Analysis of Pesticides Residues in Breast Milk of primiparous and multiparous women in Gilgit

Syyeda Urooj1, Maisoor Ahmed Nafees1, Saifud Din1,2, Shaukat Ali2

1Department of Biological Sciences, Karakoram International University Gilgit-Baltistan, Pakistan.

2Department of Environmental Sciences, Karakoram International University Gilgit-Baltistan, Pakistan.

Crossponding author email: gltsaifuddin@gmail.com

Citation |Urooj. S, Nafees. M. A, “Din. S and Ali. S, “Analysis of Pesticides Residue in Breast Milk primiparous and multiparous women in Gilgit”. International Journal of Innovations in Science and Technology, Vol 3, Isue 4, pp: 166-173, 2021                                                    

Received | Nov 19, 2021; Revised | Dec 15, 2021 Accepted | Dec 21, 2021; Published|Dec 27, 2021

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Abstract.                                                                                                         

Milk contains all the essential nutrients like fats, proteins, and minerals. The utilization of contaminated food can induce a proportion of pesticides in the body. The main purpose of the study was to determine the pesticide residues and current status of breast milk in primiparous and multiparous mothers. In a current study, a total of 50 samples were collected from different areas of District Gilgit and Astore.  The pesticides cypermethrin, deltamethrin, and chlorpyrifos were analyzed using gas chromatography. The presence of cypermethrin in 10 samples was in a range 0.00 – 0.012 mg/kg, while the detection of Deltamethrin in 07 with variation from 0.000.12mg/kg. Whereas chlorpyrifos was found in 05 samples with the ranges of 0.00-0.0062 mg/kg. Residue level was quite higher in urban areas than rural areas. The multiparous women had prominent residues level than primiparas and the concentration of Deltamethrin was higher than other pesticides. All the pesticides residues levels in the breast milk of primiparous and multiparous mothers were within the permissible limits of WHO. Yet the women of these areas are not vulnerable but prolong exposure may pose a serious threat to neonatal and maternal health and other relevant reproductive issues. To manage the risk of milk contamination in the future, the demand for public awareness campaigns and the adoption of alternative clean approaches to control pests and other disease-spreading vectors in the best interests of public health seems reasonable. 

Keywords:  Milk, Gas Chromatography, Pesticides, Primiparous. Multiparous

  1. INTRODUCTION

Pesticides are used to protect crops, control the damage, and maximize the crop production. Different pesticide groups were in use including organochlorine, organophosphorus pesticides, pyrethroids, and carbamate, pesticides which contains toxic compounds that are banned [1]. Breast milk is the only source of diet for growing children which contains all the essential nutrients required for the development of newborns such as proteins, fats, and carbohydrates. Unluckily nowadays breast milk is not pure it is contaminated with toxic substances [2, 3].

Worldwide pesticides are extensively used for enhancing food production, controlling pests, and insects, and destroying the vectors of human and animal diseases like malaria, dengue, encephalitis [4]. Consumption of toxic chemicals occurs in the field, diet, or through inhalation, and dermal contact may cause instability in daily life. OCPs are lipophilic and are more persistent in breast milk [5, 6].

In Pakistan farmers use different types of pesticides for the betterment of crops, it includes 39 types of herbicides, 108 insecticides, and 30 fungicides.  Farmer’s usually use organochlorine, organophosphorus, and alternates of urea. Residues are extensively found in different areas of Pakistan due to their unrelenting nature. When mothers consume contaminated food, level of residue rises in the body [7, 8, 9]. According to an economic survey 2012-13 of Pakistan locally 30,000 tons of pesticides were manufactured and 12665 tons were imported. In Pakistan mostly pesticides are used in fields which are an alarming situation for the presence of residues in food [10].

Human milk is thought to be the natural superior food for infants to meet their nutritional needs as they grow. Human milk, on the other hand, is an ideal matrix for the accumulation of pollutants. Chlorinated pesticides were common chemicals in the previous century's environment. They were widely used to control agricultural and public health insects all over the world, and they are still used in some countries to control the malarial insect vector. When newborns and infants are exposed to these pollutants through feeding, they may suffer from lower birth weight [11], neuro-developmental delay [12].

Pregnant women are primarily exposed to these compounds through their diet [13], particularly through fish, meat, and milk [14, 15]. Despite a decades-long global ban on the use of OCPs, residues have been discovered in breast milk all over the world [16, 17, 18, 19].

Human breast milk is an ideal marker for OCPs because it provides information on the toxic effects of these substances on mothers and newborns. Pesticide accumulation in breast milk can be influenced by a variety of factors, including diet, place of residence, smoking, maternal age and weight, and previous lactation duration; however, the literature provides conflicting information [20].

  1. Material and Methods.

Study Area. 

Samples have been collected from five areas of district Gilgit (Danyore, Chilmis, Nomal, Gilgit City, and Oshikhandas) and five areas of district Astore (Eidgah, Nowgam, Fina, Patipora, and Pakora).

Sample collections. 

50 Samples were collected from volunteer mothers primiparas and multiparas between the ages of 20-40, before they are enrolled in the study all women signed authorization. Around 40 mL milk was collected in 100mL sterilized glass bottles with identification codes and placed in an icebox during the collection period and stored in a freezer at -20 ⁰C till laboratory analysis.

Samples have been collected during a period of two months from November to October 2019 from mothers who were either nativ or lived there from last five years. A questionnaire has been designed to access the socio-demographic data, food habits during and after pregnancy, age, and residence.

The selective criteria in the present study were followed in previous similar studies. A self-administered questionnaire was also employed to acquire information about the subject’s age, parity, body mass index and  eating habits. Questions regarding demographic characteristics and feeding habits were modified from Environmental Protection Agency EPA [21].

Laborartory Analysis.

1mL of milk sample and 15mL of acetonitrile containing 0.01% acetic acid were mixed in a polytetrafluoroethylene  centrifuge tube. Samples were shaken vigorously for 02 min and then sonicated for 10 minutes. 6g of anhydrous MgSO4, 1g of NaCl, and 1g of sodium acetate trihydric were added to the sample. Again shaken the sample for 02 min and centrifuged for 05 min at 400rpm. The supernatant of 8mL was transferred into another PTFE tube and added 1g of MgSO4 and Florisil. Again centrifuged the samples and transferred the upper layer into a round bottom flask and left to evaporate until complete dehydration using a rotavapor. Then reconstituted in 1mL of n-hexane containing 10-15% acetone and finally analyzed the sample through GC.

Statistical Analysis.Obtained data were précised as mean and ANOVA and LSD were applied to check to mean the difference among samples by using SPSS 21 and Statitix 8.1 Capital letter represents LSD and Significance difference among means in column-wise.

RESULTS AND DISCUSSIONS. 

Table 1.Mean value of pesticides residue area wise in mg/kg

 

Locations

Cypermethrin

Deltamethrin

Chlorpyrifos

Astore

Pakora

0.012A

NDA

0.005A

Nowgam

0.001B

0.01B

NDB

Patipora

NDC

NDAC

NDBC

Fina

NDCD

NDACD

0.002D

Eidgah

0.011AE

0.002E

NDBCE

Gilgit

Oshikhandas

NDCDF

NDACDF

NDBCEF

Chilmis Das

NDCDFG

NDACDFG

NDBCEFG

Nomal

0.0012H

0.015H

NDBCEFGH

Danyore

0.00574I

0.00528I

0.0062I

Gilgit City

0.0005J

0.12J

NDBCEFGJ

Note: ND: Not Detected

Figure1.Mean concentration of pesticide residues in District Astore and Gilgit

Figure 2.Percentage of primiparous and multiparous mothers

In the current study, three pesticides were under observation named cypermethrin, deltamethrin and dhlorpyrifos from breast milk, 54% of mothers were multiparous and 46% were primiparous among selected mothers for sampling shown in Figure 2.  Out of 50 samples, 13were detected with pesticide residues. The residues of cypermethrin were found in 10 samples maximum value was found in Danyore 0.014mg/kg and the minimum value was obtained in Nowgam 0.001mg/kg. Deltamethrin was found in 09 samples maximum value of 0.12mg/kg was found in Gilgit and lowest 0.001mg/kg in Eidgah. Chlorpyrifos was found in 05 samples with highest value in Danyore 0.019mg/kg and lowest in Fina 0.002mg/kg. The result shows that all the samples have residues of pesticide among them cypermethrin was the major contaminant in milk samples and mean values are shown in Table 1 and Figure 1. Residue level in all samples was lower than the Maximum residue limits (MRLs) set by FAO/WHO.

In the current study, cypermethrin residues were found in 7 samples of district Gilgit and highest residual level was obtained in the samples of Danyore and the value was 0.014mg/kg and the lowest was found in Nomal and Gilgit with the residual level of 0.0012 mg/kg and 3 samples from Astore have the residues of cypermethrin maximum value was present in the sample of Pakora 0.012mg/kg and minimum value in Nowgam 0.001mg/kg. To analyze the presence of pyrethroids in human milk a study was conducted in Columbia. The residues of cypermethrin, permethrin, and fenvalerate were present in samples. Pyrethroids were useful to control dengue and the level reach28ngg-1/W. Evaluation of daily intake in nursing infants was calculated and compared to an adequate WHO level [22].  A similar study was conducted in urban and agricultural areas in Mexico on lactating women. The concentration of pp-DDT and cypermethrin in breast milk was higher in those women who are living in urban areas than in agricultural areas (p < 0.05and p = 0,001). Since pesticide levels do not exceed the acceptable daily intake (ADI) according to EPA and FAO/WHO [23]. The study investigates the residues of chlorpyrifos and other pesticides in cow’s milk from Punjab India with CPS, DDT, and HCH as the major contaminants. Also the presence of cypermethrin, cyhalothrin, fenvalerate, deltamethrin, malathion, profenofs, andehion was reported. 12 samples exceed the MRLs for lindane, 18 for DDT, 1 sample for chlorpyrifos, profenophos and cypermethrin [24].

The current study shows that the residue level of deltamethrin was presented in 09 samples among which 6 samples were collected from different areas of district Gilgit and the maximum value 0.12mg/kg was found in Gilgit-city and the minimum value detected in Danyore was 0.0011mg/kg and 3 samples were collected from district Astore with maximal value is obtained in Nowgam and Eidgah 0.01mg/kg and minimal value in Eidgah 0.01mg/kg.  A similar study was conducted for the determination of nine pesticides. The residues of deltamethrin, trifluralin, cypermethrin, endosulfan, HGH, p’p’DDE and p’p-DDT were found in breast milk. Still, mothers were subjected to feed infants because breast milk is considered as nutritive food during infancy [25]. A study was conducted in Punjab to monitor the residue level of various pesticides (DDT, DDE, cypermethrin, deltamethrin, permethrin, aldrin, and bifenthrin) 70% of samples were contaminated. DDE, DDT and endosulfan were present in minor proportion. Bifenthin residues are higher among pyrethroids with a mean concentration of 1.68µg/ml, cypermethrin 0.23µg/ml and deltamethrin 0.21µg/mL. most of the samples were polluted with cypermethrin, deltamethrin, permethrin and bifenthrin [26]. In the present study chlorpyrios residue was found in 3 samples of district Astore the highest residue level was 0.01mg/kg found in Eidgah and the lowest is 0.002mg/kg in Fina.  Only 2 samples from Gilgit were detected with the value of 0.019 mg/kg and 0.012mg/kg in Danyore.  A study was conducted to investigate OCPs, OPs, pyrethroids, and carbamate residues in human and cow milk. Hexachlorobenzene, p, p’-dicofol, and chlorpyrifos were found in all samples [27]. Shangi et al., [28] studied the presence of chlorpyrifos, malathion and methyl-parathion was monitored in human milk from Bhopal, Madhya Pradesh. The concentration of endosulfan was highest and the concentrations of chlorpyrifos and malathion were 3.5, 1.5, and 8.4 fold, respectively.

According to the study  thas was conducted to investigate the relation between pesticides contamination of feedstuffs and residues in bovine milk, chlorpyrifos is the main contaminant with a residue level of 6.01in feedstuff and 2.58ng/g in milk samples. Other pesticide residues observed in feed and milk samples endosulphan sulphate, cypermethrin, DDE, lindane, Malathion and fenvalerate. The main source of pesticide residue in milk is the feedstuff on that animal's feed [29]. The milk samples were analyzed for pesticide residues of chlorpyrios, endosulfan, profenofs, and bifenthrin. The residues in raw and ultra-heated milk were determined between the range of 0.1-30µg/g. Residues in heat-treated samples were within the range of 0.1-30µg/L. All  ultra high temperature (UHT) processed samples contain pesticide residue within the acceptable limit set by the WHO on the other hand raw milk samples, chlorpyrios and endosulfan were found above the MRL, raw milk samples show a higher prevalence than heat-treated samples [30].

Conclusion.

            It was concluded that the pesticide residues were found magnificently in the mother milk of Danyore, Gilgit and Eidgah, Astore. Mostly detected pesticides in mothers milk were using fruits and vegetables exported from urban areas of Pakistan, and use of pesticides and synthetic fertilizers to their fields extensively. The multiparous women had prominent pesticide residues than primiparas. Among the analyzed pesticides the residues of cypermethrin, deltamethrin and chlorpyrifos were present in some of the samples. The lactating mothers were mainly consuming vegetables and fruits imported from urban areas during the gestation period and after delivery. Although the residue level was low in all samples if preventive measures are not taken by people and the government it would be a threat to human health in future.

Author’s Contribution. This study was designed and directed by S. urooj and M.A.NAfees as principal investigators. Saif-Ud-Din provided all the technical and experimental Guidance. S.Ali and Saif-Ud-Din performed and participated in Statistical analysis and preparation of the manuscript draft.

Conflict of interest. No conflict of interest is associated with this publication among Authors.

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