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Aeromonas salmonicida and aeromonas hydrophila (liquefaciens) as pathogens of salmonid fish. A. Selective aeromonas medium. B. Comparative characteristics of virulent and avirulent strains of aeromonas salmonicida. C. Effect of water temperature on aeromonas infections

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Title Aeromonas salmonicida and aeromonas hydrophila (liquefaciens) as pathogens of salmonid fish. A. Selective aeromonas medium. B. Comparative characteristics of virulent and avirulent strains of aeromonas salmonicida. C. Effect of water temperature on aeromonas infections
Names McCoy, Ralph Hines (creator)
Pilcher, K. S. (advisor)
Date Issued 1973-05-03 (iso8601)
Note Graduation date: 1973
Abstract Studies were directed toward the formulation of a selective
Aeromonas medium that would permit the isolation of Aeromonas
species in the presence of other common bacterial flora. The final
composition of this medium, PBG Agar, in grams per liter, is:
Bacto-peptone, 10; Bacto-beef extract, 10; glycogen, 4; NaCl, 5;
sodium lauryl sulfate, 0. 1; brom thymol blue, 0. 1; agar, 15. The
pH of the medium after autoclaving is 6. 9-7. 1.
Any organism isolated from natural sources that gives characteristic
yellow colonies in this medium under the specified conditions
may presumptively be identified as either Aeromonas,
Vibrio, or Pleisomonas.. Aeromonas colonies are frequently surrounded by a yellow acidic halo in the green medium, and
occasionally produce a small bubble of gas under the non-nutrient
agar overlay.
Biochemical and antigenic analyses were performed in an
attempt to differentiate virulent (LD₅₀ from one to 10³ colony
forming units per fish) from avirulent (LD₅₀ greater than 10⁶ cfu)
strains of A. salmonicida. The virulent and avirulent representatives
gave similar responses in the following tests: onset and duration
of bacteremia in juvenile coho salmon after intramuscular injection;
time of recovery of the organism from fish kidney during
an acute infection; production of a leucocytolytic agent active on
juvenile steelhead trout white blood cells at a dilution of 1;3000; a
factor cytotoxic to cultured chinook embryo cells at a dilution of
1:320; formation of eight extracellular enzymes including elastase,
several proteases and hemolysins; and the failure to produce
hyaluronidase.
There were two real differences observed between a virulent
strain (As-SS-70) and an avirulent strain (As-Sil) of Aeromonas
salmonicida. When grown on 1% peptone agar containing 1. 5% NaCl,
colonies of As-SS-70 were opaque, convex, and granular while those
of As-Sil were translucent, less convex, and non-granular. An
attenuated strain of As-SS-70 gradually assumed avirulent colony
characteristics with continued passage on artificial media. Sonicates of As-SS-70 and As-Sil were concentrated about 20
fold by dialysis against Polyethylene Glycol 4000. Use of these
concentrated antigens and As-SS-70 rabbit antiserum demonstrated
a unique antigen in the As-SS-70 sonicate by immunodiffusion techniques.
This unique precipitin line was not observed in the As-Sil
sonicate, or when anti-As-Sil rabbit antiserum was used. This
was the second difference found between the virulent and avirulent
strains.
Groups of juvenile coho salmon were injected intramuscularly
with about two LD₅₀
doses of virulent A. salmonicida and held at
temperatures of 74°F, 69°F, 64°F, 59°F, 54°F, 49°F, 44°F, and
39°F. Water temperatures of 59°F and above produced high mortality
rates, with losses exceeding 40 percent at 54°F and 49°F.
Mortality rates were very low at temperatures of 44°F and 39°F.
The mean time to death was estimated to be 3.5 days at 69°F, with
steady increases as water temperature decreased, to a maximum
of 31 days at 39°F. Thus, the disease process was accelerated
progressively as water temperature increased.
When juvenile spring chinook salmon were intraperitoneally
injected with about 1.5 LD₅₀
doses of A. salmonicida, the mean
time to death was estimated to be 2.9 days at 74°F, and was found
to progressively increase as water temperature decreased, to a
maximum of 18.4 days at 39°F. The effect of temperature on the growth rate of A. salmonicida
in vitro appeared to be similar to its effect on the rate of progress
of the infection in juvenile salmonids.
The percentage of fatal infections among juvenile steelhead
trout and coho salmon injected with Aeromonas hydrophila was high
at temperatures of 64°F and above, moderate at 59°F, and zero at
49°F and below.
Genre Thesis/Dissertation
Topic Salmon -- Diseases
Identifier http://hdl.handle.net/1957/44685

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