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Post Mortem Muscle Softness in the Spotted Seatrout Cynoscion

nebulosus: Effect of the Myxozoan parasite Kudoa inornata

Candice Alge, Eric McElroy, and Isaure de Buron Department of Biology, College of Charleston, Charleston, SC 29424

Candice Alge

Department of Biology, College of Charleston

Email: candicealge@gmail.com

Website:

Contact 1. Dyková, I., de Buron , I., Fiala, I., & Roumillat, W. A. (2009). Kudoa inornata sp. n. (Myxosporea:

Multivalvulida) from the skeletal muscles of Cynoscion nebulosus (Teleostei: Sciaenidae). FOLIA

PARASITOLOGICA, 91-98.

2. Sunquist,L., McElroy, E., & de Buron, I. (2013) Temperature mediated myoliquefaction: The effect of a

myxozoan parasite Kudoa inornata on the spotted seatrout (Cynoscion nebulosus), College of Charleston,

SC

References

Kudoa inornata is a myxozoan that infects the skeletal muscles of the

spotted seatrout, Cynoscion nebulosus. A previous study in our

laboratory indicated that infected wild fish displayed increased post

mortem flesh softness when compared to non-infected fish

(mariculture raised). We hypothesized that infection by K. inornata was

at the origin of the difference in flesh softness observed. Plasmodium

density, plasmodium area, and spore density were determined from

biopsies of seatrout previously tested for muscle softness (3 biopsies

per fish, N=33). Results indicated that spore density was positively

correlated with plasmodium density and area. Although muscle

softness was not correlated with spore and plasmodia densities, data

suggest that the larger the plasmodia, the softer the muscle is. Hence,

the older the infection in a fish, the higher the post mortem muscle

softness may be. Since old infection may occur in larger (older) fish, this may be of concern to anglers.

Abstract

Introduction

Methods and Materials

• Muscle softness is observed in post-mortem infected

muscle.

• Spore density is correlated to both plasmodium density

and plasmodium area.

• Number of spores, plasmodia do not seem to be

associated with post-mortem muscle softness but

plasmodium size may be.

• Analysis of replicate samples need to be observed to

determine if plasmodium area at 24 hours post mortem

is associated with muscle softness.

Conclusions

Kudoa inornata

• Spores of K. inornata (Fig.1) are organized into

plasmodia (Fig. 2).

• The plasmodium infects the skeletal muscle of

Cynoscion nebulosus1 (the spotted seatrout) (Fig. 3).

• Previous studies show that Kudoa inornata is associated

with post mortem muscle softness2 (Fig. 4 ).

Hypothesis

• Post mortem muscle softness of infected muscle is

associated with spore density, plasmodium density, and

plasmodium area.

Results

Fig. 1 Spores of Kudoa inornata in

muscle squash

Fig. 2 Plasmodium in skeletal

muscle

Fig. 3 Cynoscion nebulosus

Materials

• 11 fish were examined

• 3 biopsies per fish (Fig. 5)- each was weighed

• Each biopsy was taken at a different post mortem

time point: 24, 72, and 144 hours

Plasmodium Density

• Standardized squashes of each biopsies were

made

• Squashes were observed via light microscopy

and counted (Fig. 6)

Plasmodium Area

• Pictures of first 10 plasmodia per biopsy were

taken

• Area was determined using NIH program Image

J (Fig. 7)

Spore density

• Biopsies were trypsinized

• Spores were counted via hemocytometer (Fig. 8)

• Total number of spores per biopsy was inferred

Fig. 6 Skeletal muscle squash

with plasmodium under light

microscopy

Fig. 7 Plasmodium outlined

to determine area

Fig. 8 Spores on a hemocytometer

Individual spores

• A significant positive correlation was found

between spore density and plasmodium density,

R2= 0.34, p<0.003 (Fig. 8).

• A significant positive correlation was found

between spore density and plasmodium area,

R2=0.36, p<0.0002 (Fig. 9).

• A positive correlation between plasmodium area

and muscle softness at 24 hours post mortem

was found to be on the verge of significance, R2=

0.28, p=0.059 (Fig. 10).

Fig. 9 Log spore density and Log plasmodium density

Fig. 10 Log spore density and Log plasmodium area

Fig. 11 Log plasmodium area compared to log force at 24, 72,

and 144 hours.

5µm

Fig. 5 Biopsies taken from seatrout filet

Figure 4. Maximum force readings over time for infected and uninfected fish

Parasite Effect: F1,60 = 5.4, p =0.023

Time Effect: F2,327 = 32.6, p < 0.001

(Sunquist et. al, 2013)