THE SUMMARY OF DOCTORAL THESIS · 2011. 9. 29. · THE SUMMARY OF DOCTORAL THESIS RESEARCH REGARDING THE CONSEQUENCES OF MINERAL POLUTANTS ON VARIOUS TISSUES AND ORGANS IN CATTLE

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THE SUMMARY OF DOCTORAL THESIS

RESEARCH REGARDING THE CONSEQUENCES OF MINERAL POLUTANTS ON VARIOUS TISSUES AND ORGANS IN CATTLE SLAUGHTERED FOR HUMAN CONSUMPTION

2

General considerations

The existence, until 2004 of a highly polluted area in Zlatna, around the

chemical plant, has made possible the release in the environment of various pollutants

such as heavy metals more or less toxic which, due to their long lasting remanence,

have negatively influenced the living organisms.

The soils, due to their stationary features, have a significant role in retaining

and accumulating pollutant factors. Mineral pollutants like Lead, Cadmium, Copper,

Zinc from the soil, are absorbed very easily by the plants, accumulating in the roots

and leaves, being as such, intaken by cattle, which constitute the link between plants

and humans, but also in products destined for human consumption. The combination

of heavy metals, minerals, and oligo-minerals results in blockants, frustrating the

organisms of these elements indispensable to life.

Regardless of the contamination source (air, soil, plants, water), the toxic efects

are as harmfull to human health, as to life in general. As a consequence, polution,

through all its forms, must be fought against, and its efects removed, because the

chemical agression on human beings and animals is oriented not just towards the

environment, but to themselves.



3

PART I

CHAPTER 1.Mineral pollutants and their implications on the enviroment,

animals and food

1.1 . Classification of mineral pollutants

1.2 .Mineral pollutants toxicity and mechanisms of toxicity.

1.3 .Transport and body action of toxins in body.

1.4 .Mineral pollutants mode of action on the environment (water, soil, plants, animals

and human body)

1.4.1.Lead. Food contamination and its action on animal and human.

1.4.1.1 .General overview

1.4.1.2 .Lead contamination of food.

1.4.1.3 .Lead action to the animals.

1.4.1.4 .Lead action to the human.

1.5.1. Cadmium. Food contamination and its action on animal and human.

1.5.1.1. General overview

1.5.1.2. Cadmium contamination of food.

1.5.1.3. Cadmium action to the animals.

1.5.1.4. Cadmium action to the human.

1.6.1. Copper. Food contamination and its action on animal and human.

1.6.1.1. General overview

1.6.1.2. Copper contamination of food.

1.6.1.3. Copper action to the animals.

1.6.1.4. Copper action to the human.

1.7.1. Zinc. Food contamination and its action on animal and human.

1.7.1.1. General overview.



4

1.7.1.2. Zinc contamination of food.

1.7.1.3. Zinc action to the animals.

1.7.1.4. Zinc action to the human.

Chapter II. Storage organs and tissues of mineral pollutants and lesions induced

by these.

2.1. Main storage organs and tissues of lead and lesions induced by it.

2.2. Main storage organs and tissues of cadmium and lesions induced by it.

2.3. Main storage organs and tissues of copper and lesions induced by it.

2.4. Main storage organs and tissues of zinc and lesions induced by it.

Chapter III. Bibliographical data regarding soil, vegetation, water, and air in

Zlatna area

3.1. Presentation of polluted area

3.1.1.The pollution source of the Ampoi basin

3.1.1.1. .Source of the air pollution

3.1.1.2. Source of the soil pollution and spontaneously vegetation



5

Structure of personal research

The thesis comprises a number of 273 pages and is structured according to

legal previsions; there are 82pictures (images, graphics, diagrams) and 77tables. In

part I there are presented the most important and conclusive data from specialty

literature referring to mineral pollutant’s toxicity and toxicity mechanisms on the

environment, the lesions they induce in the main organs and tissues, and data

regarding pollution sources in Zlatna area. In part II there are presented personal

research that had in view the manner of organizing the research (chap.IV), monitoring

mineral pollutants from soil, water, air and spontaneous vegetation in Zlatna area

(chap.V), evaluation of biological consequences of pollutant factors on tissues and

organs in cattle slaughtered for human consumption (chap.VI). The bibliographical

part comprises a number of 151 titles from foreign and Romanian specialty literature.

In chapter IV there are described the importance, the purpose and objectives

proposed for the research, as well as the materials and work methods. The importance

of the study resides in the fact that the theme gives the poossibility to open new

knowledge perspectives, and the results could be applied to asses the polution degree

in the area, to identify the lessions and malfunctions induced by polutants, and the

assesment of their gravity related to concentration in tissue and organ sampkles. The

research theme includes the results obtained in Zlatna area up to the present, regarding

the degree of accumulation of heavy metals in soil, vegetation, water and air, as well

as the results regarding polution consequences on the cattle stock in this area.

The purpose of this study as been:

- evaluation of biological consequences of poluting factors on tissues and

organs in cattle slaughteres for human consumption, raised in Zlatna area, area in wich

the concentrations of mineral polutants (Lead, Cadmium, Copper, Zinc) have been



6

raised levels compared to the maximum limits, both in the soil, and in the air and

water;

- the study of relationships between the high level of polutant factors in the

tissue, and the resulted lesions;

- coroboration of our data with the legal regulations regarding the maximum

levels of polutants, and the normal polutant concentrations mentioned in the specialty

literature from the bibliography.

The work material can be structured as follows:

1. To evaluate the degree of soil, water, air and vegetation polution in Zlatna area,

during the functioning of the polution source (up till 2004), and after the polution has

stopped (2005-2008):

• Monthly data regarding monitoring of mineral polutants (Lead, Cadmium, Copper)

in soil, water, air and spontaneous vegetation taken from The Environment

Protection Agency Alba, and poublic information we have processed and

identified the following parameters:

- the number ofsamples that have overcome the maximum allowed concentration;

- the frequency of overcoming of maximum allowed concentration;

- average concentration;

- monthly and yearly average concentration;

- maximum concentration in the respective month and year;

- data are presented as tables and processed as graphics.

The concentrations measured have been processed and assesed in accordance with

present legislation at the time, and normal concentratins mentioned in the papers from

the biblioigraphy.

2. To evaluate the biological consequences of polutant factors on tissues and organs

investigated for cattle slaughtered for human consumption:

Cattle raised in individual farms in the polluted area, which have been

slaughtered for human consumption.



7

• Organ and tissue samples in order to performe quantitative analysis for Lead,

Cadmium and Copper, and highlights of histopathological changes in organs

and tissues from the same cattle slaughteres for consumption, from the

mentioned areas; the data have been processed and used for an applied

discussion related to quantitative analysis of mineral polutants of tissues and

organs from cattle and lessions identified in the examined tissues.

Work methods: I Qantitative analysis of mineral polutents like heavy metals from soil, water, air

organs and tissues has been performed through spectophotometric methods, using the

atomic absorbtion spectrophotometer.

II The histopathological exams have been performed on the following types of tissues

and organs: miocard, liver, kidney, lung, spleen, uterus, mammary gland, to identify

microscopic lessions.

In ordewr to evaluate the degree of contamination of tissues from examined

cattle, the analysed concentrations have been compared with the maximum levels

regarding the maximum quantities of polutants according the Reg. no. 629/2008 of the

Comission, which modifies Reg. 1881/2006 which establishes the maximum degrees

for some contaminants from food, valid since December 2006, and with the normal

values specified in the biblioigraphy.

In chapter V, called Data regarding mineral polutants in soil, water,

spontaneous vegetation in Zlatna areawe describe the degree of polution given by the

polutant concentrations from soikl, water, air and spontaneous vegetation during the

functioning of the polutant source, and after it ceised its activity.

In order to evaluate the degree of pollution of the area, the data has been

processed for mineral pollutants from soil, water, spontaneous vegetation between

2005-2008.

During the functioning of the pollution source as well as after SC Ampelum

Zlatna ceased its activity, there have been measured in the soil, high levels of



8

pollutants. For Lead in the soil, between 2005-2008, the average values have been

between 688, 4 mg/kg. During the functioning of the plant the levels of concentration

of Lead, Cadmium, Copper in vegetation overcame the maximum allowed limits,

measuring overcomings in the grass of 31 times for Lead, 41 for Cadmium and 1,1 for

Copper. Between 2005-2008 the concentrations of heavy metals in spontaneous

vegetation have reached overcomings of 6 times for Lead (60,37 mg/kg) in the center

of Zlatna up to 37 times for Copper (741 mg/kg) in Izvorul Ampoiului, and up to 25

times for Cadmium (10,03 mg/kg) in Izvorul Ampoiului.

The values of the indicators measured in the air (powders in suspension in 24

h) have not overcame the maximum limits, and for Lead 24 h, Cadmium 24 h, Copper

24 h, and Zinc 24 h, the values have been within the range of maximum allowed limits

according to Ord.M.A.P.M 592/2002 for 2005-2008. Ampoi River frames in 2005 and

2006 in class II of quality due to the overcome concentrations in Cadmium and Lead,

salinity indicators and high content of nutrients. In 2007 and 2008 it has framed in

class I of quality.

In chapter VI, called Evaluation of poluting factors consequences on various

tissues and organs in cattle slaughtered for human consumption we present the

concentrations of Lead, Cadmium, Copper found in liver, kidney, muscle tissue, lung,

spleen, mamary gland and the tissue lesions we identified.

Cattle raised in private households in Zlatna-Patranjeni area, situated within the

range of direct influence of the pulutant source, and feed constantly fith foder from

this area, have been slaughtered in normal conditions, in slaughter houses in Alba

County, between 2006-2008. From these cattle, we have sampled muslce tissue, heart,

lung, liver, spleen, kidney, mamary gland, uterus.

The organs have been macroscopically examined,to identify possible lesions.

Parts of the harvested organs have been set in formalin 10% to be prepared through

the histological technique in paraffin and coloured with the coloring methods eozin-



9

hematolizine and hematolosine blue to allow identifying microscopical lesions. The

rest of the organ portions have been analysed quantitatively through spectophotometry

with atomic absorbtion to find heavy metals (Lead, Cadmium, Copper).

The quantitative results for Lead, Cadmium, Copper, found for the organs and

tissues of cattle slaughtered for human consumption have been analysed in

comparison to the maximul allowed concentrations from Regulation 629/2008 of the

Comision, to modify the Regulation 1881/2006 which sets the maximum levels for

certain contaminants from food. This establishes that: in case of Lead for meat coming

from catle, ovines, pork, poultry, a maximum level of 0,50 mg/kg, in case of

Cadmium for meat 0,050 mg/kg , for liver from cattle ovine, pork, poultry and

equines, 1,0 mg/kg.

DISCUTION OF THE RESULTS

The investigations and the results allow us some observations and discutions

LIVER Table 1

The values of Lead,Cadmium and Copper concentrations in the liver of slaughtered cattle

CATTLE

LIVER

LEAD mg/kg CADMIUM mg/kg COOPERmg/kg

Case 1 0.068 0.306 3.80 Case 2 0.589 0.235 9.84 Case 3 Ned. 0.402 2.50 Case 4 Ned. 0.584 2.84 Case 5 Ned. 1.383 1.83 Case 6 Ned. 0.327 3.30 Case 7 Ned. 0.591 2.20 Case 8 0.163 0.919 4.50

The avarage value 0,1 0,520 3,85 LMA/maximum

permitted concentration

0.5 0.5 3.0

* The normal value

0,5 0,3 5,7

Normal value * (after MITRANESCU 1990/Normal values)



10

Fig. 1. Graphic representation of concentrations for lead, cadmium in the liver at cattle

examined against maximum limit allowed

Analysing the average values for Lead, Cadmium and Copper in liver, we notice

that the average value of Copper (3,85 mg/kg ) in 8 cases, is over the maximum value

allowed by Codex Alimentarius (3,00 mg/kg) and under the normal value of Copper

in liver mentioned by the literature (5,7mg/kg) .

For Cadmium the average value is equal to the maximum limit allowed by the

regulations (0,520 mg/kg) overcoming though the normal value which, after

Mitranescu 1998, is 0,30mg/kg and under the normal value (0,75mg/kg) mentioned by

Ghergariu (1980 ).The average value measured for lead is under the maximum limit

allowed by regulations and under the normal value mentioned in the specialty

literature.

On cases, the situation present itself as follows:

Lead measured from liver from the 8 cattle analysed has registered overcomings

compared to CMA (0,5 mg/kg ) in a single case (0,589mg/kg). We can not

neglect the presence of Lead in the liver for one more case (value of 0,163

mg/kg) ;

Cd, Caz 1, 0.3 Cd, Caz 2, 0.23

Cd, Caz 3, 0.4

Cd, Caz 4, 0.58

Cd, Caz 5, 1.38

Cd, Caz 6, 0.32

Cd, Caz 7, 0.59

Cd, Caz 8, 0.91

mg/kg

Pb

Cd

LMA Pb

LMA Cd



11

• in case 2 (0,589mg/kg) ;

• in case 8 a significant value of 0,163 mg/kg ;

Cadmium from the liver o the 8 cattle has registered overcomings of CMA

(0,5mg/kg) in 4 cases and has been detected, even if there was a lower

concentration than CMA for the rest of the cases: :

• case 4 value of 0,584 mg/kg ;

• case 5 with a value of 1,383mg/kg ;

• case 7 with a value of 0,591 mg/kg ;


Copper from liver has overcomings of CMA (3mg/kg) in 4 cases and has been

detected in lower concentrations in the rest of the cases.





Analysing the values of Lead, Cadmium and Copper from the liver of the 8 cattle and

comparing the values with the maximum limits allowed by the regulations, we can

see:

• the presence of Cadmium in the liver for all the 8 cases, and in cases 4,5,7,8 the

values have overcome the maximum limit allowed by the legislation, and 7 cases

are over the normal value from the specialty literature;

• The Lead has been present in the liver of 3 cattle, in one case overcoming the

maximum limit allowed by the current legislation;

• Copper has been identified in the liver of all cattle, but just in 4 cases it has

overcame the allowed limit, and in just one case the normal value is overcame;

• In conclusion, liver is an organ of accumulation for Copper, Lead, Cadmium

showing the cummulating effect of theese and causing lesions and malfunctions.

RESEARC

Histo

taken

case

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some

3) bi

case

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(case

Fig.4. Ca

1. A

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an

CH REGARDIAND ORG

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7) or the e

ei are vesi

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ig liverid (

3) obvious

plasma with

ations, rare

e1,case 5,ca

ase 1,Liver: v

As a conclu

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Copper, Lea

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asma, some

ifferent size

case7); obv

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ase 6) hepat

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is in liverid

SUMMARY O

NSEQUENCETTLE SLAUG

he whole, th

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ith the nuc

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giving a ha

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tosis and ce

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microscopic

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2 cases in 8

dmium accu

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d cytoplasm

OF DOCTORA


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liverid pres

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th rare red

alo aspect,

lipoproteic

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image of t

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8).

umulation

tifies the di

a.

AL THESIS


e of liver is

sent small v

ted (case

posed centr

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e (case 2) o

omatic cells

ith no alter

cells (case

hepatosis;

infiltration

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Fig.3

la

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with high

iagnosis of


s kept to al

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islands – co

n in liverid

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3.Case 1, Liv

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concentrati

f lipo-protei

IOUS TISSUEN

1

l the 8 case

ase1, case 5

case 5). Th

the nuclea

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(case 3) o

vessel (cas

se 2, case

, big liverid

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13

KIDNEY Table2

The values of concentration for Lead, Cadmium and Copper in the kidney of slaughtered cattle

Fig. 5. Graphic representation of the lead and cadmium concentrations in kidneys at

examined cattle compared to LMA

Analysing the average values for Lead, Cadmium, Copper from kidneys, we

can see that in the case of Lead, the average value of 0,32 mg/kg is under the

maximum limit allowed by the regulations and under the normal value of Lead in

kidneys (0,5 mg/kg) after Mitrănescu (1998) and is equal to the normal value 0,32

CATTLE KIDNEY

LEADmg/kg CADMIUM mg/kg COOPERmg/kg

Case 1 0.104 1.38 2.83 Case2 0.300 1.0 1.97 Case 3 Ned 1.0 1.85 Case 4 0.781 1.049 2.85 Case 5 0.245 1.458 1.97 Case 6 Ned 1.321 1.77 Case 7 1.194 0.703 2.30 Case 8 Ned 1.203 2.0

The avarage value 0,32 1,13 2,19 maximum permitted

concentration 0.5 1.0 3

The normal value* (after MITRANESCU 1990)

0,5 0,5 12,6

Pb, Caz 1, 0.1

Pb, Caz 2, 0.3

Pb, Caz 3, 0

Pb, Caz 4, 0.78

Pb, Caz 5, 0.24

Pb, Caz 6, 0

Pb, Caz 7, 1.19

Pb, Caz 8, 0

Cd, Caz 1, 1.38

Cd, Caz 2, 1 Cd, Caz 3, 1

Cd, Caz 4, 1.49 Cd, Caz 5, 1.45 Cd, Caz 6, 1.32

Cd, Caz 7, 0.7

Cd, Caz 8, 1.2

Pb

Cd

LMA Pb

LMA Cd

mg/kg



14

mg/kg mentioned by the specialty literature. The normal value of Lead in the kidneys

for adult cattle is 0,42 mg/kg.

The average value of Cadmium in kidneys of 1,13 mg/kg. Is over the maximum

limit allowed by legislation (1 mg/kg) and over the normal value od Cadmium in the

works from the bibliography (0,5mg/kg). In the case of Copper, the average

concentration in the kidneys of 2,19 is under the maximum concentration allowed by

the legislation (3mk/kg) and under the normal value mentioned in bibliographical data

(12mk/kg) .

On cases, the situation is as follows:

Lead in kidneys is overcome for 2 cattle, compared to CMA (0,1 mg/kg) and

has been detected, even if with small values, under the maximum allowed limit, for

other 3 cases.


• case 7 with a value of 1,194 mg/kg , an overcoming in this case of 2,3 times;

Cadmium Cadmiul in kidneys is overcome or at the maximum allowed limit (1,0

mg/kg) fnext 7 cases:

• case 1- with a value of 1,38mg/kg;

• case 2 and case 3 with a value of 1,0 mg/kg;

• case 4 with a value of 1,049 mg/kg;

• case 5 with a value of 1,458 mg/kg in this case the value is 1,4 times overcome;

• case 6 with a value of 1,321 mg/kg , 1,3 times overcomed;

• case 8 with an overcoming of CMA of 1,2 times (a value of 1,2 mg/kg);

In a case, Cadmium had a value of 0,7 mg/kg situated under the maximul

concentration allowed by present regulations.

Copper in kidneys has no previsions for the maximum limit in the regulation.

The normal concentration of Copper in liver is 12,6 mg/kg. Copper has been detected

in all the 8 cattle. All the concentrations for the 8 cases are under this value.

In case of the kidney the values of Lead, Cadmium and Copper found in

quantitative enalysis reveal the presence of Cadmium in the kidney of all the 8cattle in

RESEARC

study, an

the valu

in 5 case

overcam

lower co

W

Cadmium

for 8 ca

their cum

H

vacuoliz

straight

microcy

the epite

epitelium

epitelium

nuclei in

of glom

epielium

FGC

CH REGARDIAND ORG

nd for 7 the

e was unde

es (case1, c

me the maxi

oncentration

We can conc

m and Cop

ases from 8

mulating ac

Histological

zations (wh

tubes, wit

ysts and lise

elium is atro

m forming c

m tubes, lik

n picnosos

meruls (case

m of the tube

ig.6.,Case 6Glomerular cCytolysis of p

THE S


e values hav

er the maxim

ase2, case4

imum allow

ns.

clude that i

pper have b

8, two elem

ctin resultin

l lesions:

hite cytopla

h cytorexic

ed cells (cas

ophied (cas

cellular cili

ke biliar co

(case 2, cas

e 6) the sam

es with vac

,Kidney: renapsule thickpodocytes

SUMMARY O

NSEQUENCETTLE SLAUG

ve overcom

mum limit.

4, case5, cas

wed limit, f

n kidney, in

been simulta

ments, Cadm

g in serious

we can se

asma) (cas

c (case 1

se 2) in som

e2, case1),

inders ( cas

olourings (c

se 7) oxiph

me content

uolae and c

nal glomerulkening, imag

OF DOCTORA


me or were e

Kidney ha

se7). In two

for the rest

n 5 cases fr

aneously fo

mium and C

s lesions.

ee alteration

e 2), vacu

, case 2, c

me of the tu

images of

se 2, case 3

case 1), or

hylic conten

t in the ma

cytorexis (c

le, Fies of

AL THESIS


equal to CM

as been the

o cases (cas

of the case

rom 8 all th

ound, in low

Copper ar s

ns of the

uolized epit

case 3, cas

ubes the lum

f catarrh wit

), fine gran

thikening o

nt with seric

ajority of

ase 1) .

ig.7.,Case 3,K epithelium


MA. In a sin

place to sto

e 7 si case 4

es Lead wa

he three elem

wer or high

simultaneou

tubular epi

telium in s

se 4 , case

menum is e

th the detac

nulatins in c

of pedocitis

c aspect in

contort tub

Kidney: vacm

IOUS TISSUEN

1

ngle case (7

ore the Lea

4) the value

as present, i

ments, Lead

her amount

usly presen

itelium, lik

some of th

e 7, case 8

enlarged an

hment of th

cytoplasm o

s cytoplasm

the majorit

bes (case 6

cuolated

ES

15

7)

ad

es

in

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ts,

nt,

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he

8)

nd

he

of

m,

ty

6)



16

1) The kidney presents lesions of tubulonephrosis type hydroprotidic and catarrh

to 5 of the 8 cases, granulo-vacuolar degeneration and intra-tubular and intra-

glomerular edema in 2 of the 8 cases.

2) The relation between the accumulation of Copper, Lead, Cadmium in kidneis in high concentrations, the cummulative and simultaneous efect and the severe lesions is more than obvious.

LUNG Table3

Values for lead and cadmium concentrations in the lung from slaughtered cattle

Analysing the average Lead value in the lung of all the 8 cattle, we can see

that this is at the normal value mentioned in bibliography, and under the maximum

allowed concentration 0,5 mg/kg. Cadmium has been detected in the lungs of all the 8

cattle, with values between 0,040 and 0,685mg/kg. The average value of Cadmium in

the lung is 0,347mg/kg .

On cases, the situation is as it follows:

Lead found in lungs has no overcomings for any of the cases, compared to

CMA (0,5 mg/kg) but has been detected even in small amounts under the maximum

limit in 4 cases. In the other 4 cases, Lead has not been detected in lungs.

CATTLE LUNG

Lead mg/kg Cadmium mg/kg

Case 1 0.032 0,040 Case 2 0,234 0,395 Case 3 Ned 0,300 Case 4 Ned 0,397 Case 5 Ned 0,307Case 6 Ned 0,120 Case 7 0,250 0,685

Case 8 0,220 0,539 The average value 0,09 0,347

Reg./ LMA/ maximum aloowed concentration

0.5 No limit established

Normal value *(after MITRANESCU 1990)

0,5 -

RESEARC

C

values w

• in ca

• in ca

• in ca

Copp

H

granulat

thickenin

emphyse

alveolae

peribron

pneumon

(case 7,

In

mg/kg a

value di

and in 4

Fw

CH REGARDIAND ORG

Cadmium f

were: la case

ase 5 with a

ase 7 with a

ase 8 with a

per has not

Histological

tions, some

ng of alveo

ema with la

e septi an

nsiolitis and

nia (case 7)

case 4).

n lungs, va

nd case 8 w

d not overc

cases Lead

ig .8, Caz 2,with alveolar

THE S


found in th

e 4 with a v

value of 0

value of 0

value of 0

t been analy

l lesions:

e fagocytes

olae septi w

arge spaces

nd formati

d bronchic

), peribrons

alues of Ca

with 0,53 m

come the m

d has not be

,Lungs: empseptums thi

SUMMARY O

NSEQUENCETTLE SLAUG

he lungs ha

value of 0,3

0,307mg/kg

0,685 mg/kg

0,539mg/kg

ysed from lu

in perib

s (case 1)

with lymph

s and fragm

ion of lar

catarrh (ca

siolic infiltr

admium we

mg/kg) and

maximul allo

een detected

physema nning

OF DOCTORA


as been det

397 mg/kg;

;

g;

;

ungs.

bronsiolic

with histo

hohistiocites

mented septi

rge cavitie

ase 2, case

ation (case

ere overcom

Lead has be

owed limit

d.

Fig.9, C and per

AL THESIS


tected to al

and periar

ocytary rea

s (case 1, c

i (case 2, ca

es (case

4), fibrotic

e 1), peribro

me in 2 ca

een detecte

(case 1, cas

Caz 2, Lungsribronchiolit


ll 8 cases.

rterial t

action aroun

case 2, case

ase 3 , case

1) limph

c and limph

onsiolic antr

ases (case

d in 4 cases

se 2,case 7

s: bronchiolutis and break

IOUS TISSUEN

1

The highe

tissue blac

nd (case 1

e 3, case 7

e 6), brocke

hohistocitar

hohistocitar

racosis spo

7 with 0,6

s, even if th

and case 8

us catarrh king

ES

17

st

ck

),

7),

en

ry

ry

ts

68

he

8)

RESEARC

1) The l

pulm

2) The t

than

hysto

M

Value

Fig. 1

tissue

C

CCCCCCCCTLcNM

CH REGARDIAND ORG

lesions cha

monary emp

toxic and ir

obvious, c

ocitary perib

MUSCLE TI

es for Lead a

10. Graphic

e from cattl

Cattle

Case 1 Case 2 Case 3 Case 4 Case 5 Case 6 Case 7 Case 8 The average valuLMA/ maximconcentration Normal vaMITRANESCU 1

THE S


aracteristic f

hysema, fi

rritating act

ausing fibr

bronsiolitis

ISSUE

and Cadmiu

c representa

le examined

ue mum allowed

alue *(after1990)

SUMMARY O

NSEQUENCETTLE SLAUG

for the histo

ibrosis due t

tion of Lea

rosis lesions

s.

m concentra

ation of conc

d against LM

Muscle tissue

LEAD mg/kg

0.1 0.266 0.301 0.10 0.011 0.402 0.380 0.102 0,207

d 0.1

r 0,25

OF DOCTORA


ological exa

to antracosi

ad and Cadm

s, alveolary

ations in mus

centrations

MA

e

g

AL THESIS


am of the lu

is, lymphoh

mium at the

y emphysem

Tscl tissue fro

for Lead, C

CADMIUM /mg/

0.024 0.354 0.434 0.457 0.017 0.307 Ned 0.548 0,26 0.05

0,1


ung to all th

histocitary p

e level of lu

ma, spots o

Table 4 om cattle slau

Cadmium in

/kg

IOUS TISSUEN

1

he 8 cattle

pneumonia.

ungs is mor

of antracosi

ughtered

n muscle

ES

18

is

.

re

s,



19

Analysing the average concentrations for Lead in muscle tissue, we can see that

the average value of 0,207 mg/kg is 2 times higher than the maximum allowed

concentration (CAM) of (0,1 mg/kg) and smaller than the value of 0,25 mg/kg normal

value (96).

The average concentration of Cadmium from muscle tissue of 0,26 mg/kg is 2

times higher from the normal value of 0,1 mg/kg mentioned in the specialty literature

and overcomes 5,2 times the maximum allowed concentration of 0,05 mg/kg(96).

On cases, the situation presents itself as follows:

The Lead in muscle tissue from the examined cattle has been overcome from CMA

(0,1 mg/kg ) or has been at the maximum limit in 7 cases: in cases 1,4 and 8 the value

of 0,1mg/kg is equal to the maximum allowed limit;

• in case 2 the value of 0,26 mg/kg is 2,6 times higher than CMA;

• in case 3 the value of Lead is 3 times higher than CMA;

• in case 6 the value of Lead is 4 times overcome compared to the maximum

allowed concentration;

• in case 7 Lead has 3,8 times overcome CMA;

Cadmium in muscle tissue has proved raised levels in the following cases:

• in case 2 Cadmium value is 7 times higher than CMA

• in cases 3 and 4 Cadmium value in muscle tissue is 8 times higher than

CMA;

• in case 6 Cadmium value in muscle tissue is 6 times higher than CMA;

• in case 8 Cadmium value in muscle tissue is 10 times higher than CMA;

The microscopic examination of muscle tissue has not proven any changes. In

specialty literature we find mentions about the chronical intoxications, there is a

deposit of Cadmium in organs, without histological changes (96). Cadmium in excess

increases the toxicity of Lead ( 96).

1) High concentrations of Lead and Cadmium in muscle tissue for the 8 cases results

from the fact that those animals come from a poluted area, and muscle tissue of

those animals is a risk factor for humans consuming it.

RESEARC

2) We

Cad

and

S

Valuslaug

CH REGARDIAND ORG

can obser

dmium polu

Lead in mu

PLEEN

ues of concghtered

CATTLE

Case 1 Case 2 Case3 Case4 Case5 Case6 Case3

Case3 The average

LMA ReThe normal valuMITRANESCU

THE S


rve a core

utants in soi

uscle tissue

centration fo

E

value eg. ue *(after U 1990)

SUMMARY O

NSEQUENCETTLE SLAUG

lation betw

il, water an

.

for Lead, C

Lead mg/kg

0,050 0,526 ned ned

0,308 0,505 ned

Ned 0,170

0.5 0,25

OF DOCTORA


ween the h

nd vegetatio

Cadmium an

Cadm

No lim

AL THESIS


high conce

on and conc

nd Copper

SPLEEN

mium mg/kg

0,031 0,430 0,518 0,028 0,303 Ned

0,336

0,237 0,234

mit is provided 0,1


ntrations o

centrations

Tin the sple

Cooper m

1,021,3 1,301,7 1,6 1,9 2,2

1,5 1,56

No limit is p

IOUS TISSUEN

2

of Lead an

of Cadmium

Table 5 een of cattl

mg/kg

2

0

6 provided

ES

20

nd

m

le



21

Fig.11. Concentrations of Lead, Cadmium and Copper in the spleen of examined

cattle Analysing the average value for Lead in spleen (0,170 mg/kg) for the 8 catle,

we can see that it is 2 times higher than the normal value mentioned in bibliography

(0,08 mg/kg). The average value of Cadmium in spleen (0,234 mg/kg) is significant,

having in view that in spleen, both Cadmium and Lead, are present in small amounts.

On cases, the situation is as follows:

Lead from spleen is 2 times overcome for 2 cattle compared to CMA (0,5

mg/kg) and has ben detected in small amounts, under the maximum allowed limit for

3 cases:



Cadmium has been detected in spleen in 7 cases, having various values. The

higher values were in the following cases:

• case 3 with a value of 0,5mg//kg ;



Copper has been detected in spleen in all the cases. For Copper and Cadmium

in spleen maximum limits have not been previsioned.

Histological lesions: spleen folicules circumcisedand structured (case 1, case

2, case 8 ) or less shaped, (case 3) small (case 6) massive red highly coloured pulp

(case 1, case 2, case 3, case 4), in red pulp fine granulations (antracosis) (case 1) non

aboundant colouring in rough parts (case 8) red pulp with hemosiderinic colouring

(case 7, case 8), small folicules, hypertrophy endotelium arterioles proliferating

towards the lumenum (case 7).



22

1) The lesions characteristic for the histological exam of the spleen for the 8 cattle

are hemosyderosis in 4 cases snd hypertrophya and hyperplasia of atrery

endotelium in 1 case.

2) High concentrations of Cadmium ald Lead found in spleen of 8 cattle proves

that spleen can be an accumulation organ alongside the liver, kidney, muscle

tissue.

Myocardium:

Cysts with sarcosporidium (case 1, case 5), arterilae with endotelial

proliferaton without lesons for the rest of the cases. In the heart there has been

identified, both the presence of Cadmium and Lead. Lead in the heart had significant

values in case 1, case 6 and case 8 and Cadmium was present in the heart in case 1, 2,

case 3, case 7 and case 8.

Mamary gland: with no content, big, with several cells or apoptotic bodies

(case 7), with no alterations for the rest of the cases.

GENERAL CONCLUSIONS

1. The study has proven that, although in Zlatna area every industrial activity has

stopped, heavy metal polution (Lead, Cadmium, Copper) resides in the soil,

vetetation, water, and air, because the soils accumulate and keep certain levels

of potentially toxic ions of heavy metals considered persistent polutants of the

environment.

2. In the area of maximum polution, between 2005-2008 the concentrations of

heavy metals in the superficial lairs of the soil overcome the maximul allowed



23

limits, according to the Order MAPPM 756/1997. The highest overcomings

have been shown in Lead, up to 13 times, Cadmium, 1,2 times, Copper, 6,7

times, and heavy metal concentrations in apontaneous vegetation have been

overcome 6 times the maximum allowed concentration in Lead, 37 times in

Copper and 25 times in Cadmium.

3. In 2005, 2006, 2007, 2008 the values for the measured indicators in the air

(suspension powders 24 h) did not registered overcomings of the maximul

allowed limits and Pb24h, Cd24h, Cu24h, Zn24h framed within the maximum

allowed limits. The values of monitored indicators in the air in 2005-2008,

after the polluting source ceased its activity framed within the standards,

according to Ord.592/2002 .

4. In 2006 Ampoi river framed in class II of quality due to its high concent of

nutrients (nitrates, nitrits), overcoming of salinity indicators (Chlorine,

Calcium, Magnesium) and to its content of hazardous substances (Lead,

Cadmium) which were unacceptable. In 2007 and 2008 Ampoi river framed

within class I of quality.

5. The quantitative measurments of mineral pollutants (Lead, Cadmium, Copper)

from tissues and organs of cattle raised in the vicinity of the pollutant source

hav proven overcomings of the maximum allowed concentrations according to

present legislation of 10 times in the liver, 9 times in the kidney and 12 times

in the muscle tissue.

6. Analysing the values of Lead, Cadmium and Copper obtained in the

quantitative measurments of the liver, we can see that Cadmium was found in

the liver of all 8 cattle and in 4 cases the measured values overcame the

maximum allowed limit from the regulations. Copper has been identified in

the liver of all cattle, but just in 4 cases overcame CMA.

7. At the level of the liver the maximum concentration was of 1,383mg/kg.

Maximum concentration of Lead - 0,589 mg/kg and maximum

concentration of Copper - 9,84mg/kg.



24

8. The microscopic image of the liver for the 8 cattle was dominated by

lipoproteic infiltration in liverid cytoplasma (3 in 8 cases) and lipoproteic

hepatosis (2 in 8 cases). Copper, Lead, Cadmium accumulation in liver in high

concentration, their cummulative efect is corelated with the diagnosis of

lipoproteic infiltration and hepatosis in liverid cytoplasma.

9. In case of the kidney, the values of Lead, Cadmium and Copper reveal the

presence of Cadmium in the kidney of all the 8 cattle, and for 7 of those, the

measured values overcame or equaled the maximum allowed concentration. In

a single case (7) the measured value was under the maximum limit.

10. The kidney is the stoking place for Lead in 5 cases. In two cases the measured

values overcame the maximum allowed limit, for the rest of them Lead was

present in small amounts. Kidneys present lesions of tubulonefrosis

hydroprotidic type and tubula catarrh to 5 of the 8 cases, granuvacuolar

degeneration and intraglomerular and intratubular edema in 2 from 8 cases.

11. The relation between the accumulation of Copper, Lead and Cadmium in

kidneys in high concentration, their cummulative and simultaneous efect and

drastic lesions of tubulonefrosis reaching necrobiotic alterations with citorexis

an citolisis is rather obvious.

12. In case of the lung high values of Cadmium were measured in 2 cases (0,68

mg/kg and 0,53 mg/kg) and Lead was measured in 4 cases, even if the values

did not overcome the maximum allowed limit, and in 4 cases Lead was not

detected within the detection limit. The toxic anr irritative action of Lead and

Cadmium in the lungs result in fibrosis lesions, alveolar emphysema,

peribronsiolic spots of antracosis, hystocitary peribroisiolitis.

13. The charateristic lesions in the histoloogic exam of the lungs for the 8 cattle

are: pulmonary emphysema, fibrosis duet o antracosis, lymphohistocitary

pneumonia.

14. The muscle tissue has been the stocking place for Cadmium, Lead, Copper,

this tissue registering values that overcame the maximum limit allowed by CE



25

Regulation 1881/2006. Thus, Cadmium was overcome in 5 cases, Lead in 4

cases, and was in within the maximum allowed limit (0,1 mg/kg) in 3 cases.

15. Raised concentrations of Lead and Cadmium in the muscle tissue of the 8

cattle are the consequence of raising the cattle in a polluted area, with high

concentrations of pollutants with Lead and Cadmium in soil, water and

vegetation. The muscle tissue of these animals present an obvious risk for the

consumers.

16. High concentrations of Cadmium and Lead in the splin of the 8 cattle prove

that the splin can be an accumulating organ for metals, alongside the liver,

kidney, muscle tissue.

17. The lesions characteristic in the histological exam of the splin for the 8 cattle

are hemosideriosis in 4 cases, and hypertrophia and hyperplazia in arteriolary

endotelia in 1 case.

18. The presence of mineral pollutants has been identified in the rest of tissues

and organs – in the heart the presence of Cadmium and Lead has been

identified. Lead in the heart was present in 5 cases.

19. We can observe that the most important lesions at the level of liver – lipo-

proteic infiltration in liverid cytoplasma, hepatosis; at the level of kidney –

tubulonephrosis, microcysts, cellular cilinders; at the level of lung –

peribronsiolitis, alveolary emphysema, and in splin – high quantity of

colouring in the red pulp, large amounts of hemosiderinic colouring, have

been identified in the same organs in which the concentration of heavy metals

were high.

20. Overcame amounts of Cadmium, Lead, Copper in cattle organs are reflected

in degenerative and vascular lesions.

Liver: the pericentroglobular liverid present vscuolar aspect, whitr cytoplasma or

fine granulated cytoplasma. The majority of the nuclei are vesicular with the

nuleolum centrally disposed. Some liverid have intensly coloured cytoplasma

(hyperchromatic cells), sometimes long shapes or compact aspect.



26

Kidney: we can observe alterations of the tube epiteliums in the shape of

vacuolizations (white cytoplasma); in some of the straight tubes the epitelium is

vaculized with rare images of citorexa, microcystic aspect and lized cells, in some

of the other tubes the lumenum is widen and the epitelium atrophyed, catarrh

images with the detachment of the epitelium and cellular cilinder formation; in the

glomerular cavities of the majority of the glomeruls oxifilic content with serous

aspect, the same content in the majority of contort tubes.

Lung: in peribronsiolic periarterial tissue black granulations, some phagocytate

in macrophagues with sorounding histocitary reaction; thickening of the alveolae

septum, vicariant emphysema with large spaces, fragmented septs, breacking of

alveolar septs and large cavities formation; lymphohistocitary peribronsiolitis;

lymphohistocitary pneumonia, peribronsiolic spots and antracosis. At the level of

the lung we noticed the thickening of alveolar septs, wich reduces the respiratory

surface, that leads to opening of the arteriolae.

Spleen: spleen folicules well circumcised and structured, or less shaped; red

massive pulp with much colouring, and in the red pulp fine granulations

(antracosis) or hemosiderinic colouring predominance; arteriolae with hypertrofic

endotelium proliferating towards the lumenum.

Myocardium: sarcosporidial cysts; endotelial proliferating arteriolae.

Mammary gland: bags with no content, large, with few cells or apoptotic bodies.

Microscopic lesions of degenerative and vascular type identified in tissues and

organs of cattle, prove the gravity of polutant toxicity, especially since they act

cumullatively ans simoultaneously.

٭٭٭٭٭

In the present study we achieved an evaluation of the degree of polution of soil,

water, air and egetation in Zlatna ares, and we have used this data to an applied

discussion related to quantitative analysis of mineral pollutants from tissues and



27

organs harvested from cattle and with histologivcal lesions identified in the studied

tissues.

The study reached its objectives and highlighted the relations between the high

tisular level of investigated polutant factors and the microscopic lesions.

The conditioning of mineral polutants action even if they are detected in small

amounts in the tissues, leads to malfunctions and major ireversible lesions in vital

organs.

Following the evaluation of the results regarding the concentrations of mineral

polutants in organs and tissues, we can say that those ones accumulate in large

amounts in liver, kidney, muscle tissue, and, in smaller but not insignificant amounts

in heart, spleen, mammary gland. Another way of action of heavy metals is

represented by their accumulative efect which makes the tissues very sensible and

thus, a small dose of toxic may start severe intoxications. The study managed to

prove the relations between the accumulation of mineral polutants in high amounts,

their cumultive and simultaneous efect and drastic lesions of the liver, kidney, and

lung, and to prove through degenerative and vascular lesions, the gravity and tixicity

of the oolutants.

The present legislation for Cadmium comprises the maximum allowed linits

just for the muscle tissue, kidney, liver, and for the heart, spleen, lungs, the legislation

does not provide the maximum allowed limits. In specialty literature there are

incomplete data about the normal values of mineral polutants in organs and tissues.

In poluted areas, in muscle tissue, large quantities of mineral polutants

accumulate in time, without causing mny histological lesions. Muscle tissue, but also

the rest of the organs coming from animals in poluted areas, present a risk for the

people consuming it.

Besides the fact that the study has revealed the inter-relationships between

the high tissular level of polutant factors under investigation, and microscopical



28

lesions, and the fact that people are exposed in a considerable degree to

environment polution and human interventions.

Animal contamination through poluted food with toxic metals can not be

totally avoided, having in view the prevalence of these polutants in the environment,

there is a clare need to reduce the contamination, in order to reduce, both the direct

effects n animal health, and indirect effects on human health.

RECOMMENDATIONS

1. Periodical monitorization of soil, water, in order to establish the concentrations for mineral pollutants;

2. Periodical monitorization of spontaneous vegetation and fodder that reach the food of animals, in order to establish mineral pollutants content;

3. Avoiding grazing in areas in which pollutant concentrations in soil and fodder are high;

4. Frequently checking of heavy metals concentrations in animal tissue coming from animals from polluted areas.

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