Abstract
The present study was conducted to determine daily intake of cadmium (Cd), copper (Cu), and lead (Pb) and to assess noncarcinogenic human health risk caused by these trace metals in the commonly consumed fish species (
Introduction
Fish is an important component of many meals around the world because it has high contents of proteins, essential fats, vitamins, and minerals.1-3 Its growing demand is evident from the vigorous growth in aquaculture and in the fact that the world’s per capita fish consumption has risen from 9.9 kg in the 1960s to 14.4 kg in the 1990s to 19.7 kg in 2013. 4 Nonetheless, increased aquatic pollution caused by industrial discharge, domestic waste, agricultural runoff, mine drainage, and accidental oil spills indicate that fish could be a serious source of human exposure to potentially hazardous substances like trace metals.2,5,6 Trace metals are persistent and nonbiodegradable, and once they enter aquatic ecosystems, they bioaccumulate in aquatic organisms and their concentrations are biomagnified as they move up the aquatic food chain.1,5,7 Trace metals in fish may be transferred and pose serious health problems to fish consumers. Cadmium, for instance, may cause cardiovascular, neurological, and reproductive disorders.7,8 Short-term exposure to high doses of Cd can cause nausea, vomiting, abdominal disturbances, and fatigue. 8 In addition, it has been classified as a probable human carcinogen by the International Agency for Research on Cancer.9-11 Exposure to Pb has been associated with renal, muscular, and cardiovascular problems. It may also cause cognitive and development deficits in children and reproductive disorders in men and women.7,11-15 Unlike Cd and Pb, Cu is an essential element required for various physiological functions in the human body. 16 Even so, Cu levels above the recommended limit may lead to brain, liver, and kidney disorders. 17 Short-term exposure to high doses of Cu can cause diarrhea, stomach pains, vomiting, and death due to liver and kidney failure or due to the depression of the central nervous system. 17
Because of the aforementioned and many other health effects associated with dietary trace metals, there is growing interest in food safety with regard to the accumulation of these substances in food. Studies around the world show that fish and other seafood are a major source of dietary trace metals.18-23 Majority of these studies reveal that human exposure to trace metals from fish and seafood is within the recommended limits. Conversely, other studies indicate that consumers of fish and seafood may be exposed to hazardous levels of these toxicants and adverse health incidents may occur as a result of this exposure.22,24-28 Vegetables and cereals are the other major dietary sources of trace metals. Vegetables and cereals are valued components of many meals around the world. Thus, the presence of trace metals in these food groups is raising concern world over as is made evident by these studies.29-34
In Kenya, various studies have assessed trace metals content in fish,35-39 beef, 40 honey,41,42 and vegetables43-45 to determine whether they exceeded internationally set standards for these substances in food. Nonetheless, exceedance does not always represent human health risk. 46 Thus, in the present study estimated daily intake (EDI) of Cd, Cu, and Pb via consumption of the commonly consumed fish species in Nakuru town, Kenya, were calculated and compared with the provisional tolerable daily intake (PTDI) established by the Joint FAO/WHO Expert Committee on Food Additives (JECFA).47-49 In addition, target hazard quotient (THQ) and hazard index (HI) were computed to assess human health risks posed by Cd, Cu, and Pb in the commonly consumed fish species in Nakuru town, Kenya.
Materials and Methods
Questionnaire survey
A questionnaire-based survey was conducted to establish the commonly consumed fish species in Nakuru town and the consumption rates of these fish species. Questionnaires were administered to 385 randomly selected participants buying fish in 5 retail outlets in Nakuru town. The questionnaire acquired basic information including gender, age, education, and income level of the respondents. It was used to acquire information on fish preferences of consumers. It was also used to determine the frequency and ingestion rates of the commonly consumed fish species in Nakuru town.
The cross-sectional survey established that
Sample collection and preparation
Four commonly consumed fish varieties (
Trace metals analysis
The composite fish samples were thawed at room temperature, oven dried for 72 hours at 105°C until constant dry weight. They were pulverized in a dry pestle-mortar then stored in sterile containers placed in desiccators awaiting trace metal analysis.
The powdered composite samples were thoroughly mixed to attain optimum homogeneity. They were then digested using Food and Agriculture Organization (FAO) procedures.50,51 Briefly, 1 g of homogenized composite sample was weighed into a digestion vessel. Ten milliliters digestion mixture consisting of 65% nitric acid (HNO3) and 30% peroxide hydrogen (H2O2) was prepared and added to the digestion vessels, kept overnight to allow various reactions to occur. The digestion vessels were heated on a hot plate to 130°C until the solutions turned clear and the volume reduced to between 2 and 3 mL. They were allowed to cool, filtered then diluted with double distilled water to 50 mL in volumetric flasks.
Analyses of Cd, Cu, and Pb were performed using flame atomic absorption spectrophotometer (Thermo Jarrell Ash). The detection limits of Cd, Cu, and Pb were 0.0015, 0.003, and 0.01 mg/L, respectively. To ensure reliability of the results, double distilled water was used throughout the study. Glass vessels were precleaned thoroughly using detergent, soaked overnight in 20% nitric acid (HNO3) then rinsed using double distilled water. The experimental reagents and standards used during the study were of analytical grade obtained from Sigma Aldrich. The accuracy of the instrument was ensured by running samples in triplicates. In addition, a reagent blank and a standard were run after every 3 samples to check instrumental drift. The AAS instrument was calibrated with a series of standard solutions prepared in the concentration range of 0 to 15 mg L−1. A calibration curve of absorbance versus concentration was obtained and used to quantify Cd, Cu, and Pb content in the commonly consumed fish species. Trace metal concentration was expressed in milligram per kilogram dry weight.
Human health risk assessment
The risk to human health as a result of consuming the commonly consumed fish varieties sold in Nakuru town, Kenya, was evaluated by calculating EDI and THQ of trace metals. In addition, literature indicates that exposure to 2 or more trace metals from the same source may result to additive effects. 52 Thus, THQs of Cd, Cu, and Pb were summed to generate hazard indices (HI) that represent overall health risk posed by the 3 trace metals in the commonly consumed fish species.

Geographic locations of the main fish retail markets in Nakuru town (Kenya).
EDI was calculated using equation (1).22,53
where EF is exposure frequency (365 days year−1); ED is the frequency duration (70 years, equivalent to the average human lifespan); FIR is the ingestion rate of fish in kilograms person−1 day−1; C is the concentration of Cd, Cu, or Pb in the commonly consumed fish species in mg kg−1; BW is the average body weight which is equivalent to 60.7 kg for an average African adult 37 ; and TA is the average exposure time for noncarcinogens (365 days year−1 × ED). Computed EDI values were compared with PTDI values of Cd (0.001 mg kg−1 day−1), Cu (0.5 mg kg−1 day−1), and Pb (0.00357 mg kg−1 day−1)47-49 to determine whether the daily recommended values were exceeded or not.
Target hazard quotient a ratio between exposure to a potentially hazardous element and its reference dose 5 was determined using equation (2). 22 Computed THQ values less than 1 indicate that the exposed population is unlikely to experience adverse effects associated with trace metals. On the other hand, computed THQ values greater than 1 suggest possible health risks for the exposed consumers.5,54
where RfDo is the oral reference dose of trace metals in mg kg−1 day−1 based on the safe upper level of element’s oral intake for an adult. The oral RfD for Cd, Cu, and Pb is 0.001, 0.04, and 0.004 mg kg−1 day−1, respectively. 55
THQs of Cd, Cu, and Pb were summed to generate a hazard index using equation (3). 56 Assuming additive effects, HI is a measure of the potential risk of adverse health effects from more than 1 element.5,54 HI greater than 1 suggests likelihood of adverse effects on human health and the necessity for further action.5,54
Statistical analysis
SPSS version 22 was used for statistical data analysis. Results are presented as arithmetic mean and standard deviation (mean ± SD). One-way analysis of variance (ANOVA) was used to assess differences among mean concentrations of Cd, Cu, and Pb in fish. It was used to examine variation in trace metal content among the commonly consumed fish species. It was also used to examine trace metal content in fish based on the markets where the fish were purchased. All statistical test were regarded significant when
Results and Discussion
Trace metal concentrations in the commonly consumed fish species
The mean concentrations of Cd, Cu, and Pb in the commonly consumed fish species, ie,
The mean concentrations of Cd in the commonly consumed fish species ranged from 0.11 ± 0.045 to 1.11 ± 0.931 mg kg−1. Similar Cd concentrations in the range of 0.05 ± 0.02 to 0.95 ± 0.09 mg kg−1 were reported by Oyoo-Okoth et al
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in
Cu content in
Pb concentrations in
Human health risk assessment
To assess human health risk associated with Cd, Cu, and Pb in the commonly consumed fish species, EDI, THQ, and HI were computed. Human health risks were calculated with the assumption that an adult weighing 60.7 kg
37
consumes 0.027, 0.012, 0.035, and 0.023 kg day−1 of
Estimated daily intake of Cd, Cu, and Pb by consuming the commonly consumed fish species in Nakuru town, Kenya.
Provisional tolerable daily intake (mg kg−1 day−1) established by the Joint FAO/WHO Expert Committee on Food Additives (JECFA).47-49 60.7 kg is the average body weight of an African adult. 37 Abbreviation: PTDI, provisional tolerable daily intake.
The highest EDI value of Cd (0.0003 mg kg−1 day−1) was much lower than the PTDI, whose values is 0.001 mg kg−1 day−1 of body weight48,49 corresponding to 0.0607 mg kg−1 for an adult weighing 60.7 kg. Daily Cd intake through consumption of the commonly consumed fish varieties was lower than or similar to the daily intake reported for adult consumers from other developing countries—Ghana: 0.000092 mg kg−1 day−1,58 Nigeria: 0.000063 mg kg−1 day−1,54 Ethiopia: 0.000029 mg kg−1 day−1,59 D. R. Congo: 0.00012 to 0.00094 mg kg−1 day−1,60 Egypt: 0.000048 to 0.00022 mg kg−1 day−1,61 Iran: 0.00012 mg kg−1 day−1,62 and Bangladesh: 0.00027 to 0.00246 mg kg−1 day−1. 63 Daily Cd intake was higher than that reported for adults in Yaoundé, Cameroon, 0.000017 mg kg−1 day−1,64 and roughly 26 times higher than that reported for adults from Dares salaam, Tanzania. 65
With respect to Cu, the highest daily intake (0.00099 mg kg−1 day−1) corresponded to consumers of
As for Pb, the greatest daily intake (0. 00438 mg kg−1) was also observed in consumers of
Target hazard quotient and hazard index
Noncarcinogenic THQs and hazard index of the 3 trace metals through consumption of the commonly consumed fish species sold in Nakuru Town, Kenya, are presented in Table 3. The trend of THQ values for consumers of the commonly consumed fish species decreased in the order of Pb > Cd > Cu. The THQ values of Cd, Cu, and Pb varied from 0.022 to 0.331, 0.005 to 0.025, and 0.230 to 1.168, respectively. Target hazard quotient is a ratio between potential exposure to a given trace metal and its oral reference dose. It is used to assess potential health risk associated with long-term exposure to dietary trace metals.5,54 If the computed ratio is greater than 1, the exposed population is likely to develop adverse health effects.55,56,66 According to the New York State Department of Health (NYSDOH), if the computed THQ value is greater than 1 but less than 5, the risk is low; if it is greater than 5 but less than 10, the risk is moderate; however, if the THQ value is greater than 10, the risk is high.
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Computed THQ values of Cd and Cu were all below 1, suggesting that health effects associated with Cd and Cu are unlikely to occur. In contrast, THQ values of Pb for consumers of
Target hazard quotient and hazard index of consumers of commonly consumed fish species from 5 retail markets in Nakuru town, Kenya.
Abbreviations: HI, hazard index; THQ, target hazard quotient.
Hazard index is the numerical sum of the computed THQ values.5,54 Like THQ, HI values should not exceed 1 otherwise they would present significant health risks to consumers.5,54 HI values of Cd, Cu, and Pb in this study ranged from 0.286 to 1.488. THQ values of Pb were significantly higher than those of Cd and Cu and contributed between 66% and 92% to the HI values. Consumers of
Conclusion
Estimated daily intake, THQ, and hazard index were used to assess consumers’ exposure to Cd, Cu, and Pb via consumption of the commonly consumed fish species (
Footnotes
Funding:
The author(s) disclosed receipt of the following financial support for the research, authorship, and/or publication of this article:
Declaration of conflicting interest:
The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.
Author Contributions
FE, WNM, and MM conceived, designed and planned this research. WNM and MM supervised the project. FE and TW carried out the experiments. FE, WNM, MM, and TW contributed to the analysis and the interpretation of the results. FE took the lead in writing the manuscript with significant and critical contributions from WNM, MM, and TM.
