Quantitative PCR for <i>Tenacibaculum dicentrarchi and T. finnmarkense</i>

Nowlan, Joseph P.; Lumsden, John S.; Russell, Spencer

doi:10.1111/jfd.13357

Cited by 5 publications

(20 citation statements)

References 23 publications

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“…Prior research into aquatic pathogens has shown that cycle quotients from qPCR assays can differentiate disease states of fish and determine thresholds of bacteria required to cause infection, where similar applications have occurred for salmon gill poxvirus [19] and bacterial coldwater disease [20]. However, the qPCR assays developed by a previous study [7], which were used in the present study, are not suitable for determining such a threshold in uncontrolled conditions as there are limitations relating to the specificity and copy number for the 16S rDNA gene in Tenacibaculum. For Tdic, mortality of exposed fish within 48 h is similar to [4] and may indicate that the results of the Tdic exposure are not solely based on the bacteria.…”

Section: Discussionmentioning

confidence: 99%

“…Prior to transport to the Centre for Innovation in Fish Health (CIFH), at Vancouver Island University in Nanaimo (BC), the salmon population was screened for and tested negative for infectious salmon anemia virus, infectious pancreatic necrosis virus, and piscine orthoreovirus. Water from the recirculating aquaculture systems (RAS) systems at the CIFH, were tested using qPCR for Tmar, Tdic, and Tfinn prior to the introduction of fish following previously developed protocols [7].…”

Section: Fish Husbandrymentioning

confidence: 99%

“…Three qPCR assays specific to Tmar [24,25], Tdic, and Tfinn [7] were used on tissue samples and bacterial isolates. The procedure and thermal profiles designed previously [7,24] were used with minor modifications: the Tmar probe (0.05 µM) the Tmar primers (0.5 µM); and the Tdic probe (0.125 µM), per reaction. Primers were obtained from SIGMA-ALDRICH (Canada) and probes were obtained from Eurofins Genomics (USA).…”

Section: Quantitative Pcrmentioning

confidence: 99%

“…For each reaction, each well received 100 ng of sample DNA. Similar to a previous study [7], a cycle quotient of 35 was used as a cut-off to represent no bacteria.…”

Section: Quantitative Pcrmentioning

confidence: 99%

“…Several Tenacibaculum bacterial species are putative pathogens associated with tenacibaculosis in fishes [1][2][3][4][5][6]. In British Columbia (Canada; BC), Tenacibaculum species are commonly isolated from Atlantic salmon (Salmo salar L.) with mouthrot; a regional, clinical presentation of tenacibaculosis, often characterized by epidermal ulcerations and development of yellow plaques around the mouth [1,[5][6][7][8].…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

Experimental Induction of Tenacibaculosis in Atlantic Salmon (Salmo salar L.) Using Tenacibaculum maritimum, T. dicentrarchi, and T. finnmarkense

et al. 2021

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There is a limited understanding of the pathogenesis of tenacibaculosis in Atlantic salmon (Salmo salar L.) and there are few reproducible exposure models for comparison. Atlantic salmon were exposed via bath to Tenacibaculum maritimum, T. dicentrarchi, or T. finnmarkense, and were then grouped with naïve cohabitants. Mortalities had exaggerated clinical signs of mouthrot, a presentation of tenacibaculosis characterized by epidermal ulceration and yellow plaques, on the mouth and less frequently on other tissues. Histopathology showed tissue spongiosis, erosion, ulceration, and necrosis ranging from mild to marked, locally to regionally extensive with mats of intralesional bacteria on the rostrum, vomer, gill rakers, gill filaments, and body surface. Exposure to T. maritimum resulted in less than a 0.4 probability of survival for both exposed and cohabitants until Day 21. Exposures to T. dicentrarchi resulted in 0 and 0.55 (exposed), and 0.8 and 0.9 (cohabitant) probability of survival to Day 12 post-exposure, while T. finnmarkense had a 0.9 probability of survival to Day 12 for all groups. This experimental infection model will be useful to further investigate the pathogenesis of tenacibaculosis, its treatment, and immunity to Tenacibaculum species.

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Section: Discussionmentioning

confidence: 99%

Section: Fish Husbandrymentioning

confidence: 99%

Section: Quantitative Pcrmentioning

confidence: 99%

“…For each reaction, each well received 100 ng of sample DNA. Similar to a previous study [7], a cycle quotient of 35 was used as a cut-off to represent no bacteria.…”

Section: Quantitative Pcrmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Experimental Induction of Tenacibaculosis in Atlantic Salmon (Salmo salar L.) Using Tenacibaculum maritimum, T. dicentrarchi, and T. finnmarkense

et al. 2021

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Development of a PCR assay for the specific identification of the fish pathogen Tenacibaculum piscium

Saldarriaga‐Córdoba

Araya‐León

Irgang

et al. 2023

Journal of Fish Diseases

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Tenacibaculosis is an emerging disease that severely affects salmonid farming in Chile, producing high mortalities and causing great economic losses. This work describes a novel PCR assay for the specific detection of Tenacibaculum piscium, a species recently described and identified in tenacibaculosis outbreaks in Norway and Chile.The designed primers amplified a 678-bp fragment of the peptidase gene (peptidase M23 family) from T. piscium. This method is specific for T. piscium; no other chromosomal DNA amplification products were obtained for other Tenacibaculum species.In pure cultures, the PCR assay detected up to 500 pg of DNA, or the equivalent of 2.44 ± 0.06 × 10 4 CFU/ml. For seeded fish samples (i.e., gills, liver, kidney, and mucus), the sensitivity limit was 4.88 ± 0.11 × 10 6 CFU/g, sufficient to detect T. piscium in acute infections in fish. Notably, this sensitivity level was 100-fold lower for DNA extracted from mucus samples. As compared to other existing methodologies (e.g., gene sequencing), the PCR approach described in this work allowed for the easiest detection of T. piscium in mucus samples obtained from challenged fish, an important outcome considering that the identification of this bacterium is difficult. Our results indicate that the designed specific primers and PCR method provide a rapid and specific diagnosis of T. piscium.

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Tenacibaculum ovolyticum 16S rDNA Quantitative-PCR Assay Development and Field Testing

Nowlan

Heese²,

Wilson³

et al. 2022

Fishes

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In British Columbia (BC; Canada) Atlantic salmon (Salmo salar L.) production, Tenacibaculum members are associated with ‘mouthrot’ and disease identification is based on gross observation and clinical data. Genomic similarities (i.e., putative virulence factors) between T. ovolyticum and other better-characterized agents of mouthrot could imply potential pathogenicity. While T. ovolyticum has not been directly linked to salmon mortality events in BC, it has been isolated from diseased marine fish. To investigate T. ovolyticum’s pathogenicity in situ, a T. ovolyticum 16S rDNA qPCR assay targeting a ~155 bp amplicon was developed. The assay was used to screen 67 biotic and 33 abiotic samples collected from a BC Atlantic salmon (Salmo salar L.) net-pen site before, during, and after a mouthrot outbreak. The assay was specific, quantifiable and detectable for T. ovolyticum over 6-log and 8-log units, respectively. However, cycle quotients differed between the BC isolate and type strain of T. ovolyticum, suggesting that qualitative use of the qPCR assay in field samples would be more accurate. Only two out of 100 samples were T. ovolyticum-positive, indicating limited involvement in this particular outbreak. However, the ecological role of T. ovolyticum and its involvement in the pathogenesis of other mouthrot outbreaks in Atlantic salmon is unknown.

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Quantitative PCR for Tenacibaculum dicentrarchi and T. finnmarkense

Cited by 5 publications

References 23 publications

Experimental Induction of Tenacibaculosis in Atlantic Salmon (Salmo salar L.) Using Tenacibaculum maritimum, T. dicentrarchi, and T. finnmarkense

Experimental Induction of Tenacibaculosis in Atlantic Salmon (Salmo salar L.) Using Tenacibaculum maritimum, T. dicentrarchi, and T. finnmarkense

Development of a PCR assay for the specific identification of the fish pathogen Tenacibaculum piscium

Tenacibaculum ovolyticum 16S rDNA Quantitative-PCR Assay Development and Field Testing

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