Determination of Heavy Metals in Cow Milk

Determination of Heavy Metals in Cow drawThe cow take out feeding concept of infants, in the larger move of Lake Victoria basin of Kenya, for about the first six months afterward birth, has immensely invigorated the interest in investigating the movement and levels of some ominous metallic elementlic elements in awe draw. Toxicity of a hefty metal depends on its fractional bioavailability and concentration in the environment, therefore its speciation is of great importance. The knowledge of concentrations of hepato venomous free weighty metals like atomic number 48, chromium, bulls eye, iron, collar and zinc in cows take out is hence very essential.Fresh milk samples from lactating cows eitherow be keeped by self milking into sterilized polye and soe bottles and labeled accord to time, date, place and replicate. Other parameters that leave behind be collected from the triple localization principles on the infrastructure of 7-day interval and thereafter inv estigated include grass feeds, sediments, urine, kingdom, and lactating cows faecal drops and urine. This information therefore gives a suitable background for assessing and determining the concentrations of enceinte metal contaminants on cows and their subsequent intake by hu soldiery. a couple of(prenominal) drops of 0.1 M trichloroacetic unpleasant exit be added to the sample for coagulation and the aqueous layer heated at 500C for matchless hour. Digestion get out be done with 0.5 M nitric erosive as heraldic bearing and concentration of concentrated metals analyzed using an Atomic Absorption Spectrophotometer, AAS.statistical abstract testament be conducted using MSTATC two factor release randomized block design, with the heavy metal concentration as the primary(prenominal) factor with the localizations as the sub treatment. The package go away perform analysis of variance (ANOVA) at P 0.05 with two factor experiment and students T-test at P 0.05. The mean, standard deviation, range and linear correlation co-efficient on the mensurable parameters go forth be determined. The study is expected to give an denotation of the depiction of m new(prenominal)s and infants in the region to the heavy metals and also ascertain the rubber of commanding milk feeding of infants.This study will be conducted in Kisumu city, at the shore of Lake Victoria, Kenya.Key words Heavy metals, infants, cows milk and contamination.2.0 dis course2.1 BackgroundHuman and animals countenance been exposed to heavy metal perniciousness for an immeasurable time. The industries have dramatically increased the overall environmental stretch of the toxins to levels that they be be in every argona of modern consumerism. Therefore, it is necessary to know the environmental fate of all xenobiotics so as to presage their persistence and possible effects on non-target organisms (Kengara F.O 2004).Anthropogenic activities appear to play an important role in this stud y since, in the past, solid wastes have been dumped by residents in their respective localities. In an app bent attempt to support the environment unclouded, in Kisumu city, the local authorities, in the past four geezerhood have collected and dumped the solid wastes at a new site, Nyalenda Kachok, Kisumu. in like manner lorry-loads from all over the citys supermarkets, industrial set-ups, petrol stations, residences and markets dump theirs solid wastes at the site. cadmium, chromium, pig bed, iron, lead and zinc be among the most common heavy metals know as contaminants in the environment and therefore come affront as hazardous substances to both human and animal wellness (Roberts J.R, 1999). This is collect to across-the-board spread environmental pollution by materials containing them like batteries, paints, pipes, soldering rods, pesticides, fungicides, gasoline, railway locomotive oils, chemical fertilizers or when they occur in steep amounts in air, soil, water, plant s and other compounded animal feeds.They therefore increase concentrations of heavy metals in air, water, soil and subsequently interpreted by plants and animals into their food chain (Ahmad, W.M.S, 2002). The presence of heavy metals in cows milk whitethorn be attri exclusivelyed to contamination of the maestro one, which may be due to film of lactating cow to environmental pollution or consumption of contaminated feeding stuffs and water (Carl M, 1991).This fact stinker lead to considerable concentrations in human body since they are not metabolized therefore poses a serious risk to human health when consumed even in small amounts (Selinger B, 1979).Most of them, like cadmium, lead and quicksilver persist in the body and exert their toxic effect by combining with one or more than reactive groups essential for conventionalism physiological single-valued functions of the cells thus causing cellular disturbances or clinical manifestation. The untoward toxic effects caused by lead, cadmium mercury are wide recognized (Friberg, L. and Elinder, C.G, 1988). The major clinical signs in animals and man for lead and copper poisoning include, among others, deviations of the hematological parameters due to their come in effects on hematopoiesis, cut back integrity of red blood cells membrane leading to intravascular haemolysis, anemia and vaporization (Radostits O. M. et al., 1994). Therefore hematological parameters have diagnostic value in animals surmise of heavy metal toxicity (Mlay P.S and Migumia Y.O, 2008). Man becomes at risk by eating food and drinking fluids contaminated with heavy metals,through air, direct contact with the metals like in people working in railcar wash or body spraying industries or factories transaction with heavy metals and their derivatives (Farr G, 2001).Kisumu city is endowed with relatively many but small enterprises dealing with metal whole shebang, car maintenance and repair (Jua Kali Sheds), construction works that po se a risk of contamination to the environment with hazardous substances including heavy metals. molarsistence farming and husbandry are quite enhanced in its neighborhoods.The gone(p) information prompts the desire to investigate the presence of some heavy metals in cows milk. The data generated will assist the concerned city planners, institutions and bodies aerated with environmental control and surveillance to formulate measures and policies that would firmly govern the toss out of solid wastes, re-locate the site and authoritatively bar animals from feeding on the wastes.2.2 arguing OF THE PROBLEMThe presence and concentration of heavy metals in water, sediments, soil and cows milk urine and faecal drops are unknown. This means that both man and animals likely assimilate the heavy metal contaminants unabated, thus endangering their lives.2.3 JUSTIFICATION OF THE queryThe toxic heavy metals from the possible sources as stated above detain to get into the environment and the biota. This phenomenon is a real threat to the human animation therefore it is an urgent issue that the study needs to address.It is important to situation that with the known concept of milk feeding of infants for the first six months after birth, either through mothers dressers or other sources like cow milk, more so in the rural set-ups, presents a possible lethal exposure route of heavy metal poisoning. The clinical manifestations that un-permissible levels of the heavy metals cause to both man and animals are fatal and expensive to treat thus jeopardise the economic progress of the affected residential district.The fling point at the present site has continued to pollute the air due to organics releasing barbarous gases when they decompose and burnt. The constant burning of the wastes has hindered visibility, caused breathing difficulty and eye-aches to the road users as the site is at the high way and the entrance to the city. It is therefore a rude welcome to the tour ists visiting the city.Therefore, the study will give an indication of the exposure of mothers and infants in the region to the heavy metals and also ascertain the safety of absolute milk feeding of infants.2.4 HYPOTHESIS(i) The solid wastes dumped at the dumping site at Kachok, Kisumu city, contain toxic heavy metals like cadmium (Cd), copper (Cu), iron (Fe), lead (Pb), manganese (Mn), and zinc (Zn) .(ii) The animals that feed on the wastes take-up the heavy metals into their body systems.3.0 LITERATURE REVIEWHeavy metals are elements with specific density of 5gcm3, at least volt times as the specific gravity of water (Florea T et al., 2006) and (Steven, D, 2003). They have been found in human breast milk and shown to affect health in infants. This may be due to mothers being susceptible to chemicals mostly in foods. intake and dermal routes are possible though they are insignificant. Heavy metals, mercury, lead, arsenic, cadmium, bismuth, antimony most often disrupt immune func tion, neurological and endocrine functions.some(a) common effects of heavy metal toxicity include fountainhead fogginess. Insomnia in children, memory loss, dementia tremors delay development (Molin J, 2000). Due to their toxic nature, the human body upon assimilation begins to get rid of them through the organs such as the skin, liver, kidney and through urine and sweat. However, this process is quite straining thus burdens and damages the organs (Bentum J.K, et al., 2010).Unfortunately human milk is one of the routes of elimination this burden, and therefore a source of exposure to infants (Oskarsson A, 1998). Some of these metals are stored in the mothers bones and are extracted from her to provide calcium for the development of the childs bones. As a result, they enter the maternal blood and breast milk during pregnancy and lactation, thus exposing the fetus and infants to risk (Sonawane R.B, 1994). However, at permissible levels, some of them are essential for normal physiol ogical functions in animal tissues (Ahmed, E.E.K, et al., 1999).Dietary deficiencies of copper, zinc, calcium, iron, protein and wasted fats cause an increase in the absorption and toxicity of lead (Goldfrank, L.R. et al., 1990). period copper is a delimitate element in various metabolic functions in the body, lead and other heavy metals have no function in the body and can be highly toxic due to interference directly in metabolic pathways or indirectly by causing deficiencies of other trace metals (Farr G, 2004). Excessively higher levels of the metals in milk and tissues of animals suggest an exposure either from the air, soil, water or feeds or all of these sources (Farr G, 2001) and (Dupler D, 2001).Animals can tolerate elevated levels of these metals though at certain levels clinical signs of toxicity manifest which can be acute or inveterate when there is low exposure for a long time since these metals bio-accumulate in the body (IARC, 1997) and (Allcroft R, 1951).Heavy met als like cadmium, lead and mercury have been sight in breast milk in many parts of the macrocosm and have different means and ranges (Appendix 6.1).In many parts of the world, they pass by the recommended limits (Oskarson A et al., 1995) composition in others lead has been found in breast milk between 5-20 ppb (Rabinowitz M et al., 1985). This may be attributed to the fact that sources of lead exposure are numerous ranging from ceramic and pottery glazed with lead, electronic works, welding and solders, jewelry making and repairing, certain hair dyes, automobile repairs (ATSDR, 1990).The presence of cadmium has been detected in breast milk as 0.28 g/litre. It is found in many components of vehicles and in electrical and electronic equipment (Honda R et al., 2003).Cadmiums levels in breast milk have also been associated with cigarette smoking. arsenous anhydride has not been thoroughly studied in breast milk but is however known to cause cancer in humans (Radisch B and passel W , 1987).METHODOLOGY4.1 Sampling and Sampling DesignA twain factor completely randomized block design will be employed in sampling 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 septenary days. Table2 shows the experimental design detailing the number of samples per location and the sampling intervals of 7 days, 14 days and 21 days (Table 2)Key D= days Re= Replicates Lo= Locations4.2 Study AreaThe study sphere of influence will be at the shore of Lake Victoria, Kisumu city and its environs within the area limits of 00 51 South and Longitude 0041 North and longitudes 330 20- 35020 East and an altitude of 528m above the sea level. The sideline locations will be picked for the studyLocation 1 Nyalenda-Kachok whose animals feed and snip at the dumping site- suspected to be polluted with the metals.Location 2 Mamboleo 8 km north eastern outskirts of the city with relative high anima l husbandry.Location 3 Chiga 8 km- eastern outskirts of the city with subsistence and swingy animal husbandry.The choice of the sampling areas 2 and 3 is based on the fact that the cows graze freely in their areas but can not r all(prenominal)(prenominal) the dumping site where only those from location 1 access for herbage. All samples will be collected from 5 (hence 5 replicates) indiscriminately selected points from each of the three locations 1, 2 and 3 and immediately taken to the laboratory for preparation, digestion and analysis at Chemistry Laboratory, Maseno University, Kenya.4.3 SAMPLES COLLECTION4.3.1 Cow take out tryBy self milking into sterilized polyethylene bottles, about 50ml good milk samples will be collected from five lactating cows from randomly selected homes in each location on a three day milking interval in the morning (i.e 50ml x 5cows x3 locations x 3 milking intervals = 2,250 ml will be collected in total.The samples will so be packed into ice-ba gs and labeling will be done with respect to time, date, location and replicate.4.3.2 Urine Sample50ml urine sample will be randomly collected from each lactating cow (whose milk is sampled) from its shed. The samples will then be wrapped and tied with sterilized polythene news reports, packed into ice-bags and labeled according to time, date, location and replicate.4.3.3 Faecal Drops SampleIn each location, nigh 100g of faecal drops will be randomly collected from each of the five lactating cows (whose milk is sampled) from their sheds, wrapped and tied with sterilized polythene news report, packed into ice-bags and labeled according to time, date, location and replicate.4.3.4 body of water Sample just about 50ml of water samples from five different points in each location will be placed in clean unused 100ml plastic bottles with keister caps The points are across the swamp and dam for locations 1 and 3 while five equidistant spots along the stream that passes through the bro wse area of location 2.All the samples will then be packed into ice-bags and labeling will be done with respect to time, date, location and replicate.4.3.5 Grass Feeds and kingdom SampleGrass feeds will be cut at approximately1 cm height from the ground in an area of 1x1m2, (within five randomly selected cows grazing ground per location), bundled, labeled and packed in clean polythene bags.At the centre of the 1x1m2 area where grass feeds are taken, the topsoil will be dug to 12 cm depth at an area of 24x24cm2.The soil will then be put in clean polythene bags and labeled according time, date, location and replicate.4.3.6 Sediment Sample five sediment sub-samples will be randomly taken in each location. Approximately top 2 cm surface layer will be collected with a strainer and the samples packed and labeled with respect to time, date, location and replicate.4.4 PREPARATION, DIGESTION, ANALYSIS AND QUANTITATION4.4.1 Cow Milk SamplesFive drops of 0.1 M trichloroacetic acid will be a dded to the cow- milk sample to precipitate the proteins, and the aqueous layer of the milk separated by centrifugation. 5 ml of the aqueous layer will be placed in porcelain crucible and heated in a furnace at a temperature of 500 0C for about 45 minutes.Thereafter, 3 ml of 0.5M nitric will be added and then filtered through Whatman filter paper (No 40) into a 10ml measuring cylinder. Further 0.5M nitric acid will be added to the 10 ml mark of the measuring cylinder. The concentrations of Cd, Cr, Cu, Fe, Pb, and Zn in dumbbell and the milk samples will be analyzed with an AAS.4.4.2 Urine and Water Samples100ml of each sample will be boiled till complete temperance. 10ml of conc. nitric acid will be added to the sample and boiled close to dryness then diluted to 20 ml with de-ionized water. 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 water samples assiduousnesss of Cd, Cu , Fe, Pb, Mn, Se and Zn in examined samples will be calculated according to the following equation-Mg/kg in examined samples = AxB/WA= mg/kg of metal in prepared samples (obtained by calibration).B= final volume of prepared sample in ml.W= weight of samples in grams.4.4.4. Quantitation of Heavy Metals in Faecal Drops, Grass, Sediments Soil SamplesThe samples will be rinsed with de-ionized water several times and separately air-dried on sacrifice plastic bags for 24 hours, ground in a mortar to obtain small particles of uniform size, thus large surface area. Conventional cobalt blue regia digestion will be performed in 250ml glass beakers covered with watch glasses. A well-mixed sample of 0.50 g each of the samples will be digested in 12ml of cobalt blue regia on a hot plate for 3 h at 110C.After evaporation to near dryness, the sample will be diluted with 20 ml of 2% (v/v with H2O) nitric acid and transferred into a 100-ml volumetric flask after filtering through Whatman filter paper grade 40 and diluted to 100 ml with de-ionized distilled water thereafter analyzed for levels of Cd, Cu, Fe, Pb, Mn, Se and Zn using AAS.5.0 REFERENCES1. Ahmad, W.M.S. (2002) Studies on heavy metal pollution in poultry farms in relation to production capital punishment Ph.D. dissertation-Faculty of Vet. Medicine. Zag. University.2. Ahmed, E.E.K, Haleem, H.H. and Aly, A.A. (1999) Effect of copper and ascorbic acid in restriction of cadmium toxicity. J. Egypt. Vet. Med. Ass., 59 (5) 1549-1573.3. Allcroft R. 1951 Lead poisoning in cattle and sheep. Veterinary Record 63583-593.4. ATSDR Case study in environmental medicine Cadmium toxicity U.S Department of wellness and Human Services. Atlanta G.A, 1990.5. Roberts J R, 1999 Metal toxicity in children. In upbringing Manual on Pediatric environmental wellness Putting It into manage 1999 Jun. Emeryville, CA Childrens 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 breast milk of lactating mothers in Odumanse-Atua community in Manya Krobo district of eastern region of Ghana.7. Carl, M. (1991) Heavy metals and other trace elements. Monograph on residues and contaminants in milk and milk products. Special Issue 9101, pp. 112-119. outside(a) Dairy Federation IDF, Belgium.8. Dupler D. 2001 Heavy metal poisoning Gale Encyclopedia of ersatz Medicine. Farmington Hills, MI Gale Group.9. Farr G 2001 The Hair Tissue Mineral abridgment.10. Farr G 2004 why Heavy Metals are a Hazard to Your Health.11. Florea T, Sarolta O.B and Gheorghe C, 2006 Heavy metals in odoriferous cow-milk and cheese.12. Friberg, L. and Elinder, C.G. 1988 Cadmium toxicity in humans. Essential and toxic trace elements in human health 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. (Ed s.) Goldfranks Toxicological Emergencies. quaternary 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 2003186(3) 255-259.15. IARC (International Agency for search on cancer) 1997 Monograph of carcinogenic risk to human. Lyon. Supplement. 7230-231.16. Kengara F.O, 2004 epitome of organo-chlorine pesticides in Nyando catchments of Lake Victoria and fate studies of atrazine and glyphosate in soil using the radioisotope tracer bullet technique MSc Thesis-Faculty of Science, Department of Chemistry, Maseno University, Kenya.17. Mactaggart D.L and Farewell S.O Analytical use of regression. fragment 1 Regression procedures for calibration and quantitation, 1992, Journal of AOA International, 75 594-606.18. Mlay P.S and Mgumia Y.O, 2008 Levels of lead and copper in plasma of dairy cows, pastures, soil and water from selected areas of Morogoro suburbs. (Department of Physiology, Biochemistry, Pharm acology and Toxicology, Tanzania).19. Molin J Journal of occupational and environmental medicine 2000 42(11) 1070-1075.20. Ongeri, D.M.K, 2008 Physicochemical parameters, heavy metal residue levels and their speciation studies 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 health 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 intrepid C. C, 1994 Veterinary Medicine A Textbook of the Disease of Cattle, Sheep, Goat and Horses eighth Edition. Paston press ltd, London, Norfolk, UK 1469-1499p.27. Roberts J R, 1999 Metal toxicity in Children. In Training Manual on Pediatric Environmental Health Put ting It into Practice 1999 Jun. Emeryville, CA Childrens Environmental Health Network.28. Selinger B, 1979 Chemistry in the market place.29. Sonawane R.B Envronmental Health Perspective, 1994 196.30. Tsoumbaris, P. and Papadopoulou, T.H. 1994 Heavy metals in common food stuffs Quantitative analysis. Bulletin Environ. taint Toxicology, 53 61-66.31. Stevens, D. 2003. CSIRO Land and waters Methods Manual. Impact of Heavy Metals on Sustainability of impregnation and Waste Recycling in Peri-Urban and Intensive Agriculture in southeastward Asia. Australian Centre for International Agricultural Research (ACIAR).29. World Health Organization, (WHO, 1993).6.0 APPENDICES6.1 Table 1- WHO Selected Concentration Mean Ranges of Heavy Metals, 1993.Heavy MetalConcentration, ppbConcentration Range, ppbArsenic0.30.10 -0.80Cadmium0.10.10 3.80Lead5.00.00 41.10Mercury2.70.64 257.10Manganese18.07.00 102.006.2 Table 2 Experimental Design and Sampling Record TableKey D= Days Re= Replicates Lo= Locati ons and F/D- Faecal DropsRe.LoMilkUrineF/DropsWaterSoilGrassSediments123123123123123123123112321233123412351236.3 Table 3- TIME SCHEDULEACTIVITY geological periodDURATIONProposal Writing and PresentationJan March 201112 weeksResearch send Survey and PreparationApril 20114 weeksAcquisition of Chemical Reagentswhitethorn 20114 weeks1st Sample Collection and stemmaJune 20111 week1st Experimentations and synopsis AASJune 20113 weeks2nd Sample Collection and ExtractionJuly 20111 week2nd Experimentations and Analysis AASJuly 20113 weeks3rd Sample Collection and ExtractionAugust 20111 week3rd Experimentations and Analysis AASAugust 20113 weeksDiscussion and Statistical InterpretationSeptember, 20114 weeksThesis Writing and SubmissionOct-Nov, 20118 weeksTotal1 year44 weeks6.4 Table 4 BUDGETItemQuantityUnit set (KSh)Total Cost (KSh)ChemicalsUniversal Indicator1 litre1,350.001,350.00Distilled Water40 litres1,200.0048,000.00Aqua Regia5 litres3,500.0017,500.00Nitric sharp2.5 litres3,500.008,750.00Sulphuric Acid2.5 litres3,500.008,750.00Hydrochloric Acid2.5 litres3,500.008,750.00Ammonium treat500g3,500.003,500.00Ferrous Nitrate500g3,500.003,500.00Lead (II) Nitrate500g3,500.003,500.00Unhydrous Sodium Sulphate500g2,850.002,850.00Copper (II) Nitrate500g3,500.003,500.00Zinc Nitrate500g3,500.003,500.00Trichloricacetic acid2.5 litres4,500.0011,250.00Chromium (II) sulphate500g3,500.003,500.00Cadmium Nitrate500g3,500.003,500.00Sub Total131,200.00Apparatus, Equipment and OthersAAS Analysis Lamps127,500.0090,000.00Polythene Bags61,000.006,000.00Brown PVC Bottles180100.0018,000.00Whatman strain Papers grade 4061,000.006,000.00Thesis Preparation and back20,500.00Goggles21,000.002,000.00Disposable Gloves4 Boxes3,500.0014,000.00Ice Box19,000.009,000.00Labels1 Packet750.00750.00Spade11,200.001,200.00Sickle1600.00600.00Sub Total168,050.00Travels and SubsistenceSubsistence duringSampling53,500.0017,500.00Analysis20 Days500.0010,000.00Site Visits and Sampling10 Tri ps3,000.0030,000.00Enumerators,3loc.x5Repsx3Intervals3x3x51,000.0045,000.00Sub Total102,500.00Grand Total401,750.00