Qualitative and Quantitative Analyses of Antibiotics, Heavy Metals, Mycotoxins, and Pesticides Residues in Milk, Eggs and Meat Meant for Human Consumption by Dr. Muhammad Younus, College of Veterinary and Animal Sciences, Jhang

College of Veterinary and Animal Sciences , Jhang (UVAS, Lahore) 1

INTERIM REPORT
Qualitative and Quantitative Analyses of
Antibiotics, Heavy Metals, Mycotoxins, and
Pesticides Residues in Milk, Eggs and Meat
Meant for Human Consumption

Dr. Muhammad Younus-Principal Investigator
Dr. Tariq Abbas -Co-Principal Investigator

3/18/2013 12:49
PM

Outline
2

 Introduction/ Background
 Health effects of chemical contaminants in milk
 Problem statement
 Review of literature at a glance
 Methodology AND hypotheses
 Progress to date and future plans

College of Veterinary and Animal Sciences , Jhang
3/18/2013 12:49 PM
(UVAS, Lahore)

Introduction/Background
3

 Pakistan is the fourth largest milk producing country
located in a milk deficient region.
 It has rapidly increasing demand and competition in
international market.
 Milk consumers in Pakistan are often faced with low-
quality, adulterated milk.
 Lack of hygiene, adulteration with various agents, and
absence of cold chains are primary contributors to this
low quality.

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(UVAS, Lahore)

4

Residues in milk may come
through : i) feed and water ,
ii) drugs , iii) post harvest
contamination & adulteration

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(UVAS, Lahore)

Introduction/Background
5

 Malicious chemicals in various farm inputs
(e.g. concentrates), agriculture, drinking
water, environmental pollution, and use of
veterinary drugs lead to excretion of residues
in milk.
 As such there are no data on the content of
these chemicals in milk and a system for their
effective surveillance.

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(UVAS, Lahore)

Health effects of residues in milk-
Antibiotics
6

 Antibiotics are widely used in dairy sector, both for
treatment and prophylactic purposes.
 Antibiotics residues in milk can be potential hazard
and may cause serious illness to people, such as
hypersensitivity and allergic reactions.
 They may also increase the resistance to pathogenic
bacteria
 In addition, antibiotic residues in milk cause
inhibition of bacterial starter cultures used in dairy
production (losses !) .

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(UVAS, Lahore)

Drugs prohibited for extra-label use in
animals being prudently used without
7 observing codal formalities

College of Veterinary and Animal Sciences , Jhang (UVAS,
3/18/2013 12:49 PM
Lahore)

Pesticides
8

 Multiple studies indicate a wide range of pesticide-
related clinical and subclinical effects, including
significant positive associations between pesticide
exposure and solid tumors, hematological
cancers, and genotoxic effects.
 In addition, pesticides were found to impact on
mental and emotional functioning, the nervous
system – causing Parkinson’s disease and other
neurological diseases, and the reproductive system
diseases – causing birth defects, fertility, fetal
death, and intrauterine growth retardation.
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(UVAS, Lahore)

Mycotoxins
9

 Aflatoxin B1 (AFB1) produced by moulds in the
contaminated cattle feed get transformed to 4-
hydroxylated metabolite known as “milk toxin”, i.e.,
aflatoxin M1 (AFM1) in mammals (B1 to M1 conversion
in stomach of cow).
 The fungal toxin is resistant to high temperature
treatments including pasteurization and sterilization.
 The presence of AFM (1) in milk, pose a major risk for
humans especially kids as it can have
immunosuppressive, mutagenic, teratogenic and
carcinogenic effects

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(UVAS, Lahore)

Mouldy feed- a source of
mycotoxins for dairy animals and
10 their milk

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(UVAS, Lahore)

Heavy metals
11

 A variety of exposure routes allow toxic heavy metals
predominantly lead and cadmium to enter the food
chain of dairy farm animals, the commonest being
come through :
 The contaminated animal feed and water,
 Atmospheric deposition ,
 Land application of inorganic
fertilizers, biosolids, agrochemicals,
 Urban , industrial effluents and animal manures
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(UVAS, Lahore)

Urban and industrial wastes from Jhang
city being used in Agriculture and falling
12 into river

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Lahore)

Problem statement
13

 “There are few studies on contamination
level of various chemical residues in
unprocessed, non-branded liquid milk
available at conventional milk shops in
various cities of the country such as
Jhang”.
 The objective of this project is to quantify the
residues of selected mycotoxin, heavy
metals, pesticides and antibiotics in milk being
sold on the street shops in Jhang city.
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(UVAS, Lahore)

Unprocessed , unbranded liquid milk
being sold on traditional street shops in
14 Jhang city

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(UVAS, Lahore)

Review of exiting studies
15

 Given the importance of chemical residues in milk and
dairy products, different countries have conducted
studies related to antibiotics, pesticides, Aflatoxin M1
and heavy metals. The review of such studies is given
as a separate chapter in the report.
 “In short ; results of the studies
provided a base-line information for
veterinary authorities, drug
administration and quality control,
other concerned organizations and
professionals to take measures in
controlling and preventing of
occurrence of drug residues”. 12:49 PM
3/18/2013
(UVAS, Lahore)

16
Endnote library !

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(UVAS, Lahore)

Methodology :
Description of the study area
17

 Jhang (Urdu: , Punjabi: ), is the capital city
of District Jhang which is located at the gateway of
central Punjab - the milk belt of Pakistan. It is situated
at place where two rivers Jhelum and Chenab join each
other (Head Trimu)
 It is a neglected area, rich in livestock and focus for
improvement in dairies particularly Sahiwal breed of
cattle.
 The city has been purposively selected because its
dairy production and marketing is representative of the
whole country.
 In year 2006, College of Veterinary and Animal
Sciences, Jhang was established. The3/18/2013 12:49 PM
(UVAS, Lahore) college intends to

Methodology : Selection of the chemical
contaminants
18

 After review of literature, field visits and informal
meetings with the dairy experts , a list of chemical
contaminants was prepared for inclusion in the
survey.
 The chemicals were selected on the basis of
following two criteria:
1. Possibility level in milk through farm inputs (e.g.
concentrates etc) , agriculture, drinking
water, medication, environmental contamination
and/adulteration, and
2. Public health effects Sciences , Jhang 3/18/2013 12:49 PM
College of Veterinary and Animal
(UVAS, Lahore)

Methodology :
Selection of the chemical contaminants
19 Category Chemical Name
Heavy metal Lead
Chromium
Copper
Cadmium
Mycotoxins Aflatoxin M1
Antibiotics Enrofloxacin
Penicillin G
Streptomycin
Gentamycin
Oxytetracycline

Chloramphenicol
Pesticides Acephate
Chlorpyrifos
Cypermethrin
Lambda Cyhalothrin
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(UVAS, Lahore)

Hypotheses maintained and to
be tested
20

1. The mean concentration of residues of any
selected chemical contaminant in unprocessed, non-
branded liquid milk available at conventional retail
shops of Jhang city is more than maximum residue
level(MRL)

2. For any selected chemical contaminant, there is
an association between proportion of positive
samples and seasons [positive sample means
samples with concentration of more than MRL ]

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Lahore)

Methodology :
Sampling strategy
21

 From the street-level map of the Jhang city,
100 streets were randomly selected and geo -
referenced in Quantum GIS version 1.8.0
Lisboa(Roads>Streets>shops)
 In each street, every second shop (right/left)
was selected through systematic random walk
from entrance to the street .

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Lahore)

100 streets randomly selected followed by
systematic random walk sampling
22

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Lahore)

Methodology:
23
Sampling
 From each selected shop, 250 ml of unprocessed
liquid milk was purchased, out of which 25 ml was
sampled (in duplicate) in clean sterilized screw
capped glass tubes.
 The sampling was done from November to
December, 2012.
 The sampling will be repeated during mid summer
and moonson of 2013
 The sample size was decided from already
published journal articles on same topic and
available resources.
 The sample size will be re-assessed later based
on initial findings
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Methodology :
24
Data analyses
 The data shall be analyzed in SPSS version
21. Beside descriptive analyses, one sample
t-test will be used for hypothesis testing with
maximum residue level (MRL) as test value.
 The proportion of samples with more than
MRLs will be compared across seasons using
Chi square test.

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Lahore)

Progress to date
25

 Review of literature : completed (Endnote
Library given in methodology)
 Selection of chemical of contaminants :
Decided (Table given in methodology)
 Devising a strategy to sample shops through a
probability sampling : Multi stage cluster
sampling through GIS (Google Earth file &
shapefile created)
 First round of sampling : completed and being
analyzed and Animal Sciences , Jhang (UVAS,
College of Veterinary
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Lahore)

Future plans
26

 The sampling will be repeated during summer
and moonson to assess seasonal trends if any.
 The same samples shall further be analyzed to
quantify antibiotics, and pesticides mentioned
in the table.
 The results will be discussed with dairy
association to plan recommendations to
control the problem using measures feasible in
local settings.
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Lahore)

27

Thanks

3/18/2013 12:49 PM
(UVAS, Lahore)

Qualitative and Quantitative Analyses of Antibiotics, Heavy Metals, Mycotoxins, and Pesticides Residues in Milk, Eggs and Meat Meant for Human Consumption by Dr. Muhammad Younus, College of Veterinary and Animal Sciences, Jhang

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Qualitative and Quantitative Analyses of Antibiotics, Heavy Metals, Mycotoxins, and Pesticides Residues in Milk, Eggs and Meat Meant for Human Consumption by Dr. Muhammad Younus, College of Veterinary and Animal Sciences, Jhang