World Applied Sciences Journal 16 (7): 903-906, 2012

ISSN 1818-4952
© IDOSI Publications, 2012

Microbiological Status in a Fully Export-Oriented Shrimp Processing Plant

M. Yusuf Ali, M. Belal Hossain and M. Shamsuddin

Department of Fisheries and Marine Science,

Noakhali Science and Technology University, Sonapur, Noakhali. Bangladesh

Abstract: The present study was carried out in the period from 1st April 2010 to 1st August 2010 to asses the
microbial load of frozen shrimps processed for exporting to different countries of the world. For the microbial
analyses, samples were collected from cooked IQF (Individual Quick Freezer) shrimp and raw block frozen
shrimp. Total bacterial load of water, ice, contact surfaces at different stages were also investigated. The mean
total coliforms observed in Cooked IQF shrimp was <3 ±0.00 MPN/g, while it was 23.50 ± 13.72 MPN/g in raw
block frozen shrimp. Fecal coliforms for both raw block frozen and cooked IQF shrimp were <3 MPN/g. In the
current investigation the SPC of normal water, UV radiated water, ice, receiving table, grading table and panning
tray and worker’s hand (average of 3 samples ) were 8.34, 0, 5.14 44, 40, 31 and 23 cfu/cm2 respectively, which
were under limit of international standard. So the present findings indicate that the hygienic condition of the
investigated fish processing plant was good and the quality of Cooked IQF shrimp was excellent for export.

Key words: Coliforms % IQF % Block Frozen Shrimp

INTRODUCTION implementation of HACCP system as well as poor

established of GMPs (Good Manufacturing Practices),
Fisheries items are the major protein contributing GHPs (Good Hygienic Practices) and inadequate
source of Bangladesh. Fisheries sector contributes 3.74% sanitation procedures (Pers. Com.). On July 30, 1997, the
of our GDP and 22.23% of agricultural sector [1]. EU (European Union) banned imports of fishery products
Bangladesh is a sea food exporting country and exports from Bangladesh after the inspection of Bangladesh’s
mainly frozen shrimps, fresh water fishes and marine water seafood processing plants. Inspection found serious
fishes to Japan, USA, Europe, Saudi Arabia, the UAE and deficiencies in the infrastructure and hygiene in
Gulf States [2]. Export earnings from fisheries sector have processing establishments and insufficient guarantees of
increased from 1283 Taka core in 1995-1996 to Taka core quality control by Bangladeshi government inspectors.
3025.93 in 2009-2010 [2]. The ban was estimated to cost the Bangladesh shrimp-
By the year 1985, a good number of processing processing sector nearly US$15 million in lost revenues
plants has developed without biological survey between from August to December 1997 [4].
capture and culture fishery. In the past time most of the Bangladesh frozen shrimp exporters continue facing
factory led the emphasis on quantity rather than quality. problems with buyers in the U.S. the EU and Japan;
Processed products quality depends on the quality of raw concerning the safety and quality of their products
materials but it is too difficult to retain freshness of raw because many fish processing plants in Bangladesh did
materials due to long period of time between harvesting not follow the HACCP system and EU hygienic
and processing periods. Inadequate processing has regulations. Recently, Bangladesh government has been
resulted in microbial growth which deteriorates food taking serious action against the shrimp and fish
products [3]. Now export market of Bangladesh is processing plants which are not following importers
threatened for inadequate processed foods which may conditions and not implementing HACCP system for
be contaminated by decomposition, high bacterial load, ensuring quality products [4]. Therefore the present study
filth, unexpected foreign matters as well as pathogenic was undertaken to asses’ microbial load of frozen shrimps
microbes (E. coli, Salmonella, V. cholerae etc). These processed for exporting to different countries of the
activities occur occasionally due to improper world.

Corresponding Author: M. Belal Hossain, Department of Biology, FOS, Universiti Brunei Darussalam, Brunei.
Tel: +6738665470.
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MATERIALS AND METHODS by the respective dilution factor. The total numbers of
bacteria found from each Petridis for each dilution were
Study Area: The investigation was carried out in the averaged to find a reliable standard plate count (SPC).
microbiology laboratory of the processing plant (ARK Sea
Food Ltd. Chittagong Bangladesh) for 4 months during 1st Enumeration of Total Coliforms: Samples were prepared
April 2010 to 1st August 2010. as described in section serial dilution. Dilution blanks
remaining from the determination of the plate count were
Sampling Procedure and Processing: For microbial used. One ml of each of the decimal dilutions (10G1, 10G2,
analysis four samples were taken from different stages of 10G3) was poured into each of the three separate tubes of
processing e.g. from normal water, UV radiated water, ice, lauryl tryptose broth (LTB) containing Durhams Tube.
receiving table, grading table, panning tray and worker’s The tubes were incubated at 37°C for 48 hrs. The
hand. Shrimp samples were collected from different grades formation of gas after 48 hrs was considered a sufficient
of processed shrimp in the plant e.g. Sample 1: shrimp IQF evidence of the presence of coliforms. The positive gas
(Individual Quick Freezer) (tail-on), Sample 2: shrimp IQF production was recorded and the result was computed
(tail-off), Sample 3: Shrimp Peel Deveining Tail on and using MPN chart [7].
Sample 4: Shrimp Peel Deveining Tail off.
Representative samples of shrimps/ice/water were Test of Fecal Coliforms: Tubes of lauryl tryptose broth
collected at specific steps of processing and were that was positive for gas production were selected and a
assessed for microbial analyses soon after sampling. The loopful of Broth from each positive culture were
samples were carried to the laboratory in a sterile wide inoculated into a tube of Brilliant Green Bile (2%) Broth
mouthed bottle within 5 minutes. Frozen shrimps were and a tube of tryptone water tube were tasted with
stored at -20°C and unfrozen perishable samples held at 0- Kovac’s reagent to determine the presence of indole [5].
4°C for not longer than 6 hrs. A positive indole reaction in a broth that has produced
gas at 44°C indicates the presence of E. coli. The positive
Swab Test: A solution of peptone (1 g/L) and sodium gas production tubes were recorded and the results were
chloride (8.5 g/L) was made up. The solution was computed using MPN chart [7].
distributed in bottles and was sterilized for 15 min at
121°C. Bacteria from a known area of a surface were Detection of Salmonella: The presence of Salmonella
removed by passing a sterile cotton wool moistened with was detected by homogenizing a 25 gm portion of the
sterile peptone water, from which bacterial counts were composite sample in 225 ml (pH 7.5) sterile buffered
made as the procedure adopted for standard plate count peptone water aseptically and incubating for 24-48 hrs at
(colony/cm2) [5]. 37°C in an incubator. After incubation 1 ml sample was
transferred to duplicate tubes of tetrathionate (9 ml) and
Microbial Analysis: selenite cysteine broth (9 ml), incubated for 24 hrs at 37°C
Enumeration of Total Bacterial Load (SPC): Twenty gm and sub-cultured into xylose lysine deoxycholate (XLD)
of the sample were blended for 1 min with 180 ml of sterile and brilliant green agar (BGA) [8]. After incubation for
dilute (0.1% peptone) in an automatic blender (dilution of 24-48 hrs at 37°C the characteristics colonies (on XLD;
10G1); 1 ml of the 10G1 dilution was transferred to a screw black centered, convex entirely glossy colonies and on
cap vial containing 10 ml of sterile dilute to give a dilution BGA-pink, red, convex, entirely glossy colonies
of 10G2. The containing screw cap vial was shaken gently. surrounded by brilliant red zones in the agar) were
This process was repeated, using the progressively streaked with sterile platinum wise loop and incubated at
increasing dilution to prepare dilutions of 10G3, 10-4 and 10- 37°C for 6 hrs. If black color and H2S gas were found, that
5
[6]. Each dilution was plated by pipette 1 ml into a sterile indicated the presence of Salmonella [7].
plate containing 15 ml agar which was melted and brought
to 45°C and poured into the plates. Fewer than 15 minutes Detection of V. Cholerae: Twenty-five gm portion of the
were elapsed between making the dilution and pouring the composite sample was added in 225 ml sterile alkaline
agar. After solidification of the media, the plates were peptone water aseptically and incubated at 37°C for 24
inverted and incubated at 37°C for 18-24 hrs [7]. The total hrs. After incubation a loopful from the alkaline peptone
number of bacteria per gm of sample was obtained by water was streaked on the surface of separate plates of
multiplying the average number of colonies on paradises thiosulfate citrate bile salts (TCBS) agar in such a manner

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to obtain individual colony and incubated at 37°C for 24

hrs. The colonies of V. cholera were plain, yellow in color
and of very big size (generally 2-3 mm). From TCBS the
selected colony was transferred to triple sugar iron (TSI)
agar slant with streaking. Then TSI agar tubes were
incubated at 37°C for 24-48 hrs. The presence of gas and Fig. 1: Standard plate count in water.
black color in TSI slants, indicated that V. cholerae was
absent. If the media became yellow in color then V.
cholerae is present [7].

RESULTS AND DISCUSSION

Total Bacterial Load/Standard Plate Count (SPC):

The SPC of normal water, UV radiated water, ice, receiving
table, grading table and panning tray, worker’s hand
(average of 3 samples) were 8.34, 0, & 5.14 cfu/ml, 44, 40,
31 and 23 cfu/cm2, respectively (Fig. 1& 2).
Fig. 2: Standard plate count in SWAB test at different
Total and Faecal Coliforms: Total and fecal coliforms stages.
were nil from normal water, ice, receiving table, grading
table and panning tray and worker’s hand (Fig. 2). Total Table 1: MPN/g (mean ± SD) counts of total coliform detected in different
samples of different processes
coliforms of cooked IQF <3 MPN/g and in raw block
Sample name Raw block frozen shrimp Cooked IQF shrimp
frozen 23.50 MPN/g. But faecal coliform is <3 MPN/g both
Sample 1 9 <3
of cooked IQF and raw block frozen (Table 1&2). Sample 2 23 <3
Sample 3 20 <3
Salmonella and V. Cholera: Salmonella and V. cholerae Sample 4 42 <3
were totally absent from all sorts of sample viz. normal Mean 23.50 ± 13.72 <3±0.00
water, UV radiated water, ice, receiving table, grading
table, panning tray, worker’s hand as well cooked IQF and Table 2: MPN/g counts of fecal coliform observed in different samples of
different processes
raw block frozen shrimp.
Sample name Raw block frozen shrimp Cooked IQF shrimp
According to ICMSF the acceptable upper limit of
Sample 1 <3 <3
total bacterial load, total and fecal coliforms is 106 cfu/g, Sample 2 <3 <3
100 and <3 MPN/g, respectively, while Salmonella and/ Sample 3 <3 <3
or V. cholera should not present. In the present study the Sample 4 <3 <3
total coliforms was under the limit of ICSMF [9] and
presence of coliform is not permitted in the food products IQF shrimp. They observed total coliforms in Cooked IQF
in importing countries [10]. Besides, Salmonella and V. shrimp <3 MPN, while it was 21.00 ± 0.25 and 4.20 ± 1.20
cholera were not detected in the Raw Block Frozen and MPN in Raw Block Frozen shrimp and Raw IQF shrimp
cooked IQF shrimp. Thus the samples of Raw Block respectably. Fecal coliforms, V. cholerae and Salmonella
Frozen and cooked IQF shrimp were under the acceptable were not detected in any of the samples. Huq et al. [6]
limit according to ICMSF and FDA guidelines [9, 3]. worked on quality aspect of frozen shrimp product in
Hossain, et al. [11] carried out an experiment on processing industry: a case study in Khulna Bangladesh.
microbiological quality of processed frozen black tiger They reported that the highest SPC was 7 CFU/cm2 in the
shrimps in fish processing plant. The study was overhead tank and lowest was 3 CFU/cm2 in the panning
conducted to evaluate the effectiveness of processing tape water. SWAB samples were collected from worker
techniques of shrimps by microbiological quality hand, where mean SPC before and after working, were 20
assessment. The abundance of total aerobic bacteria, total and 30 per cm2 respectively. During the processing
coliforms, fecal coliforms, Vibrio cholerae and technique, the samples of Cooked IQF Shrimp showed the
Salmonella were determined in Raw Block Frozen shrimp, lowest total coliforms. In Cooked IQF shrimp elimination
Cooked IQF (Individual Quick Freezer) shrimp and Raw of bacteria occurs in two steps first during cooking and

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then freezing. On the other hand, in raw block frozen, 7. USFDA, 1984. Bacteriological Analytical Manual
abolition occurs only during freezing. The lowest count in Association of Official Analytical Chemist (6th
Cooked IQF shrimp might be because of this reason. From edition). AOAC Intl. Arlington, VA, pp: 529.
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are highly qualified for export purpose. action (1st edition). Taraporeavala Sons & Co. Private
Ltd, Mumbai, pp: 108-110 and 142-147.
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