College of Veterinary and Animal Sciences , Jhang (UVAS, Lahore)   1    INTERIM REPORT         Qualitative and Quantitativ...
Outline2       Introduction/ Background       Health effects of chemical contaminants in milk       Problem statement  ...
Introduction/Background3       Pakistan is the fourth largest milk producing country        located in a milk deficient r...
4                                     Residues in milk may come                                     through : i) feed and ...
Introduction/Background5       Malicious chemicals in various farm inputs        (e.g. concentrates), agriculture, drinki...
Health effects of residues in milk-    Antibiotics6       Antibiotics are widely used in dairy sector, both for        tr...
Drugs prohibited for extra-label use in    animals being prudently used without7   observing codal formalities         Col...
Health effects of residues in milk-    Pesticides8       Multiple studies indicate a wide range of pesticide-        rela...
Health effects of residues in milk-    Mycotoxins9       Aflatoxin B1 (AFB1) produced by moulds in the        contaminate...
Mouldy feed- a source of     mycotoxins for dairy animals and10   their milk          College of Veterinary and Animal Sci...
Health effects of residues in milk-     Heavy metals11        A variety of exposure routes allow toxic heavy metals      ...
Urban and industrial wastes from Jhang     city being used in Agriculture and falling12   into river        College of Vet...
Problem statement13        “There are few studies on contamination         level of various chemical residues in         ...
Unprocessed , unbranded liquid milk     being sold on traditional street shops in14   Jhang city           College of Vete...
Review of exiting studies15        Given the importance of chemical residues in milk and         dairy products, differen...
16      Endnote library !     College of Veterinary and Animal Sciences , Jhang                                           ...
Methodology :     Description of the study area17        Jhang (Urdu:                 , Punjabi:               ), is the ...
Methodology : Selection of the chemical     contaminants18         After review of literature, field visits and informal ...
Methodology :      Selection of the chemical contaminants19   Category                                 Chemical Name     H...
Hypotheses maintained and to     be tested20     1. The mean concentration of residues of any     selected chemical contam...
Methodology :     Sampling strategy21        From the street-level map of the Jhang city,         100 streets were random...
100 streets randomly selected followed by     systematic random walk sampling22        College of Veterinary and Animal Sc...
Methodology:23     Sampling        From each selected shop, 250 ml of unprocessed         liquid milk was purchased, out ...
Methodology :24     Data analyses        The data shall be analyzed in SPSS version         21. Beside descriptive analys...
Progress to date25        Review of literature : completed (Endnote         Library given in methodology)        Selecti...
Future plans26        The sampling will be repeated during summer         and moonson to assess seasonal trends if any.  ...
27                           Thanks     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

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Presented on February 10th, 2013 at the Second Research Competitive Grants Conference in Islamabad, Pakistan.

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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

  1. 1. 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 Investigator3/18/2013 12:49 PM
  2. 2. Outline2  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)
  3. 3. Introduction/Background3  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. College of Veterinary and Animal Sciences , Jhang 3/18/2013 12:49 PM (UVAS, Lahore)
  4. 4. 4 Residues in milk may come through : i) feed and water , ii) drugs , iii) post harvest contamination & adulteration College of Veterinary and Animal Sciences , Jhang 3/18/2013 12:49 PM (UVAS, Lahore)
  5. 5. Introduction/Background5  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. College of Veterinary and Animal Sciences , Jhang 3/18/2013 12:49 PM (UVAS, Lahore)
  6. 6. Health effects of residues in milk- Antibiotics6  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 !) . College of Veterinary and Animal Sciences , Jhang 3/18/2013 12:49 PM (UVAS, Lahore)
  7. 7. Drugs prohibited for extra-label use in animals being prudently used without7 observing codal formalities College of Veterinary and Animal Sciences , Jhang (UVAS, 3/18/2013 12:49 PM Lahore)
  8. 8. Health effects of residues in milk- Pesticides8  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. College of Veterinary and Animal Sciences , Jhang 3/18/2013 12:49 PM (UVAS, Lahore)
  9. 9. Health effects of residues in milk- Mycotoxins9  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 College of Veterinary and Animal Sciences , Jhang 3/18/2013 12:49 PM (UVAS, Lahore)
  10. 10. Mouldy feed- a source of mycotoxins for dairy animals and10 their milk College of Veterinary and Animal Sciences , Jhang 3/18/2013 12:49 PM (UVAS, Lahore)
  11. 11. Health effects of residues in milk- Heavy metals11  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 College of Veterinary and Animal Sciences , Jhang 3/18/2013 12:49 PM (UVAS, Lahore)
  12. 12. Urban and industrial wastes from Jhang city being used in Agriculture and falling12 into river College of Veterinary and Animal Sciences , Jhang (UVAS, 3/18/2013 12:49 PM Lahore)
  13. 13. Problem statement13  “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. College of Veterinary and Animal Sciences , Jhang 3/18/2013 12:49 PM (UVAS, Lahore)
  14. 14. Unprocessed , unbranded liquid milk being sold on traditional street shops in14 Jhang city College of Veterinary and Animal Sciences , Jhang 3/18/2013 12:49 PM (UVAS, Lahore)
  15. 15. Review of exiting studies15  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 College of Veterinary and Animal Sciences , Jhang 3/18/2013 (UVAS, Lahore)
  16. 16. 16 Endnote library ! College of Veterinary and Animal Sciences , Jhang 3/18/2013 12:49 PM (UVAS, Lahore)
  17. 17. Methodology : Description of the study area17  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 College of Veterinary and Animal Sciences , Jhang Sciences, Jhang was established. The3/18/2013 12:49 PM (UVAS, Lahore) college intends to
  18. 18. Methodology : Selection of the chemical contaminants18  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)
  19. 19. Methodology : Selection of the chemical contaminants19 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 College of Veterinary and Animal Sciences , Jhang 3/18/2013 12:49 PM (UVAS, Lahore)
  20. 20. Hypotheses maintained and to be tested20 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 ] College of Veterinary and Animal Sciences , Jhang (UVAS, 3/18/2013 12:49 PM Lahore)
  21. 21. Methodology : Sampling strategy21  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 . College of Veterinary and Animal Sciences , Jhang (UVAS, 3/18/2013 12:49 PM Lahore)
  22. 22. 100 streets randomly selected followed by systematic random walk sampling22 College of Veterinary and Animal Sciences , Jhang (UVAS, 3/18/2013 12:49 PM Lahore)
  23. 23. 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 College of Veterinary and Animal Sciences , Jhang (UVAS, 3/18/2013 12:49 PM Lahore)
  24. 24. 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. College of Veterinary and Animal Sciences , Jhang (UVAS, 3/18/2013 12:49 PM Lahore)
  25. 25. Progress to date25  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 3/18/2013 12:49 PM Lahore)
  26. 26. Future plans26  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. College of Veterinary and Animal Sciences , Jhang (UVAS, 3/18/2013 12:49 PM Lahore)
  27. 27. 27 Thanks College of Veterinary and Animal Sciences , Jhang 3/18/2013 12:49 PM (UVAS, Lahore)

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