Republic of the Philippines

Department of Agriculture
Bureau of Fisheries and Aquatic Resources
NATIONAL INTEGRATED FISHERIES TECHNOLOGY DEVELOPMENT CENTER
MICROBIOLOGY AND FISH HEALTH LABORATORY
Bonuan Binloc, Dagupan City, Pangasinan, Philippines
Tel. Nos. (075) 649-6086; (075) 649-6068
Email address: bfarniftdc@yahoo.com / bfarniftdc.microlab@gmail.com

CLINICAL LABORATORY RECORD

Case Code No : AAHL-04-06-2021-006 Date & Time Submitted:04/06/21|02:00 PM

Sample Code No. : LA-FHL-009-010 Date & Time Examined:04/06/21|02:30 PM
Sample(s) : 1 brackishwater
1 pooled Macrobrachium larvae
Date &Time Collected: April 6, 2021 | 02:30 PM
Place Collected/Source: Macrobrachium Hatchery larval rearing tank
Submitted by : Mr. Jomar DV. Parana
Company Name : BFAR – NIFTDC – Macrobrachium Hatchery Section
Address : BFAR – NIFTDC Compound, Bonuan Binloc, Dagupan City, Pangasinan, 2400
Analysis Conducted : Aerobic Plate Count (cfu/ml) | Total (Presumptive) Vibrio Count
(cfu/ml) | Luminous Bacterial Count (cfu/ml) and bacterial identification

RESULT OF BACTERIOLOGICAL ANALYSIS

A. Result of Bacterial Enumeration

Total
Percentage of Percentage of
Sample Aerobic Plate (Presumptive) Luminous
Sample Code Source Green Vibrio Yellow Vibrio
Description Count (cfu/ml) Vibrio Count Bacterial (cfu/ml)
Colonies (%) Colonies (%)
(cfu/ml)
LA-FHL-009-BW water Larval rearing 1.2 x 104 1.8 x 103 37.93% 62.07% ND
LA-FHL-010-LV larvae tank #08 <2.5 x 102 1.4 x 103 2.70% 97.39% ND
METHODOLOGY: Serial Dilution Method (Spread Plate)

STANDARD BACTERIAL PROFILE (Bacterial Analysis For Shrimp and Water: W1-BA-01 – BFAR CO)
Total Bacterial Count <5.0 x 106 - NORMAL
5.1 x 106-8.9 x 106 - Slightly high
9.0 x 106-5.0 x 107 - High
5.1 x 107-9.0 x 107 - Very high
9.1 x 107-1.5 x 107 - Extremely high

Note: For shrimp and pond water, percentage of yellow Vibrio colonies should be more than 60%
ND – means Not Detected
TNTC – means Too Numerous to Count

B. Result of Bacterial Identification

Specimen Bacterial Isolate

Macrobrachium dacqueti larvae Aeromonas sobria
Vibrio cholerae
Larval rearing water Vibrio mimicus
 Remarks:

All samples were found to have Aerobic Plate Count (cfu/ml) and Total Vibrio Count
(cfu/ml) within the normal range based on standard bacterial profile for shrimp and water: W1-
BA-01-BFAR CO. In addition, the predominant bacterial isolates from the larvae and water were
identified as Aeromonas sobria, Vibrio cholerae and Vibrio mimicus using Analytical Profile
Index 20E.

Result(s) as per sample(s) received and analyzed. This report shall not be reproduced except in full and with written
consent from BFAR-NIFTDC Microbiology and Fish Health Laboratory Section

Examined by: Checked by: Noted by:

JOHN CARLO G. GOSILATAR EDITH C. ROXAS CORDELIA B. NIPALES

Laboratory Analyst Senior Aquaculturist Supervising Aquaculturist
Head, Microbiology and Officer-In-Charge, BFAR -
RIZALYN J. BAUTISTA Molecular Pathology NIFTDC
Laboratory Analyst Laboratory
Head, Microbiology and Aquatic
GEELYN DV. CAROLINO
Laboratory Analyst
Literature(s):

Vibrio species are Gram negative bacteria, possessing a curved-rod morphology and test positive
for oxidase. These bacteria are part of the natural microflora of wild and cultured prawns and become
opportunistic pathogens when natural defense mechanisms of the prawn are suppressed (Brock and
Lightner, 1990).

The non-halophilic species Vibrio cholerae and V. mimicus persist autochthonously in a diverse
range of aquatic environments. These bacteria have been isolated from marine, estuarine, brackish, and
freshwater environments (Davis et al . 1981 ; Colwell 1984; Nair et al . 1988) . The distribution of
nonhalophilic Vibrio in confined brackish and freshwater environments is facilitated by conditions of
warmer water temperatures and high levels of pH and organic nutrients (Singleton et al . 1982 ; Nair et
al . 1991).

Vibrio species exist in the water used in prawn culture facilities. Bacteria enter prawns via
wounds or cracks in the cuticle and are ingested with food (Lavilla-Pitogo et al., 1990; de La Pena et al.,
1995).

Vibriosis is one of the major secondary bacterial diseases affecting mariculture, though less
attention has been paid to the implications of nonhalophilic vibrios for the freshwater aquaculture
industry (Egidius 1987 ; Ruangpan and Kitao 1991). Vibrio cholerae non-O1 and V. mimicus have been
isolated from apparently healthy marine and freshwater fish (Lupiani et al . 1993) and have been
associated with brackish and freshwater prawns in Bangladesh and India (Chowdhury et al . 1986; Nair et
al . 1991). The bacteria isolated in these cases were not associated with disease.

Furthermore, Aeromonas species are Gram-negative bacteria that are ubiquitous in aquatic
environments.

the resident bacteria in the culture water of freshwater prawn identified in this study were mostly
opportunistic pathogens which can proliferate beyond the threshold levels and cause disease
outbreaks in prawn larvae especially if the host is stressed. It is, therefore, necessary to regulate
the bacterial load in the freshwater prawn aquaculture system by maintaining good water quality
and good management practices through disinfection of culture facilities, rearing water and
Artemia cysts, the practice of good hygiene of hatchery personnel, regular water exchange and
feed regulation among others. These practical safety measures managed the resident bacterial
microbiota in the rearing water, eggs, and larvae and consequently prevented the proliferation of
potential pathogens. The use of antibiotics may also be explored but with regulation and caution
because of known negative health implications (Asian Fisheries Science 33 (2020):241–248)

References: Kh.M. Sharshar and E.A. Azab, 2008. Studies on Diseased Freshwater
Prawn Macrobrachium rosenbergii Infected with Vibrio vulnificus. Pakistan Journal of
Biological Sciences, 11: 2092-2100.

F. Y. K. Wong , K. Fowler & P. M. Desmarchelier (1995) Vibriosis Due to Vibrio mimicus in

Australian Freshwater Crayfish, Journal of Aquatic Animal Health, 7:4, 284-291, DOI:
10.1577/1548-8667(1995)0072.3.CO;2

M.A. MONGHIT-CAMARIN1,2, E.R. CRUZ-LACIERDA1, *, R.V. PAKINGKING, JR.3 ,

M.L. CUVIN-ARALAR4 , R.F. TRAIFALGAR1 , N.C. AÑASCO5 , F.W. AUSTIN6 , M.L.
LAWRENCE6 Bacterial Microbiota of Hatchery-Reared Freshwater Prawn Macrobrachium
rosenbergii (de Man, 1879)
Bower, S.M. (1997): Synopsis of Infectious Diseases and Parasites of Commercially Exploited
Shellfish: Vibrio spp. (Vibrio Disease) of Cultured Shrimp. December 1997
 Result(s) as per sample(s) received and analyzed. This report shall not be reproduced except in full and with written
consent from BFAR-NIFTDC Microbiology and Fish Health Laboratory Section

Examined by: Checked by: Noted by:

JOHN CARLO G. GOSILATAR EDITH C. ROXAS CORDELIA B. NIPALES

Laboratory Analyst Senior Aquaculturist Supervising Aquaculturist
RIZALYN J. BAUTISTA Head, Microbiology and Officer-In-Charge, BFAR - NIFTDC
Laboratory Analyst Molecular Pathology Laboratory
Head, Microbiology and Aquatic
GEELYN DV. CAROLINO Animal Health Laboratory
Laboratory Analyst