Food Control, Vol. 8, No. 2, PP.

87-89, 1997
0 1997Elsevier Science Ltd
All rights reserved. Printed in Great Britain
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PAPER

Total plate count and coliform

levels in Nigerian periwinkles
from fresh and brackish water
E. 0. Ekanem* and B. N. Otti

Three species of fresh& harvested Nigerian periwinkles- Tympanotonus fuscatus,

T. fuscatus var radula (both of brackish water habitat) and Pachymelinia
bryonensis (a fresh water species) - were evaluated for bacteriologicalquality
using aerobic plate counts and colifonn counts. Results showed that all the
periwinkles contained unacceptable levels of bacteria with P. bryonensis carrying
up to 8.90 f 1.93 mean log cfulg meat. The levels of coliforms in the shellfish were
generally high, ranging from 89.6% of the total load in T. fuscatus var radula to
90.5% in T. fuscatus No signi’cant difference (P>O.O5) was detected between
the periwinkles with respect to both SPC and colifonn count. Bacterial genera
identified in the molluscs included Bacillus. Salmonella, Klebsiella, Micro-
coc&, Shigella, Pseudomonas, Escherichia and Staphylococcus 0 1997
Elsevier Science Ltd.

Keywords: periwinkles; total plate count; coliform density; shellfish

INTRODUCTION the pollution of the waters in which the shellfish grow

Periwinkles represent one of the most important
shellfish resources of the world. In West Africa,
periwinkles are the most dominant of the aquatic
molluscs, occurring widely in both fresh and brackish
water (Buchaan, 1954; Okon, 1987). Periwinkles are
widely consumed as food in many countries and
regions. In areas where they are available, consumer
acceptance is independent of religion, socio-economic
status, ethnic group or sex (Ekanem and Udoidiong,
submitted). The shellfish is therefore a mass-
consumer product.
Shellfish have been widely implicated in outbreaks
of food-borne disease in many parts of the world.
These illnesses, which include typhoid fever, hepatitis
and similar disorders of the digestive system (Cutting
and Spencer, 1968; Metcalf et al., 1973) are due to
Food Technology Unit, Department of Animal Science,
University of Uyo, PMB 1017, Uyo-Nigeria. *To whom
correspondence should be addressed.
(Hackney et al., 1980; Ekanem and Adgoke, 1995).
The shellfish can concentrate and accumulate high
levels of pathogens when grown in such polluted
waters. For example, dangerous levels of pathogenic
organisms, including enteroviruses and faecal
coliforms have been found in Louisiana oysters and
their growing waters (Cole et al., 1986a, b). Similarly
unacceptable levels of pathogens have been detected
in shellfish beds in Massachusetts (Khalifa et al.,
1986) in the West African clam (Ekanem, 1990;
Ekanem and Adegoke, 1995), in the Sydney Rock
oyster (Rowse, 1981), in Long Island oysters (Tepen-
dino, 1982), in fresh Nigerian market shrimps (Lawal
and Avhurhi, 1990), in Indian paddy field-cultured
prawns (Nair et al., 1991) and in the New Hamsphire
shellfish waters (Metcalf et al., 1973).
The present study was undertaken as a preliminary
attempt to evaluate the bacteriological quality of
three Nigerian periwinkle species, Tympanotonus
fixates, Tympanotonusfuscatus var. radula and
Pachymelania bryonensis. There is a dearth of

Food Control 1997 Volume 8 Number 2 87

Bacterial contamination of periwinkles: E. 0. Ekanem and B. N. Otti
published data regarding this and many other aspects
of these popular food delicacies.
MATERIALS AND METHODS
The periwinkle (freshly harvested) samples used in
this study were purchased from periwinkle markets at
Itu (I! bvonensis) and Mbo (Tympanotonus spp), both
of which are famous periwinkle-producing areas in
Akwa Ibom State, Nigeria. Samples were collected in
sterile plastic containers and taken to the laboratory
for analysis within 3 h. The periwinkles were
scrubbed, rinsed and the meat aseptically extracted as
described by APHA (1970).
Bacteriological analyses were carried out in tripli-
cate on 50 g raw samples which were blended with
450 ml of sterile 0.1% peptone water as described in
the Bacteriological Analytical h4anual (FDA, 1984).
Pour plates were prepared from lo-fold dilutions in
nutrient agar (Oxoid) for SPC enumeration, and
MacConkey agar (Oxoid), using the 11-tube most-
probable number (MPN) technique (APHA, 1970)
for coliforms. Counts were made after incubation at
37°C for 48 h. Tubes showing acid gas within 48 h
were considered positive for coliforms.
Six typical and atypical colonies were selected
randomly from the incubated nutrient agar and
MacConkey agar plates for identification. Bacterial
cultures were characterized on colonial morphology,
cellular morphology and by gram stain, catalase,
motility, coagulase and MR-VP tests, indole produc-
tion, carbohydrate fermentation tests and capsule
stain as appropriate.
Significance of differences between means was
evaluated by analysis of variance according to
Walpole (1974).
RESULTS AND DISCUSSION
Table I shows the levels of bacterial load in the
periwinkles studied. Mean log total bacterial popula-
tions varied from 5.81 to 9.41 cfu/g meat for T
fuscatus var. radula to 6.97-10.83 cfulg meat for R
bryonensis. I! bryonensis, the fresh water species,
therefore had the highest level of bacterial contami-
nation. Mean coliform density was also highest (mean
log count = 8.05 cfu/g meat) in I! bryonensis. Differ-
ences between species were, however, insignificant
(P>O.O5) with respect to both total plate and
liable 1 Bacteriological characteristics of three Nigerian species
of periwinkles: mean log cfu/g meat + SW
Total plate Coliform
Speciesh count count
1: fuscatus 8.86+ 1.92 8.02 + 1.84
?: fuscotus var. radula 7.61* 1.80 6.82 L-3.97
l? btyonensis 8.90 + 1.93 8.05 + 1.82
“Values shown are means of three determinations.
*Differences between species are insignificant (E-0.05).
88 Food Control 1997 Volume 8 Number 2
coliform counts. In general, the proportions of total
bacteria that were coliforms were very high, ranging
from 89.6% in lYfuscatus var radula to 90.5% in ?:
fuscatus.
The higher level of bacteria in E bryonensis could
be due to the fact that it grows in fresh water which
supports the proliferation of a broader spectrum of
microorganisms than brackish water in which the
Tympanotonus species live (Nester et al., 1983). Also,
there are greater pollution-causing activities like
bathing, washing and sewage discharge in the fresh
water environment where E bryonensis is harvested
than in the brackish water environment. Jay (1978)
and Ekanem and Adegoke (1995) have reported that
the level of contamination of shellfish depends on the
extent of pollution in the growing waters.
Bacterial contaminant levels in the three periwinkle
species evaluated exceeded the acceptable limits for
shellfish. The International Commission on Microbio-
logical Specifications for Foods (Jay, 1978) and the
US Food Drug and Administration (FDA, 1991) have
suggested a maximum SPC of +=lOOOOO and coliform
level of +100/g of shellfish for consumer safety. The
results of the present study corroborate the earlier
reports of Ekanem et al. (1994) and Ekanem and
Adegoke (1995) who observed unacceptable levels of
bacterial contaminants (including pathogens) in clams
obtained from the waters from which the periwinkles
in this study were harvested.
The bacterial genera identified in the periwinkles
included Bacillus, Salmonella, Klebsiella, Micrococcus,
Shigella and Pseudomonas. Escherichia coli and
Staphylococcus aureus were also specifically identified.
The occurrence of enteric organisms in the periwin-
kles was an indication of the pollution of their
overlying waters with untreated faecal wastes and
sewage. More detailed studies are necessary to
achieve better understanding of these preliminary
observations.
CONCLUSIONS
The three periwinkle species evaluated contained
unacceptable levels of bacteria, with the fresh water
species I! bryonensis carrying the highest level of
contaminants. Coliforms accounted for very high
proportions of the total bacterial load in all species.
Considering the public health implications of the
poor bacteriological quality of these products, strin-
gent attention should be paid to their safety through
proper processing, storage and handling procedures.
The introduction of enforcable microbiological guide-
lines as a way of protecting consumers appears to be
highly desirable. This should, however, wait until
more elaborate and detailed evaluation of the bacter-
iological characteristics of this seafood has been
carried out, and the relationship between the
envisaged criteria and the hazards they are meant to
offer protection againts properly examined.
 Bacterial contamination of periwinkles: E. 0. Ekanem and B. N. Otti
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Food Control 1997 Volume 8 Number 2 89