Minimally Invasive Detection of
            <i>Piscirickettsia salmonis</i>
            in Cultivated Salmonids via the PCR

Marshall, Sergio H.; Heath, Sekou; Henríquez, Vitalia; Orrego, Cristián

doi:10.1128/aem.64.8.3066-3069.1998

Cited by 57 publications

(45 citation statements)

References 14 publications

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“…ITS A sequences, in both isolates, were almost the same as their respective ITS B sequences, but they were interrupted by an insert that contained two tRNA genes: tRNA-Ala and tRNA-Ile (GenBank access AF205384 and AF205385). This contra- dictory result with respect to those obtained previously [15,20] can be explained by analyzing the experimental design of these studies. Marshall et al [20] ampli¢ed the ITS using primers complementary to the available P. salmonis ITS sequence (corresponding to our ITS B).…”

Section: Discussionmentioning

confidence: 47%

“…This contra- dictory result with respect to those obtained previously [15,20] can be explained by analyzing the experimental design of these studies. Marshall et al [20] ampli¢ed the ITS using primers complementary to the available P. salmonis ITS sequence (corresponding to our ITS B). When analyzing the ITS sequences obtained in our study (ITS A and B) we veri¢ed that sequences complementary to Marshall's primers were present only in ITS B but not in ITS A.…”

Section: Discussionmentioning

confidence: 47%

“…When we analyzed the 16S^23S rDNA spacer region of P. salmonis obtained by PCR using G1 and L1 primers, a di¡erence was found from that described previously [15,20], i.e. two major ampli¢cation products were obtained instead of one.…”

Section: Discussionmentioning

confidence: 74%

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tRNA genes were found in Piscirickettsia salmonis 16S–23S rDNA spacer region (ITS)

Casanova¹

2001

FEMS Microbiology Letters

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Piscirickettsia salmonis is the etiological agent of Salmonid Rickettsial Septicemia, a disease affecting salmon aquaculture industry. We analyzed the 16S^23S rDNA spacer region (internal transcribed spacer, ITS) of Chilean P. salmonis isolates LF-89 and EM-90. Two main ITS amplification products were obtained by PCR using L1 and G1 primers, differing from that described where only one ITS region was found. By Southern blot, it was established that these two amplification products contained sequences related to P. salmonis ITS. Sequence analysis confirmed that P. salmonis had two ITS regions: ITS A and ITS B. In both isolates, the smaller (ITS B) corresponded to ITS sequences previously described for each one, and the larger (ITS A) were almost the same as their respective ITS B sequences interrupted by an insert which contained two tRNAs genes: tRNA-Ile and tRNA-Ala. ß

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Section: Discussionmentioning

confidence: 47%

Section: Discussionmentioning

confidence: 47%

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tRNA genes were found in Piscirickettsia salmonis 16S–23S rDNA spacer region (ITS)

Casanova¹

2001

FEMS Microbiology Letters

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“…For sampling, fish were exposed to a lethal dose of 2-phenoxyethanol (1 ml/L water), and liver and muscle tissues were isolated immediately and snap-frozen in liquid N 2 prior to being stored at −80°C for long-term storage. Samples were utilized to determine the presence of bacteria by using specific primers for P. salmonis (Contreras-Lynch et al, 2015;Marshall, Heath, Henriquez, & Orrego, 1998).…”

Section: Exposure Of E Maclovinus To P Salmonismentioning

confidence: 99%

Intermediary metabolic response and gene transcription modulation on the Sub‐Antarctic notothenioid Eleginops maclovinus (Valenciennes, 1930) injected with two strains of Piscirickettsia salmonis

Soto‐Dávila

Martínez

Oyarzún

et al. 2019

Journal of Fish Diseases

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Pathogen interactions with cultured fish populations are well studied, but their effects on native fishes have not been characterized. In Chile, the disease caused by bacterial species Piscirickettsia salmonis represents one of the main issues and is considered to be one of the important pathogens in the field of aquaculture. They have been found to infect native fish. Therefore, it is necessary to understand the impact of P. salmonis on native species of local commercial value, as well as the potential impact associated with the emergence of antibiotic‐resistant strains of P. salmonis. Due to this purpose, the native fish Eleginops maclovinus was used in our study. Fish were randomly distributed in tanks and intraperitoneally inoculated with two strains of P. salmonis. No mortality was recorded during the experiment. Cortisol, glucose and total α‐amino acid levels increased in fish injected with AUSTRAL‐005 strain compared to sham‐injected and LF‐89‐inoculated fish. Moreover, results showed an increase in the activity of carbohydrates and lipids metabolism in liver; and an increase in the carbohydrates, lipids and total α‐amino acid metabolism in muscle after injection with AUSTRAL‐005. Our results suggest that P. salmonis modulates the physiology of E. maclovinus and the physiological impact increase in the presence of the antibiotic‐resistant strain AUSTRAL‐005.

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“…In Chile, piscirickettsiosis is the most important disease that a¡ects salmon mariculture [1]. The causative agent corresponds to a Gram-negative intracellular bacterium called Piscirickettsia salmonis, isolated in our country [26 ]. Although this disease has also been reported in Canada, Norway and Ireland, its pathogenesis in ¢sh farms in our country seems to be more severe [7^10].…”

Section: Introductionmentioning

confidence: 99%

Electrophoretic analysis of ITS fromPiscirickettsia salmonisChilean isolates

Casanova

Gaggero

et al. 2003

FEMS Microbiology Letters

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Piscirickettsia salmonis is the most important pathogen in salmonid mariculture in Chile. Since it was reported numerous piscirickettsiosis outbreaks have occurred differing in virulence and mortality. Genetic variability of P. salmonis isolates has been suggested as one factor to explain this. However until now isolates obtained from outbreaks have not been analyzed. Knowledge of genetic variability of P. salmonis is very limited and also a useful screening method for genetic variations in isolates without sequencing is not available. Here we report an electrophoretic analysis of internal transcribed spacer region (ITS) of eleven P. salmonis isolates obtained from different salmon species and places in southern Chile. When PCR products were submitted to polyacrylamide gel electrophoresis (PAGE) a characteristic electrophoretic pattern was observed, distinguishable from ITS of other bacteria, including fish pathogens. Even though this pattern is conserved in all isolates, a difference in ITS electrophoretic mobility was observed, determining clearly two groups: ITS with higher or with lower electrophoretic mobility, including LF-89 and EM-90 isolates, respectively. A higher ITS sequence homology inside each group was shown by heteroduplex mobility assay (HMA). Our results show that genetic variability between Chilean P. salmonis isolates allows the differentiation of two groups with similar behavior observed previously when six P. salmonis isolates from three geographic origins were analyzed by 16S, 23S and ITS sequencing. PAGE analysis of ITS and HMA could be a basis to develop an assay for screening genetic variability between P. salmonis isolates. ß

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Minimally Invasive Detection of Piscirickettsia salmonis in Cultivated Salmonids via the PCR

Abstract: The attributes of the PCR allowed implementation of an assay for specific detection of Piscirickettsia salmonis from a few microliters of fish serum. This opens the way to less invasive modes of sampling for this microbial pathogen in salmonids.

Cited by 57 publications

References 14 publications

tRNA genes were found in Piscirickettsia salmonis 16S–23S rDNA spacer region (ITS)

tRNA genes were found in Piscirickettsia salmonis 16S–23S rDNA spacer region (ITS)

Intermediary metabolic response and gene transcription modulation on the Sub‐Antarctic notothenioid Eleginops maclovinus (Valenciennes, 1930) injected with two strains of Piscirickettsia salmonis

Electrophoretic analysis of ITS fromPiscirickettsia salmonisChilean isolates

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