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Meri-Rastilantie 3 B, FI-00980 Journal of Food, Agriculture & Environment Vol.12 (2): 1260-1265. 2014 www.world-food.net
Helsinki, Finland
e-mail: info@world-food.net

Bacteriological study of pond water for aquaculture purposes

Adedayo Olajide Ajayi * and Anthony Ifeanyin Okoh
Applied and Environmental Microbiology Research Group (AEMREG), Department of Biochemistry and Microbiology,
University of Fort Hare, Private Bag X1314, Alice 5700, South Africa. *e-mail: AAjayi@ufh.ac.za
Received 12 February 2014, accepted 3 April 2014.

Abstract
Bacteriological analysis of fish pond water is very important in aquaculture as this gives insight to the likely hazard that may occur to the fishes, the
farmer and the consumer. Some physico-chemical properties of this study show that the values of pH range from 7.82 in sample source 2 to a high
range of pH 8.15 in sample source 1, and the temperature ranges from 27°C to 31°C in sample sources. Similarly, the Cd content and the total hardness
of the pond water sources varies from low of 0.98 mg/L and 9.31 mg/L in Academic planning office (a) to a high of 7.80 mg/L in Araromi area 2 and
128.92 mg/L in Araromi area 1, Akungba-Akoko, Nigeria, respectively. The bacterial load during this study ranged from low of 16×103 cfu/ml in
Araromi (1) to a high of 12.4×105 cfu/ml in Academic planning pond source (Sample 6). Similarly, the coliform count ranged from 3.5×103 cfu/ml in
Araromi (1) to 9.0×105 cfu/ml in Adefarati sampling area. This study shows diversified forms of bacterial species populating the pond water sources.
This includes Staphylococcus spp., Streptococcus spp., Bacillus spp., Pseudomonas spp., E. coli, Enterobacter spp., Proteus spp., Citrobacter spp.
and others. The presence of these organisms shows lack of qualitative pond management services which could become harmful to both fishes and
humans in the food web systems. Hence, the need to protect our water resources for aquaculture purposes and sustainable development through the
detection of aquatic aetiologic agents and possible control of this microbes.

Key words: Aquaculture, bacteriological, pond, water.

Introduction
Pond water sources are useful for diversified purposes including ponds, whereby, some are beneficial, others are not.
aquaculture and other related uses at the domestic level. Ponds Beneficial ponds bacteria are natural and safe for fish, pets and
are naturally formed by a depression in the ground filling and people. Beneficial bacteria are microorganisms that occur naturally
retaining water. Streams or spring water is usually fed into these in water gardens, streams, ponds, etc. They are responsible for
bodies. A fish pond is an artificial lake (reservoir, pond) intended maintaining crystal clear healthy water, breaking down organic
for fish breeding. Fishes are the most popular animal cultured in waste, breaking down ammonia from fish waste, reducing nitrite
the pond. Fishes are among the edible food sources naturally and nitrate, reducing nutrient load in ponds and balancing the
living in water, consumed by man and containing many nutrients ecosystem. Aquatic bacteria, through the process of decomposition
such as protein, minerals, fat, oil, etc. Fish ponds are constructed and as sources of food, play an important role in pond ecosystems
where fishes are fed and their growth are easily observed and and also in fish production. Previous researches 3-5 show that the
monitored. Fishes that are commonly produced in fish ponds are production levels based on aquatic bacteria are greater, by 1.8 to
catfish, tilapia and codfish, to mention a few 1. In Nigeria, catfish 9.3 times than in system based on phytoplankton. Non-beneficial
is produced in 98% of our fish ponds. The species of catfish that bacteria cause offensive odour to ponds and also diseases in
can be produced include: Clarias anguillaris, Clarias gariepinus, fishes. However, fishes have evolved a number of protective
Heterobranchus congifilis, Heterobranchus bidorsalis, etc. adaptations with several unique traits against bacteria and other
Water is very essential in fish pond, water plays a vital role in organisms in their outside coverings including scales, fins and in
the proper functioning of earth ecosystem and also essential for particular, protective mucous which cover the body, and also
fish and living creatures for metabolism. The temperature of water secretions from various organs (gills, liver), possesses antibiotic
supplied to a fish pond ranges from 25°C to 35°C as this supports properties 6, 7.
the growth of the microorganisms and fishes found in the pond. The distribution of heterotrophic bacteria and total aquatic
There are various sources of water, including well water, borehole bacteria vary with the water layers. According to previous research 8,
water, stream water, river water, etc., that can be supplied to the in the 50 cm water sources, which is near the water surface and
fish pond. Some bacteria coliform groups like E. coli, in the ponds contains abundant quantities of dissolved oxygen and organic
are transported from these sources of water or the media of water, the aerobic bacteria reproduce very quickly and the result
transportation into the ponds. There are several microorganisms is a large number of heterotrophic bacteria in this layer. In the 100
found in ponds including bacteria, fungi, algae, protozoa, cm middle layer, where most of the filter feeding fish takes food,
nematodes and viruses. Bacteria has a unique characteristics, they the number of bacteria is lowest of the three in this water region.
are ubiquitous in every habitation on earth, growing in soil, acidic The number of bacteria is higher of the silt layer because the
hot springs, radioactive wastes, water and the live bodies of plants sediment contains organic matter such as food and fish manure.
and animals 2. Thus, bacteria are important microorganisms in The dissolved oxygen content is low, however. There is also

1260 Journal of Food, Agriculture & Environment, Vol.12 (2), April 2014
difference in the levels of bacteria produced during the morning Bacteriological analysis of samples: Triplicates of the sample
and relatively 40% higher level in afternoon probably due to change sources were serially diluted and appropriate diluents of 1 ml
in water temperature and dissolved oxygen content (DO) 8. sample size were inoculated with pipette into sterile Petri dish.
There are various factors affecting the distribution of bacteria Sterile culture medium was introduced and swirled for mixture.
in fish pond which includes predatory protozoa present in water. The Petri dishes were inverted to prevent condensation dropping
This has significant impact in decreasing the number of bacteria. from the lid into the agar and incubated in the incubator at 37°C
Protozoa require living or dead bacteria for food and easily engulf for 24-48 h 10. The total bacteria and coliform counts were
large number of these organisms, provided the water contains enumerated from these sources and Gram stained.
sufficient dissolved oxygen. In a water supply, the toxicity of
ultraviolet rays is inversely proportional to turbidity. Similarly, Sub-culture by streak method: Eosin methylene blue (E. M. B)
increasing temperature exerts a harmful effect upon the survival and nutrient agar (NA) were generally used during the study.
of some organisms in water, especially those capable of producing Subsequently sub-culture was carried out until pure isolates were
disease. On the other hand, multiplication of certain soil and transferred onto agar slant in McCartney or super bottles and
intestinal forms actually occur when the temperature of the water kept in the refrigerator at 4°C to serve as stock culture for
is increased, e.g. E. coli is capable of multiplying when inoculated subsequent test during identification. This process was carried
at 37°C. An increase in food supply also usually results in an out aseptically to prevent contamination.
increase in bacterial number. On the other hand, certain toxic
substances such as acids and bases produced a marked reduction Identification of microbial isolates: Isolates were picked from
in the number of viable organisms 9. stock culture and sub-cultured by streaking on Nutrient Agar,
In this study, we determined various types of microorganisms and incubated at 37°C for 24 h, followed by the various biochemical
including coliforms that are commonly associated with fish pond tests.
water and clarify protective ecological conditions favourable for
the survival, multiplication, and growth of stock fishes. This is Indole test: A pure bacterial culture was grown in a sterile
valuable in environmental monitoring and also helps to know tryptophan or peptone broth for 24-48 h, following incubation 5
the necessary management systems required for microbiological drops of Kovacs reagent (isoamyl alcohol, P-dimethylami-
quality maintenance of pond water sources. nobenzaldehyde, concentrated hydrochloric acid) that was added
to the culture broth. Positive result is shown by the presence of
Materials and Methods a red or red-violent colour in the surface alcohol layer of the
The materials used for this study were sterilized by appropriate broth, while negative result appears yellow.
technique. All glass wares such as Petri dishes, conical flasks,
test tubes, beakers, McCartney bottles, etc., were thoroughly Carbohydrate utilization test: Bacteria produce acidic products
washed and sterilized in the hot air oven at 170°C for about 2 h. when they ferment certain carbohydrates signified with change
The media were sterilized at 121°C for about 15-30 min in autoclave. in pH of the medium. If gas is produced as a by-product of
The inoculation loop was sterilized by flaming in the Bunsen burner fermentation, then the Durham tube will have a bubble in it. The
until it turns red hot. Similarly, working surface was sterilized by carbohydrate tests conducted include glucose test, lactose test,
the application of disinfectant solution (95% ethanol). fructose, sucrose test, etc. 11.

Sample collection: Pond water samples for the study were Citrate utilization test: Citrate is vital in Krebs cycle, the only
collected from different fishponds in Akungba Akoko, Ondo carbon source available to the bacteria in the media. If it cannot
state. Analytical study of these samples was carried out in the use citrate, such organism will not grow. If it can use citrate,
microbiological laboratory of Adekunle Ajasin University. then the bacteria will grow and the media will turn a bright blue
Samples were collected from Surulere Street, Adefarati, AAUA as a result of an increase in the pH of the media 11. The test was
fishpond, Araromi area 1 and Araromi area 2. These samples were carried out by using 5 ml Simmons citrate medium distributed into
collected aseptically and appropriately labeled. test tubes and subsequently plugged with non-absorbent cotton
wool. It was sterilized by autoclaving at 121°C for 15 min and
Physico-chemical analysis: The pH of each water sample was cooled. Isolates were incubated at 37°C for 24 h; utilization of
measured using pH meter, and the temperature of the samples was citrate was indicated by a change of colour of the medium from
measured with the help of a mercury thermometer by immersion green to deep blue.
into the water sample. In determining the total hardness of water
samples, 2 mm of Hydra pH buffer powder solution meant for Catalase test: Catalase enzyme protects bacteria from hydrogen
hardness determination was added to 100 ml of the sample and peroxide (H2O2) accumulation, which can occur during aerobic
swirled. Then 0.1 g scoop of ManVer 2 powder used as hardness metabolism. If hydrogen peroxide accumulates, it becomes toxic
indicator powder was also added to the solution and swirled. The to the organism. Catalase breaks down H2O2 into water and
mixture was titrated against 0.800 M EDTA until a change from red oxygen 11. In this test, small amount of the test organism was
to pure blue was observed as an end point. It is read and value is smeared onto the head of a Petri plate or culture dish. Then a drop
expressed as mg/L (CaCO3) standard 500 mg/L. The cadmium (Cd) of hydrogen peroxide was added to the smear. If bubbles become
components in the water sources were also determined, and the visible, this concludes that the organism produces catalase. Lack
resultant reading was taken on the atomic absorption of bubbles indicates negative result.
spectrophotometer (AAS).

Journal of Food, Agriculture & Environment, Vol.12 (2), April 2014 1261
Oxidase test: Some of the test culture was swabbed into one of Results
the bones on an oxidase dry slide. Colour changes to purple or This study helps to determine the microbiological properties and
blue after 30 s to 1 min is an evidence that the result is positive. some ecological parameters of fish pond water sources from the
The lab test is based on detecting the production of the enzyme area (Table 1). Pond water samples were collected from different
cytochrome oxidase by Gram- negative bacteria. fish ponds in the study area. Table 1 shows some physico-chemical
properties including the values of pH which range from pH 7.82
Motility test: The motility test is not a biochemical test since we (sample 2) to a high range of pH 8.15 (sample 1), and the temperature
are not looking at the metabolic properties of the bacteria. Rather, ranges from low unit of 27°C (sample 3) to a high temperature of
this test can be used to check for the ability of bacteria to migrate 31°C (sample 4). Similarly, the Cd content and the total hardness
a way from a line of inoculation using physical features like flagella 11. of the pond water sources vary from low of 0.98 and 9.31 mg/L in
Here, the bacterial sample was inoculated into motility media using Academic planning office (a) to a high of 7.80 mg/L in Araromi
a sterile needle. The media was stabbed in a straight line as possible area 2 and 128.92 mg/L in Araromi area 1, respectively.
and the needle was withdrawn very carefully to avoid destroying Table 2 shows the total bacterial and coliform counts of the
the straight line. After incubating the sample for 24-48 h, observation samples. The bacterial load ranged from low of 16×103 cfu/ml in
was made. It there is migration away from the line of inoculation, Araromi 1 to a high of 12.4×105 cfu/ml in Academic planning pond
it is evident that the test organism is motile (positive test). Lack of source (sample 6). Similarly, the coliform count ranged from 3.5×103
migration away from the line of inoculation indicates a lack of cfu/ml in Araromi 1 to 9.0×105 in Adefarati area (Table 2b).
motility (negative test). Morphological characteristics and some biochemical characteristics
including Gram staining reaction were used to give probable
Methyl red-Voges-Proskauer test (MRVP): This test is used to identity of the isolate (Table 3). Different bacterial species obtained
determine two things. The MR portion (methyl red) is used to during the study include Staphylococcus aureus, Escherichia coli,
determine if glucose can be converted to acidic products like Pseudomonas spp., Streptococcus spp., Bacillus spp., Enterobacter
lactate, acetate and formate. The VP portion (Voges- Proskauer) is spp., Proteus spp., Citrobacter spp. and Shigella spp.
used to determine if glucose can be converted to acetone. In
doing this, a single tube of MRVP media was prepared and Table 2a. Bacteria count of pond water
inoculated using a transfer loop, after sterilizing in the autoclave. sources on solid media.
The culture was allowed to grow for 3-5 days. After the culture is NA EMB MAC M.R.S
Sample MPN
10-3 10-2 10-2 10-3
grown, about half of the culture was transferred to a clean sterile 1 33 2 1 7 75
tube. One tube containing the culture was used to conduct the 2 14 5 2 12 150
MR test, and the second tube was served as the VP test. 3 16 35 5 58 460
4 30 36 12 17 210
Legend: NA - Nutrient Agar (Total bacterial count); EMB - Eosin
MR (Methyl red) test: Methyl red was added to the MR tube. Red Methylene Blue Agar (Coliforms count); MAC - Mac Conkey agar
colour indicates positive test result, i.e. glucose can be converted (Gram negative enteric bacillus); M.R.S. - De Man, Rogosa, Sharpe
Agar (Anearobic Lactobacillus spp); MPN-Most Probable Number.
into acids and products such as lactate, acetate and formate. Yellow
colour indicates negative test result, that is, glucose is converted Table 2b. Tota1 bacterial and coliform
into neutral end product 11. counts of pond water samples.
Total bacterial count Coliform count
Sample
VP (Voges-Proskauer) test: First alpha-napthrol (also known as (x 104 cfu/mL) (x 103 cfu/mL)
Barritt’s reagent A) and the potassium hydroxide (Barritt’s reagent 1 33 210
2 28 110
B) were added to the VP tube. The culture was allowed to stay for 3 42 340
about 15 min for colour development to occur. If culture turns red 4 76 410
colour, it indicates a positive result. If culture appears yellow to 5 107 824
copper in colour, it means a negative result. 6 124 34
7 120 900
8 85 640
9 1.6 3.5
10 3.0 3.6

Table 1. Physico-chemical properties of pond water sources.

Type Pond water Total hardness Cd
Sample Location pH
(water) temperature (°C) (mg/L) (mg/L)
1 EBF Fish farm (AAUA) Unfertilized 25.80 6.9 31.72 4.65
2 EBF Fish farm (AAUA) Unfertilized 26.00 6.7 19.43 3.67
3 EBF Fish Farm (AAUA) Unfertilized 25.60 6.9 31.72 6.43
4 EBF Fish Farm Unfertilized 26.40 6.2 32.55 4.73
5 Academic planning office (a) Fertilized 27.80 6.0 9.31 0.98
6 Academic planning office (b) Fertilized 28.20 5.8 12.10 2.10
7 Opposite AAUA gate Unfertilized 27.10 6.8 59.80 5.20
Surulere street, Adefarati area.
8 Pa Jinad Compound Fertilized 26.90 4.7 115.42 7.40
9 Araromi area 1 Fertilized 270C 8.04 128.92 2.46
10 Araromi area 2 Fertilized 310C 8.03 59.80 7.80
Legend: EBF - Environmental biology and fisheries; AAUA - Adekunle Ajasin University; Akungba - Akoko, Nigeria.

1262 Journal of Food, Agriculture & Environment, Vol.12 (2), April 2014
 Table 3. Boichemical characteristics of isolates.

Isolates Cultural characteristics Cell shape Identification

Indole
Lactose

Sucrose

Glucose
Motility
Oxidase

Catalase
Fructose
Mannitol
Ornithine

Coagulase
Methyl red

Gram reaction
Voges proskauer

Starch hydrolysis
Hydrogen sulphide
PD 1 Creamy, Raised, Circular, Entire + Rod + - + - AG AG -G AG AG + + + - + + Bacillus spp.
PD 2 Light purple, Raised, Circular, Entire - Short rod in cluster + - + - AG -G -G AG AG - + - - + + Pseudomonas spp.
PD 3 Transparent, Flat, Irregular, Lobate + Cocci + - + - AG -G -G AG AG + - + - + - Staphylococcus spp.
PD 4 Pale pink with dark center,Flat, Circular, Entire - Short rod . - + - AG -G -G AG AG - + + - + - Enterobacter spp.
PD 5 Grey like pale pink, Convex, Circular, Entire - Short rod in cluster + + + - AG -G -G A -G + + + + - - Edwardsiella spp.
PD 6 Whitish grey, Flat, Circular, Entire - Short rod - + + - AG AG AG A AG + - + + - - Escherichia coli
PD 7 Translucent, Flat, Irregular, Lobate + Cocci in cluster + - + + AG -G -G AG AG + - + + + - Staphylococcus spp.
PD 8 Green metallic sheen, Convex, Circular, Entire - Short rod - + + - AG -G -G AG A + + + + - - Escherichia coli
PD 9 Yellowish Raised, Circular, Entire - Short rod + + + - AG -G -G AG AG + - + - + + Flavobacterium spp.
PD 10 Translucent, Flat, Irregular, Lobate + Cocci + - + - AG AG -G AG AG + - + + + - Staphylococcus spp.
PD 11 Pale purple, Umbonate, Circular, Entire + Cocci in chain + - - - AG -G -G AG AG + - + + + - Streptococcus spp.
PD 12 Pale purple, Umbonate, Circular, Entire + Cocci in chain - - - - A- A- - AG AG + - + - + - Streptococcus spp.
PD 13 Pale pink with dark center, Convex, Circular, Entire - Short rod + - + + AG AG -G AG AG - + + - + - Enterobacter spp.
PD 14 Pink, Convex, Circular, Entire - Short rod + - + + AG -G -G AG AG - - - - + - Klebsiella spp.
PD 15 Translucent, Flat, Irregular, Serrated - Short rod in cluster + - + - AG -G -G AG AG + + - - - - Salmonella spp.
PD 16 Translucent, Flat, Irregular, Serrated - Short rod in cluster + - + + AG AG AG AG AG + + - - - - Salmonella spp.

Journal of Food, Agriculture & Environment, Vol.12 (2), April 2014

PD 17 Purple with metallic sheen, Umbonate, Circular, - Short rod - + + - AG AG -G AG AG + - + + - - Escherichia coli
Entire
PD 18 Pale purple, Raised, Circular, Entire + Cocci - - + - AG -G -G AG AG + - + - - - Enterococcus spp.
PD 19 Yellowish, Raised, Circular, Entire - Short rod + - + - -G -G -G AG AG - - + - - + Flavobacterium spp.
PD 20 Yellowish, Raised, Circular, Entire - Short rod - - + - AG AG -G AG AG - - + - - + Flavobacterium spp.
PD 21 Pink, Convex, Circular, Entire - Short rod + - + - -G -G -G AG AG - - - - + - Klebsiella spp.
PD 22 Pink, Convex, Circular, Entire - Short rod - + + - AG -G -G AG AG + - + + + - Escherichia coli
PD 23 Pink, Convex, Circular, Entire - Rod in cluster + - - + AG -G -G AG AG - - - - + - Klebsiella spp.
PD 24 Transparent, Flat, Irregular, Lobate + Cocci + - + - AG -G -G AG AG + - + - + - Staphylococcus spp.
PD 25 Pink, Convex, Circular, Entire - Rod + - + - AG -G -G AG AG - - - - + - Klebsiella spp.
PD 26 Pale pink, Raised, Circular, Entire - Rod + - + + -G -G -G AG AG + - - - + + Pseudomonas spp.
PD 27 Pink, Convex, Circular, Entire - Rod in chains + - + - AG AG AG AG AG - - - + + - Klebsiella spp.
PD 28 Milky, Raised, Circular, Entire + Cocci in long chains + - - - AG AG -G AG AG + - + - + - Streptococcus spp.
PD 29 Milky, Raised, Irregular, Lobate + Rod + - - - -G AG -G AG AG + - + - + - Lactobacillus spp.
PD 30 Creamy, Raised, Circular, Entire + Rod in long chains + - - - -G AG -G A AG + + + + + - Bacillus spp.
PD 31 Milky, Raised, Irregular, Lobate + Short rod in cluster + - - - A A A A AG + - + - - - Lactobacillus spp.
PD 32 Creamy, Raised, Circular, Entire + Rod in long chain - - + + AG -G -G AG AG + + + + + - Bacillus spp.
PD 33 Milky, Raised, Irregular, Lobate + Short rod + - - - -G -G -G A AG + - + + + - Lactobacillus spp.
PD 34 Translucent, Flat, Irregular, Lobate - Short rod + - + - A A A A AG - - - - + - Klebsiella spp.
PD 35 Whitish opaque, Raised, Circular, Entire - Short rod + - + - AG -G -G AG AG - + + - + - Enterobacter spp.
PD 36 Creamy, Raised, Circular, Entire + Long rod + - + - AG AG -G AG AG + + + - + + Bacillus spp.
PD 37 Milky, Raised, Irregular, Lobate + long rod + + + - A A A A - - + + - - + Bacillus spp.
PD 38 Creamy, Raised, Irregular, Lobate + Short rod in cluster + - + - - - - AG AG + + + + + + Bacillus spp.
PD 39 Translucent, Flat, Irregular, Rhizoid + Long rod in cluster + - + - - - - AG - + + + + + - Bacillus cereus
PD 40 Creamy, Raised, Circular, Undulate - Short rod in cluster + - + - - - - A AG - + - - + + Pseudomonas spp.

1263
 Discussion
This study shows the ecological nature and complexity of the fish
pond water sources (Tables 1 and 2). It further shows that the

Streptococcus spp.

Streptococcus spp.

Enterobacter spp.
Micrococcus spp.

Escherichia coli
Escherichia coli
Identification

Bacillus spp.

Bacillus spp.

Bacillus spp.

Bacillus spp.
Bacillus spp.
values of pH in the study zone range from 7.82 in sample 2 to a
high range of pH 8.15 in sample 1, and the temperature ranges
from low unit of 27°C in sample 3 to a high temperature of 31°C in
sample 4. This result is in consistence with Frederickson et al. 2,
that the range at which fish has optimum growth is at temperature
Oxidase 24°C-35°C and pH range 6.5-9.0 (Table 1). The high observable

+
+

-
-
-
-

-
-

-
-
total hardness of 128.92 mg/L in Araromi area 1 is still within the
maximum allowable level of 500 mg/L for domestic water 12. However,
Voges proskauer

+
+

+
+
-
-
-

-
-

-
Methyl red the Cd content range of 0.98 mg/L in Academic planning office (a)
+
+
+

+
+

+
+

-
-

- and 7.80 mg/L in Araromi area 1 of the sampled pond water sources
exceed the allowable limit of 0.01 mg/L 13-15.
Ornithine
+
+
+
+

+
+
+

+
+
+

Motility A detailed comparative study was made using microbiological

+
+
+

+
+
-
-
-

-
-

examination on selected fish pond water in Akungba-Akoko

Hydrogen sulphide
community for the detection of various bacteria and their
+
+

+
+

+
+
+

+
+
-

population that could be found in these different ponds. Microbial

AG
AG

AG

Mannitol
-G

-G

-G

load of fish pond sample sources ranges from 24×10-4 CFU/ml

-

-
-

-
-

in Sample 4 (Araromi area 2 location) to 8×10-4 CFU/ml in

AG

AG
AG

AG
AG

AG

Fructose
Sample 2 (AAUA location) (Table 2). This is consistent with the
A

A
A
A
A

study of Jun et al. 5, who reported microbial load of aerobic

AG

Lactose
-G

-G

-G
-G
-G

-G

-G

heterotrophic bacteria in the pond water which fluctuated

-

-
-

between 0.01 and 8.7×105 ind/ml. Though microbes can serve as

AG
AG
AG

AG

Sucrose
-G

-G

-G
-G
A
-

food source to fishes, some nutrients can also be obtained through

-

the sediment sources, hence, high microbial load can be inimical

AG

AG

AG
AG
AG

AG

Glucose
-G
-G
-

-
-

to health. Sun and He 16 and Jun et al. 5 show that the contact
Coagulase layer between pond mud surface and water is the major source of
-
-

-
-

-
-

-
-
-

Catalase nutrition.
Coliform counts per unit sample sources show some levels of
+

+
+

+
+

+
+
-

Indole contamination. Though fishes feed on some microorganisms, high

+
+
-

-
-

-
-

-
-

Starch hydrolysis
level of contamination with the presence of these indicator
organisms could be alarming and could be linked to neglecting
+

+
+

+
+

+
-

-
-

good fishpond management practices. It could also be as a result

Short rod in chain

of increase in the rate of microbial infiltration possibly due to fecal

Cocci in cluster

Chain in chain
Cocci in chain
Cell shape

contamination either of animal or human origin. These conditions

Short rod

Short rod
Short rod
Short rod

Short rod
Long rod
Cocci

therefore necessitate good water quality for fish pond management

practices in order to get improved quality fish yields, less outbreak
of diseases, decreased mortality rate of fishes and also reduced
infections to humans 17.
In this study, seven genera of Gram-negative bacteria were
Gram reaction
+
+

+
+

+
+
+
+

-
-
-

encountered. These include Shigella sp., Escherichia coli,

Pseudomonas spp., Proteus spp., Citrobacter spp., Klebsiella
spp. and Enterobacter spp. Similarly, three genera of Gram-positive
bacteria obtained during the study are Bacillus spp.,
Whitish grey, Umbonate, Circular, Entire

Whitish opaque, Raised, Circular, Entire

Staphylococcus aureus and Streptococcus spp. (Table 4). The

Cultural characteristics

bacteria species isolated in this course of study were identified

Yellowish, Convex, Circular, Entire
Yellowish, Raised, Circular, Entire
Whitish grey, Flat, Circular, Entire

using the Bergey’s manual of determinative bacteriology 18.

Milky, Convex, Irregular, Lobate
Whitish, Raised, Circular, Entire
Milky, Raised, Irregular, Lobate
Milky, Raise, Irregular, Lobate

The bacteriological examination of fish pond water is important

Milky, Flat, Irregular, Lobate
Milky, Flat, Irregular, Lobate

to detect the presence of microorganisms that might constitute

health hazard and death of fish in the fish pond. This can serve as
a guide to monitor and protect our fish ponds as intensified in this
+ - Positive; G - Production of gas only;
- - Negative; A - Production of acid only;

study. Also, the bacteriological examination of the source of water

Table 3. Continued.

AG - Production of acid and gas.

to the fish pond is very necessary in order to detect the kind of

bacteria being transferred into the fish pond. The presence of
some bacteria species such as Pseudomonas and streptococci
indicates a contamination which may affect the growth and survival
in the fish pond environment. From research the presence of
Isolates

PD 41
PD 42

PD 47
PD 43

PD 45

PD 49
PD 44

PD 46

PD 48

PD 50
PD 51
Legend:

bacteria such as Pseudomonas fluorescence, P. anquilliseptica

1264 Journal of Food, Agriculture & Environment, Vol.12 (2), April 2014
Table 4. Bacterial isolates from the pond sources.
Coliforms/ Common oil Pathogens and other
Genus Bacillus Coccus forms
Enterobacterieace degraders bacterial species
Bacillus spp. Staphylococcus spp. Escherichia coli Pseudomonas spp. Salmonella spp.
Bacillus cereus Streptococcus spp. Klebsiella spp. Flavobacterium spp.
Micrococcus spp. Enterobacter spp. Edwardsiella spp.
Enterococcus spp. Lactobacillus spp.

and Streptococcus iniae had been discovered to cause diseases F. (eds). The Studies of the Ecosystem in the Chinese Fish Pond.
in fishes. Pseudomonas fluorescence causes lesion with Science and Technology Press, Shanghai, pp. 29-36.
hemorrhagic septicemia resulting in hemorrhage of the fins and
5
Jun, X., Xiuzheng, F. and Tongbing, Y. 2000. Physico-chemical factors
tail and ulceration of the skin. Also, Streptococcus iniae affects and bacteria in fish ponds. Naga, The ICLARM Quarterly 23(4):16-
20.
fishes in the same manner. Therefore, in order to prevent harmful 6
Foster, R. and Smith, M. 2013. Fish Anatomy and Physiology.
bacteria from the fish pond, a lot of work should be done to LiveAquaria.com. Veterinary and Aquatic Services Department, pp. 1-
reduce ponds’ microbial load and also to ensure that the water 2. http://www.peteducation.com/article.cfm?c=16+2160&aid=583.
used in supplying the pond is free from harmful bacteria. Some Accessed on 8th March, 2013.
of the factors that may contribute to infiltration of microbial 7
Bailey, M. and Burgess, P. 2000. Tropical Fishlopedia. Howell Book, New
contamination are improper location of the pond, very close to York, 1 p.
latrine and introduction of contaminated food or material into
8
Mathias, J. A., Charles, A. T. and Baotong, H. (eds). 1994. Integrated
the fish pond water 19, 20 and use of bucket or container directly Fish Farming. CRC Press, Boca Raton, pp. 140-143.
from the soil into the fish pond.
9
Salle, A. J. 2002. Fundamental Principles of Bacteriology. 7th edn. Tata
McGraw-Hill Publishing Company limited, New Delhi, India, 688 p.
10
APHA 1995. Standard Methods for the Examination of Water and
Conclusions Wastewater. 19th edn. American Public Health Association, Washington,
This study shows the presence of some imminent contaminants DC.
in fish pond water sources that are of ecological threat and inimical 11
MacFaddin, J. F. 1980. Biochemical Tests for Identification of Medical
to health. Based on these observations, some recommendations Bacteria. 2nd edn. Williams and Wilkins, Baltimore, Maryland, pp. 173-
are made to reduce the problem of microbial contamination of fish 183.
pond water. This includes the knowledge and observation of the
12
Roberge, P. R., Amercoat, C. V. and Folick, L. L. P. 1981. Drinking
following factors that could affect the microbial quality of fish Water Standards. Notification of the Ministry of Public Health, No.
pond water. Therefore, it is recommended that proper construction 61 B.E. 2524 (1981), issued under the Food Act B.E. 2522 (1979),
published in the Royal Gazette, Vol. 98, Part 157 (Special Issue),
of fish pond should be ensured. The environment where the fish
dated September 24, B.E. 2524 (1981). Assessed 16 November, 2012.
ponds are located should be protected from pollutants and weeds Available: http://www.corrosion-doctors.org/NaturalWaters/
which can harbour microorganisms that find ways into fish pond Drinking.htm.
by themselves or by passive process through wind, rainfall, etc. 13
WHO-UNICEF 2005. Joint Monitoring Programme: Worldwide
thereby affecting the fishes negatively. Similarly, water supply to Sanitation. http://www.who.int
the fish pond should be clean and free of contamination. Sample 14
US EPA (US Environmental Protection Agency) 1996. Integrated Risk
of the fish pond should be taken and examined in the laboratory Information System (IRIS). Washington, DC. http://cfpub.epa.gov/ncea/
for its microbiological quality before stocking. This will also give iris/index.cfm.
an insight to the possible presence of certain types of micro-
15
EPA (US Environment Protection Agency) 2002. Safe Drinking Water
Act Amendment. Available: http:// www. epa.gov/safe water /mcl. Html.
organisms, hence provide enabling environment for aquaculture 16
Sun, Y. and He, Z. Y. 1997. Shrimp pond settlement - the quantity of
purposes. nutrients dispersion and seasonal changes between water and mud
contacting surfaces. Mar. Fish. Res. 18(1):60-66.
Acknowledgements 17
Carballo, E., van Eer, A., van Schie, T. and Hilbrands, A. 2008. Agrodok
We thank the fish pond managers from where samples were 15 Small-scale Freshwater Fish Farming, pp. 6-23. http://
collected in Akungba-Akoko, Nigeria, for granting accessibility. journeytoforever. org/farm_library/AD15.pdf.
We also acknowledge the University of Fort Hare, Alice, South
18
Buchanan, R. E. and Cibbons, N. E. 1974. “Bergey’s manual of
Africa, for the opportunity to complete this research. determination bacteriology” Baltimore. The Williams and Wilkings
C., USA, pp. 529-549.
19
Ogedengbe, M. O. and Aina, P. O. 1980. Coexisting well water and pit
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