Research & Reviews: A Journal of Toxicology

ISSN: 2231-3834 (Online), ISSN: 2349-1264 (Print)

Volume 10, Issue 2
www.stmjournals.com

Toxicological Evaluation of Two Nigerian Refineries’

Effluents using Natural Bio-monitors
Samson Ogagaoghene Egurefa1, Micheal Uchenna Orji2, Bright Obidinma Uba3,*
1
PhD Student, Department of Applied Microbiology and Brewing, Nnamdi Azikiwe University,
P.M.B. 5025 Awka, Anambra State, Nigeria
2
Professor, Department of Applied Microbiology and Brewing, Nnamdi Azikiwe University, P.M.B.
5025 Awka, Anambra State, Nigeria
3
PhD, Department of Microbiology, Chukwuemeka Odumegwu Ojukwu University, P.M.B. 02 Uli,
Anambra State, Nigeria

Abstract
The present study aimed to evaluate the toxic effects of two Nigerian refineries’ effluents using
natural bio-monitors. Standard physicochemical analysis, toxi-chromo test using mutant E.
coli and sub-chronic animal toxicity testing with emphasis on tissue heavy metal
accumulation, biochemical parameters and histopathology, were adopted during the study.
The physicochemical results revealed that the two effluent samples are highly polluted
although PH sample was more polluted than the Warri sample. Toxi-chromo testing revealed
that the positive control sample (HgCl2) exhibited the most harmful effect on 50% population
of mutant E. coli with EC50 value of 1.535 mg/L. The mice model revealed that iron (1.616
mg/kg) and lead (0.169 mg/kg) had the highest accumulation in the Warri exposed liver
tissues of the mice. PH sample had the highest values of the biomarkers but statistical analysis
showed a significant difference (p<0.05) between the samples and control with R2 value of
0.304. There were non-cancerous changes in the architectural structure and integrity of the
observed liver and kidney tissues in comparison to their controls with normal architecture.
Hence, adequate and strict measures should be established by government and environmental
policymakers to ensure proper treatment and monitoring of these liquid wastes before they are
discharged into our terrestrial and aquatic environments.

Keywords: Animal model, E. coli, environmental health, pollution, refinery effluents

*Author for Correspondence E-mail: ubabright4real@yahoo.com; bo.uba@coou.edu.ng

INTRODUCTION hydrocarbon, indicating the presence of

Produced water characterizes the largest hydrocarbons in the effluent.
volume of liquid waste during oil and gas
production and operations on most offshore For these reasons, several assays particularly
platforms. It is mostly composed of mono and chemical and biological, have been established
poly aromatic compounds such as benzene, for screening of toxic substances from these
toluene, xylenes, anthracene, inorganics, liquid wastes. Among the commonly used
heavy metals, organic acids, phenols, and biotests, Toxi-ChromoTestTM have been
additives [1, 2]. Due to complex organic and applied by EBPI [4] and Uba et al. [5] for
inorganic nature of produced water, there is detection of active toxins and do not usually
growing concern about the environmental and require a lengthy process of sample
animal health problems that may be associated preparation. The principle of the assay is based
with the indiscriminate discharge of produced on the capability of pollutants to prevent the
water into the aquatic ecosystems [1]. Odinga synthesis of an inducible β-galactosidase
et al. [3] reported that the effluent from Port enzyme in a highly permeable mutant of
Harcourt Refining Company contained levels Escherichia coli. On further addition of a
of polycyclic aromatic hydrocarbon, total chromogenic substance, produces a
hydrocarbon, and total polycyclic colorimetric endpoint that evaluates β-

RRJoT (2020) 22-31 © STM Journals 2020. All Rights Reserved Page 22
Toxicological Evaluation of Two Nigerian Refinery Effluents Egurefa et al.

galactosidase activity in the test bacterium Egurefa et al. [10]. The samples were mixed
after exposed period of time [4]. together to obtain a total of two representative
samples and were immediately transported to
Liver is the major organ site for transformation the laboratory in ice packed coolers, analyzed
and cleansing of xenobiotics, and so for physicochemical parameters and stored at
particularly vulnerable to these agents [6]. 4°C for further analysis.
Also, kidney, as the main organ for the
removal of xenobiotics and the metabolites, is Preparation of Industrial Effluent
primarily susceptible to their damaging effects The refinery industrial effluents samples were
[7]. Damage to these organs frequently results prepared by centrifuging and filtering of the
in elevated levels of clinical liver and kidney produced water suspensions prior to the
serum enzyme biomarkers such as aspartate physicochemical and toxicological assays [5,
transaminase (AST), alanine transaminase 10–13].
(ALT), alkaline phosphatase (ALP), total
protein, urea and creatinine, as indicators of Chemical Analysis
the impairment of hepatic and renal functions The chemical parameters such as oil and
[8]. A study by Andjelkovic et al. [9] showed grease content, total petroleum hydrocarbon
that the accumulation of metals in tissues (TPH), total hydrocarbon content (THC) and
accompanied the disturbances of both total phenol were determined by adopting the
hematological and biochemical parameters. methods of Kiepper [14], Akpveta et al. [15],
Odinga et al. [3] reported that the renal Nwineewii and Azuonwo [16] and Leouifoudi
function test showed a significant increase in et al. [17]. The standard method of APHA [18]
the level of the renal biomarkers when was adopted for heavy metals analysis using
exposed to Port Harcourt refinery effluent. atomic absorption spectrometry.
Odinga et al. [9] reported that the hormonal
and hepatic function test showed significant Toxicity Bioassay
increase (P<0.05) in ALT, AST and ALP Bacterial Toxicity Test (Toxi-Chromo Test)
levels, indicating impaired liver function and The toxi-chromo test was carried out
hepatotoxicity when exposed to Port Harcourt according to method described by EBPI [4]
refinery effluent. and as previously by Uba et al. [5]. The
samples were diluted and incubated with the
There are a lot of literatures on the toxic test genetically modified E. coli and reagents
effects of refinery or produced water on the for 1 hour 50 minutes in a microplate. The
aquatic and terrestrial ecosystems using microplate was read quantitatively at
bioassays. These literatures limited their scope absorbance at 600±20 nm and result were
on single effluent. Among the natural monitors recorded and after which toxicity factor (TF),
applied, there is paucity of information coefficient of variation (CV) and median
regarding the application of mutant of E. coli effective concentration was determined for the
on the refinery effluent and hence necessitate individual samples. The median effective
this study. The present study aimed to evaluate concentration (EC50) was calculated and
the toxic effects of two Nigerian refineries determined by the non-linear equation: Y =
effluents using natural bio-monitors. Bottom + (Top-Bottom)/(1+10^ ((LogEC50-X)
*Hillslope)).
MATERIALS AND METHODS
Description of the Studied Area and Sample Mice Toxicity Test
Collection Test Animal
Samples of untreated produced water (refinery All animal experiments were also conducted in
effluents) were aseptically collected at accordance with standard guidelines on
different discharge points through funnels into chemical safety and animal welfare of
sterile and clean 2 L plastic containers from Organization for Economic Cooperation and
both, Port-Harcourt and Warri Refineries Development (OECD) [19]. The experimental
Facilities in the Niger Delta region of Nigeria mice were obtained from Green Stone Farm
in September 2019 as previously described by Okohio, Otolo Nnewi, Anambra State.

RRJoT (2020) 22-31 © STM Journals 2020. All Rights Reserved Page 23
Research & Reviews: A Journal of Toxicology
Volume 10, Issue 2
ISSN: 2231-3834 (Online), ISSN: 2349-1264 (Print)

Animal Adaptation (UR). The blood biochemistry levels were

A total of thirty (30) healthy adult Wister albino analyzed at Dominion Care Diagnostic
mice of equal sexes weighing between 23.2– Laboratory Nnewi, Anambra State, Nigeria.
27.9 g were housed in large, spacious
aluminum wire cages at ambient room Histopathological Investigation
temperature with 12:12 hour light and darkness As described by Odinga et al. [3, 20], the fixed
cycles. The mice where allowed ad libitum to liver and kidney were dehydrated in alcohol,
grower feed of poultry (TOP VITAL FEEDS, cut sagittally and embedded in paraffin wax
Kano State, Nigeria) and water and adapted for mounted on slides. The slides were stained
two weeks before carrying out the experiment. with hematoxylin, counter stained with eosin
(H and E stains) and viewed under a
Experimental Procedure compound microscope (CX 23 Olympus,
After the two weeks of acclimatization, the Japan). All aberrations from the normal
mice were randomly allotted into 3 groups of structure were recorded. Photomicrographs
10 mice each and the experiment lasted a were obtained at different magnifications to
period of 14 days. Animals used as control show the differences in tissues of the liver and
were fed with normal rat chow and 2 mL oral kidney of the mice for each experimental
administration of normal saline ad libitum group after 14 day experimental period at
throughout the period of the experiment and Proklass Diagnostic Laboratory, Nnewi,
were termed group 1. Animals were fed with Anambra State, Nigeria.
feed and 2 mL of 100% Port Harcourt effluent
and water ad libitum throughout the period of Analysis of Data
the experiment and were termed group 2. The data obtained in this study were subjected
Animals were fed with feed and 2 mL of 100% to descriptive statistics, one or two way
Warri effluent and water ad libitum throughout ANOVA with Dunnet multiple comparison,
the period of the experiment and were termed Paired two tailed T-test, linear and non-linear
group 3. All the test animals were sacrificed regressional analysis and Pearson coefficient
after day 14 of the experiment. of variation among the samples, controls and
concentrations at 95% confidence intervals
Harvesting of Tissue and Blood such that values lower that 5% are considered
On day 14 of the experiment, animals were statistically significant (P<0.05).
sedated with chloroform vapour and whole
blood was collected by direct cardiac puncture RESULTS
from each animal into labeled sample bottles Chemical Profile
and centrifuged. The animals were dissected, The result of the chemical qualities of the
the liver and kidney tissues were collected, refinery produced discharge water, is
and used for histopathological examination. presented in Table 1. From the result, the Port
Harcourt (PH) produced water had the highest
Tissue Heavy Metal Analysis values of oil and grease (51, 900.00 mg/L),
Metal accumulation in the organs were TPH (18, 270.00 mg/L), THC (11,390.00
determined using atomic absorption mg/L), copper (0.982 ppm), mercury (0.029
spectrophometer (FS240AA- Agilent, USA) at ppm), lead (0.683 ppm) and phenol (594.52
Springboard Laboratory, Awka, Anambra GAE) while Warri sample had the highest
State by adopting the method described by values of iron (3.765ppm).
Ekaye et al. [13].
Bacterial Toxicity Profile
Determination of Biochemical Indices The result of microtitre plate absorbance
By adopting the method of Odinga et al. [3, reading using toxi-chromo test is presented in
20], the biochemical parameters observed in Table 2. From the Table 2, negative control
this study include alanine transaminase (ALT), had the highest absorbance value of 0.093
aspartate transaminase (AST), alkaline while Warri sample had the lowest absorbance
phosphatase (ALP), creatinine (CRE) and urea value of 0.017, both in bold red.

RRJoT (2020) 22-31 © STM Journals 2020. All Rights Reserved Page 24
Toxicological Evaluation of Two Nigerian Refinery Effluents Egurefa et al.

The results of toxicity factor and 1.5 h median toxicity factor (1702.04) while PH sample had
effective concentration of refinery discharge the lowest toxicity factor (1686.5). In the same
effluents and mercury chloride (HgCl2) after vein, positive control (HgCl2) and Warri
exposure to genetically modified E. coli are samples had the highest and lowest EC50
shown in Figures 1 and 2. From the results, the values of 1.535 mg/L and 0.852%,
positive control (HgCl2) had the highest respectively.

Table 1: Chemical Qualities of the Refinery Produced Discharge Water.

Parameters PH sample Warri sample DPR standards
Oil and grease (mg/L) 51,900.00 7,900.00 40.00
TPH (mg/L) 18,270.00 17,590.00 -
THC (mg/L) 11,390.00 9,296.00 40.00
Phenol (GAE) 594.52 441.85 -
Iron (ppm) 3.13 3.77 1.00
Chromium (ppm) 0.07 0.03 0.50
Mercury (ppm) 0.03 0.02 0.00
Lead (ppm) 0.68 0.79 -
Copper (ppm) 0.98 0.94 -
Legend: PH = Port Harcourt, TPH = Total petroleum hydrocarbon,
THC = Total hydrocarbon content, GAE = Gallic acid equivalent, ppm = Part per
million, mg/L = Milligram per liter, DPR = Department of Petroleum Resource.

Table 2: Microtitre Plate Absorbance Reading Using Toxi-Chromo Test.

Sample Positive control Warri sample PH sample Negative control
0.041 0.047 0.044 0.051 0.061 0.015
0.041 0.058 0.019 0.051 0.031 0.048
0.049 0.053 0.017 0.054 0.021 0.051
0.035 0.031 0.036 0.053 0.039 0.043
0.042 0.088 0.078 0.085 0.085 0.070
0.054 0.040 0.092 0.055 0.058 0.093
0.044 0.030 0.043 0.039 0.049 0.043
Blank 0.044 0.040 0.043 0.094 0.141 0.145
CV (%) 14.41 23.80 16.89 46.51

1702.04
1686.5
PH
Warri
Positive control

1694.8

Fig. 1: Toxicity Factor of Refinery Discharge Effluents and Mercury Chloride (HgCl2) After
Exposure to Genetically Modified E. coli.

RRJoT (2020) 22-31 © STM Journals 2020. All Rights Reserved Page 25
Research & Reviews: A Journal of Toxicology
Volume 10, Issue 2
ISSN: 2231-3834 (Online), ISSN: 2349-1264 (Print)

Positive control 1.535

Warri 0.852

PH 1.231

0 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8

1.5 h median effective concentration (mg/L/%)

Fig. 2: 1.5 h Median Effective Concentration of Refinery Discharge Effluents and Mercury Chloride
(HgCl2) After Exposure to Genetically Modified E. coli.

Control Liver Warri Liver PH Liver

Lead

Iron

Copper

0 0.5 1 1.5 2

Fig. 3: Heavy Metal Accumulation (Mg/Kg) in the Liver Tissue of Mice Exposed to Refinery
Industrial Effluents.

Mice Toxicity Profile values of AST (24 µ/L), ALT (29 µ/L), ALP
Figures 3 and 4 showed heavy metal (89µ/L), urea (14.6 mmol/L) and creatinine
accumulation (mg/kg) in the liver and kidney (146 mmol/L) while the control sample had
tissues of mice exposed to refinery industrial the lowest values of AST (12 µ/L), ALT (10
effluents. From the results, Warri liver and µ/L), ALP (43 µ/L) urea (2.6 mmol/L) and
control liver had the highest and lowest iron creatinine (75 mmol/L).
concentration values of 1.616 mg/kg and
1.084 mg/kg, respectively. Warri liver and The results of the haematoxylin and eosin stain
control liver had the highest and the lowest sections of the liver and kidney tissues of the
copper concentration values of 0.167 mg/kg mice exposed to refinery industrial effluents
and 0.066 mg/kg, respectively. Warri liver are shown in Plates 1a–c and 2a-c. From the
and the control liver had the highest and plates, control liver revealed a normal tissue
lowest lead concentration values of 0.169 architecture; central vein (arrow) and radiating
mg/kg and 0.099 mg/kg, respectively. hepatic plates, PH liver showed a portal
inflammation (arrows), Warri liver showed a
Table 3 showed the biochemical indices of diffuse hyperemia (arrow), control kidney
mice exposed to refinery industrial effluents. revealed a normal tissue architecture;
From the result, PH sample had the highest glomeruli inside the Bowman’s capsule

RRJoT (2020) 22-31 © STM Journals 2020. All Rights Reserved Page 26
Toxicological Evaluation of Two Nigerian Refinery Effluents Egurefa et al.

(arrow) and tubules, PH kidney showed an Warri kidney showed a more constricted
evidence of focal inflammation (circle) and glomerulus (arrow) and a normal one.

PH Kidney Warri Kidney Control Kidney

2.341

0.947

0.802

0.433
0.123

0.21
0.065

0.057

0.054
Copper Iron Lead

Fig. 4: Heavy Metal Accumulation in the Kidney Tissue of Mice Exposed to Refinery Industrial
Effluents.

(a) (b)

(c)
Plate 1A–C. Haematoxylin and Eosin Stain Section of the Liver Tissue of the Mice Exposed to
Refinery Industrial Effluents. A. Control liver- Liver section revealing a normal Tissue Architecture;
Central vein (Arrow) and Radiating Hepatic Plates (x40) B. PH Liver - Liver Section showing a
Portal Inflammation (arrows) (x4) C. Warri liver - Liver Section showing a Diffuse Hyperemia
(arrow) (x4).

RRJoT (2020) 22-31 © STM Journals 2020. All Rights Reserved Page 27
Research & Reviews: A Journal of Toxicology
Volume 10, Issue 2
ISSN: 2231-3834 (Online), ISSN: 2349-1264 (Print)

Table 3: Biochemical Indices of Mice Exposed to Refinery Industrial Effluents.

Sample AST (5–18 µ/L) ALT (3–25 µ/L) ALP (21–92 µ/L) Urea (1.7–1.9 mmol/L) Creatinine (8–127 mmol/L)
PH 24 29 89 14.6 146
Warri 15 20 64 5.8 94
Control 12 10 43 2.6 75
Key: PH = Port Harcourt, AST = Aspartate transaminase, ALT = Alanine transaminase, ALP = Alkaline phosphatase

(a) (b)

(c)
Plate 2A–C. Haematoxylin and Eosin Stain Section of the Kidney Tissue of the Mice Exposed to
Refinery Industrial Effluents. A. Control kidney- tissue section reveals a normal tissue architecture;
Glomeruli inside the Bowman’s Capsule (Arrow) and Tubules (x4) B. PH Kidney - renal section
showing an Evidence of focal Inflammation (Circle) (10). C. Warri Kidney - renal section showing a
more Constricted Glomerulus (Arrow) and a Normal One (x10).

DISCUSSION to avert future environmental consequences.

The wanton discharge of untreated The result in Table 1 revealed that the samples
petrochemical and refinery effluents into the contain high contents of linear and branched
Niger Delta ecosystems with its attendant chains of aromatic and aliphatic hydrocarbon
environmental and animal health dangers has fractions. The samples contain high oil and
been a growing concern in recent and past grease as well as phenol content which may be
times. The present study was undertaken to attributed to the high THC, TPH and phenolic
evaluate the toxic effects of two Nigerian compounds synthesized during refining
refineries’ effluents using natural bio-monitors processes. There were higher values of the
and to recommend the likely steps to be taken heavy metals analyzed. Comparatively, PH

RRJoT (2020) 22-31 © STM Journals 2020. All Rights Reserved Page 28
Toxicological Evaluation of Two Nigerian Refinery Effluents Egurefa et al.

sample had higher values of oil and grease, and AST in the plasma of the effluent
TPH, THC, copper, chromium, mercury and exposed mice. These biomarkers increased in
phenol while Warri sample had a higher iron, the blood of the animals when treated with
and lead metals, respectively. The results the effluents. These increases signify that the
revealed that the two effluent samples are liver and kidney tissues have been penetrated
highly polluted although PH sample was more leading to functional disruptions of the
polluted than the Warri sample. Most of the membranes of the effluent exposed mice liver
sample parameters exceeded most DPR [21] and kidney. These phenomenals could be
limits for produced wastewater which means attributed to leakage from the liver and
that the discharge of these liquid wastes into kidney due to the inflammation caused by the
aquatic or terrestrial environments would create 14 days mice exposure to the effluents. PH
structural and environmental disparity among sample had the highest values of the
living organisms. biomarkers but statistical analysis showed a
significant difference (p < 0.05) between the
The results in Table 2 and Figure 1 revealed samples and control with R 2 value of 0.304.
that the samples inhibited the biosynthesis of β- This shows that the samples had a significant
galactosidase which is an inducible enzyme in effect on the organs resulting to damage
the genetically modified E. coli as depicted by unlike the control set up. It is worthy to note
the high toxicity factor of the samples. The that the damages could also be attributed to
result in Figure 2 showed that the positive the presence of heavy metals, hydrocarbons
control sample (HgCl2) exhibited the most and other pollutants emitted during refining
harmful effect on 50% population of mutant E. processes and is in line with the published
coli with EC50 value of 1.535% in accordance works of Odinga et al. [3, 20] who reported
with the GESAMP [22] and CSP [23] toxicity that higher levels of these markers were
classification system as described before. The observed when rats were exposed to the Port
reason for this high toxic impact could be as a Harcourt refinery untreated effluents. Lead
result of the influences of the hydrocarbons, and iron had the highest accumulation in the
heavy metals, organic and inorganic liver and kidney tissues of the mice. The
contaminants that were present in those samples metals were carried via red blood cells to
as presented in Table 1. There were no different parts of the body after absorption.
statistically significant differences detected (P > When they reached the liver and kidney,
0.05) among the samples and control with R2 damage and physiological malfunctions
value of 0.022. The coefficient of variation occurred. These damages led to a significant
(CV) of all the samples were found to be less increase in the blood enzymes and a reduction
than 25% showing the validity standard of this in protein synthesis as shown in Figures 3 and
study and substantiate the recommendation 4. Andjielkovic et al. [9] reported that the
value (≤ 25%) of EBPI [4] that for Toxi- accumulation of lead and cadmium metals in
Chromo TestTM quantitative results to be liver and kidney tissues of rats resulted into
considered valid, the CV between the an imbalance in the biochemical parameters
absorbance values of negative controls and of the heavy metal exposed rats. In order to
sample replicates must be less than 25%. The reveal the extent of these damages by the
results of this study collaborate with the refinery industrial effluents, histopathological
research work of Uba et al. [5] who reported examinations were carried out on the
that wastewaters from the crude oil polluted dissected liver and kidney tissues and the
Niger Delta marine ecosystem had their CV results are illustrated in Plates 1a–c and 2a–c,
values less than 25%. Ajuzieogu et al. [12] respectively. From the results, there were
reported 15-minute EC50 values of the untreated evidences of structural abnormalities,
and treated produced water samples for Vibrio alteration of metabolites biosynthesis and
fischeri was 1.0% and 23.27%, respectively. excretory malfunctions in both, PH and Warri
effluents exposed mice in comparison to the
The result in Table 3 revealed the increase in control with normal structural architectures of
concentration of urea, creatinine, ALT, ALP the liver and kidney tissues. These alterations

RRJoT (2020) 22-31 © STM Journals 2020. All Rights Reserved Page 29
Research & Reviews: A Journal of Toxicology
Volume 10, Issue 2
ISSN: 2231-3834 (Online), ISSN: 2349-1264 (Print)

could be termed non-cancerous changes of effluent from Port-Harcourt refining

although carcinogenicity could result if the company and its effect on renal function of
mice were exposed for a very long time. albino rats. J. Res. Env. Sci. Toxicol. 2015;
Odinga et al. [3, 20] reported that the Port 4(2): 28–31.
Harcourt refinery untreated effluents induced 4. EBPI. Biotoxicity/microbiotest products
proximal degeneration of the integrity of the handout and manual. Ontario, Canada:
hepatic cells, inflammation, hepatocyte Environmental Bio-detection Products
degeneration, partial architectural distortion and Incorporation; 2016.
hemorrhage, and a renal damage resulting to a 5. Uba BO. Effects of aromatic hydrocarbons
proximal tubular degeneration and and marine water from Niger delta on the
haemorrhage. The observations of these β-galactosidase activity of mutant
researchers support the findings of the present Escherichia coli. Arch. Curr. Res. Internat.
study. 2019; 16(3): 1–16.
6. Sturgill MG, Lambert GH. Xenobiotic-
CONCLUSION induced hepatotoxicity: mechanisms of
The study showed that untreated effluents liver and methods of monitoring hepatic
from refineries could constitute and pose great function. Clin. Chem. 1997; 43(8 Pt 2):
danger to the environment and animal health 1512–1526.
when released indiscriminately. Hence, 7. CBMUT. Subcommittee on biologic
adequate and strict measures should be markers in urinary toxicology. Washington
established by government and environmental DC, USA: National Academy Press; 2002.
policymakers to ensure proper treatment and 8. Ferguson MA, Vaidya VS, Bonventre JV.
monitoring of these liquid wastes before they Biomarkers of nephrotoxic acute kidney
are discharged into our terrestrial and aquatic injury. Toxicol. 2008; 245(3): 182–193.
environments. 9. Andjelkovic M, Djordjevic AB,
Antonijevic E, Antonijevic B, Stanic M,
ACKNOWLEDGEMENTS Kotur-Stevuljevic J, Spasojevic-
We wish to thank Mr Collins Obi of Iyienu Kalimanovska V, Jovanovic M, Boricic N,
Hospital, Ogidi Anambra State, Nigeria for his Wallace D, Bulat Z. Toxic effect of acute
technical support towards the success of this cadmium and lead exposure in rat blood,
work.
liver, and kidney. Int. J. Env. Res. Public
Health. 2019; 16(2): 274.
REFERENCES 10. Egurefa SO, Orji MU, Uba BO. Toxic
1. Lee K, Cobanli SE, Robinson BJ, effect of refinery industrial effluent using
Wohlgeschaffen G. Application of
three toxicity bioassays. S. Asi. J. Res.
microbiological methods to assess the
Microbiol. 2020; 6(2): 10–23.
potential impact of produced water
11. Aruoja V, Sihtmäe M, Dubourguier HC,
discharges. In: K. Lee, J. Neff (eds.)
Kahru A. Toxicity of 58 substituted anilines
Produced Water: environmental risks and
and phenols to algae Pseudokirchneriella
mitigational advances in mitigational
subcapitata and bacteria Vibrio fischeri:
technologies. New York: Springer Science
Business Media; 2011. pp. 353–373. comparison with published data and
2. Manfra L, Moltedo G, Virno Lamberti C, QSARs. Chem. 2011; 84(10): 1310–1320.
Maggi C, Finoia G, Giuliani S, Onorati F, 12. Ajuzieogu CA, Odokuma LO, Blaise CC.
Gabellini M, Di Mento R, Cierco AM. Toxicity assessment of produced water
Metal content and toxicity of produced using microtox rapid bioassay. S Asi J. Res.
formation water (PFW): study of the Microbiol. 2018; 1(4): 1–9.
possible effects of the discharge on marine 13. Ekaye S, Uwagie-Ero EA, Aghayedo CO.
environment. Arch. Environ. Contam. Ameliorative effects of coconut oil on the
Toxicol. 2007; 53: 183–190. ovaries of refinery effluent intoxicated
3. Odinga T, Ayalogu EO, Essien EB. Norwegian rats. Nig. J. Environ. Sci. Tech.
Physicochemical and hydrocarbon content 2019; 33(1): 1–8.

RRJoT (2020) 22-31 © STM Journals 2020. All Rights Reserved Page 30
Toxicological Evaluation of Two Nigerian Refinery Effluents Egurefa et al.

14. Kiepper B. Understanding Laboratory 20. Odinga T, Ayalogu EO, Essien EB. Effect
Wastewater Tests. USA: University of of effluent from Port Harcourt refining
Georgia Cooperative Extension Circular company on hepatic and reproductive
922; 2013. functions of wistar albino rats. J. Nat. Sci.
15. Akpoveta OV, Egharevba F, Medjor O, Res. 2016; 6(4): 112–117.
Weltime O, Ize-iyamu K, Enyemike ED. 21. DPR. Environmental Guidelines and
Microbial degradation and its kinetics on Standards for the Petroleum Industry in
crude oil polluted soil. Res. J. Chem. Sci. Nigeria (EGASPIN). Revised Edition.
2011; 1(6): 8–14. Nigeria: Department of Petroleum
16. Nwineewii JD, Azuonwo K. Total Resources; 2002.
hydrocarbon concentrations (THC) in 22. GESAMP. Towards Safe and Effective
surface water, sediments and biota from Use of Chemicals in Coastal Aquaculture:
Otamiri River, Rivers State, Nigeria. Int. Reports and Studies. 65th edition. Rome:
J. Chem. Stud. 2018; 6(3): 2743–2748. Group of Experts on the Scientific Aspects
17. Leouifoudi I, Zyad A, Amechrouq A, of Marine Environmental Protection;
Oukerrou MA, Mouse HA, Mbarki M. 1997.
Identification and characterisation of 23. CSP. Chemical risk assessment: aquatic
phenolic compounds extracted from toxicity. Pro L. (Editor) Chem Safety Pro
Moroccan olive mill wastewater. F. Sci. Publication; 2016. (Accessed on
Tech. 2014; 34(2): 249–257. 01/03/1997).
18. APHA. Standard Methods for
Examination of Water and Wastewater.
22nd edition. Washington, DC, USA: Cite this Article
American Public Health Association; Samson Ogagaoghene Egurefa, Micheal
2012. Uchenna Orji and Bright Obidinma Uba.
19. OECD. Chemical Safety and Animal Toxicological Evaluation of Two Nigerian
Welfare. Paris, France: Organization of Refinery Effluents using Natural
Economic Cooperation and Development Biomonitors. Research & Reviews: A
Publishing; 2009. Journal of Toxicology. 2020; 10(2): 22–31p.

RRJoT (2020) 22-31 © STM Journals 2020. All Rights Reserved Page 31