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Online ISSN: 2383-4501

University of Tehran Press
https://jpoll.ut.ac.ir/

National Survey of Stream Water Quality Revealing Threats to

Antibio-Resistant Bacteria, Antibiotic Residues and Heavy Metals
in Benin
Elodie Gbotche1 | Marie Camille Houssou Quenum2 | Tamègnon Victorien Dougnon1 |
Aemere Ogunlaja3 | Jean-Robert Klotoe1 | Kafayath Fabiyi1 | Alidehou Jerrold Agbankpe1
| Edna Hounsa1 | Kévine Vodounnon1 | Wassiyath Mousse4 | Sophonia Ahouandjinou1 |
Curiace Hinnilo1 | Eskyl Togbe1 | Nelly Kelome5 | Clément Agbangla6 | Honoré Sourou
Bankole1 | Lamine Baba-Moussa4 | Iyayi E. Unuabonah3
1. Research Unit in Applied Microbiology and Pharmacology of Natural Substances, Research Laboratory in Applied Biology,
Polytechnic School of Abomey-Calavi, University of Abomey-Calavi, 01 P.O. Box 2009, Cotonou, Benin
2. Laboratory of Monitoring and Environmental Studies, Ministry of the Living Environment and Sustainable Development,
Benin
3. Department of Chemistry, University of Ibadan, Ibadan, Nigeria
4. Laboratory of Biology and Molecular Typing in Microbiology, Faculty of Sciences and Techniques, University of Abomey-
Calavi, P. O. Box 1604, Cotonou, Benin
5. Laboratory of Geology, Mines and Environment, Faculty of Sciences and Techniques, University of Abomey-Calavi, Benin
6. Laboratory of Genetics and Biotechnology, Faculty of Sciences and Techniques, University of Abomey-Calavi, Benin

Article Info ABSTRACT

Article type: Benin’s waterways are affected by several forms of pollution that are linked in particular
Research Article to anthropic activities. This study aims to detect the presence of antibiotic residues,
the frequency of antibiotic resistant bacteria and the levels of heavy metals in Benin’s
Article history:
waterways. 160 surface water samples from streams in Benin were collected. They were
Received: 13 Sep 2022
Revised: 28 Nov 2022
filtered by the membrane filtration method, then incubated on different media. The
Accepted: 17 Jan 2023 isolated bacterial species were identified by API 20E gallery and specific biochemical
tests. After detection of the resistance profile of the latter, the antibiotic residues were
Keywords: quantified in the samples by the ELISA technique on plate and the physicochemical
Antibiotic resistant analyses were performed by Multi 3630 IDS SET KS2 multimeter. Finally, heavy metal
Bacteria levels were detected by the MERCK test kit method specific to each metal. The bacterial
Antibiotic residues species mostly identified were Klebsiella pneumoniae (56.59%), Klebsiella spp. (18.68%),
Heavy metals Enterobacter spp. (12.63%). The most abundant resistance of bacterial strains was to
Streams
amoxicillin + clavulanic acid (92%), followed by metronidazole (86%). Metronidazole
Benin
was the antibiotic with the highest residue concentration in the samples (6.578 to 6.829
µg/L), followed by ciprofloxacin (2.142 to 9.299 µg/L). Benin streams contain heavy
metals such as mercury (0.454±0.129 µg/L), lead (0.040±0.50 mg/L), zinc (6.120±16.017
mg/L), nickel (0.155±0.233 mg/L) and cadmium (0.154±0.132 mg/L). The analysis of
the physico-chemical parameters showed that, apart from electrical conductivity, all
parameters comply with Beninese and World Health Organization standards. Actions
must be taken to clean up these rivers to preserve the integrity of aquatic ecosystems
in Benin.
Cite this article: Gbotche, E., Camille Houssou Quenum, M., Victorien Dougnon, T., Ogunlaja A., Klotoe, J.R., Fabiyi,
K., Jerrold Agbankpe, A., Hounsa, E., Vodounnon K., Mousse, W., Ahouandjinou, S., Hinnilo, C., Togbe, E., Kelome, N.,
Agbangla, C., Sourou Bankole, H., Baba-Moussa, L., & Unuabonah, I.E. (2023). National Survey of Stream Water Quality
Revealing Threats to Antibio-Resistant Bacteria, Antibiotic Residues and Heavy Metals in Benin. Pollution, 9 (2), 678-692.
http://doi.org10.22059/POLL.2022.348629.1629

© The Author(s). Publisher: University of Tehran Press.

DOI: http://doi.org10.22059/POLL.2022.348629.1629

*Corresponding Author Email: [email protected]

679 Gbotche, E. et al.

INTRODUCTION

For several decades, streams have played an important role in the development of nations
through their importance in agriculture and transportation, without forgetting that they are
also the receptacle of municipal wastewater (Kubera, 2021). Thus, they are often exposed to
physical, chemical and biological contamination (Tissera et al., 2013). The continual release of
antibiotics into the aquatic ecosystem is due to their uncontrolled and anarchic use (Tshibanda
et al., 2021). The main origin of these substances in natural waters being runoff, livestock and
effluent discharges from wastewater treatment plants (Zhang et al.,2020). Thus, reservoirs of
multidrug resistant bacteria and resistance genes are formed through the interaction between
anthropogenic effluents (hospital effluents, wastewater from slaughterhouses, industries or
communities) and streams (rivers and lakes) (Marti et al.2014). In addition, the presence of
antibiotic residues and multidrug resistant bacteria in human and animal feces leads to their
introduction into wastewater and then into streams (rivers and/or lakes) through direct
discharge or wastewater treatment systems (Zhang et al.,2020). All these interactions promote
the growth of multidrug resistant bacteria in streams, through the contact of high bacterial
density with high therapeutic concentrations of antibiotics under nutritionally favorable
conditions (Rodriguez-Mozaz et al.,2015). In low- and middle-income countries (LMICs) such
as Benin, the situation of contamination of streams by antibiotic residues and the development
of multi-drug resistant bacteria is still dramatic. This is due to the high level of poverty of
the population, the absence of inadequacy of health care facilities and poor sanitation and
water treatment infrastructure (Collignon et al., 2018; Croix, 2019). This situation increases
the risk of bacterial infection and therapeutic failures in humans due to the consumption
of water (drinking) and fish containing antibiotic residues and contaminated by multi-drug
resistant bacteria. Apart from the contamination of streams with antibiotic residues and multi-
drug resistant bacteria, their pollution with heavy metals is one of the greatest threats to the
aquatic ecosystem due to their persistence and possible bioaccumulation and biomagnification
in food chains (Jovanović et al.,2017). Heavy metal contamination of the aquatic ecosystem
can originate from industrial or community wastewater, pesticide and inorganic fertilizer
use, runoff, landfill leaching, marine and port activities, as well as geological alteration of the
earth’s crust and atmospheric deposition (Ajima et al.,2015). The capacity of bioaccumulation
of heavy metals in several aquatic organisms, and thus in fish has been demonstrated by several
authors (Verep et al.,2012; Janjić et al.,2015; Jovičić et al., 2015). Similarly, studies have shown
that the accumulation of heavy metals in fish at given concentrations, can be considered as
indicators of public health problems for human who are fish consumers (Abdel-Baki et al.,2011;
Montalvo et al.,2014). The level of bioaccumulation and bioavailability of heavy metals in fish
waters and tissues is under the influence of several factors, such as fish biological habitats and
physico-chemical parameters of waters (water temperature, pH, oxygen concentration in water,
electrical conductivity, turbidity, redox potential and total dissolved solids) (Kehrig et al.,2013;
Jovanović et al.,2017). A few studies in Benin have focused on microbial contamination of water
bodies (Ichola et al.,2021; Koudokpon et al.,2021), but very few have provided information on
the antibiotic residues and heavy metal. In order to prevent the health of aquatic environment,
of animals and of Beninese’s, it is important to evaluate the level of contamination of the main
streams of Benin with multidrug resistant bacteria, antibiotic residues, heavy metals and to
understand the influence of physicochemical factors of the water on the bioaccumulation of
heavy metals. It is in this same context that the present study was initiated. Its objectives are
to identify multidrug resistant bacteria, to detect and quantify antibiotic residues, to measure
heavy metals and to measure the physicochemical parameters of the samples of Benin’s streams.
680 Pollution 2023, 9(2): 678-692

MATERIALS AND METHODS

Stream surface water samples were collected from all over Benin (Ganvié, Grand-Popo, Bopa,
Tori, Porto-Novo, Adjohoun, Tokpa, Zangnannando, Kota, Tanougou, Koumagou, Malanville,
Okpara, Sota, Mékrou, Pendjari) (Figure 1) from 16th April to 28th June 2021. The samples
were collected aseptically in 1L bottles and stored in a cooler with ice packs and processed
in a laboratory within 24 hours. At each stream surface, 10 points were randomly selected
and samples were taken at these points (Table 1). Thus, 10 stream surface water samples were
collected at each stream surface. In total, of 160 stream surface water samples were collected and
analyzed.
After homogenization, 600 ml of each sample was filtered twice at a rate of 300 ml per
filtration. The filtrations were performed on membranes with a pore size of 0.20 µm. For each
sample, the two membranes were incubated on eosin methylene blue (EMB) and Chapman agar
plates, respectively. Then, the plates were incubated at 37°C for 24 hours. These both culture
media were respectively used for the isolation of colonies of enterobacteria (Gram negative
bacilli) and cocci. Two or three characteristic colonies obtained on the both culture media used
were selected. These colonies were streaked on Mueller-Hinton agar plates for purification and
then Gram staining, biochemical tests (catalase and oxidase), seeding of the API 20E gallery
(for only Gram-negative bacilli), and free staphylocoagulase and DNAse tests (for only Gram-
positive cocci) were performed for the identification of Enterobacteriaceae and Staphylococcus
aureus strains (Ichola et al.,2021).
All identified bacteria were subjected to antimicrobial susceptibility testing on Muller-

Fig. 1. Sample collection sites

Figure 1: Sample collection sites

 681 Gbotche, E. et al.

Table 1. Summary of stream samples by region and location of origin and stream type
Table 1. Summary of stream samples by region and location of origin and stream type

Benin regions Location or name of the streams Types of streams Number of samples
Kota waterfalls 10
Tanougou waterfalls 10
Koumagou River 10
Malanville River 10
Northern
Okpara River 10
Sota River 10
Mékrou River 10
Pendjari River 10
Ganvié River 10
Grand-Popo River 10
Bopa Lake 10
Tori River 10
Southern
Porto-Novo River 10
Adjohoun Lake 10
Tokpa Lake 10
Zangnannando Lake 10
Total 160

Hinton agar plates using the Kirby–Bauer disc diffusion method to the following antibiotics:
ciprofloxacin (5 µg), chloramphenicol (30 µg), metronidazole (5 µg), aztreonam (30 µg),
ceftriaxone (30 µg), ertapenem (10 µg), imipenem (10 µg), amoxicillin + clavulanic acid (30
µg), gentamicin (10 µg) and nalidixic acid (30 µg). Measured inhibition zone diameters were
interpreted according to EUCAST guidelines (EUCAST,2019). The reference strain Escherichia
coli ATCC 25922 was used as a control.
Seven antibiotics were searched in this study. These were amoxicillin (AMO) and ampicillin
(AP) of the β-lactam family, chloramphenicol (CAP) a phenicol, ciprofloxacin (CPFX) a
fluoroquinolone, metronidazole (MNZ) an imidazole, sulfamethoxazole (SMZ) a sulfonylurea
and neomycin (NEO) an aminoglycoside. They were chosen in view of their frequency of
use in hospitals in West African countries (Dougnon et al.,2020). Both the detection and the
quantification were carried out according to the instructions of the MyBioSource® ELISA kit
specific to each antibiotic sought. The plate in each kit was read at 450nm on a microplate reader
and then the calibration curve of the standards was plotted with the percentages of absorbances
(A/A0x100 with A: average absorbance of the standard or sample and A0 the average absorbance
of the 0ppb standard) as a function of logC of the standards contained in each kit. The
determination of sample concentrations from the linear equation (y=ax+b) of the calibration
curves (Figures 2 and 3) obtained for each antibiotic was performed.
The determination of five metals, namely mercury, lead, zinc, cadmium and nickel, was
carried out by the method of the MERCK test kits specific to each metal. The concentrations
were determined using a photo Lab 6600UV-VIS molecular absorption spectrophotometer.
The principle is that in the presence of (pyridyl-2’-azo)-4-resorcinol (PAR), the metal forms a
complex which is determined photometrically. The concentrations of mercury, zinc, cadmium
and nickel are directly determined with the Photo Lab 6600UV-VIS. For lead, the process
also depends on the purity of the water and requires a total hardness test. With the hardness
determination, the lead analysis is done in two steps. A first reading with a molecular absorption
spectrophotometer after addition of reagent A and a second one after addition of reagent B. The
682 Pollution 2023, 9(2): 678-692

Fig. 2. Calibration curves for Amoxicillin, Ampicillin, Chloramphenicol and Metronidazole

Figure 2. Calibration curves for Amoxicillin, Ampicillin, Chloramphenicol and Metronidazole

Fig. 3. Calibration curves for Ciprofloxacin, Sulfamethoxazole and Neomycin

Figure 3. Calibration curves for Ciprofloxacin, Sulfamethoxazole and Neomycin
683 Gbotche, E. et al.

final lead concentration in the sample is the difference between the values obtained after the
different readings.
The physicochemical characterization was performed to determine the following parameters:
temperature, hydrogen potential (pH), salinity, electrical conductivity (EC), resistivity, redox
potential (P-redox) and total dissolved solids (TDS). They were analyzed according to the
protocols recommended by (Rodier et al.,2009) by direct measurement with a multimeter Multi
3630 IDS SET KS2.
Data were recorded in Excel 2016 spreadsheet; the same software was used to perform the
different antibiotic calibration curves. R software was used for data analysis. Two-way ANOVA
test was used for multiple comparison between the mean concentrations of antibiotic residues,
values of physicochemical parameters and concentrations of heavy metals of the river samples
from Northern and Southern Benin. The graphs were produced using GraphPad Prism 7
software.

RESULTS AND DISCUSSION

Rivers are dynamic, ever-changing ecosystems that undergo many changes due to natural
processes and human activity. Anthropo-pression has a significant impact on water quality
in aquatic environments and on human health (Gotkowska-Plachta et al.,2015). Thus, a
characterization of multi-drug resistant bacteria and antibiotic residues, an assessment of
heavy metal contamination of streams and an understanding of the relationship between
physicochemical water parameters, heavy metals and multi-drug resistant bacteria would be
important and urgent in order to reduce the risks of infections and heavy metal contamination.
This is what this study was dedicated to. From the 160 surface water samples analyzed, we
isolated 283 bacterial strains of which 64.31% (n = 182) were identified to the species level and
35.69% (n = 101) were identified as non-enterobacteria. Of One hundred eighty-two bacterial
species identified, Klebsiella pneumoniae was the most represented species with 56.59% (n =
103), followed by Klebsiella spp. (18.68%; n = 34) and Enterobacter spp. (12.63%; n = 23) (Figure
4).
We’ve observed an abundance of non-enterobacterial and Klebsiella pneumoniae strains but
also the presence of other bacterial species such as E. coli, Enterobacter spp, Salmonella spp and
Klebsiella spp. In Benin, Ichola et al. (2021) also observed an abundance of the genus Klebsiella
and the presence of non-enterobacteria, E. coli species, and Enterobacter spp in water samples
from the Cotonou-Lac Nokoué hydrographic complex. Studies in Algeria and Nigeria have

Fig. 4. Percentage of different bacteria species identified

Figure 4. Percentage of different bacteria species identified
684 Pollution 2023, 9(2): 678-692

Fig. 5. Percentage of resistance of bacterial strains to antibiotics used

Figure 5. Percentage of resistance of bacterial strains to antibiotics used
reported presence of non-enterobacteriacea, and Klebsiella spp in water samples as was detected
also in our study (Olaoye et al.,2009; Djenadi et al.,2018). The same species (resistant) were
also found in some wastewater samples collected in Seme Krake, a town in southern Benin
(Koudokpon et al.,2021).
Ten different antibiotics belonging to several different families were used in order to determine
the resistance level of the identified bacterial isolates. The most abundant resistance of the
bacterial strains was to amoxicillin + clavulanic acid (92%), followed by metronidazole (86%),
ceftriaxone (44%), ertapenem (32%), ciprofloxacin (24%), and aztreonam (24%) (Figure 5). It
should be noted that all bacterial strains are resistant to more than three antibiotics belonging to
different families, thus multidrug-resistant.
The bacterial strains isolated from the stream samples were all multidrug resistant with a
low resistance to phenicol, aminoglycosides and more especially fluoroquinolones has been
observed. Sani et al.,2018 have also obtained similar results. A strong resistance to beta-lactams
and imidazole has been recorded. These results are similar to those of (Azaiez et al.,2019 ;
Khalfallah et al.,2021) who have reported a resistance to beta-lactams. The resistance of strains
of enterobacteria and non-enterobacteria to beta-lactams is an emergence reported by the World
Health Organization (WHO,2017), as priority resistance in human health. This shows that the
rivers of Benin constitute a reservoir of pathogens indicators of risk to public health. Resistance
to imidazole, more precisely metronidazole observed in our study has also been shown by some
studies (Gombima,2021). Our result is different from those obtained by (Azaiez et al.,2019) in
which strains were sensible to metronidazole. This resistance could be explained by the fact that
this antibiotic is considered as a dewormed and an antispasmodic by the populations of Benin
and taken without control to treat bacterial infections (Ichola et al.,2021).
The detection of antibiotic residues in the surface water of the 16 collected streams showed
that all the searched antibiotics were detected, except for sulfamethoxazole and neomycin.
Chloramphenicol and metronidazole were detected in all streams sampled. Amoxicillin was
detected in 4 streams, 2 in the north (12.5%) and 2 in the south (12.5%). As for ampicillin
residues, they were detected in 8 rivers in the south (50%) and in 4 in the north (25%). Finally,
ciprofloxacin residues were detected in 50% (6/16) and 37.5% (8/16) of the streams sampled in
the south and north, respectively (Table 2).
The concentrations of antibiotic residues varied from one course sample to another.
 685 Gbotche, E. et al.

Table 2. Distribution of samples according to the frequencies of detection of antibiotic residues

Table 2. Distribution of samples according to the frequencies of detection of antibiotic residues

Streams samples
(n = 32)
Antibiotics detected
Streams from Northern Streams from Southern
(n = 16) (n = 16)
Amoxicillin (AMO) 2 (12.5 %) 2 (12.5%)
Ampicillin (AP) 4 (25%) 8 (50%)
Chloramphenicol (CAP) 16 (100%) 16 (100%)
Metronidazole (MNZ) 16 (100%) 16 (100%)
Ciprofloxacin (CPFX) 6 (37.5%) 8 (50%)
Sulfamethoxazole (SMZ) - - - -
Neomycin (NEO) - - - -

Table 3. Mean concentration of antibiotic residues according to the two regions of Benin where the streams were
Table 3. Mean concentration of antibiotic residues according to the two regions of Benin where the streams were
collected
collected
Mean concentrations of antibiotic residues detected (μg/L)
Antibiotics residues
Northern Southern
Amoxicillin (AMO) 0.111 ± 0.016 0.065 ± 0.005
Ampicillin (AP) 1.550 ± 0.050**** 2.415 ± 0.065****
Chloramphenicol (CAP) 0.104 ± 0.004 0.106 ± 0.003
Ciprofloxacin (CPFX) 1.395 ± 0.045**** 3.010 ± 0.003****
Metronidazole (MNZ) 6.635 ± 0.035 6.700 ± 0.050
****: p-value < 0.0001

Metronidazole was the antibiotic with the highest residue concentration in the samples (6.578
to 6.829 µg/L), followed by ciprofloxacin with residue concentrations ranging from 2.142 to
9.299 µg/L and ampicillin (1.272 to 4.235 µg/L). The comparison of the average concentrations
of the different antibiotic residues obtained in the northern and southern streams showed that
the concentrations of chloramphenicol and metronidazole residues in the southern streams
(0.106 ± 0.003 µg/L; 6.700 ± 0.050 µg/L) are higher than those obtained in the northern streams
(0.104 ± 0.004 µg/L; 6.635 ± 0.035 µg/L). In contrast, the concentration of amoxicillin residues
in northern streams (0.111 ± 0.016 µg/L) is higher than those in southern streams (0.065 ± 0.005
µg/L). However, the difference between these different antibiotic residue concentrations is not
statistically significant. On the other hand, there is a statistically significant difference between
the mean concentrations of ampicillin and ciprofloxacin residues obtained in the northern and
southern streams of Benin, with higher concentrations of these two antibiotics in the southern
(2.415 ± 0.065 µg/L; 3.010 ± 0.003 µg/L) streams than in the northern (1.550 ± 0.050 µg/L; 1.395
± 0.045 µg/L) one (p < 0.0001) (Table 3).
Various antibiotic residues have been detected in stream surface water samples from
Benin. Aquatic organisms are thus exposed to cocktails of antibiotics. Even if their individual
concentrations measured in the environment are low, when combined, they can be significant
and cause significant toxicity to aquatic organisms (Everaert et al.,2020) by interacting with
each other having either a large effect (synergistic effect) or a smaller effect together than alone
(antagonistic effect) (Danner et al.,2019). Metronidazole, ciprofloxacin and ampicillin were also
detected at high concentrations in the rivers of northern and southern Benin. This could be
explained by the frequent use of these molecules in human and animal health but also by their
686 Pollution 2023, 9(2): 678-692

low biodegradation in the environment. The concentrations found in the environment are highly
variable in relation to variations in consumption patterns (type of antibiotic and quantity) and
with the persistence of the different compounds disseminated (Peralta et al.,2016). On the other
hand, the conditions of the receiving environments and the intrinsic chemical characteristics of
the molecules condition their persistence (Michael et al.,2013). Therefore, the study of processes
governing persistence such as sorption, mobility and degradation of antibiotics is an essential
step. The most important mechanisms conditioning sorption and thus mobility of antibiotic
compounds are interactions with organic matter and mineral constituents in the soil, as well as
ion exchange, hydrogen bonding and complex formation with metal ions (Morel et al.,2014).
In addition, the lack of sanitation and wastewater treatment facilities in low-income countries
means that the average concentration of antibiotic residues is generally much higher.
The concentration of heavy metals varies from one sample to another. The average
concentration of zinc (6.126 ± 16.017 mg/L) was the highest of the other metals tested in
southern Benin streams, followed by mercury (0.454 ± 0.129 mg/L), cadmium (0.140 ± 0.159
mg/L) and nickel (0.126 ± 0.163 mg/L). In terms of heavy metal concentrations in the northern
streams, mercury (0.277 ± 0.147 mg/L) was the highest, followed by nickel (0.155 ± 0.233
mg/L), cadmium (0.154 ± 0.132 mg/L) and zinc (0.139 ± 0.114 mg/L). The difference between
the average concentrations of heavy metals in the northern and southern Benin streams is not
statistically significant (Table 4).
All the metals searched for were found in concentrations largely superior to the standards
tolerated in Benin with a predominance of zinc (Zn). These results highlight the environmental
danger and the health risks to which the surrounding populations are exposed when we know
that in Benin, these waterways represent a source of water supply in certain localities. Various
studies including those of (Dimon et al.,2014; Hounkpè et al.,2017) have also reported high
concentrations and above the standards illustrating the toxicological pollution of Beninese
waterways. Anthropogenic activities as well as runoff water constitute a non-negligible source
of pollution of this aquatic ecosystem (Hounkpè et al.,2017)
The values of the different physicochemical parameters vary from one river to another.
The conductivity of Benin’s rivers varies from 7.55 to 22300 µS/cm. The mean conductivity of
the southern streams (8770.081 ± 12611.220 µS/cm) was significantly higher than that of the
northern streams (111.237 ± 51.205 µS/cm) (p = 0.0073) (Table 5). Total dissolved salts (TDS)
had the same values as conductivity in each stream. Like conductivity, its mean value in the
southern streams (8770.081 ± 12611.220 mg/L) is significantly higher than in the northern
streams (111.237 ± 51.205 mg/L) (table 5). Temperature is one of the essential parameters
governing chemical equilibrium and the development of flora and fauna in our waterways.
The average temperature of the rivers in southern Benin was 26.75±0.434°C with a maximum
temperature of 27.3°C and 26.4°C as minimum temperature. The average temperature of the
northern rivers was 27.237±0.292°C with 27.5°C and 26.6°C as the maximum and minimum
temperature respectively (table 5). The pH is a measure of the acidity of the water, i.e. the
concentration of hydrogen ions (H+). The pH of Benin’s rivers varies from 5.73 to 7.67. Streams
in the south have a mean pH of 6.604±0.658. With a slight difference, the mean pH of the
northern streams was 6.579±0.256 (table 5). Our results showed that the salinity of rivers in
northern Benin was 0%. Similarly, of the 8 streams sampled in southern Benin, 4 had zero
salinity. The salinity of the other 4 southern streams varied from 4.2 to 19.2% with a mean of
5.825±7.475% (table 5). The mean of redox potential of the rivers in northern and southern
Benin was 21.438±39.048 mV and 22.938±11.193 mV, respectively. It should be noted that the
redox potential of three rivers in the south was negative (Ganvié river, Grand-Popo river and lake
Ahémé) as well as the Sota river in the northern region of Benin (table 5). The mean resistibility
of streams in southern Benin was 6.490±9.008 KΩ/cm. While that of the northern streams was
15.225±6.238 KΩ/cm. However, this difference in stream resistibility between these two regions
687 Gbotche, E. et al.

Table 4. Concentrations of heavy metals in streams by region and sampling location in Benin
Table 4. Concentrations of heavy metals in streams by region and sampling location in Benin

Streams Heavy metal concentration in streams samples

Suspended
Mercury Lead Zinc Nickel Cadmium
solids
(μg/L) (mg/L) mg/L (mg/L) (mg/L)
(mg/L)
Southern
Ganvié river 0.09 0.5076 0.13 0.229 - 0.033
Grand-Popo river 0.02 0.6046 - 0.014 - 0.143
Lake ahémé (Bopa) 0.12 0.6632 0.03 0.109 0.23 0.073
Tori river 0.02 0.4272 - - 0.07 0.125
Porto-Novo river 0.04 0.4098 0.08 0.148 - 0.04
Lake tovègbamé
0.06 0.3094 - 0.096 0.4 0.106
(Adjohoun)
Lake nokoué
0.19 0.4036 0.08 45.7 0.31 0.075
(Tokpa)
Lake azili
0.5 0.3065 0.0 2.71 - 0.522
(Zangnannando)
0.130 ± 0.454 ± 0.040 ± 6.126 ± 0.126 ± 0.140 ±
Mean ± SD
0.160a 0.129b 0.050c 16.017d 0.163e 0.159f
Northern
Falls of kota 0.02 0.3365 - - - 0.06
Falls of tanougou 0.32 0.1822 - 0.043 0.05 0.032
Koumagou river 0.04 0.6204 0.11 0.091 - 0.055
Niger river 0.27 0.2337 0.0 0.213 0.46 0.335
Okpara river 0.13 0.2138 - 0.226 0.58 0.182
Sota river 0.02 0.2188 - 0.156 0.15 0.137
Mekrou river 0.06 0.1924 0.0 0.336 - 0.369
Pendjari river 0.02 0.1917 - 0.051 - 0.061
0.110 ± 0.277 ± 0.014 ± 0.139 ± 0.155 ± 0.154 ±
Mean ± SD
0.121a 0.147b 0.039c 0.114d 0.233e 0.132f

of the country is not statistically significant (table 5).

The analysis of the physicochemical quality of the streams through some parameters revealed
that these last ones vary in space. Apart from the electrical conductivity, all other parameters
are in conformity with the standards. The mean concentrations of suspended solids (SS) are all
in compliance with WHO and Beninese standards respectively (20mg/l and 35mg/l). Indeed, as
a result of anthropogenic activities, chemicals released into the environment can enter aquatic
ecosystems and become incorporated into suspended solids (Dimon et al.,2014). Their presence
in groundwater prevents light penetration and poses a risk to fish species in that it causes their
death through gill plugging (Dimon et al.,2014). Electrical conductivity, resistivity and total
dissolved solids (TDS) are parameters that allow us to assess the overall mineralization of a
water. Thus, a low conductivity for a stream is also synonymous with a low mineralization of
salts present in the environment (Ben Moussa et al.,2012). External inputs of water into the
aquatic environment contributes to the dilution of these parameters with the corollary of low
values of these parameters (Zinsou et al.,2016). In the rivers, only those in the South showed a
mean conductivity (87700.81µS/cm) very much higher than the norm with high levels in the So
(22300µS/cm) and Ouémé (16440µS/cm) rivers as well as in Lake Nokoué (31000µS/cm). The
 688

Table 5. Values of physicochemical parameters of streams

Tablesamples
5. Values of physicochemical parameters of streams samples

Physicochemical parameters
Streams samples Conductivity Temperature Salinity Total dissolved salts Redox potential Resistibility
pH
(μS/cm) (° C) (%) (mg/L) (mV) (KΩ/cm)
Southern
Ganvié river 22300 27.2 6.99 13.5 22300 -1.2 0.0448
Grand-Popo river 148.2 26.6 6.98 0 148.2 -0.9 6.74
Lake ahémé (Bopa) 7.55 26.4 7.67 4.2 7.55 -41.9 0.1322
Tori river 144.8 26.4 6.33 0 144.8 37.7 6.91
Porto-Novo river 16440 27.3 6.45 9.7 16440 30.6 0.0608
Lake tovègbamé
81.2 27.3 5.73 0 81.2 73.1 12.3
(Adjohoun)
Lake nokoué (tokpa) 31000 26.4 6.89 19.2 31000 4.3 0.0323
Lake azili
38.9 26.4 5.79 0 38.9 69.8 25.70
(zangnannando)
8770.081 ± 26.75 ± 6.604 ± 5.825 ± 8770.081 ±
Mean±SD 21.438 ± 39.048 6.490 ± 9.008
12611.220** 0.434 0.658 7.475 12611.220**
Northern
Falls kota 89.8 27.4 6.75 0 89.8 12.7 11.16
Falls tanougou 185.2 27.5 6.37 0 185.2 35.4 5.4
Koumagou river 40.5 27.2 6.25 0 40.5 42.6 24.7
Niger river 114.7 27.1 6.4 0 114.7 33.4 8.73
Okpara river 120 27.3 6.4 0 120 33.5 8.33
Sota river 182.4 27.3 6.98 0 182.4 -0.8 5.48
Mékrou river 87.1 27.5 6.77 0 87.1 11.4 11.48
Pendjari river 70.2 26.6 6.71 0 70.2 15.3 14.26
Mean±SD 111.237 ± 27.237 ± 6.579 ±
0 111.237 ± 51.205** 22.938 ± 11.193 15.225 ± 6.238
51.205** 0.292 0.256
**: p < 0.01
Pollution 2023, 9(2): 678-692
689 Gbotche, E. et al.

rivers in the South have a mean resistivity of 6.49 KΩ/cm. On the other hand, the So, Ouémé
and Nokoué streams showed significant mineralization. The high TDS contents of these three
streams reflect their character translate the presence of salts, minerals and metals. This result is
corroborated by the mean salinity (5.82%). In sum, the rivers studied in the South, notably the
So River, Ouémé and Lake Nokoué, have high mineralization, are loaded with total dissolved
solids and therefore have a high salt content and are therefore unpleasant waters. On the other
hand, in the North, the mean values of conductivity, total dissolved solids and resistivity allow
us to conclude that they are weakly mineralized with a low or zero content of salt (0%), minerals
and metals.

CONCLUSION

The analysis of streams in northern and southern Benin revealed the presence of antibiotic
residues in significant concentrations and of multi-drug resistant bacteria. The results show that
the concentrations of antibiotic residues obtained are likely to have direct or indirect effects not
only on the microbial component of aquatic populations but also on human health. This situation
must lead us to evaluate the risk presented by these molecules but also to improve techniques
and develop methods adapted to limit the release of these residues into the environment because
they also participate in the increase of the antibiotic resistance phenomenon. Our study showed
the presence of several heavy metals (mercury, lead, zinc, nickel and cadmium) in Benin’s
streams. The sources of the presence of these metals in the streams are domestic and industrial
wastes and the excessive use of phosphatic fertilizers. Apart from lead, the average concentration
of mercury, zinc, nickel and cadmium was above the norm in Benin. This justifies that there is
a risk of heavy metal contamination in aquatic organisms and in humans who consume water
and fish.

ACKNOWLEDGEMENTS

The authors are very grateful to The World Academy of Sciences (TWAS) and the Islamic
Development Bank (IsDB), which funded one of them for this study. They are finally grateful to
the Minister of the Living Environment and Sustainable Development in Benin, his Excellency
his Excellency M. Jose ́ TONATO and all his staff.

AUTHORS’ CONTRIBUTIONS

EG, MCHQ, TVD, AO, NK, CA and HB wrote the protocol. WM and IEU got the funding.
KF, AJA, EH, KV, SA, CH and ET processed the samples. TVD, JRK performed the statistical
analyses. TVD, AJA and JRK wrote the draft of the manuscript. All authors read and approved
the final manuscript.

GRANT SUPPORT DETAILS

The authors are very grateful to The World Academy of Sciences (TWAS) and the Islamic
Development Bank (IsDB). These two institutions have made this research possible through
research funding allocated to the research team under the IsDB-TWAS Grants for Research
Collaboration in Sustainability Sciences (506808). They reviewed the research protocol and
validated the design of the study and collection, analysis, and interpretation of data.
690 Pollution 2023, 9(2): 678-692

CONFLICT OF INTEREST

The authors declare that there is not any conflict of interests regarding the publication of this
manuscript. In addition, the ethical issues, including plagiarism, informed consent, misconduct,
data fabrication and/ or falsification, double publication and/or submission, and redundancy
has been completely observed by the authors.

LIFE SCIENCE REPORTING

No life science threat was practiced in this research.

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