Growth of
            <i>Piscirickettsia salmonis</i>
            to High Titers in Insect Tissue Culture Cells

Birkbeck, T. H.; Griffen, A. A.; Reid, Helen I.; Laidler, L. A.; Wadsworth, Simón

doi:10.1128/iai.72.6.3693-3694.2004

Cited by 31 publications

(30 citation statements)

References 13 publications

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“…However, Birkbeck et al (2004) showed that P. salmonis replicates in higher titers in an insect cell line than in the CHSE-214 cells that are normally used to culture the organism and that P. salmonis retains virulence for Atlantic salmon Salmo salar after repeated culture in insect cells. On the other hand, the tissue chosen for the isolation of P. salmonis during active infection in salmonids is kidney, and SHK-1 is a cell line from S. salar head kidney, which exhibits macrophage properties (Dannevig et al 1997).…”

Section: Resultsmentioning

confidence: 99%

Broth medium for the successful culture of the fish pathogen Piscirickettsia salmonis

Yáñez¹,

Valenzuela²,

Silva³

et al. 2012

Dis. Aquat. Org.

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Piscirickettsiosis or salmonid rickettsial septicaemia (SRS) caused by Piscirickettsia salmonis constitutes one of the main problems in farmed salmonid and marine fishes. Since the first reports of the disease, it has been successfully isolated and maintained in eukaryotic cellculture systems, but these systems are time-consuming, the media are costly, and eliminating heavily contaminated host cell debris is difficult. In this report, we describe a marine-based broth supplemented with L-cysteine, named AUSTRAL-SRS broth, that facilitates superior growth of P. salmonis strains. Strains reached an optical density of approximately 1.8 when absorbance was measured at 600 nm after 6 d incubation at 18°C. Several passages (n = 6) did not alter the culture kinetics. We report for the first time the purification of DNA, lipopolysaccharide (LPS) and whole membrane protein obtained from P. salmonis grown in this liquid medium, and thus provide a suitable platform to simplify the preparation of P. salmonis cells for genetic and serological studies. Moreover, the results of the cytopathic effect test showed that P. salmonis grown in AUSTRAL-SRS broth maintained their virulence properties, inducing apoptosis after 3 d. This makes the medium a good candidate for the successful growth of P. salmonis and an excellent basis for the development of low cost vaccines. KEY WORDS: Piscirickettsia salmonis · Broth · CultureResale or republication not permitted without written consent of the publisher

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Section: Resultsmentioning

confidence: 99%

Broth medium for the successful culture of the fish pathogen Piscirickettsia salmonis

Yáñez¹,

Valenzuela²,

Silva³

et al. 2012

Dis. Aquat. Org.

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“…Birkbeck et al (2004) showed that P. salmonis can be cultured in Sf21 insect cells with a yield approximately 60 to 100 times greater than that obtained in CHSE-214 cells and that the virulence for Atlantic salmon was unchanged. P. salmonis was cultured in four 175 cm 2 flasks containing confluent monolayers of Sf21 cells in 100 ml TC100 medium + 10% foetal calf serum (Invitrogen).…”

Section: Methodsmentioning

confidence: 97%

“…A significant difference was defined as a p-value of < 0.05. Birkbeck et al (2004) showed that intraperitoneal injection of Piscirickettsia salmonis SCO-95A is virulent for Atlantic salmon, with an LD 50 of < 2 × 10 3 TCID, but that only 1 of 20 cohabitant fish held in the same tank succumbed to SRS. Therefore, we exposed fish to 1 × 10 5 TCID ml -1 P. salmonis for 1 h in a bath challenge.…”

Section: Methodsmentioning

confidence: 99%

Infectivity of a Scottish isolate of Piscirickettsia salmonis for Atlantic salmon Salmo salar and immune response of salmon to this agent

Birkbeck¹,

Rennie²,

Hunter³

et al. 2004

Dis. Aquat. Org.

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A Scottish isolate of Piscirickettsia salmonis (SCO-95A), previously shown by intraperitoneal injection to have a lethal dose (LD 50 ) of < 2 × 10 3 infectious rickettsial units, was tested for virulence by bath challenge, surface application to the skin, or dorsal median sinus injection. Atlantic salmon Salmo salar post-smolts were used in all experiments, and exposure to 1 × 10 5 tissue culture infective doses (TCID) of P. salmonis ml -1 for 1 h in a bath challenge resulted in only 1 mortality, 18 d later, in 10 exposed fish. Application of 2.5 × 10 6 TCID of P. salmonis SCO-95A to paper discs on the skin failed to induce any mortalities within 42 d. Intraperitoneally, fish were administered vaccines containing 10 9 heat-inactivated (100°C, 30 min) or 10 9 formalin-inactivated P. salmonis SCO-95A in adjuvant, with a control group receiving phosphate-buffered saline (PBS) in adjuvant. After an induction period of over 6 mo fish were challenged by injection of P. salmonis into the dorsal median sinus. Mortalities in the control group reached 81.8% and the heat-inactivated and formalin-inactivated vaccines gave significant protection from P. salmonis, with relative percentage survivals of 70.7 and 49.6%, respectively. The nature of the protective antigen is unknown, but could be lipopolysaccharide or a heat-stable outer membrane protein. Fish that survived a dorsal median sinus challenge of P. salmonis or were cohabitants showed a strong immune response to P. salmonis.

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“…More recently, growth of R. felis in Drosophila melanogaster S2 cells was reported, with a cell infection rate of 100% after 19 days of incubation at 25°C (331). Other arthropod-borne bacteria, such as Diplorickettsia massiliensis (332), Wolbachia sp., and Bartonella bacilliformis (40), as well as other bacteria such as Piscirickettsia salmonis (333) and M. ulcerans (334) can also be cultivated more effectively by using this cell line.…”

Section: Cell Culturementioning

confidence: 99%

Current and Past Strategies for Bacterial Culture in Clinical Microbiology

et al. 2015

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SUMMARY A pure bacterial culture remains essential for the study of its virulence, its antibiotic susceptibility, and its genome sequence in order to facilitate the understanding and treatment of caused diseases. The first culture conditions empirically varied incubation time, nutrients, atmosphere, and temperature; culture was then gradually abandoned in favor of molecular methods. The rebirth of culture in clinical microbiology was prompted by microbiologists specializing in intracellular bacteria. The shell vial procedure allowed the culture of new species of Rickettsia . The design of axenic media for growing fastidious bacteria such as Tropheryma whipplei and Coxiella burnetii and the ability of amoebal coculture to discover new bacteria constituted major advances. Strong efforts associating optimized culture media, detection methods, and a microaerophilic atmosphere allowed a dramatic decrease of the time of Mycobacterium tuberculosis culture. The use of a new versatile medium allowed an extension of the repertoire of archaea. Finally, to optimize the culture of anaerobes in routine bacteriology laboratories, the addition of antioxidants in culture media under an aerobic atmosphere allowed the growth of strictly anaerobic species. Nevertheless, among usual bacterial pathogens, the development of axenic media for the culture of Treponema pallidum or Mycobacterium leprae remains an important challenge that the patience and innovations of cultivators will enable them to overcome.

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Growth of Piscirickettsia salmonis to High Titers in Insect Tissue Culture Cells

Cited by 31 publications

References 13 publications

Broth medium for the successful culture of the fish pathogen Piscirickettsia salmonis

Broth medium for the successful culture of the fish pathogen Piscirickettsia salmonis

Infectivity of a Scottish isolate of Piscirickettsia salmonis for Atlantic salmon Salmo salar and immune response of salmon to this agent

Current and Past Strategies for Bacterial Culture in Clinical Microbiology

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