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Comparative Analysis of Microbiological Status Between Raw and Ready-to-eat Product of Black Tiger Shrimp (penaeus Monodon)

Md. Shakir Hossain
Department of Fisheries and Marine Science, Noakhali Science and Technology University, Sonapur 3814, Bangladesh

Mehedi Mahmudul Hasan
Department of Fisheries and Marine Science, Noakhali Science and Technology University, Sonapur 3814, Bangladesh

Md. Golam Sarwer
Department of Fisheries and Marine Science, Noakhali Science and Technology University, Sonapur 3814, Bangladesh

Shuva Bhowmik
Department of Fisheries Technology, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh

Abstract/ Executive Summary:

The investigation was carried out to detect the health hazard microbiological status in raw (head on and shell on) and ready-to-eat product of black tiger shrimp (Penaeus monodon). The abundance of total bacterial load, total coliform, faecal coliform, Salmonella spp., Vibrio cholerae were determined with three different samples of raw and ready-to-eat product of black tiger shrimp. The study represents that raw (head on and shell on) shrimp contains more microorganisms, because in raw shrimp elimination of head and shell was not occurred which contains maximum number of microorganisms of the total body. Total bacterial load (Aerobic Plate Count) of raw black tiger shrimp (head on and shell on) was 1.8967× 105±0.187× 105 CFU/g whereas in ready-to-eat product of shrimp was 0.17×10⁵CFU/g ± 0.0088× 10⁵CFU/g respectively. However total coliform of raw black tiger shrimp and ready-to-eat product of shrimp were 66.67±14.50 MPN/g and <3 MPN/g respectively, and faecal coliform of raw black tiger shrimp (head on and shell on) and ready-to-eat product of black tiger shrimp were 2.20±1.11 MPN/g and <3MPN/g, respectively. Furthermore Salmonella spp. and Vibrio cholerae were totally absent in raw and ready-to-eat product of shrimp. The above mentioned data were under the limit of international standard. This study implies that the quality of ready-to-eat product of black tiger shrimp was excellent for exporting and presented no harm to human health.

Keywords: Microbiological status, Raw products, Ready-to-eat product, Salmonella spp. and Vibrio cholerae

Location: Noakhali Science and Technology University

Start Date: 00-01-2014

End Date: 00-00-2014

Program Area: Animal Health and Management

Commodity: Shrimp

Objectives:

To detect the health hazard microbiological status in raw (head on and shell on) and ready-to-eat product of black tiger shrimp (Penaeus monodon).

Methodology:

Sample selection for Microbial Analysis: Three samples of black tiger shrimp (head on and shell on) and three samples of ready-to-eat products were collected randomly from three different lots in receiving hall at the beginning of processing. Microbial Analysis: Microbial analysis was performed according to the standard procedure for the enumeration and identification of microorganisms. Enumeration of Total Bacterial Load (APC): 20g of the raw shrimp sample was blended for 1 min with 180 ml of sterile dilute 0.1% peptone in an automatic blender. That provided a dilution of 10^-1. This process was repeated, using the progressively increasing dilution to prepare dilution of 10^-2, 10^-3, 10^-4, and 10^-5. The same procedure was followed for the ready-to-eat product samples.1ml 10^-1 solution was added with 9ml, 0.1% peptone water (10^-2) and 1ml with 9ml LTB (Lauryl Tryptose Broth) in the Durham’s tube (10^-1). Then 10^-2 solution was converted to 10^-3, 10^-4, 10^-5 solution with the 0.1% peptone solution. Poured 1ml of solution from each test tube was the sterile Petri dish. Approximately 15 ml agar which has been melted and brought to 45°C was poured into the plates. Plates were rotated by hand 5 times in the clockwise direction, 5 times in the counter clockwise direction and several times crosswise for equal distribution of the media. Fewer than 15 minutes were elapsed between making the dilution and pouring the agar. After solidification of the media, the plates were inverted and placed in incubator to incubate at 37°C for 18 hrs. After 48 hours, the number of colonies which were developed in the Petri dishes was properly counted by colony counter machine. The total number of bacteria per gram of sample was obtained by multiplying the average number of colonies on Petri dishes by the respective dilution factor. The total number of bacteria found from each Petri dish for each dilution was averaged to find a reliable aerobic Plate Count (APC). Enumeration of Total Coliform: 20g of the sample was blended for 1 min with 180 ml of sterile dilute 0.1% peptone in an automatic blender. That provided a dilution of 10^-1. 1ml 10^-1 solution was added with 9ml. 0.1% peptone water (10^-2) and 1ml with 9 ml LTB (Lauryl Tryptose Broth) in the Durham’s tube (10^-1). Then 10^-2 solution was converted to 10^-3, 10^-4, 10^-5 solution with the 0.1% peptone solution. Solution of 10^-1, 10^-2, and 10^-3 were incubated Durham’s tubes at 37°c for 48 hours. The formation of gas after 48 hours was considered sufficient evidence of the presence of coliform. Gas formation was recorded and the result was computed by using MPN chart. Detection of Faecal Coliform: Tubes of lauryl tryptose broth that was positive for gas production were selected and a loopful of Broth from each positive culture were inoculated into a tube of Brilliant Green Bile (2%) Broth and a tube of tryptone water tube were tasted with Kovac’s reagent to determine the presence of indole. A positive indole reaction in a broth that has produced gas at 440C indicates the presence of E. coli. The positive gas production tubes were recorded and the results were computed using MPN chart. Detection of Salmonella: The presence of Salmonella spp. was detected by homogenizing a 25 g portion of the composite sample in 225 ml (pH 7.5) sterile buffered peptone water aseptically and incubating for 24-48 hr. at 37°c in an incubator. After incubation 1 ml sample was transferred to duplicate tubes of Tetrathionate (9 ml) and Selenite Cysteine Broth (9 ml), incubated for 24 hours at 37°c and sub-cultured into Xylose Lysine Deoxycholate (XLD) and Brilliant Green Agar (BGA). After incubation for 24 hr at 37°c, characteristics colonies (on XLD- black centered, colony, convex, entries, glossy and on BGA-pink, red, convex, entree glossy colonies surrounded by brilliant red zones in the agar) were streaked with sterile platinum wise loop incubated at 37°c for 6 hrs. After incubation characteristics change may help about presence or absence of Salmonella. H₂S gas was not found which indicate that Salmonella was absent in the sample. Detection of V. cholerae: 25g portion of the composite sample was added in 225 ml. sterile alkaline peptone water aseptically and incubated at 37°c for 24 hrs. Loopful alkaline peptone water was streaked on the surface of separate plates of Thiosulfate Citrate Bile Salts (TCBS) agar in such a manner to obtain individual colony and incubated at 37°c for 24 hrs. After 24 hours, V. cholerae colony was tested. The colony of V. cholerae was plain, yellow color and very big size (generally 2-3 mm). From TCBS the selected colony was transferred to the butt of Triple Sugar Iron Agar (TSIA) Slant with streaking. Then, TSIA tubes are incubated at 37°C for 24 hrs. Black color gas was observed in TSIA indicated that V. cholerae was absent. Data Analysis: All data were analyzed with Microsoft Excel 2007. Data were presented as mean ± SEM.

Source of Information: International Journal of Biosciences Vol. 6, No. 8, p. 43-49, 2015

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

Good quality of sea food depends on the careful manipulation of every stage from initial production to processing, storage, marketing and consumption. As it ensured the reduce amount of microbial load so it may be said that is ready for eat as well as export. The GMPs and sanitation procedures of the selected fish processing plant were excellent; hence their product performance (quality) earned a better place in the foreign markets. It is revealed that ready-to-eat product of black tiger shrimp a reduced amount of microbes product than raw (head on and shell on) tiger shrimp for export purpose.

Knowledge Management: Journal