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Assessment of Plankton Diversity in Relation To Water Quality Variables of Musgola Fish Farm at Lapai - Gwari, Minna, Niger State, Nigeria

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Volume 8, Issue 2, February – 2023 International Journal of Innovative Science and Research Technology

ISSN No:-2456-2165

Assessment of Plankton Diversity in Relation to


Water Quality Variables of Musgola Fish Farm at
Lapai - Gwari, Minna, Niger State, Nigeria
Link Between Fish Farm Water Quality and Plankton Diversity at Lapai-Gwari

*
Auta, Y. Iliya1,
Shehu,Abubakar1.,
Mohammed, A.Z.1,
Hamzat , Aliyu2.,
Mohammed,C.Dangana3.,
Gilbert, Ndatimana1.
1
Animal Biology Department, Federal University of Technology, Minna. Nigeria.
2
Ibrahim Babangida Badamasi University, Lapai, Niger State. Nigeria.
3
Plant Biology Department, Federal University of Technology, Minna. Nigeria.
*
Corresponding Author

Abstract:- This study was carried out to determine the community, on which the whole aquatic population
plankton interaction with the physico-chemical depends, is largely affected by an interaction of a
parameters of four earthen fish ponds in Musgola Fish number of limiting factors such as, moderate sulphate,
Farm Minna, Niger state. The study was conducted from nitrate, phosphate and other factors. Variations of the
March to June; 2021. Samples of Water were analyzed physical and chemical parameters in a fish pond showed
using standard methods. One-way analysis of variance that the limnological conditions in the fish pond are not
with Duncan multiple tests was used to analyze the water spatially homogenous.Thus, water quality management
quality parameters. PAST (Multivariate: Canonical in the fish pond expressed direct influence on planktonic
correspondence analysis) was used to evaluate population, since fish ponds are shallow and constantly
relationship between plankton communities and receive large nutrient loads (feed, fertilizing, and fish
environmental variables. Physico-chemical Parameters waste).
determined include: Biochemical Oxygen Demand,
Alkalinity, Dissolve Oxygen, Nitrate (NO3), Phosphate Keywords: Planktons Diversity, Water Quality, Physico-
(PO4), Sodium(Na),potassium(k), Electricalconductivity, Chemical Parameters, Musgola Fish Farm.
Air temperature and potential hydrogen. There was no
significant temporal variation in physico-chemical I. INTRODUCTION
variables. However, there were slightly change in
parameters throughout the study period. The study The plankton encompasses the phytoplankton and
revealed thatzooplanktons were dominated by Cladocera zooplankton inhabitants. Phytoplankton are important
species (MoinaMicrura) with (43.02%), followed by organisms which act as primary producers in any aquatic
Copepod (Cydopoid copepods) with (26.99%) and ecosystem (Bwala et al., 2009). They are the early biological
Rotifers (Brachionuscalyciflorus)representing7.45%.The constituent by which energy is conveyed to complex
abundance of zooplankton observed in this study can be organisms along the food chain (Babatunde and Saifullah,
attributed to the anthropogenic perturbations on-going 2014). Phytoplankton are the biological resources of the
in and around the Fish farm as well as the farmed fish. water body and form the bases of food chain in ponds
Moreover, the presence of suitable zooplankton species is (Otene and Nnadi, (2019). On the other hand, zooplanktons
essential for successful farming. Similarly, seventeen are the animal portion of the planktons (Nasir et al, 2017).
species of phytoplankton were identified,including They are defined as pelagic animals which are unable to
Bacillariophytes, Chlorophytes and cyanophytes. Chlorella maintain their position by swimming against the physical
species of the class Chlorophyceae was the most movement of water.Zooplanktons are a principal component
prevalent group in the four ponds over Bacillariophyceae of food for omnivorous fish that are usually farmed in
and Cyanophyceae.Canonical correspondence Analysis extensive aquaculture (Oteneet al., 2019).Further, Brummett
revealed that physico-chemical variables have positive and Noble, (2013) emphasized on the fact that zooplanktons
influence on the composition and diversityof plankton in are very important in the food web of open water ecosystem.
earthen fish ponds at Mosgola Fish Farm. The plankton Zooplanktons are heterotrophic animals that are incapable of

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Volume 8, Issue 2, February – 2023 International Journal of Innovative Science and Research Technology
ISSN No:-2456-2165
synthesizing organic matter by themselves They play key farms has economic value due to the fishing activities
role in the pelagic food web by controlling phytoplankton’s carried out by individuals and some irrigation farming by
production and shaping pelagic ecosystem(Nasir, et al, some of the community in vicinity. Other anthropogenic
2017). Furthermore, Otene and Nnadi, (2019) observed that activities that affect directly the stream are bathing,
lack of zooplanktons causes poor survival of spawn in swimming, washing and sand mining.
nursery ponds.
 Water Sample Collection:
They are commonly divided into Rotifers (Rotifera), Water Samples were collected from the fish ponds
Cladocerans (Cladocera) and Copepods (Copepoda). The located in Musgola fish farm for 4 months from March to
composition of zooplanktons has vital role for successful June, 2021. Sampling was done once in a month (i.e; mid-
fish farming (Oteneet al., 2019), since they play important week) for all of the fish ponds.
role in the food chain, ad being in the second trophic level as
primary consumer and also as contributors to the next  Physico-Chemical Parameters Analysis:
trophic level (Arazu and Ogbeibu, 2017).Zooplankton Water analysis was conducted within a day to five days
population get improved with the application of manure to of collection of the water samples. Air and water
maintain the water quality favourable for fish production. As temperatures were measured in situ using Mercury-in-glass
such, poultry manures are found to release soluble salts thermometer, (APHA, 2014). pH was determined using dip-
continuously, resulting in high production of zooplankton in mobile battery-operated pH meter, Electrical conductivity
(Arazu and Ogbeibu, 2017). Moreover, Manoharan,et al, was determinedusing EC/TDS meter in whichthe meter was
(2015) suggested that duck excreta were good source of calibrated in μS/cm. Turbidity was determined using
nutrients, easily soluble in water and available for plankton HANNA LP 2000 model turbidity meter this was done after
production. In his study, Roy(2014) revealed that poultry the meter was calibrated at HI 93703-0 and HI 93703-10
droppings produce more zooplanktons compared to cow NTU standards. Alkalinity and total hardness were
dung and pig dung.Zooplanktonoccupyboth freshwater and determined titrimetrically. Dissolved oxygen was
saline water; and found in almost all water bodies, including determined using 200 model DO meter. The biological
river, stream, lakes, reservoir, ponds, irrigation canals, rice- oxygen demand (BOD) was carried out after 5 days
field and temporary water bodies(Nasir, et al, 2017). The incubation in the dark at 280C. Alkalinity, phosphate-
zooplankton community is a dynamic system that responds phosphorous, Nitrate-nitrogen and calcium were determined
quickly to environmental changes (Roy, 2014). The titrimetically following the methods in APHA (2014).
relationship between the physico-chemical parameters of
water quality and plankton production in fish ponds is of  Planktons Sampling And Analysis:
great importance and essential for fish culture (Oteneet al., Zooplankton’s sample were collected horizontally and
2019). vertically with 30µm mesh size plankton net. The samples
were preserved in 100ml 10% Formalin. The samples were
Phytoplankton sorts are bio-indicators of water quality also recounted after 24 hours. Identification, estimation and
due to their reaction which bring active response to counting using Goswami, (2012) as a guide. Both vertical
alterations in the surrounding environment (Siddika, 2012). and horizontally, the volume filtered was calculated using:
Qualitative and abundance of phytoplankton signify the
prolific condition of a water body (Chowdhury,et al., 2008), 𝑉 = 𝜋𝑟 2 ℎ
thus a thorough awareness of abundance of phytoplankton r = radius of diameter of sampling net.
and its quality in time and space in relative to environmental h = distance travel horizontally or vertically
conditions is essential for fish production. The awareness
on how water quality through physico-chemical variables Samples for phytoplankton study were collected using
affect plankton would bring new discernment on future a cone curved, silk securing cloth net of 20 μm mesh size, a
management of ponds for ecological monitoring. However, mouth radius of 20 cm and a 50 mL use bottle. The 50 mL
there is no well-known information on the work of plankton concentrates were transferred to distinctly labeled 100 mL
in earthen ponds in Minna. Therefore, this evaluation glass jars and fixed instantly with Lugol solution to preserve
focused on measurement of some physico-chemical green algal cells (APHA 2014). Handling and examination
parameters and to assess the richness and variety of of algal samples were study according to the procedures of
planktons in relation to these factors in fish earthen ponds in APHA (2014). Microscopic cell counts using the drop count
Musgola fish farm, Minna, Niger state. method (Bartram & Rees 2000) was used to regulate the
green algal cell mass (no of cells per mL). A drop of the
II. MATERIALS AND METHODS distillate was placed on a glass slide and the total number of
characters in that drop counted. Prior to these counts, the
 Study Area: glass dispenser used was adjusted to limit the number of
Musgola farms, is located at Lapai-Gwari, Bosso local drops that provided one milliliter.
Government Area Northern region of Minna, Niger State,
the area falls between latitude 90 31ʹ N and longitude 60 31ʹ  Data Analysis:
E (Fig.1).These farmsarebanded by a stream which flows Water quality and plankton’s analysis were done using
through Minna, the Niger state capital and carry domestic Microsoft Excel. One-way Analysis of Variance (ANOVA)
waste-water from this town and its surrounding. Musgola was used to test for statistical variances between the means

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Volume 8, Issue 2, February – 2023 International Journal of Innovative Science and Research Technology
ISSN No:-2456-2165
of the physical and chemical constraints of the fish ponds. species. However, Total hardness (TH), Total Dissolved
The weakness and pattern of association between the water Solids (TDS), Phosphate (PO4), Dissolved Oxygen (DO),
quality variables and plankton abundance in each pond Biological Oxygen Demand (BOD) and Electrical
within the sampling period was correlated using Canonical Conductivity (EC) had negative correlation with the
Correlation Analysis in PAST software. planktons (Fig.4)

III. RESULTS IV. DISCUSSION

 Physico-Chemical Variables of Lapai Gwari Fish The variations in physico-chemical properties of the
Earthen Pond: four selected ponds investigated in Musgola fish farm
Air temperature and pH shows no substantial Minna, Niger State may be due to the influence of climatic,
difference (P>0.05). The highest pH value was recorded in topographic, and edaphic condition of water quality of fish
June (7.79±0.58) and the lowest in the month of March ponds in the area. It is similar to the reported study by
(6.65±0.22). Electrical conductivity (EC) revealed that in Ayanwale et al., (2012), which argued the contribution of
the month of June there was low mean value edaphic factors to variation in physico-chemical properties
(180.50±10.92µs/cm) and the highest mean value was in the of the ponds. The water temperature in the four ponds was
month of April (425.50±25.49µs/cm), Biochemical Oxygen not significantly different which may be as a result of
Demand (BOD) was low in the month of June (3.83±0.28 dominant influence of atmospheric temperature on water
mg/l) while, the month of April have the highest mean value temperature. The average temperature observed from the
(4.90±0.20mg/l), NO3, PO4, Na and K showed no significant four fish ponds ranged from (21.77±0.31). This is within the
difference (P>0.05), while alkalinity (96.50±9.07mg/l), recommended temperature range for fish activities
Dissolve Oxygen (DO) (7.98±0.46 mg/l) were the highest (Kurniawan et al., 2022).Roy, (2014) recommended
(Table 1). temperature between 25°C to 32°C for good performance of
fishes. The highest temperature (31.34°C) recorded in
 Composition of Zooplankton in the Sampled Ponds: Musgola fish ponds was expected since the stations had the
Composition of zooplankton showed 12 species which highest exposure to sunlight with the least shading
include rotifers (5), Cladocera (5), and copepod (2) all from effects.Generally, the dissolved oxygen requirement for fish
the four sample sites. The results illustrated that Moina ponds varies with species.However, a minimum constant
Micrura belonging to the class of Cladocerawas more value of 2.0mg/L of DO is satisfactory for the most species
abundant, representing 43.02%, followed by Cyclopoid and stages of aquatic life(Rajagopal, et al., (2010).The
copepods with 26.99% belonging to the class of Copepoda. average dissolved oxygen variation in Musgola fish pond
Brachionus calyciflorus(7.45%) was the most abundant in may also be connected to atmospheric impurities by wind,
the class of Rotifera and ranked the third in whole samples, wave and human disturbance and a continual discharge of
whereas the rest of zooplankton species were foundwithin water from the inlet pipes. This reflects the conducted study
the range of 0.70% to 4.22% (Fig. 2) about the factors affecting the variation of DO in stagnant
water especially ponds (Long et al., 2018). pH was
 Composition of Phytoplankton in the Sampled Ponds: significantly different in the selected ponds, thus the average
The researched confirmed seventeen phytoplankton pH recorded were 6.65±0.22; 7.03±0.18); 6.99±0.05 and
sorts, appropriate to the classes Chlorophyceae (7), 7.79±0.58 for first to fourth pond respectively. According to
Bacillariophyceae (2), and Cyanophyceae. from the four Sterling and Philips (2007), the pHof these ponds falling in
sampling sites. Chlorella species had the highest proportion the range of recommended for good fish production (6.5-
with 51.82% belonging to the class 8.5). The highest BOD value was recorded in the month of
Chlorophyceae.Anacystis species was the second most April (7.98±0.46), these could be attributed to the presence
abundant species with 7.30% belonging to the class of decomposition of organic matter in the fish ponds. This is
Cyanophyceae. Other species of the phytoplankton ranged not far from the study revealed that BOD reach maximum
between 6.50% and 0.80%for Zygnema species and due to the activities taking place in water (Susilowati et al.,
Scenedis musquadricauda species respectively (Fig. 3). 2018; Prambudy et al., 2019); basically due to the organic
matter degradation from increased weeds decomposition.
 Correlation of Physico-Chemical Variables and Nitrate level range from 9.8-49mg/L and phosphate 3.2-
Plankton Interaction in Mosgola Fish Pond: 6.30mg/L.(Adeniji, 2005) observed the same results in more
Canonical Correspondence Analysis (CCA) showed productive inland lakes and ponds of Africa; this might
that Temperature(WT), Total alkalinity (TA), pH and indicate that it supports aquatic productivity. Total dissolved
Nitrate (NO3) had positive correlation with the most of solid (TDS) showed significant difference (P<0.05) between
phytoplankton of cyanophyceae group such as antrospira the months of April and May. This observation may be
species, Analytis species, Oscillatoria species and found in the management practice, being that artificial fish
Microspora species. These physico-chemical variables habitual organic particles were not allowed to accumulate.
expressed positive correlation to the zooplankton species, This may be due to the tendency of the fish reared in the
mostly of the genus Branchianus including Branchianus pond to feed on such particles. Electrical Conductivity and
angularis, Branchiarus falcatus and Branchianus alkalinity varied significantly among the ponds, these may
Calyciflorus. Moreover, Sodium (Na) and Potassium (K) be due to the influence of edaphic factor especially the
ions expressed the same correlation to these plankton substratum, even though the four ponds were located closed

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Volume 8, Issue 2, February – 2023 International Journal of Innovative Science and Research Technology
ISSN No:-2456-2165
to each other. Bird et al., (2009) argued that the ideal range variables correlate positively with some plankton species
of alkalinity to be between 20-30 mg/L. Thus, the average whereas other were inversely associated (Liu et al., 2010).
alkalinity for the four ponds did not fall within the Given the fact that CCA of the present study is reporting
recommended range. This may be as a result of the positive association with physical chemical variables; it is in
topography and climate change. Conductivity varied confluence with the reported study by Coelho et al., (2007)
significantly between the months, (393.25±22.42), for temperature and PH; and Kalin et al., (2001) for Sodium
(425.50±25.49), (323.00±25.05) and (180.50±10.92) for and Potassium. Moreover, the same results have been
March, April, May and June respectively. Sodium (Na) obtained on positive correlation between plankton and total
showed no significant difference (p<0.05) in all the months. alkalinity (Ferdoushi et al., 2021; Malik and Rathi, (2022)
for Total alkalinity. Our findings revealed inversely
Most of the plankton encountered in the study area association between plankton and DO, BOD, TH and EC.
appears to be normal inhabitants of natural lakes, ponds, Although, this result fall to agreement with the report study
streams and artificial impoundment in the tropics and by Simantiris et al., 2021 about inversely correlation of
subtropics regions (Usman, 2015).Cladocera were the most plankton with DO as well as BOD and Tufail et al., (2022)
abundant collection of zooplankton documented in all the for Electric conductivity; It is in contrast with the study by
stations. The capacity of Cladocera to experience vertical Malik and Rathi, (2022), where they found that positive
immigration, which minimizes opposition through niche correlation of DO and BOD with phytoplankton. Therefore,
operation and food utilization, could probably be the there are evidence that the physico-chemical variables have
purpose for their control. The dominant status of Cladocera slight influence on composition and abundance of plankton
species in the pond comparative to the Rotifers and species in water body such as pond.
copepods is characteristic of tropical lakes and rivers
(Thirupathaiah, et al., 2012). The number of Rotifers in V. CONCLUSION
Musgola fish farm was relatively low; this may be attributed
to the absence of aquatic macrophytes, which may have There are evidence to conclude that the dominant
accelerated the rate of predation by fish (Usman, 2015). This zooplanktons compositions of Mosgola Fish Farm, Bosso
is in agreement with the findings of Arimoro and Oganah, Local Government in Niger State were rotifers, copepods,
(2010) in a perturbed tropical stream in the Niger Delta. cladocerans and microscopic protozoans, while the
Phytoplankton’s are dominated by Chlorella species
Seven species of class Chlorophyceae were recorded Signifying seasonal and monthly variations in the physico-
throughout the period such as Spirogyra species, Ulothrix chemical variation in the distribution ofplanktons.
species, Scenedesmus quadricauda, Scenedis musincrasa Conclusively, it was observed that there was a good
tulus, Pediastrum species, Chlorella species and Zygnema interaction between the phytoplankton species and physico-
species. The minimum units of algae of class Chlorophyceae chemical parameters and a moderate water quality with the
were recorded during dry season and maximum units were dominance of Chlorella species. This indicate that the
recorded during rainy season between April and May. physicochemical variables restricted the spreading and
Chlorophyceae was widespread and most dominant group abundance of the phytoplankton. The results of the physio-
among other phytoplankton. It is not doubtful, since the chemical parameters showed that all the values recorded
Chlorella species has been used as a source of dietary were within the guideline limit and the recommended safety
protein for freshwater fish as it causes an improvement in limit for survival of aquatic organisms except for Dissolved
weight and meat quality. Therefore, considering these fish Oxygen which exceeded the WHO standards in the month of
ponds, there would be the reason for high abundance. This April. Variations of the physical and chemical parameters in
reflected the study by Nirmal and Cini, (2011) where they a fish pond reveal that the limnological conditions in the fish
opined that high temperature favors the growth of blue- pond are not spatially homogenous, since differences have
greens. This is clear from the result that Cyanophyceae were been observed not only between the different sampling
lowest during dry months, then when the water column was months, but also between the different sites analyzed. Based
remarkably stratified to a large extent because of heavy on the results of the current analysis, water quality
rainfall, reduced salinity, decreased temperature and pH, management in the fish pond showed a direct influence on
overcast sky and cool conditions. Contrary, Roy, (2014) planktonic population, since fish ponds are shallow and
reported minimum density of phytoplankton during dry constantly receive large nutrient loads (feed, fertilizing, and
season and maximum during rainy season in Chatla Lake, fish waste).
Assam; as the same as the result reported by Rajagopal, et
al., (2010). Generally, the plankton community, on which RECOMMENDATION
the whole aquatic population depends, is largely affected by
an interaction of a number of limiting factors such as, The present study recommends research on how the
moderate sulphate, nitrate, phosphate and other factors, the plankton species together with the physico-chemical
physico-chemical variables affect the partial spreading and properties of the water quality should be carried out in an
richness of plankton (Baird and Telfer, 2009). algal cultured environment to see the effectiveness on how
this interaction can benefit the ecosystem, since the
As many studies illustrated the effects of phytoplankton species are essential natural feed for higher
physicochemical parameters to the composition of planktons organisms which includes fishes. The regular analysis, either
(Chandran et al., 2021, Heneash et al., 2022); the specific monthly or bi-monthly should be carried out to either

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Volume 8, Issue 2, February – 2023 International Journal of Innovative Science and Research Technology
ISSN No:-2456-2165
supplement the fish feeds with the plankton community or to [12]. Brummett R E & Noble R (2013). Aquaculture for
replace entirety the synthetic foods as well, to prevent the African Small Holders. ICLARM – The World Fish
ponds from algal bloom (eutrophication).In addition, this Center, Penang, Malaysia pp. 1–69.
study recommends the application of the water quality index [13]. Bwala, R.L. &Omoregie, E. (2009). Organic
as a very helpful tool that will enable the public and decision enrichment of fish ponds: application of pig dung vs
makers to estimate water quality of fish ponds. Finally, the tilapia yield. Pakistan Journal of Nutrition 8:1373-
qualitative study and the plankton abundance and delivery in 1379.
the ponds; there is a need to work methods to improve the [14]. Chowdhury, M.M.R, Shahjahan M., Rahman, M.S.,
value of the ponds water for sustainability of the cultured &Sadiqul Islam M. (2008). Duckweed (Lemna
fishes. minor) as supplementary feed in monoculture of Nile
tilapia, Oreochromis niloticus.Green algal
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Volume 8, Issue 2, February – 2023 International Journal of Innovative Science and Research Technology
ISSN No:-2456-2165

 Figures:

Fig 1 Vector Map of Musgola Farm, Lapai-Gwari with the Sample Sites

Fig 2 Abundance of Zooplankton of Mosgola Fish Farm in Lapaigwairi, Minna

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Volume 8, Issue 2, February – 2023 International Journal of Innovative Science and Research Technology
ISSN No:-2456-2165

Fig 3 Abundance of Phytoplankton in Mosgola Fish Farm

Fig 4 Correlation Of Physico-Chemical Variables and Plankton Interaction in Mosgola Fish Pond

(Wt.)=Water Temperature; (At)=Air Temperature; (BOD)=Biochemical Oxygen Demand; (EC)= Electrical Conductivity;
(TDS)=Total Dissolved Solid; (TA)=Total Alkalinity; (TH)=Total Hardness; (DO)=Dissolved Oxygen; (Na)=sodium;
(NO3)=Nitrate; (PO4)=Phosphate; (K)=Potassium

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Volume 8, Issue 2, February – 2023 International Journal of Innovative Science and Research Technology
ISSN No:-2456-2165
 Tables:

Table 1 Physico-Chemical Variables of Lapai Gwari Fish Earthen Pond from March to June 2021
PARAMETERS MARCH APRIL MAY JUNE WHO NIS
Wt (℃) 21.77± 0.31a 21.53±0.27a 31.43±0.52b 31.40±0.17b 30-32 - -
At (℃) 29.10±0.00a 29.30±0.00b 37.10±0.00c 37.30±0.00d - -
b b a
TDS(mg/L) 138.50±22.07 200.00±37.02 1.00±0.41 22.50±5.04a 600 600
a b ab
pH 6.65±0.22 7.03±0.18a 6.99±0.05 7.79±0.58b 6.5-85 6.5-8.5
c c b
EC(µs/cm) 393.25±22.42 425.50±25.49 323.00±25.05 180.50±10.92a 1000 01000
ab b a
DO(mg/L) 7.48±0.36 7.98±0.46 6.43±0.28 6.15±0.68a 7.5 -
b b ab
BOD(mg/L) 4.88±0.36 4.90±0.20 4.38±0.26 3.83±0.28a 6 -
a ab b
TA(mg/L) 67.00±2.12 82.25±9.33 96.50±9.07 79.50±9.17ab 100 100
b b b
TH(mg/L) 89.25±1.49 103.50±8.30 95.50±4.27 67.00±7.23a 150 15O
b b a
PO4 (mg/L) 1.56±0.11 1.95±0.10 0.55±0.06 0.45±0.05a - -
a a a
NO3(mg/L) 0.59±0.11 0.62±0.20 0.87±0.15 1.84±0.55b 22 50
a a b
Na(mg/L) 4.99±0.94 5.04±0.84 7.59±0.49 8.13±0.82b - -
a a b
K(mg/L) 2.11±0.08 2.02±0.46 3.88±0.17 3.87±0.17c - - -

(Wt)=Water Temperature; (At)=Air Temperature; (BOD)=Biochemical Oxygen Demand; (EC)= Electrical Conductivity;
(TDS)=Total Dissolved Solid; (TA)=Total Alkalinity; (TH)=Total Hardness; (DO)=Dissolved Oxygen; (Na)=sodium;
(NO3)=Nitrate; (PO4)=Phosphate; (K)=Potassium ; Superscripts with the same letter in each role are not significantly different
(P≥0.05). The measurement were compared to the standard by World Health Organization (2014) and Nigerian Industrial
Standard- (NIS, 2015).

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