The cow – milk feeding construct of babies, in the larger parts of Lake Victoria basin of Kenya, for about the first six months after birth, has vastly invigorated the involvement in look intoing the presence and degrees of some heavy metals in cow ‘s milk. Toxicity of a heavy metal depends on its fractional bioavailability and concentration in the environment, therefore its speciation is of great importance. The cognition of concentrations of toxic heavy metals like Cd, Cr, Cu, Fe, lead and Zn in cow ‘s milk is hence really necessary.
Fresh – milk samples from breastfeeding cattles will be obtained by ego milking into sterilized polyethene bottles and labeled harmonizing to clip, day of the month, location and replicate. Other parametric quantities that will be collected from the three locations on the footing of 7-day interval and thenceforth investigated include: grass provenders, deposits, H2O, dirt, and breastfeeding cow ‘s fecal beads and piss. This information therefore gives a suited background for measuring and finding the concentrations of heavy metal contaminations on cattles and their subsequent consumption by homo.
Few beads of 0.1 M trichloroacetic acid will be added to the sample for curdling and the aqueous bed heated at 500 0C for one hr. Digestion will be done with 0.5 M azotic acid as presence and concentration of heavy metals analyzed utilizing an Atomic Absorption Spectrophotometer, AAS.
Statistical analysis will be conducted utilizing MSTATC two factor complete randomized block design, with the heavy metal concentration as the chief factor with the locations as the bomber intervention. The bundle will execute analysis of discrepancy ( ANOVA ) at P a‰¤ 0.05 with two factor experiment and pupils T-test at P a‰¤ 0.05. The mean, standard divergence, scope and additive correlativity co-efficient on the measured parametric quantities will be determined. The survey is expected to give an indicant of the exposure of female parents and babies in the part to the heavy metals and besides ascertain the safety of absolute milk eating of babies.
This survey will be conducted in Kisumu metropolis, at the shore of Lake Victoria, Kenya.
Cardinal words: Heavy metals, babies, cow ‘s milk and taint.
Human and animate beings have been exposed to heavy metal toxicity for an unmeasurable clip. The industries have dramatically increased the overall environmental burden of the toxins to degrees that they are present in every country of modern consumerism. Therefore, it is necessary to cognize the environmental destiny of all xenobiotics so as to foretell their continuity and possible effects on non-target beings ( Keng’ara F.O: 2004 ) .
Anthropogenetic activities appear to play an of import function in this survey since, in the past, solid wastes have been dumped by occupants in their several vicinities. In an evident effort to maintain the environment clean, in Kisumu metropolis, the local governments, in the past four old ages have collected and dumped the solid wastes at a new site, Nyalenda – Kachok, Kisumu. Similarly lorry-loads from all over the metropolis ‘s supermarkets, industrial set-ups, gasoline Stationss, abodes and markets dump theirs solid wastes at the site.
Cadmium, Cr, Cu, Fe, lead and Zns are among the most common heavy metals known as contaminations in the environment and hence come insult as risky substances to both human and carnal wellness ( Roberts J.R, 1999 ) . This is due to broad spread environmental pollution by stuffs incorporating them: like batteries, pigments, pipes, soldering rods, pesticides, antifungals, gasolene, engine oils, chemical fertilisers or when they occur in high sums in air, dirt, H2O, workss and other compounded carnal provenders.
They therefore increase concentrations of heavy metals in air, H2O, dirt and later taken by workss and animate beings into their nutrient concatenation ( Ahmad, W.M.S, 2002 ) . The presence of heavy metals in cow ‘s milk may be attributed to taint of the original one, which may be due to exposure of breastfeeding cow to environmental pollution or ingestion of contaminated eating materials and H2O ( Carl M, 1991 ) .
This happening can take to considerable concentrations in human organic structure since they are non metabolized hence poses a serious hazard to human wellness when consumed even in little sums ( Selinger B, 1979 ) .
Most of them, like Cd, lead and quicksilver persist in the organic structure and exercise their toxic consequence by uniting with one or more reactive groups indispensable for normal physiological maps of the cells therefore doing cellular perturbations or clinical manifestation. The inauspicious toxic effects caused by lead, cadmium quicksilver are widely recognized ( Friberg, L. and Elinder, C.G, 1988 ) . The major clinical marks in animate beings and adult male for lead and Cu toxic condition include, among others, divergences of the haematological parametric quantities due to their direct effects on haematopoiesis, reduced unity of ruddy blood cells ‘ membrane taking to intravascular hemolysis, anaemia and desiccation ( Radostits O. M. et al. , 1994 ) . Therefore haematological parametric quantities have diagnostic value in animate beings suspected of heavy metal toxicity ( Mlay P.S and Migumia Y.O, 2008 ) . Man becomes at hazard by eating nutrient and imbibing fluids contaminated with heavy metals, A through air, direct contact with the metals like in people working in auto wash or organic structure spraying industries or mills covering with heavy metals and their derived functions ( Farr G, 2001 ) .
Kisumu metropolis is endowed with comparatively many but little endeavors covering with metal plants, auto care and fix ( Jua Kali Sheds ) , building works that pose a hazard of taint to the environment with risky substances including heavy metals. Subsistence agriculture and farming are rather enhanced in its vicinities.
The foregone information prompts the desire to look into the presence of some heavy metals in cow ‘s milk. The information generated will help the concerned metropolis contrivers, establishments and organic structures charged with environmental control and surveillance to explicate steps and policies that would firmly regulate the dumping of solid wastes, re-locate the site and magisterially saloon animate beings from feeding on the wastes.
2.2 STATEMENT OF THE PROBLEM
The presence and concentration of heavy metals in H2O, deposits, dirt and cow ‘s milk piss and fecal beads are unknown. This means that both adult male and animate beings likely assimilate the heavy metal contaminations unabated, therefore jeopardizing their lives.
2.3 JUSTIFICATION OF THE RESEARCH
The toxic heavy metals from the possible beginnings as stated above continue to acquire into the environment and the biology. This phenomenon is a existent menace to the human life therefore it is an pressing issue that the survey needs to turn to.
It is of import to observe that with the known construct of milk eating of babies for the first six months after birth, either through female parents ‘ chests or other beginnings like cow milk, more so in the rural set-ups, presents a possible lethal exposure path of heavy metal toxic condition. The clinical manifestations that un-permissible degrees of the heavy metals cause to both adult male and animate beings are fatal and expensive to handle therefore endanger the economic advancement of the affected community.
The dumping point at the present site has continued to foul the air due to organics let go ofing toxicant gases when they decompose and burnt. The changeless combustion of the wastes has hindered visibleness, caused external respiration trouble and eye-aches to the route users as the site is at the high manner and the entryway to the metropolis. It is hence a ill-mannered welcome to the tourers sing the metropolis.
Therefore, the survey will give an indicant of the exposure of female parents and babies in the part to the heavy metals and besides ascertain the safety of absolute milk eating of babies.
( I ) The solid wastes dumped at the dumping site at Kachok, Kisumu metropolis, contain toxic heavy metals like Cd ( Cd ) , Cu ( Cu ) , Fe ( Fe ) , lead ( Pb ) , manganese ( Mn ) , and Zn ( Zn ) .
( two ) The animate beings that feed on the wastes take-up the heavy metals into their organic structure systems.
3.0 LITERATURE REVIEW
Heavy metals are elements with specific denseness of 5gcm3, at least five times as the specific gravitation of H2O ( Florea T et al. , 2006 ) and ( Steven, D, 2003 ) . They have been found in human chest milk and shown to impact wellness in babies. This may be due to female parents being susceptible to chemicals largely in nutrients. Inhalation and cuticular paths are possible though they are undistinguished. Heavy metals, quicksilver, lead, arsenic, Cd, Bi, Sb most frequently disrupt immune map, neurological and endocrinal maps.
Some common effects of heavy metal toxicity include encephalon murk. Insomnia in kids, memory loss, dementedness shudders delay development ( Molin J, 2000 ) . Due to their toxic nature, the human organic structure upon assimilation begins to acquire rid of them through the variety meats such as the tegument, liver, kidney and through urine and perspiration. However, this procedure is rather strenuous therefore loads and amendss the variety meats ( Bentum J.K, et al. , 2010 ) .
Unfortunately human milk is one of the paths of riddance this load, and hence a beginning of exposure to babies ( Oskarsson A, 1998 ) . Some of these metals are stored in the female parent ‘s castanetss and are extracted from her to supply Ca for the development of the kid ‘s castanetss. As a consequence, they enter the maternal blood and chest milk during gestation and lactation, therefore exposing the foetus and babies to put on the line ( Sonawane R.B, 1994 ) . However, at allowable degrees, some of them are indispensable for normal physiological maps in animate being tissues ( Ahmed, E.E.K, et al. , 1999 ) .
Dietary lacks of Cu, Zn, Ca, Fe, protein and extra fats cause an addition in the soaking up and toxicity of lead ( Goldfrank, L.R. et al. , 1990 ) .
While Cu is a hint component in assorted metabolic maps in the organic structure, lead and other heavy metals have no map in the organic structure and can be extremely toxic due to interference straight in metabolic tracts or indirectly by doing lacks of other hint metals ( Farr G, 2004 ) . Excessively higher degrees of the metals in milk and tissues of animate beings suggest an exposure either from the air, dirt, H2O or provenders or all of these beginnings ( Farr G, 2001 ) and ( Dupler D, 2001 ) .
Animals can digest elevated degrees of these metals though at certain degrees clinical marks of toxicity manifest which can be acute or chronic when there is low exposure for a long clip since these metals bio-accumulate in the organic structure ( IARC, 1997 ) and ( Allcroft R, 1951 ) .
Heavy metals like Cd, lead and quicksilver have been detected in chest milk in many parts of the universe and have different agencies and scopes ( Appendix 6.1 ) .
In many parts of the universe, they exceed the recommended bounds ( Oskarson A et al. , 1995 ) while in others lead has been found in chest milk between 5-20 ppb ( Rabinowitz M et al. , 1985 ) . This may be attributed to the fact that beginnings of lead exposure are legion runing from ceramic and pottery glazed with lead, electronic plants, welding and solders, jewellery devising and repairing, certain hair dyes, car fixs ( ATSDR, 1990 ) .
The presence of Cd has been detected in chest milk as 0.28 I?g/litre. It is found in many constituents of vehicles and in electrical and electronic equipment ( Honda R et al. , 2003 ) .
Cadmium ‘s degrees in chest milk have besides been associated with coffin nail smoke. Arsenic has non been exhaustively studied in chest milk but is nevertheless known to do malignant neoplastic disease in worlds ( Radisch B and Luck W, 1987 ) .
4.1 Sampling and Sampling Design
A Two factor wholly randomised block design will be employed in trying where one of the locations will be considered as a block. They will be spread out within 8 km apart. The samples will be taken within an interval of seven yearss. Table2 shows the experimental design detailing the figure of samples per location and the sampling intervals of 7 yearss, 14 yearss and 21 yearss ( Table 2 ) :
Key: D= Days ; Re= Replicates ; Lo= Locations:
4.2 Study Area
The survey country will be at the shore of Lake Victoria, Kisumu metropolis and its environments within the country bounds of 00 51 ‘ South and Longitude 0041 ‘ North and longitudes 330 20’- 35020 East and an height of 528m above the sea degree. The undermentioned locations will be picked for the survey:
Location 1: Nyalenda-Kachok: whose animate beings feed and graze at the dumping site- suspected to be polluted with the metals.
Location 2: Mamboleo: 8 km – north eastern outskirts of the metropolis with comparative high animate being farming.
Location 3: Chiga: 8 km- eastern outskirts of the metropolis with subsistence and light animate being farming.
The pick of the trying countries 2 and 3 is based on the fact that the cattles graze freely in their countries but can non make the dumping site where merely those from location 1 entree for pasturage. All samples will be collected from 5 ( hence five replicates ) indiscriminately selected points from each of the three locations ; 1, 2 and 3 and instantly taken to the research lab for readying, digestion and analysis at Chemistry Laboratory, Maseno University, Kenya.
4.3 SAMPLES COLLECTION
4.3.1 Cow – Milk Sample
By ego milking into sterilized polythene bottles, approximately 50ml fresh milk samples will be collected from five breastfeeding cattles from indiscriminately selected places in each location on a three – twenty-four hours milking interval in the forenoon ( i.e 50ml ten 5cows x3 locations x 3 milking intervals = 2,250 milliliter will be collected in entire.
The samples will so be packed into ice-bags and labeling will be done with regard to clip, day of the month, location and replicate.
4.3.2 Urine Sample
50ml urine sample will be indiscriminately collected from each breastfeeding cow ( whose milk is sampled ) from its shed. The samples will so be wrapped and tied with sterilised polyethylene documents, packed into ice-bags and labeled harmonizing to clip, day of the month, location and replicate.
4.3.3 Faecal Drops Sample
In each location, about 100g of fecal beads will be indiscriminately collected from each of the five lactating cattles ( whose milk is sampled ) from their sheds, wrapped and tied with sterilized polyethylene paper, packed into ice-bags and labeled harmonizing to clip, day of the month, location and replicate.
4.3.4 Water Sample
About 50ml of H2O samples from five different points in each location will be placed in clean fresh 100ml fictile bottles with screw caps: The points are across the swamp and dike for locations 1 and 3 while five equidistant musca volitanss along the watercourse that passes through the grazing country of location 2.
All the samples will so be packed into ice-bags and labeling will be done with regard to clip, day of the month, location and replicate.
4.3.5 Grass Feeds and Soil Sample
Grass provenders will be cut at approximately1 centimeter tallness from the land in an country of 1x1m2, ( within five randomly selected cowss croping land per location ) , bundled, labeled and packed in clean polyethylene bags.
At the Centre of the 1x1m2 country where grass provenders are taken, the surface soil will be dug to 12 centimeter deepness at an country of 24x24cm2.
The dirt will so be put in clean polyethylene bags and labeled harmonizing clip, day of the month, location and replicate.
4.3.6 Sediment Sample
Five sediment sub-samples will be indiscriminately taken in each location. Approximately top 2 cm surface bed will be collected with a strainer and the samples packed and labeled with regard to clip, day of the month, location and replicate.
4.4 PREPARATION, DIGESTION, ANALYSIS AND QUANTITATION
4.4.1 Cow – Milk Samples
Five beads of 0.1 M trichloroacetic acid will be added to the cow- milk sample to precipitate the proteins, and the aqueous bed of the milk separated by centrifugation. 5 milliliter of the aqueous bed will be placed in porcelain crucible and heated in a furnace at a temperature of 500 0C for approximately 45 proceedingss.
Thereafter, 3 milliliter of 0.5M nitric will be added and so filtered through Whatman filter paper ( No 40 ) into a 10ml measurement cylinder. Further 0.5M azotic acid will be added to the 10 ml grade of the measurement cylinder. The concentrations of Cd, Cr, Cu, Fe, Pb, and Zn in space and the milk samples will be analyzed with an AAS.
4.4.2 Urine and Water Samples
100ml of each sample will be boiled till complete waterlessness. 10ml of conc. azotic acid will be added to the sample and boiled near to dryness so diluted to 20 milliliters with de-ionized H2O. The solution will be filtered and the filtrate taken for AAS analysis for Cd, Cr, Cu, Fe, Pb, and Zn.
4.4.3 Quantitation of heavy metals in milk, urine and H2O samples:
Concentrations of Cd, Cu, Fe, Pb, Mn, Se and Zn in examined samples will be calculated harmonizing to the undermentioned equation: –
Mg/kg in examined samples = AxB/W
A= mg/kg of metal in prepared samples ( obtained by standardization ) .
B= concluding volume of prepared sample in milliliter.
W= weight of samples in gms.
4.4.4. Quantitation of Heavy Metallic elements in Faecal Drops, Grass, Sediments & A ; Soil Samples
The samples will be rinsed with de-ionized H2O several times and individually air-dried on unfastened plastic bags for 24 hours, land in a howitzer to obtain little atoms of unvarying size, therefore big surface country. Conventional aqua regia digestion will be performed in 250ml glass beakers covered with ticker spectacless. A well-mixed sample of 0.50 g each of the samples will be digested in 12ml of aqua regia on a hot home base for 3 H at 110A°C.
After vaporization to near waterlessness, the sample will be diluted with 20 milliliters of 2 % ( v/v with H2O ) azotic acid and transferred into a 100-ml volumetric flask after filtrating through WhatmanA® filter paper grade 40 and diluted to 100 milliliters with de-ionized distilled H2O thenceforth analyzed for degrees of Cd, Cu, Fe, Pb, Mn, Se and Zn utilizing AAS.
1. Ahmad, W.M.S. ( 2002 ) : Surveies on heavy metal pollution in domestic fowl farms in relation to production public presentation ; Ph.D. Thesis-Faculty of Vet. Medicine. Zag. University.
2. Ahmed, E.E.K, Haleem, H.H. and Aly, A.A. ( 1999 ) : Consequence of Cu and ascorbic acid in limitation of Cd toxicity. J. Egypt. Vet. Med. Ass. , 59 ( 5 ) : 1549-1573.
3. Allcroft R. 1951: Lead toxic condition in cowss and sheep. Veterinary Record 63:583-593.
4. ATSDR “ Case survey in environmental medical specialty: Cadmium toxicity: U.S Department of Health and Human Services. Atlanta G.A, 1990.
5. Roberts J R, 1999: Metallic toxicity in kids. In Training Manual on Pediatric Environmental Health: Puting It into Practice 1999 Jun. Emeryville, CA: Children ‘s Environmental Health Network.
6. Bentum J.K, Sackitey O.J, Tuffuor J.K. , Essumang D.K, Koranteng-Addo E. J, and Owusu-Ansah E. , 2010: Cadmium and Arsenic in chest milk of breastfeeding female parents in Odumanse-Atua community in Manya Krobo territory of eastern part of Ghana.
7. Carl, M. ( 1991 ) : Heavy metals and other hint elements. Monograph on residues and contaminations in milk and milk merchandises. Particular Issue 9101, pp. 112-119. International Dairy Federation “ IDF ” , Belgium.
8. Dupler D. 2001: Heavy metal poisoning Gale Encyclopedia of Alternative Medicine. Farmington Hills, MI: Gale Group.
9. Farr G 2001: The Hair Tissue Mineral Analysis.
10. Farr G 2004: Why Heavy Metallic elements are a Hazard to Your Health.
11. Florea T, Sarolta O.B and Gheorghe C, 2006: Heavy metals in fresh cow-milk and cheese.
12. Friberg, L. and Elinder, C.G. 1988: Cadmium toxicity in worlds. Essential and toxic hint elements in human wellness and disease, edited by A.S. Prasad ( New York: A.R.Liss ) , pp. 559-587.
13. Goldfrank, L.R. ; Osborn, H. and Hartnett, L, 1990: Lead. In: Goldfrank, L.R. ; Flomentbaum, N.E. ; Lewin, N.A. ; Weisman, R.S. and Howland, M.A. ( Eds. ) : Goldfrank ‘s Toxicological Emergencies. 4th edition. pp. 627-637. Prentice-Hall International Inc. New Jersey, USA.
14. Honda R ; Tawara K ; Nishyo M ; Nakagawa H ; Tanebe K ; Saito S, Toxicology 2003 ; 186 ( 3 ) 255-259.
15. IARC ( International Agency for Research on malignant neoplastic disease ) 1997: Monograph of carcinogenic hazard to human. Lyon. Supplement. 7:230-231.
16. Kengara F.O, 2004: Analysis of organo-chlorine pesticides in Nyando catchments of Lake Victoria and fate surveies of atrazine and glyphosate in dirt utilizing the radioisotope tracer technique: MSc Thesis-Faculty of Science, Department of Chemistry, Maseno University, Kenya.
17. Mactaggart D.L and Farewell S.O: Analytic usage of arrested development. Part 1: Arrested development processs for standardization and quantitation, 1992, Journal of AOA International, 75 594-606.
18. Mlay P.S and Mgumia Y.O, 2008: Degrees of lead and Cu in plasma of dairy cattles, grazing lands, dirt and H2O from selected countries of Morogoro suburbs. ( Department of Physiology, Biochemistry, Pharmacology and Toxicology, Tanzania ) .
19. Molin J: Journal of occupational and environmental medical specialty ; 2000 ; 42 ( 11 ) 1070-1075.
20. Ongeri, D.M.K, 2008: Physicochemical parametric quantities, heavy metal residue degrees and their speciation surveies in Lake Victoria basin ; Ph.D. Thesis-Faculty of Science, Department of Chemistry. Maseno University, Kenya.
21. Oskarson A. , Palminger H.I, and Sundberg: J. Analyst: 1995 ; 120 ( 3 ) 765-770.
22. Oskarsson, A, Analyst 1998 123 ( 1 ) ; 19-23.
23. Osweiler D. G, 1996: Toxicology. Williams and Wilkins USA 491pp.
24. Rabinowitz, M. , Leviton A. , and Needleman H. , Archives of environmental wellness 1985 ; 40 ( 5 ) 283-286.
25. Radisch B and Luck W: Nav H Toxicology letters 1987 ; 36 147-152.
26. Radostits O. M, Blood D. C and Gay C. C, 1994: Veterinary Medicine A Textbook of the Disease of Cattle, Sheep, Goat and Horses 8th Edition. Paston imperativeness ltd, London, Norfolk, UK 1469-1499p.
27. Roberts J R, 1999: Metallic toxicity in Children. In Training Manual on Pediatric Environmental Health: Puting It into Practice 1999 Jun. Emeryville, CA: Children ‘s Environmental Health Network.
28. Selinger B, 1979: Chemistry in the market topographic point.
29. Sonawane R.B: Envronmental Health Perspective, 1994 ; 196.
30. Tsoumbaris, P. and Papadopoulou, T.H. 1994: Heavy metals in common nutrient materials: Quantitative analysis. Bulletin Environ. Contamination Toxicology, 53: 61-66.
31. Stevens, D. 2003. CSIRO Land and H2O ‘s Methods Manual. Impact of Heavy Metallic elements on Sustainability of Fertilization and Waste Recycling in Peri-Urban and Intensive Agriculture in South-East Asia. Australian Centre for International Agricultural Research ( ACIAR ) .
29. World Health Organization, ( WHO, 1993 ) .
6.1: Table 1- WHO: Selected Concentration Mean & A ; Ranges of Heavy Metallic elements, 1993.
Concentration Range, ppb
0.10 – 3.80
0.00 – 41.10
0.64 – 257.10
7.00 – 102.00
6.2: Table 2: Experimental Design and Sampling Record Table
Key: D= Days ; Re= Replicates ; Lo= Locations and F/D- Faecal Drops
6.3: Table 3- TIME SCHEDULE
Proposal Writing and Presentation
Jan – March – 2011
Research Site Survey and Preparation
April – 2011
Acquisition of Chemical Reagents
May – 2011
1st Sample Collection and Extraction
June – 2011
1st Experiments and Analysis – Associate in applied science
June – 2011
2nd Sample Collection and Extraction
July – 2011
2nd Experiments and Analysis – Associate in applied science
July – 2011
3rd Sample Collection and Extraction
August – 2011
3rd Experiments and Analysis – Associate in applied science
August – 2011
Discussion and Statistical Interpretation
Thesis Writing and Submission
6.4: Table 4 – Budget
Unit Price ( KSh )
Entire Cost ( KSh )
Lead ( II ) Nitrate
Unhydrous Sodium Sulphate
Copper ( II ) Nitrate
Chromium ( II ) sulfate
Sub – Sum
Apparatus, Equipment and Others
AAS Analysis Lamps
Brown PVC Bottles
Whatman Filter Papers grade 40
Thesis Preparation and Binding
Disposable Baseball gloves
Ice – Box
Sub – Sum
Travels and Subsistence
Site Visits and Sampling
Sub – Sum
Table of Content
Cover and Title
Introduction: Background/Statement of the Problem
Statement of the Problem and Justification of the Study
Methodology and Study Area and Sample Experimental Design.
Collections: Cow-Milk and Urine Samples
Faecal Drops and Water Samples
Grass Feeds, Sediment and Soil Samples
Preparation, Digestion Analysis and Quantitation
Cow-Milk, Urine and Water Samples
Preparation, Digestion, and Analysis
Faecal Drop, Grass Feed, Sediment and Soil Samples
Cite this Heavy Metals In Cows Milk Biology
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