Isolation of a new rickettsia-like organism from Atlantic salmon in Chile. M. Recommended methods for inspection of fish for the salmonid rickettsia. Eur.. Speilberg.N.E. &rate N.L. J. Cubillos. (1995). Fryer. 11(3):5. Andrade V. Seattle. Evaluation of the in vitro sensitivity of Piscirickettsia salmonis to diffetent antimicrobial compounds. Animal Viruses. 0. Cubillos. In: Kent. (1991).P. Newsletter. J. Sci.. Bull. T. Sept+8. Sept. Lannan. Olsen. T. Piscirickettsiosis. Septicemia in farmed Atlantic and chinook salmon due to a rickettsia-like agent. M. Aquat.D.. Palma de Mallorca. Speare.. 135-140. R.. WA. W. (Abstract) Western Fish Disease Workshop. 1240-41.H. Davies. Seattle. salmonis. Fish.D. J. 1996. A. Oyanedel. H..N. Finding of Pisciricketrsia salmonis on Cerarhorhoa gaudichaudii. P. “Diseases of Fish and Shellfish”.N. S. P. a newly described intracellular pathogen of salmonid fish.R. Larenas. Berlin. 16(1):3&33. (1996). Bravo. (Abstract) Eur. M. Agric. Spain Garces. Lannan. T. A rickettsia-like organism from dragonets. J. Alvarado. (1990). Fryer.A. Smith S. (1995). 11:135-136. (Abstract) Int.. (1996). Schafer. Symp. France.. Fryer. Patologfa Animal 4:14-17. (Abstmct) In: VIII* International Congress of Virology. pp. Developments in Industrial Microbiology Extended Abstract Series Vol.. Fish Pathol. Mauel... (1993). C. Fish. The emergence of rickettsia as important pathogens of fish. J. FHSlAFS Newsletter 20(l): 1. Sci. A. Aquat. May 10-13.P. Spain Cvitanich. Sept.D.. Eur. Salmonid rickettsial septicemia.. p. Comps. FHS/AFS Newsletter 17(3):3.. (ed.A.. Nanaimo.B.L. G. Rickettsia-like organism infecting juvenile sea-bass Dicentrarchus labrax. Office/ American Type Culture Collection International des Epizooties Paris. Lannan. <<NY>> laksesykdom for&r&et av rickettsie.L. R. Eur. 7th Intemat. (1995). Vet.T. Moscow. (1994). (Abstract) Int. The discovery of a rickettsia in a fish. J.N. May 10-13. Monras.J. Norsk Fiskeoppdmtt NR. 1 lpp. 49. L. Figueroa P. Campos.Caractetisticas anatomopatolbgicas de1 sindrome de1 salmon coho (SSC). J. M. 7th Intemat. Bull.. C... Fisheries and Oceans. “Diseases of Fish and Shellfish”.. A. C. Bravo. Melby. Conf. (Abstract) American Society for Rickettsiology and Rickettsial Diseases 12th Sesqui-Annual Meeting. T. Garate N. FHS/AFS Newsletter 23(3):1-3. Fryer and C. 1st edition. D. (1996). Fryer. Lannan. Dept... Garate N. CA. V.O. Lannan.J. Conf. C.. Enriquez.. Assoc. 6pp. (1991). Tech.4-8. Fish Pathol. Raymond.L. 14(4):137-138. (1990). A. (1995). Spain Evelyn.J.C.J. Hhtein. Preliminary characterization of a rickettsia-like pathogen of salmonids. Evelyn. Far& C.N. Callionymus lyra L.. J.. (1989). Cvitanich.18-19. P. Cairo. C. Etiological agent in a Chilean coho disease isolated and confirmed by Koch’ s postulates. pp. Piscitickettsiosis: Histopathology of Atlantic salmon (Salmo salar) and coho salmon (Oncorhynchus kisurch) inoculated with P.6 J. (1992). Mohamed. Pathol. 10-15. (Teleostei: Callionymidae) in Wales.. Rickettsial disease of salmonids. Cvitanich. P. II.. Minist. Fish Pathol. Evelyn. Garc&. C.J. S..567-572. Assoc. 2. IO. (1994). Intemat. Lamnas. Gar&.T. 1995. Pub.) Diseases of Seawater Netpen-reared Salmonid Fishes in the Pacific Northwest. 1994. Smith.D.. Monras.H.N. J. Cvitanich. Assoc. Smith. (1995) “Salmonid rickettsial syndrome agent determination (SRSAJA) in wild fishes from the province of Palena.. J. (conrinued) . (1939). Piscirickettsiosis in freshwater.H.C.E.. Wm... L. lC-15. Assoc. Assoc.. J... 6(4):103. Palma de Mallorca. Assoc. lA... Can. Coho salmon syndrome in Chile. 0.L. nueva enfermedad de 10s salmonfdeos. Contreras. Fish. V. Brown Pub.L. (Abstract) Eur. J. CA. Fryer... Fryer. Armstrong. L. J.. M.. Bravo. Sect. Cormal. (1992). Fish Pathol. Piscirickerrsia salmonis. Lannan. Anonymous (1995). Bull. Eur. Smith. (1990). Sept. FHSlAFS Newsletter 22( 1):3. In: OIE Diagnostic Manual for Aquatic Animal Diseases. J.O. 1995.

Sot. (16) isolated J? salmonis from diseased coho salmon and rainbow trout held only in fresh water. (Conrinued) Schafer. reared in sea water.5). S. Khoo et al. It has been deposited with the American Type Culture Collection as ATCC VR 1361. It is taxonomically placed in the order Rickettsiales. Fish Pathol. Although all of these isolates are morphologically similar to LF-89 (Fig. (17) reportedly does not react with polyclonal antibodies against E salmonis.. A disease with liver swelling and nodule formation in cultured tilapia. this disease resulted in excessive mortality. Larenas. The rickettsia-like organism isolated by Cvitanich et al. shipped to the United States from Colombia...A.N. and a new genus and species described by Fryer et al.2-0.. J. P. (199. like P salmonis. however.. monoclonal antibodies differentiate among them (unpublished data). Taxonomic placement of these agents awaits further study. M. Shiomitsu. 10:130. Fryer. Monds. Bull. Assoc. Smith. J. 15(4):137-141. 1995. PISCIRICKETTSIA SALMONIS Taxonomic position Piscirickertsiu sulmonis is the best chyacterized of the rickettsial agents observed in and/or isolated from fish. Japan Wolf. and the remaining rickettsia-like organisms observed in and/or isolated from fish have not been characterized sufficiently to determine their relationship to LF-89.. Many of the rickettsial agents observed in or isolated from fish since 1989 have been serologically identified as E sulmonis (Table 1). was isolated from a diseased coho salmon collected from a seawater netpen in southern Chile where an epizootic was in progress. In Chile. March 30-31. Fish Pathol. Chlamydia and rickettsia of fish. Comps et al. LF-89. Gaggero et al.Rickettsial infections of fish 7 Table 3. Whipple. the family Rickettsiaceue. It was considered by the authors to be rickettsia-like. V.) and react with polyclonal antibodies against the type strain..5pm in diameter). K.H. Games. C. Eur. we have tested isolates from Chilean coho salmon and rainbow trout and from Atlantic salmon reared in Chile. Also in Chile. a different and unidentified bacterium was isolated from diseased Atlantic salmon reared exclusively in fresh water. Fish Health News 10(3):1-5. (15) found rickettsia-like organisms in the brain of moribund juvenile sea-bass (Dicentrurchus l&-ax) exhibiting abnormal swimming behavior.. Caswell-Reno. Alvarado. Wada. (1990). OS-1.L. Jarpa. J. .. Enriquez. (Abstract in Japanese) 1995 Spring Meeting Japan. Assoc. the blue-eyed plecostomus Punuque suttoni. Canada and Norway.. M. 1. L. The “coho salmon syndrome” (CSS): a new disease in Chilean salmon. (3). The type strain. Eur.. In France. reaching 95% in some cases. because it was observed within the cytoplasm of kidney and spleen cells of infected fish and could not be cultured on seven unnamed bacteriological media (17). The organism. Bull.W. and showed no reaction when tested with FITC-labelled polyclonal antiserum prepared against e salmonis. K. Piscirickettsiosis: A bacterin field trial in coho salmon (Oncorhynchus kiturch). In our laboratory. (1995). (1981). P. Hatai. (14) observed rickettsia-like organisms in moribund specimens of a tropical freshwater fish.8pm vs ca. Fish Pathol... Lannan. Tokyo. T. M. R. 0. was isolated in fish cell cultures but was reportedly smaller (ca.

N. Lannan Fig.8 J. 1.L. Fryer and C. Caption opposite. .

LF-89.5-l . retarded above 20°C and below 10°C and does not occur above 25°C. Varied clinical signs accompany p sulmonis infection. It is sensitive to a broad range of antibiotics but not to penicillin. C. Isolate EM-90 from Atlantic salmon in Chile. It peaks in the fall of the year and rises again the following spring (19). obligately intracellular bacterium.20). but few are specific to piscirickettsiosis.5 pm in diameter. The titer is decreased by 99% or more by one cycle of freeze-thaw at -70°C but the addition of 10% DMSO to the freezing medium provides a cryopreservative effect. The type strain. When Garcts et al. 0. Isolate ATL-4-91 from Atlantic salmon Salmo salar in Canada. skin hemorrhages and lesions are among the first signs observed. Internally&e kidney is swollen and the spleen enlarged. Mortality in the seawater netpens begins 6-12 weeks after the transfer of fish from fresh water (2). (2). but the greatest losses are registered in coho salmon cultured in salt water. Characteristics of the type strain LF-89 Piscirickettsiu salmonis type strain LF-89 is a non-motile. D. Piscirickettsiu safmonis is cultivable in fish cell lines where it produces a cytopathic effect. All species of salmonids cultured in Chile are affected by this disease. Areas of necrosis and inflammation are found throughout. it displays a typical Gram-negative cell wall and the protoplasmic structure of a procaryote. B. anorectic. It is Gimenez-negative (21) but retains basic fuchsin when stained by Pinkerton’ s method for rickettsia and chlamydia (22). It replicates within membrane-bound cytoplasmic vacuoles in cultured fish cells (Fig. 1A) and in the cells of tissues throughout infected fish.06%) in Canada (10) and Norway (11). The gills are pale and hematocrits low (25% or less). The mortality attributed to piscirickettsiosis in salmonids ranges from extremely high (up to 90%) in Chile (18) to low (up to O-. Petechial hemorrhages are found on the swim bladder and viscera. but it does not replicate on any known cell-free media. A. liver. I. and dark in color (10). kidney. Epithelial hyperplasia results in lamellar fusion in the gills. Isolate SLGO-94 from rainbow trout Oncorhynchus mykiss in Chile. isolated from Chilean coho salmon Oncorhynchus kisutch. Diagnostic ring-shaped. but in acute cases. It is predominately coccoid. These individuals are lethargic. In thin section. cream-colored lesions are present on the livers of chronically infected fish. mortality ranged from 88 to 100% over the 42 days of the experiment. The in vitro growth characteristics of this agent were described by Fryer et al. Bar = 1 &km. 18. 19. These lesions range from small areas of raised scales to shallow ulcers up to 2 cm in diameter (18). Rickettsiae are commonly observed within macrophages and in the cytoplasm of other infected cells. Note dividing cells (arrows). especially adjacent to blood vessels. E. Briefly. . Piscirickefrsia salmonis replicating within cytoplasmic vacuoles in cultured CHSE-214 cells. Gram-negative. mortality may be the only gross sign of disease observed. with the most severe changes in intestine. Replication is optimal at 15-18°C. It replicates to titers of lo6 to 10’TCIDs. ca. Moribund fish collect at the surface of the water along the edges of the sea cages. piscirickettsiosis produces pathological changes in most internal organs of affected fish. (5) injected ten-fold dilutions of spent medium from an LF-89-infected cell culture into groups of 40 juvenile coho salmon and groups of 30 juvenile Atlantic salmon.Rickettsial infections of fish Piscirickettsiosis 9 Piscirickettsiu salmonis produces an epizootic fish disease. Isolate NOR-92 from Atlantic salmon in Norway. and it stains dark blue with Giemsa. Frequently. The histopathology associated with /? sulmonis infection has been described by various authors (10. It does not react with a monoclonal antibody made against the group-specific chlamydial LPS antigen (2). All of the typical disease Fig. piscirickettsiosis. but also occurs as rings or pairs of curved rods. and spleen. ml-’ in fish cell cultures.

provide the source and reservoir of the rickettsiae infecting cultured salmonids in these diverse areas. With the exception of the genus Coxiella. ml-‘ ) immediately after suspension in fresh water. It must also be noted that the extended extracellular survival time of this organism in salt water (23) may be sufficiently long to permit horizontal transmission in the marine environment without the requirement for a vector. No common thread. salmonis (25). Games et al.N. Although piscirickettsiosis has been observed in fish over a widespread geographic area (Table l). isolation of an agent sensitive to low levels of .L.g. dying of piscirickettsiosis. Mode of transmission The source. reservoir. an intermediate host or vector is required for transmission of rickettsial disease agents in the terrestrial environment (24). beyond the marine environment itself. the almost immediate deactivation of the rickettsia in fresh water makes direct horizontal transmission unlikely under those conditions. Detection and identification Inoculation and observation of susceptible cultured fish cells provides the most sensitive method of detection of P. Limited studies designed to demonstrate horizontal transmission have had varied outcomes. (19) reported horizontal transmission between injected and sham-injected individuals in a low number of coho salmon held in static freshwater and seawater aquaria. recurring epizootics in southern Chile. but the identity of the alternate host(s) remains unknown. It seems likely that one or more naturally occurring aquatic animals. Norway. few fish farms are affected in each region where they occur. and at times. perhaps transiently present. reservoir. and LF-89 was reisolated from moribund fish in each inoculated group. and the low number of reported infections in fresh water would indicate that vertical transmission. and normal mode of transmission of R salmonis are open to conjecture. (5) detected no evidence of horizontal transmission between injected coho salmon. vertical.5”C vs. Lannan signs were present in the inoculated coho salmon. Fryer and C. but Cvitanich et al. has not been demonstrated. Titers dropped below the level of detection (lo* TCID. However. must be a rare event. and transmission of l? salmonis remain questions of pressing importance. if it occurs. Although rickettsiae are found in the gonads of infected fish. but an intermediate may not be required for delivery of the pathogen in the aquatic environment. However. horizontal transmission can take place. the infections are sporadic.10. the mortality in both species followed a clear dose-response pattern. and no such host or vector has been identified for I! salmonis. salmonis has not been demonstrated. but infectivity was rapidly reduced in preparations suspended in fresh water. and Ireland. the questions of source. flowing water at a mean temperature of 10. mortality was the only gross sign of disease observed. whether horizontal. with the exception of the extensive.10 J. Until definitive studies are conducted. 19) but the normal mode(s) of transmission for this agent. Piscirickettsiosis has been experimentally induced by injection (5. e. In experiments designed to test the extracellular survival of LF-89 under varied environmental conditions. static water at 15°C making the results difficult to compare. Lannan and Fryer (23) found that infectivity remained for at least 14 days in preparations of semipurified LF-89 suspended in high salinity sea water at 5. and 15”C. or through a vector. but they suggest that under some circumstances. but in the Atlantic salmon. vertical transmission of P. However. and uninjected individuals held in a cage in the same tank of flowing fresh water.. Environmental conditions differed between the two sets of experiments. and no infectious material could be recovered from these preparations at that time or at any time thereafter during the two-week experimental period. is readily apparent between sites in Canada.

Definitive studies must be conducted to clarify the normal mode(s) of transmission of P salmonis if an understanding of the pathogenesis of piscirickettsiosis is to be attained. It is unknown whether the rickettsia-like agent from tilapia is a new biotype of I? salmonis. erythromycin.g. In addition. e. Future research Many vital questions concerning ?? salmonis and the disease it causes remain unanswered. or a new species of rickettsial fish pathogen. and development of an efficacious vaccine to protect valuable stocks of fish is the subject of considerable research effort. careful adherence to good fish culture procedures is the only tool left to fish culturists for control of this disease. Giemsa. immunofluorescence (26) or immunohistochemistry (27). with confirmation of identity provided by serological methods. thereby enhancing the likelihood of contamination. or acridine orange-stained kidney imprints or smears. With no vaccines. and the infectivity of F! salmonis for native.Rickettsial infections of fish 11 many of the antibiotics routinely used in cell culture presents a considerable challenge. methylene blue. no efficacious preparations have yet been developed to protect fish against this infection. Beyond the expansion of basic knowledge about I? salmonis and piscirickettsiosis. aseptic collection of diagnostic specimens in the field is not readily accomplished. preliminary diagnosis of piscirickettsiosis is normally made by examination of Gram. These methods must be easier than isolation but more sensitive and accurate than current staining procedures. The apparent difference in virulence of P salmonis in Chile and that observed in salmonids elsewhere is intriguing. the difficulties associated with isolation of P salmonis in cell culture or detection of the agent in stained preparations require that improved methods of detection and diagnosis be developed. It is critical that the source of the infectious agent be identified if the spread of piscirickettsiosis is to be controlled. Antibiotic levels may not reach lethal concentrations within host cells in vivo. nonsalmonid fish should be investigated. Much effort must be directed toward a search for the reservoir of infection in both the marine and freshwater environments. It must be determined if intrinsic differences in the rickettsial isolates. attention is necessarily centered on prevention of the disease. Finally. tetracycline. as all cultures must be maintained in antibiotic-free medium or medium containing only penicillin. It is apparent that rickettsiae are responsible for fish diseases that span a broad .g. no effective chemotherapeutants. In the absence of effective methods for treatment of piscirickettsiosis. and the relationships between the rickettsial isolates from salmonid fish and the isolate from tilapia should be determined. the search for other rickettsial fish pathogens in established and new hosts should continue. as yet undescribed. For this reason. and little information on the source and transmission of the infectious agent. practical research is needed to find chemotherapeutants and/or treatment regimes effective in the control of the disease. e. or some combination of factors is responsible for this phenomenon. 19). Vaccines have been successfully used in the control of other fish diseases (28). In addition. but unfortunately. these preparations have been of limited efficacy in the treatment of piscirickettsiosis. The rickettsial isolates from salmonid fish in the freshwater and marine environments should be compared. the environment. oxolinic acid (3. Disease control Although P salmonis is sensitive in vitro to many of the antibiotics commonly used to combat other infectious diseases of fish. the host fish. permitting intracellular replication of the pathogen to continue despite therapy.

Oslo. Oncorhynchus mykiss (Walbaum). C. Games. I8:277-279. Egtved-Virus (VHS) der Regenbogenforelle (S&no gairdneri) und Rickettsien&nliche Organismen. Castro. 6. A. Z. Cvitanich. (1994). Egypt. Ozel. M. aquat. 42( 1):120-126. 13.. Fryer. (1996). The discovery of a rickettsia in a fish. 18:157-164. Hyg. (1994) Rickettsial and chlamydial infections of freshwater and marine fishes. Winton. We are just beginning to understand the scope and impact of these agents.B.. Fryer. British Columbia. (1990) Isolation of a rickettsiales-like organism from diseased coho salmon Oncorhynchus kisurch in Chile.E... Isolation of a new . J. J. Assoc. 230:1-14. Septicemia suspected to be caused by a rickettsia-like agent in farmed Atlantic salmon. 3. 12:40+1..A.. salmonis isolates for our study and comparison: G. Salmo salor L. (1994). Piscirickettsiosis in freshwater. and rainbow trout.D..S. Lannan. J. J. (1939). Sandino. 10. 12.J. Can. 29(2):61-71. Zool. Fryer. fanaque suttoni (Eigemnann & Eigenmann). Mohamed.. 16:361-369. E. PM. nov.12 J. PA. CONCLUSIONS A previously unsuspected class of fish pathogens was introduced with the isolation and identification of E salmonis from Chilean salmonids in 1989 (2).L.. Smith. Gaggero. Comps. Puerto Montt. Melby. (1995).N. in Ireland..L. A rickettsia-like organism from dragonets. Fish Pathol.N. (1992).J. M. Rickettsia-like organism infecting juvenile sea-bass Dicenrrarchus labrax.. 8. Assoc. project R/FSD-22. 1. Garc& L. Fish Dis. PA. Wood. Canadian Department of Fisheries and Oceans. Pathol. Larenas. C.. (1995). Fryer and C. 11:93-97. Vergleichende elektronenmikroskopische Untersuchungen an Rhabdoviren pflanzlicher und tierischer Herkunfi: ill. Fish. A. J. 9.. S.. Lannan..M. Bull. Garate N.C. G. Oncorhynchus kisurch (Waibaum). Traxler..N. In Vitro 20:671-676. Fish Dis. C. and the growing awareness of the potential impact of the rickettsiae on fish health ensures that more of these agents will be found. Marine Harvest Chile.. Acknowledgemenrs . and further study can only emphasize their role as major factors in the infectious diseases of fish. 5.. bivalves. Bacterial.. 2. C. L.R. Outbreaks of a disease caused by a rickettsia-like organism in cultured tilapias in Taiwan.. J. 14(4):137-138. P A. Larenas. Piscirickcrrsia sobnonis gen. Fryer. Serv. 4.H. Chem. (1984). Smith. University of Chile. Silva P. Chao. 25(2):107-l 14. and crustaceans.. H. 33407-408. J..L. Zbl..J. 7.N. J. Speare. Dennis.. Bravo. Nanaimo. L. Rickettsia-like organisms in the blue-eyed plecostomus. it is now apparent that the rickettsiae are an important group of fish pathogens with the potential for a significant effect on the health of both freshwater and marine species.D. A. C. Cairo. J. Evelyn T.J. S. (1995) First isolation of Pisciricketrsia salmonis from coho salmon. Tech. Lannan.D. J. sp. Schwanz-Pfitzner. Lannan host and geographic range. Org.. I. Department of Commerce. Dis. N. <<NY>> laksesykdom for&&et av rickettsie. Eur. Norwegian National Veterinary Institute. 16.B. Bull.. Sci. C.. Ministry of Agriculture. C. Fish. L. Cdionymus lyru L. Armstrong. Observation of a rickettsia-like organism in Atlantic salmon. the causative agent of an epizootic disease in salmonid fishes.. Lannan. Eur.. M.L.L. 214. 14.We thank the following individuals for providing P. Santiago. Norsk Fiskeoppdrett NR. H&stein. Raymond. Fish Pathol. Sandino.. Rodger. Lannan. R. Sect. Khoo. C.. Andrade V..D. Syst. Assoc. D. Speiltnxg. Fish Pathol. Infectivity of a rickettsia isolated from coho salmon (Oncorhynchus kisutch).N. Abt Orig. Figueroa P. Brocklebank. O. Studies 33(2):95-107. (Teleostei: Calhonymidae) in Wales. 17. 6pp.. 16(1):30-33. II. Bull. Evensen.M. Pathol. Smith. Bull.H. With the subsequent recognition of the rickettsial etiology of fish diseases in a variety of locations and host species.N. T. Davies. J. Bakt. Vet. C. (1975). Lewbart. Fish Dis. Drinan.. (1991). S. P Melby. (1993). 0. 6(4): 103. College of Veterinary Sciences. This work was supported in part by Oregon Sea Grant with funds from the National Oceanic and Atmospheric Administration Office of Sea Grant. G.R. Giovannoni. 15. nov. HP..L.. J. J. Eur.N. Vet... H. J. Plassiart. Fish cell lines: establishment and characterization of nine cell lines from salmonids.J.. Int. Smith. J. Madrid. A.... under grant NA36RGO45I. R. H.. REFERENCES 1. E. Olsen. J.. (1993). during the freshwater stage of their life cycle. Fryer. (1986). (1992).

(ed. 14:121-145. Fryer. Williams and Wilkins. Extracellular survival of Piscirickerrsia solmonis. 135-J 36. Smith S.A.N. Rodger. C.J. T. (Abstract in Japanese) 1995 Spring Meeting Japan. S. Cvitanich.) Bergey’ s Manual of Systematic Bacteriology.. Branson. (1944).(ed. Oncorhynchus kisutch (Walbaum).C. C.Rickettsial infections of fish 13 18.) 1988.. Academic Press. .S. T. 26. Games A. In: Kent. Piscirickettsiosis: Lesiones en salm6n de1Athintico (.. C. Eur. Richards.. J. I. Philadelphia. Larenas H.%&nos&r) infectados naturalmente con Piscirickerrsia solmonis..F. Fish Pathol.. BaJtimoreILondon. M. 25AL. Ellis. 25. K. An immunohistochemical diagnostic test for rickettsial disease.. Description of a new disease condition occurring in farmed coho salmon. (1964). 1995. Adams. J.18-19. N.. Bull. Fish Dis. Fish Vaccination. (1991).R. Fish Dis.. Lea and Febiger.) Diseases of Seawater Netpen-reared Salmonid Fishes in the Pacific Northwest.L...T. Alday-Sanz. pp. Nanaimo. pp. rickettsia-like organism from Atlantic salmon in Chile. Fish Dis. Aquat. 23. Order I. E. 20. (1984).L. S. Fish Dis. B.L. Japan. J. Fish. Can. J.E. (1991). Recommended methods for inspection of fish for the salmonid rickettsia. J. Fisheries and Oceans.H. 28. Moulder. Animal Hlth 3:229-234..J. Fish Pathol.. Ewing. Staining rickettsiae in yolk-sac cultures. Giminez.. 687-729. Fryer. C.. J. (1994). 5th ed. Fryer.. 27. O. March 30-31. P.. J. Sci. (1991). (1995). 22.W. T..P. J. Fryer. J. J.. Lannan. 24. A fluorescent antibody test for detection of the rickettsia causing disease in Chilean salmonids. 39:135-140. Nieto Diaz-Munoz. A. 30. V. PA. Assoc.. Hatai. Dept. Salmonid rickettsial septicemia. Aquat. Lannan. 14:147-156. L. 19. in South America. C. Vol. Stain Technol. Garate N. Simmons. (1994). H.. 116. D. Wada. 255 pp. 17:545-548. 823 pp. Shiomitsu.L. (1991). Pub. (ed. J. Laboratory methods of the United States Army.D. R.N. In: Krieg. J. H. (1992). Weiss. Smith.... A. D. Hidalgo V. E. The isolation of a rickettsia-like organism causing disease and mortality in Chilean salmonids and its confirmation by Koch’ s postulates. (In Spanish with English abstract) Avarices en Ciencias Veterinarias lO( 1):53-58.N. J. San Diego..E. (1995). Turnbull. 21. 17:189-191. Gentzkow. 11: Lannan.. CA. 29.L. Spec. Sot. A disease with liver swelling and nodule formation in cultured tilapia. Newsletters of the Fish Health Section of the American Fisheries Society 23(3):1-3. Rickettsiales Gieszczkiewicz 1939. Evelyn. Tokyo.