Characterization of Listeria monocytogenes isolated from channel catfish (Ictalurus punctatus)

Chinling Wang From the Research Program and Diagnostic Laboratory Services, College of Veterinary Medicine, Mississippi State University, Box 9825, Mississippi State, MS 39762.

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 DVM, PhD
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Li-Feng Zhang From the Research Program and Diagnostic Laboratory Services, College of Veterinary Medicine, Mississippi State University, Box 9825, Mississippi State, MS 39762.

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Frank W. Austin From the Research Program and Diagnostic Laboratory Services, College of Veterinary Medicine, Mississippi State University, Box 9825, Mississippi State, MS 39762.

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Carolyn R. Boyle From the Research Program and Diagnostic Laboratory Services, College of Veterinary Medicine, Mississippi State University, Box 9825, Mississippi State, MS 39762.

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 PhD

Abstract

Objective

To characterize Listeria monocytogenes from tissues of channel catfish for their ability to cause hemolysis and grow intracellularly in mouse macrophages.

Samples

15 isolates from processed fillets and 15 isolates from the brain, spleen, and kidneys.

Procedure

Serotype and hemolytic activity of L monocytogenes isolates were evaluated, using plate agglutination and CAMP tests, respectively. Invasiveness of L monocytogenes was determined by inoculating each strain or isolate on J774A.1 macrophage cells. Infected cells were incubated for 0 or 3 hours and lysed; then 100 μΙ of the lysate was plated onto a brain heart infusion agar plate. Colony counts for each strain or isolate were analyzed statistically.

Results

Of 30 isolates, 19 were serotype 1 and 11 were serotype 4. Mouse J774A.1 macrophages were inoculated with catfish isolates, a wild-type (EGD) or a nonhemolytic strain of L monocytogenes. Seventy-three percent (11/15) of isolates originating from catfish organs and 100% (15/15) of isolates originating from fillets were not significantly different from the wild-type EGD strain. The nonhemolytic L monocytogenes strain used as a negative control failed to replicate. Intracellular growth of all L monocytogenes isolates decreased after an additional 3-hour incubation period with medium containing 50 μg/ml of gentamicin.

Conclusions

Similar to the wild-type EGD strain, most channel catfish L monocytogenes isolates were hemolytic, serotype 1 or 4, and were invasive for mouse J774A.1 macrophages.

Clinical Relevance

L monocytogenes growth in mouse macrophages may serve as an in vitro model for determining virulence of isolates from food products or environments. (Am J Vet Res 1998;59:1125-1128)

Abstract

Objective

To characterize Listeria monocytogenes from tissues of channel catfish for their ability to cause hemolysis and grow intracellularly in mouse macrophages.

Samples

15 isolates from processed fillets and 15 isolates from the brain, spleen, and kidneys.

Procedure

Serotype and hemolytic activity of L monocytogenes isolates were evaluated, using plate agglutination and CAMP tests, respectively. Invasiveness of L monocytogenes was determined by inoculating each strain or isolate on J774A.1 macrophage cells. Infected cells were incubated for 0 or 3 hours and lysed; then 100 μΙ of the lysate was plated onto a brain heart infusion agar plate. Colony counts for each strain or isolate were analyzed statistically.

Results

Of 30 isolates, 19 were serotype 1 and 11 were serotype 4. Mouse J774A.1 macrophages were inoculated with catfish isolates, a wild-type (EGD) or a nonhemolytic strain of L monocytogenes. Seventy-three percent (11/15) of isolates originating from catfish organs and 100% (15/15) of isolates originating from fillets were not significantly different from the wild-type EGD strain. The nonhemolytic L monocytogenes strain used as a negative control failed to replicate. Intracellular growth of all L monocytogenes isolates decreased after an additional 3-hour incubation period with medium containing 50 μg/ml of gentamicin.

Conclusions

Similar to the wild-type EGD strain, most channel catfish L monocytogenes isolates were hemolytic, serotype 1 or 4, and were invasive for mouse J774A.1 macrophages.

Clinical Relevance

L monocytogenes growth in mouse macrophages may serve as an in vitro model for determining virulence of isolates from food products or environments. (Am J Vet Res 1998;59:1125-1128)

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