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828 Animal Science
REGULAR ARTICLE
Influence of food allowance in heavy metal’s concentration in raw milk
production of several feed animals
N. Gougoulias1*, S. Leontopoulos2and Ch. Makridis2
1
Technological Educational Institute of Thessaly, Department of Agriculture, Laboratory of Agricultural Chemistry, TEI of Thessaly, Ring road of Larissa, T.K. 411 10, Larissa, Greece
2
Technological Educational Institute of Thessaly, Department of Animal Production, TEI of Thessaly, Ring road of Larissa, T.K. 411 10, Larissa, Greece
Abstract
Milk and dairy products may contain varying amounts of different toxic contaminants such as heavy metals. The survey was conducted to study the rate of excretion of heavy metals such as Cu, Zn, Cd, Cr, Ni and Pb in the milk of cows, sheep and goats correlated with the corresponding quantities of metals flowing into the diet for 60 consecutive days. 300 samples of milk from five cows, were analyzed with average content (mg/L) of Cu = 0.29, Zn = 1,21, Cd <0.02, Cr <0.006, Ni <0.1 and Pb <0.02, 300 milk samples from five sheep were also analyzed with average content (mg/L) of Cu = 0.06, Zn = 0.40, Cd <0.02, Cr <0.006, Ni <0.1 and Pb <0.02, while 300 milk samples from five goats had an average content (mg/L) of Cu = 0.03, Zn = 1.38, Cd <0.02, Cr <0.006, Ni <0.1 and Pb <0.02.
Key words: Forage conventional agriculture, Heavy metals, Cow milk, Sheep milk, Goat milk
Introduction
Milk production and alteration is a dynamic and growing industry which has been recognized all over the world for its beneficial influence on human health as well as their economic importance (Tasneem et al., 2009; Silanikove et al., 2010; Rahimi, 2013). However, milk and dairy products may contain various amounts of different substances and microorganisms affecting food safety and human health. The major aspects of food safety are usually source of infection by zoonoses and human specific pathogens, presence of undesirable substances from the animal’s diet or inappropriate use of antibiotic (Silanikove et al., 2010), and other toxic substances such as heavy metals (Abdul et al., 2009; Maas et al., 2011; Tona et al., 2013; Temiz and Soylu, 2012). Although heavy metals may enter the human body through
inhalation of dust, consumption of contaminated drinking water, direct ingestion of soil (Cambra et al., 1999; Dudka and Miller, 1999; Charya et al., 2008; Jameel et al., 2009; Tabinda et al., 2013; Dhanalakshmi and Gawdaman, 2013), consumption of food plants grown in metal-contaminated soil and animal products such as milk and dairy products is in great importance (Licata et al., 2004; Mohammad et al., 2008; Rehana et al., 2009). The potential toxicity of heavy metals to human and animal health is the object of several studies indicating the importance of presence of heavy metals in milk and other dairy products (Caggiano et al., 2005; Soylak et al., 2005; Ataro et al., 2008; Tuzen et al., 2008; Kazi et al., 2009; Solis et al., 2009; Gradinaru et al., 2011; Maas et al., 2011; Rahimi, 2013; Salah et al., 2013; Vahid and Masoome, 2013). Furthermore, according to Munoz and Palmero (2004), the effects of some elements are accumulative and it is necessary to control their level concentration in consumed food. The regular absorption of small amounts of certain heavy metals may cause serious effects on the health of human being especially in growing children (Salma et al., 2000; Ataro et al., 2008). This is due to the intake of trace elements which occurs mainly through the respiratory system or Received 16 October 2013; Revised 16 February 2014;
Accepted 27 February 2014; Published Online 01 July 2014 *Corresponding Author
N. Gougoulias
through the food chain. Several methods for detection of heavy metals in milk and dairy products have been used. Among them atomic absorption spectroscopy has mostly been used for carrying out these determinations (Conti, 1997; Bag et al., 1999; Tasneem et al., 2009). According to Caggiano et al. (2005), the concentration variations from milk to products depend both on the kind of diary product and on production processes. It is important to be mentioned that according to Licata et al.(2004), the determination of the residual concentrations of metals in milk could be an important ‘‘direct indicator’’ of the hygienic status of the milk and/or of its derived products, as well as an ‘‘indirect indicator’’ of the degree of pollution of the environment in which the milk was produced.
Materials and Methods
The experiment was conducted during autumn months (October-November) of 2011 at the Technological Educational Institute of Thessaly; Department of Agriculture. Samples of raw milk from five cows of local breed (Vrachykeras), five sheep indigenous breed (Karagouniki) and five goats of local breed (Egchoria Ega), were collected daily for the first 60 days of the lactation period. The 300 milk samples of cows, sheep and goats replicated three times were analyzed in the laboratory of Agricultural Chemistry for determination of several heavy metals concentration: copper (Cu), zinc (Zn), nickel (Ni), chromium (Cr), lead (Pb) and cadmium (Cd). The average age of cows was 4 years old and their daily diet included dry matter at 3% of the animal weight, while sheep and goats were 3 years old with diet dry matter corresponding to 4% of their animal weight. The feedstuffs were collected from conventional local farms from the region of Larissa. The types of feedstuff used to feed cows were wheat, barley, maize, maize silage, alfalfa meal, wheat straw, soya bean meal, lactation feed, and basic mixture while the feedstuff types used to feed sheep and goats were wheat, barley, maize, wheat straw, alfalfa meal, lactation feed and basic mixture. The involvement of each type of feedstuff used in the diet of this experimental work was equal. Before the start of the experiment, water used for drinking animal was analyzed in order to determine and eliminate effects of heavy metals in the results obtained. Statistical analyses were performed by the use of statistical program MINITAB (Ryan et al., 2005) and Tukey’s test were used in order to detect and separate the mean treatment differences at P = 0.05.
For the determination of heavy metals, 1 g of dry matter of feed, 50 ml of drinking water and 50 ml of milk sample were homogenized and were analyzed by digestion at 350◦C. According to the method described by Allen et al. (1974) and Varian (1989), the homogenized sample containing 10 ml HNO3 + 5 ml HCLO4 were analyzed by Atomic
Absorption (Spectroscopy Varian Spectra AA 10 plus, Victoria, Australia).The detection of Cd, Ni, Cr, Cu and Zn in feedstuff was determined with the use of flame and air-acetylene mixture, while Pb bin feed stuff, metals in the water and in milk were determined using graphite furnace.
Results and Discussion
From the results obtained presented in from Table 1, it was clearly demonstrated that 5 samples of feedstuff used for feeding cows were contained in average Cd = 1.22 mg/Kg, 2 samples were contained an average of Cr = 0.1 mg/L. As regards the feedstuff used for the feeding of sheep and goats, from the results presented in Table 2 it was clearly demonstrated that none of the examined samples contained Cd in concentration greater than 0.02 mg/Kg. Moreover, 2 samples were found with an average of Pb = 3.43 mg/Kg and 3 samples were contained an average of Cr = 3.38 mg/Kg.
Milk concentrations of heavy metals in sheep were presented in Table 3, where 54 samples contained an average of Cd = 0.06 mg/L. Furthermore, 39 samples of goat’s milk were found to contain an average concentration of Cd = 0.12 mg/L. Also, from the results of Table 3, it is demonstrated that all milk samples obtained from sheep and goats were contained Pb <0.02 mg/L. Regarding the Cr concentration in the milk of sheep, all samples contained less than 0.006 mg/L, while 39 samples of goat milk was found to contain an average of 0.12 mg/L.
Table 2. Heavy metal concentrations (mg/Kg dry matter) in sheep and goats feedstuff
Cd Pb Cr Ni
<0.02 (140)
<0.02 (138)
<0.06 (137)
<0.1 (139) 3.43±0.10
(2)
3.38±0.10 (3)
5.13±0.50 (1) Data represent average means and SE deviation.(n): Number of samples
Table 3. Heavy metal concentrations (mg/L) in milk of cow, sheep and goats.
Cd Pb Cr Ni
Cow
<0.02 (300)
<0.02 (300)
<0.006 (270)
<0.1 (300) 0.10±0.02
(30)
Sheep
<0.02 (246)
<0.02 (300)
<0.006 (300)
<0.1 (300) 0.06±0.02
(54)
Goat
<0.02 (261)
<0.02 (300)
<0.006 (261)
<0.1 (300) 0.12±0.02
(39)
0.12±0.02 (39) Data represent average means and SE deviation.(n): Number of milk samples
Table 4. Heavy metal concentrations (mg/L) in used drinking water.
Cu Zn Ni Cd Cr Pb
0.09±0.02 2.02±0.05 0.0134±0.001 0.0025±0.001 0.0241±0.003 0.0051±0.002 Data represent average means and SE deviation for 5 water samples
From the results presented in Figure 1 it was demonstrated that there was no statistically important difference on the average content (mg/Kg) of Cu in the feedstuff remained at 14.42 for cows and 14,26 for sheep and goats. However, in Figure 2 the Cu concentration in milk samples especially those obtained from sheep and goats were decreased significantly, at 0,06 and 0,03 (mg/L) respectively while in cow’s the Cu concentration was obtained at 0,29 (mg/L).
Regarding the content of Zn (mg/Kg) in feedstuff used to feed cows, it was observed that this concentration was lower than the one observed in sheep and goats which was 55.52 and 78.63 respectively. The content of Zn in the milk samples of cows and goats decreased in levels where there was no statistically significant difference. It was also noted that while the feedstuff of sheep and goats contained the same level of Zn in the sheep’s milk samples, the concentration of Zn was lower than that observed in goat milk.
According to Caggiano et al. (2005) many dangerous elements or compounds, such as dioxins, pesticides, metals, and metalloids, accumulate
which the lactating ruminants were nourished. Moreover, findings by Tabinda et al., (2013) shown that in livestock milk, meat and blood concentration of Cu were greater than Ni>Pb>Cd. It was also reported that the daily intake of Cd and Pb by milk and meat was higher than their permissible limits. The toxicity of heavy metals in food and consequently in human’s health is well known (Licata et al., 2004, Caggiano et al., 2005, Duruibe et al., 2007).
Thus, maximum levels of heavy metals in foods have been determined by EU regulation 1881/2006 and FAO/WHO, (2011). These levels in milk are for Pb<20, Cu<500 and Zn<3000 (μg/L). Furthermore, according to the EU regulation 98/83 the higher amounts of heavy metals for drinking water and environment was for Cd<5, Pb<10, Ni<20, Cr<50, and for Cu<2000 (μg/L).
Figure 1. Heavy metal concentrations in feedstuff of cows, sheep and goats. Columns in each characteristic of each graph with the same letter do not differ significantly according to the Tukey’s test (P=0.05).
0 1 2 3
N
i
-(
m
g
/
K
g
)
Concentration-Ni in feedstuffs
a
b b
A)
0 5 10 15
C
u
-(m
g
/
K
g
)
Concentration-Cu in feedstuffs
B) a a a
0 30 60 90
Z
n
-(
m
g
/
K
g
)
Different species of animals Concentration-Zn in feedstuffs
Cow Sheep Goat
a a
Figure 2. Heavy metal concentration in milk of cows, sheep and goats. Columns in each characteristic of each graph with the same letter do not differ significantly according to the Tukey’s test (P=0.05).
Conclusions
In this scientific work is has been shown that the rate of excretion of Cu in the milk of cows is greater than that of sheep and goats. While, the rate of excretion of Zn in the goat milk is higher than that of sheep. Moreover, the study shows the unsafe taking of milk, due to the large number of samples of fresh milk that have exceeded safe limits content of heavy metals. This can result in multiple organ cancer and reproductive toxicity. The higher level of heavy metals contained in fresh milk is mainly explained by the pollution of the environment and feedstuff’s contamination around the industrial area of Larissa prefecture.
References
Abdul, Q. S., G. K. Tasneem, B. A. Mohammad, A. B. Jameel, I. A. Hassan, A. K. Ghulam, K. Sumaira and K. J. Mohammad. 2009. Hazardous impact of arsenic on tissues of same fish species collected from two ecosystem. J. Hazard. Mat. 167:511–515. Allen, S. E., H. M. Grimshaw, J. A. Parkinson and
C. Quarmby. 1974. Chemical analysis of Ecological materials. Blackwell Scientific Publications.
Aslam, B., I. Javed, F. H. Khan and Z. Rahman. 2011. Uptake of heavy metal residues from sewages sludge in the milk of goat cattle during summer season. Pak. Vet. J. 31:75-77. Ataro, A., R. I. McCrindle, B. M. Botha, C. M. E.
McCrindle and P. P. Ndibewu. 2008. Quantification of trace elements in raw cow’s milk by inductively coupled plasma mass spectrometry (ICP-MS). Food Chem. 111:243–248.
Bag, S., T. Vora, R. Ghatak, I. Nilufer, D. D’Mello, L. Pereira, J. Pereira, C. Cutinho and V. Rao. 1999. A study of toxic effects of heavy metal contaminants from sludge-supplemented diets onmale wistar rats. Ecotoxicol. Env. Safe. 42:163–170.
Bratakos, M. S., E. S. Lazos and S. M. Bratakos. 2001. Chromium content of selected Greek foods. Sci. Total Env. 290:47– 58.
Caggiano, R., S. Sabia, M. D’Emilio, M. Macchiato, A. Anastasio and M. Ragosta. 2005. Metal levels in fodder, milk, dairy products, and tissues sampled in ovine farms of southern Italy. Env. Res. 99:48–57.
0 0.1 0.2 0.3 0.4
C
u
-(
m
g
/
L
)
Concentration-Cu in milk A)
a
b
b
0 0.5 1 1.5 2
Z
n
-(
m
g
/
L
)
M ilk from differently species of animals
Concentration-Zn in milk B)
Cow milk sheep's m ilk Goat milk
a a
Cambra, K., T. Martınez, A. Urzelai and E. Alonso. 1999. Risk analysis of a farm area near a lead-and cadmium-contaminated industrial site. J. Soil Contam. 8:527–540.
Campillo, N., P. Vinas, I. Lopez-Garcıa and M. Hernandez-Cordoba. 1998. Direct Determination of copper and zinc in cow milk, human milk and infant formula samples using electrothermal atomization atomic absorption spectrometry. Talanta 46:615–22. Charya, N. S., C. T. Kamala and D. S. Raj. 2008.
Assessing risk of heavy metals from consuming food grown on sewage irrigated soils and food chain transfer. Ecotoxicol. Env. Safe. 69:513-524.
Conti, M. E. 1997. The content of heavy metals in food packaging paper boards: An atomic absorption spectroscopy investigation. Food Res. Internat. 30(5):343–348.
Dhanalakshnmi, B. and G. Gawdaman. 2013. Determination of heavy metals in goat milk through ICP-OES. Asian J. Food Resid. 32(3):186-190.
Dudka, S. and W. P. Miller. 1999. Permissible concentrations of arsenic and lead in soils based on risk assessment. Water Air Soil Poll. 113:127–132.
Duruibe, J. O., M. O. Ogwuegbu and J. N. Egwurugwu. 2007. Heavy metal pollution and human biotoxic effects. Internat. J. Phys. Sci. 2(5):112-118.
EC, 1998. Commission regulation 98/83/EC of 3 November 1998 on the quality of water intended for human consumption. Official Journal of the European Union, L330, 32-54. EC. 2006. Commission regulation (EC) No
1881/2006 of 19 December 2006 setting maximum levers for certain contaminants in foodstuffs. Official J. Eur. Union, L364, 5-24. FAO/WHO, 2011. Food standards programme codex committee on contaminants in foods. Fifth session. The Hague, The Netherlands, 21-25 March 2011.
Gradinaru, A. C., O. Popescu and C. Solcan. 2011. Variation analysis of heavy metal residues in milk and their incidence in milk products from Moldavia, Romania. Env. Eng. Manage. J. 10(10):1445-1450.
Iyengar, G. V. 1984. Reference values for elemental concentrations in some human samples of clinical interest: A preliminary evaluation. Sci. Total Env. 38:125–131. Jameel, A. B., G. K. Tasneem, B. A. Muhammad,
I. A. Hassan, A. K. Ghulam, A. S. Raja, K. J. Muhammad and Q. S. Abdul. 2009. Evaluation of arsenic and other physico-chemical parameters of surface and ground water of Jamshoro, Pakistan. J. Hazard. Mat. 166:662–669.
Jorhem, L., S. Slorach, B. Sundstrom and B. Ohlin. 1991. Lead, cadmium, arsenic and mercury in meat, liver and kidney of Swedish pigs and cattle in1984– 88. Food Addit. Contam. 2:201 – 12.
Kazi, T. G., N. Jalbani, J. A. Baig, G. A. Kandhro, H. I. Afridi, M. B. Arain, M. K. Jamali and A. Q. Shah. 2009. Assessment of toxic metals in raw and processed milk samples using electrothermal atomic absorption spectrophotometer. Food Chem. Toxicol. 47:2163–2169.
Kincaid, R. L. and J. D. Cronrath. 1992. Zinc concentration and distribution in Mammary secretions of peripartum cows. J. Dairy Sci. 75:481–490.
Krachler, M., E. Rossipal and K. J. Irgolic. 1998. Trace elements in formulas based On cow and soy milk and in Austrian cow milk determined by inductively coupled plasma mass spectrometry. Biol. Trace Elem. Resid. 65:53– 7.
Licata, P., D. Trombettab, M. Cristanib, F. Giofre, D. Martinoa, M. Calo and F. Naccaria. 2004. Levels of ‘‘toxic’’ and ‘‘essential’’ metals in samples of bovine milk from various dairy farms in Calabria, Italy. Env. Internat. 30:1–6. Maas, S., E. Lucot, F. Gimbert, N. Crini and P.
M. Badot. 2011. Trace metals in raw cows’
milk and assessment of transfer to Comté
cheese. Food Chem. 129:7-12.
Munoz, E. and S., Palmero. 2004. Determination of heavy metalsinmilk by potentio metric stripping analysis using a home-made flow cell. Food Cont. 15:635–641.
Rahimi, E. 2013. Lead and cadmium concentrations in goat, cow, sheep, and buffalo milks from different regions of Iran. Food Chem. 136:389–391.
Rehana, A., G. K. Tasneem, K. J. Muhammad, B. A. Muhammad, D. W. Muhammad, J. Nusrat, I. A. Hassan and Q. S. Abdul. 2009. Variation in accumulation of heavy metals in different verities of sunflower seed oil with the aid of multivariate technique. Food Chem. 115:318– 323.
Ryan, B. F., B. L. Joinerand and J. D. Cryer. 2005. MINITAB Handbook: Updated for release 14, 5th edition.
Salah, F. A., I. A. Esmat and A. B. Mohamed. 2013. Heavy metals residues and trace elements in milk powder marketed in Dakahlia governorate. Internat. Food Res. J. 20(4):1807-1812.
Salma, I., W. Maenhaut, S. Dubtsov, E. Z. Papp and G. Zaray. 2000. Impact of phase out of leaded gasoline on the air quality in Budapest. Microchem. J. 67:127–133.
Salvato, N., G. Banfi, F. Taccani, A. Rottoli and G. Banderar. 1992. Direct analysis of essential elements (chromium, copper, iron, manganese, selenium) inhuman milk by Zeeman GFAAS. Appl Zeeman Graphite Furn At Absorpt Spectrom. Chem. Labor. Toxicol. 305– 323.
Silanikovea, N., G. Leitnerb, U. Merinc and C. G. Prosserd. 2010. Recent advances in exploiting goat’s milk: Quality, safety and production aspects. Small Rum. Res. 89:110–124.
Simsek, O., R. Gultekin, O. Oksuz and S. Kurultay. 2000. The effect of environmental pollution on the heavy metal content of raw milk. Nahrung-Food 44:360–371.
Solis, C., K. Isaac-Olive, A. Mireles and M. Vidal-Hernandez. 2009. Determination of trace metals in cow’s milk from waste water irrigated areas in Central Mexico by chemical treatment coupled to PIXE. Microchem. J. 91:9–12.
Soylak, M., S. Saracoglu, M. Tόzen and D. Mendil. 2005. Determination of trace metals in
mushroom samples from Kayseri, Turkey. Food Chem. 92:649–652.
Tabinda, A. B., S. Zafar, A. Yasar and S. Minur. 2013. Metals concentration in water, fobber, milk, meat, blood, kidney and liver of livestock and associated health impacts by intake of contaminated milk and meat. Pak. J. Zool. 45(4):1156-1160.
Tasneem, G. K., Nusrat J. Jameel, A. B. Ghulam, A. K. Hassan, I. A. Mohammad, B. A. Mohammad and K. J. Abdul. 2009. Assessment of toxic metals in raw and processed milk samples using electrothermal atomic absorption spectrophotometer. Food Chem. Toxicol. 47:2163–2169
Temiz, H. and A. Soylu. 2012. Heavy metal concentrations in raw milk collected from different regions of Samsun, Turkey. Internat. J. Dairy Technol. 65(4):516-523.
Tona, G. O., V. O. Adetunji, S. A. Ameen and A. O. Ibikunnle. 2013. Evaluation of lead and cadmium heavy metal residues in milk and milk products in Ogbomoso, Southewestrn Nigeria. Pak. J. Nutr. 12(2):168-171.
Tuzen, M., S. Saracoglu and M. Soylak. 2008. Evaluation of trace element contents of powdered beverages from Turkey. J. Food Nutr. Res. 47:120–124.
Vahid, N. and A. Masoome. 2013. Heavy metals in raw cow and ewe milk from north-east Iran. Food Addit. Contam. 6(3):158-162.
Varian, M. 1989. Flama Atomic Absorption Spectroscopy. Analytical Methods. Varian Australia. Publ. N0: 85-100009-00.
