USFWS, California-Nevada Fish Health Center09/15/2006

Sample Summary – 2008Winter Chinook Adults

Hatchery:Livingston Stone NFH

Stock:2008 Winter Chinook Adults-Wild (WCS-SRW-2007-Adult)

I. Wild WCS Summary

Samples collected from Livingston NFH, May 26 through July 23, 2008(laboratory case numbers: 08-074, 075, 076, 080, 082, 089, 091, 092, 094, 095, 097, 098, 102, 105, 107, 109, 110, 113, 114, 118, 119, 123, and 124.)

1. Infectious Hematopoietic Necrosis Virus (IHNV) – ovarian fluid and kidney:

  1. 64/78, 82% Prevalence of Infection (POI)

i. Females:46/49 93%

ii. Males: 18/2962%

2. Renibacterium salmoninarum (Rs) – kidney:

a. 0/78, 0% POI

i. Females: 0/490%

ii. Males: 0/290%

3. Bacteriology – male and female kidney:

  1. 0/78Fish tested positive for Yersinia ruckeri.
  2. 7/78 Fish tested positive for Aeromonas salmonicida.

4. Parasitology – male and female intestine (Cs) and kidney (Pm)

  1. 6/78(8%) Fish tested positive for Ceratomyxa shasta.
  2. 69/78(88%) Fish tested positive for Parvicapsula minibicornis.

Table 1. Summary of Prevalence of Infection (POI) by pathogen type for Wild Adults

Assay
(Method) / # Samples
(pool size) / Total Fish / Pathogens Screened / # Positive
(% Positive)
Virology
(Tissue Culture) / 78(1) / 78 / IHNV1 / 64/78 (82%)
IPNV1 / 0
VHSV1 / 0
Culturable Bacteria
(BHIA culture) / 78 (1) / 78 / Aeromonas salmonicida1
Yersinia ruckeri1 / 7/78 (9%)
0
Renibacterium salmoninarum
(Rs- QPCR) / 78 (1) / 78 / Renibacterium salmoninarum2 / 0/78 (0%)
Parasitology
(Cs-QPCR) / 78 (1) / 78 / Ceratomyxa shasta3 / 6/78 (8%)
(Pm-QPCR) / 78 (1) / 78 / Parvicapsula minibicornis3 / 69/78 (88%)

Notes: 1 These pathogens are listed in the USFWS Fish Health Policy(713 FW 1-5) as reportable pathogens associated with

salmonid disease.

2 Quantitative PCR (QPCR) did not detect viable Rs cells in 0/78 samples.

3 Ceratomyxa shasta and Parvicapsula minibicornis screened by QPCR, which is more sensitive than

histology used in prior years.

II. Laboratory Detection Methods

Virology (IHNV, IPNV, VHSV: Inoculation of kidney and ovarian fluid in 1-fish tissue pools, onto EPC

and CHSE-214 cell lines. Observed for viral cytopathic effects (CPE) over an18 day incubation period (15°C). Blind passage of negative sample sets observed for 10-14d ((15°C).

Culturable Bacteria: Aeromonas salmonicida (AS) and Yersinia ruckeri (YR) assayed by direct culture of individual kidney tissue onto appropriate growth media (BHIA) and subsequent biochemical testing (OF, TSI) of all presumptivebacterial isolates.

Non-culturable or fastidious Bacteria: Renibacterium salmoninarum (Rs) assayed by Quantitative Polymerase Chain Reaction (QPCR), which detectsthe specific p57 DNA of Renibacterium salmoninarum bacteria, indicating the presence of presumably viable Rs cellsin the kidney tissue.

Parasitology: Ceratomyxa shasta (Cs) assayed by QPCR, which detectsthe specific 18S ribosomal DNA of Ceratomyxa shasta, indicating the presence of presumably viable parasites in intestine tract of adult fish. Parvicapsula minibicornis (Pm) assayed by QPCR, which detects the specific 18S ribosomal DNA of Parvicapsula minibicornis, indicating the presence of presumable viable parasites in the kidney tissue.

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