Romanian Biotechnological Letters Vol. 22, No. 6, 2016 Copyright © 2016 University of Bucharest Printed in Romania. All rights reserved

ORIGINAL PAPER

12062 Romanian Biotechnological Letters, Vol. 22, No. 6, 2016

Garlic, cilantro and chlorella’s effect on liver histoarchitecture changes

in Cd-intoxicated Prussian carp (Carassius gibelio)

Received for publication, February 16, 2015

Accepted, May 3, 2015

MARIOARA NICULA1, GABI DUMITRESCU1, NICOLAE PACALA1,

LAVINIA ŞTEF1, CAMELIA TULCAN1, MONICA DRAGOMIRESCU1,

DOREL DRONCA1, ADRIANA GHERBON2, LILIANA PETCULESCU

CIOCHINA1

1Banat’s University of Agricultural Sciences and Veterinary Medicine “King Michael

I of Romania” from Timişoara, 300645-Timişoara, Calea Aradului 119, Romania; 2University of Medicine and Farmacy “Victor Babeş” Timişoara, Department of

Physiology, 300173-Timişoara, Splaiul Tudor Vladimirescu 14, Romania; *Corresponding author: E-mail: mnicula@animalsci-tm.ro

Abstract Bioactive compounds from natural sources can act as oxygen free radical scavengers or metal chelators,

which enables them to be used as natural antagonists to heavy metals toxicity. So the purpose of this study was

to compare histologically the aspect of liver tissue of Prussian carps’ specimens, subjected to chronic Cd

intoxication with and without garlic, cilantro and chlorella dietary supplementation. 150 Prussian carps, with

weight of 10-12 g, were divided according to the following treatments for 21 days: C (without treatment), E1 (10

ppm Cd into water), E2 (10 ppm Cd into water+2% lyophilized garlic in feed), E3 (10 ppm Cd into water+2%

lyophilized cilantro in feed), E4 (10 ppm Cd into water+2% lyophilized chlorella in feed). Fragments of liver

were removed at the end of experimental period and analyzed in light microscopy and a specific QuickPHOTO

Micro 2.2 software has been used for the histological study. Mentioned epithelium suffered evident histo-

morphological alterations under the action of Cd while the active principles from garlic, cilantro and chlorella

powder have showed chelating and antioxidant potential, cilantro and garlic being the most effective.

Key word: cadmium (Cd) toxicity, fish, liver, histopathology, garlic, cilantro, chlorella

1. Introduction The metals are considered as major toxic pollutants that entering into biogeochemical

cycles they accumulate in natural and artificial aquatic ecosystems. Terrestrial sources that

generate heavy metals are natural - volcanic eruptions, natural weathering leading to rocks’

erosion and anthropogenic - wastewater treatment plants, industries, mining and agriculture

[1]. Non-essential metals (lead, arsenic, mercury, cadmium) are highly toxic, even at very low

levels [2], especially if they accumulate in metabolically active sites. Among them, cadmium

is considered one of the most toxic environmental substances due to its ubiquity, toxicity and

long half-life [3]. Cadmium not having an efficient excretory mechanism, its half-life in the

liver is between 4 and 19 years and, in the kidneys is between 6 and 38 years [4]. There are

three major routes to uptake heavy metals by aquatic organisms: respiratory system (gills),

digestive system and overall body surface (skin) [5]. In benthic aquatic ecosystems, where pollutants are associated with sediments, feed is an important source of their uptake [6]. Thus,

uptake of the contaminants leads to their concentration into aquatic organisms’ tissues,

including fish. Fish are ideal indicators of heavy metal contamination in aquatic ecosystems

Garlic, cilantro and chlorella’s effect on liver histoarchitecture changes in Cd-intoxicated Prussian carp (Carassius gibelio)

Romanian Biotechnological Letters, Vol. 22, No. 6, 2016 12063

[6]. The contaminant can be sequestered and then neutralized, and/or may have specific

interactions with endogenous molecules (inhibition of enzyme) and/or can be metabolized by

enzymes of the biotransformation system. All these interactions may result in: long-term

storage (neutralized fraction); direct or indirect toxic effects (after biotransformation);

excretion of contaminants or their metabolites [7]. The cell membrane is the first target

structure when metals penetrate the cell. It has been shown that metals can bind to membrane

proteins and phospholipids altering their structure and functions [8]. Heavy metals can

stimulate lipid peroxidation processes, a complex sequence of biochemical reactions, defined

as "polyunsaturated lipid oxidative deterioration". The entire process results in production of

highly toxic compounds for the cell, because of high reactivity compared to other cell

components (membrane proteins, DNA). Toxic metals with high affinity to the active groups

of proteins may affect the structure and function of these molecules and finally, the cell

physiology [9]. Chelation therapy is the most commonly used therapeutic strategy for heavy

metal poisoning [10]. Synthetic chelating agents, such as DMPS, DMSA, EDTA and others,

bind poisonous metals rendering them inert and promoting their excretion from the body

without further interaction. There are advantages and disadvantages to using these ones. One

advantage is the power of their mobilizing activity –they are quick to mobilize and eliminate

certain metals in the body, but this may place a huge burden on the body’s detoxification

systems [11]. But natural chelators from some vegetal sources as chlorella [12], garlic [13]

and coriander [14] have been shown their detox ability in human and animals. Garlic contains

many active sulfur compounds derived from cysteine with metal-chelating potential roperties;

these garlic constituents may also protect from metal-catalyzed oxidative damage [15]).

Garlic increases antioxidant action by scavenging reactive oxygen species (ROS), enhancing

the cellular antioxidant enzymes and increasing glutathione in the cells [16]. Cilantro

(Coriandrum sativum) can bind and immobilize cadmium chloride from liver and kidney [17]

and decrease toxic tissue cadmium accumulation in cultured rainbow trout Oncorhynchus

mykiss [18]. One of its principal constituents is linalool. Several animal studies provide

evidence that coriander seeds can promote the hepatic antioxidant system [19]. Chlorella, a

unicellular green algae with the ability to bind cadmium (in animal models) [20], has been

used to detoxify wastewater of metal contaminants as well [21]). The aim of the present study

was first to highlight severity of histological liver damages and secondary to assess

comparatively the chelating and antioxidant property of chlorella, coriander and garlic

compounds in Prussian carp’s cadmium-induced intoxication.

2. Material and Methods Biological material - 150 Prussian carp fingerlings (Carassius gibelio Bloch) were

chosen as test species on the basis of accessibility and ecologically representation. Individuals

belonging to this species are easily acquired and adapt to captivity. Fresh fish samples were

collected from a local pond and were gravimetrically selected. Thus, individuals of 10-12 g

weight were acclimated for two weeks under laboratory conditions prior to experiment,

removing those ones with suspicions on their health. Then, Prussian carp fingerlings were

transferred in 5 glass aquariums equipped with aeration system and 60 L capacity at a

stocking density of 30 fishes per aquarium. The control and test organisms were randomly

distributed in test solutions as follows: the control ones were maintained in Cd-free

freshwater, the others four groups receiving: E1 (10 ppm Cd into water), E2 (10 ppm Cd into

water+2% lyophilized garlic in feed), E3 (10 ppm Cd into water+2% lyophilized cilantro in

feed), E4 (10 ppm Cd into the water+2% lyophilized chlorella in feed), respectively.

MARIOARA NICULA, GABI DUMITRESCU, NICOLAE PACALA, LAVINIA ŞTEF, CAMELIA TULCAN, MONICA

DRAGOMIRESCU, DOREL DRONCA, ADRIANA GHERBON, LILIANA PETCULESCU CIOCHINA

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Chemicals -Fish in groups E1, E2, E3 and E4 were exposed for 21 days to chronic

cadmium intoxication in concentration of 10 ppm from a CdCl2 x ½H2O stock solution. The

sub lethal treatment was calculated from percentage mortalities of fish as described by B.

VEENA & al. [22].

Fish nutrition - Fish were fed twice a day with commercial dry pellets. Freeze-dried

garlic, cilantro and chlorella’s incorporation in fish feed has involved the following steps: 1)

bringing the dry pellets to fine size particles; 2) adding the lyophilized material in the

proportion determined by the experimental protocol setting the daily feed amount needed by a

specimen, according to the calculation model in fish proposed by L. OPREA & R.

GEORGESCU [23]; 3) wetting and homogenization of the obtained mixture; 4) regranulation

and drying of the final pellets. Chlorella pyrenoidosa powder was produced by Bucharest

Medical Laboratories; garlic (figure 1) and coriander (figure 2) powders were obtained by

lyophilization to Interdisciplinary Training and Research Platform "Sustainable Ecological

Agriculture, and Food Safety" from USAMVB Timisoara, Romania. Lyophilization was

carried out at a temperature of -53°C and a pressure of 0.05 mTorr and lyophilization

efficiency was 38.07% for garlic and 10.55% for cilantro, respectively.

Monitoring of physical-chemical parameters of the water in aquariums - Physical-

chemical indicators of water-dissolved oxygen, temperature, NO2-, NO3

-, pH, hardness were

daily measured (water temperature and dissolved oxygen with a movable oxygen-meter with

water resisting microprocessor Hanna HI 9145; pH, NO2-, NO3

-, pH, toughness of water with

a Germany termatest kit).

Histopathological examination - Fragments of liver were removed at the end of a 21

days experimental period after fish euthanasia with an overdose of anesthetic tricaine

methanosulfonate (MS-222) (>250 mg/L). The material was subjected to histological analysis,

being fixed in a formalin solution 10%, dehydrated, clarified, impregnated and embedded in

histological paraffin, according to standard routine methods in the Histology laboratory. Five-

micrometer-thick sections were obtained and stained by Mallory’s trichrome method. Liver

was estimated for 10 specimens from each population. Microscopic examination was

performed using a CX41 Olympus light microscope equipped with a digital camera and

specific QuickPHOTO Micro 2.2 software has been used for the histopathological study.

Tissue alterations were assessed using the histopathological score [24] ranging from – to +++

depending on the degree and extend of lesions: (-) none, (+) mild occurrence, (++) moderate

occurrence, (+++) severe occurrence. Statistical analysis was performed using SPPS IBM 22

software. Data were reported as a significant level of p<0.05. Test of differences between

means was realized by ANOVA completed with post-hoc Tukey test.

3. Results and Discussion Histopathological study of the liver

Histopathological alterations can be used as indicators for the effects of various

anthropogenic pollutants on aquatic biota and they are a reflection of the overall health of

entire population in the ecosystem [25]. The investigation of histological changes in organs of

fish is an accurate way to assess the effects of xenobiotics compounds in experimental studies

[26]. The liver performs many essential functions related to digestion, metabolism, immunity,

and the storage of nutrients within the body. At the same time, the liver is associated with

detoxification and biotransformation processes, and due to these functions combined with its

location and access to the blood supply, it is one of the organs most affected by water

contaminants [25]. But, liver ability to degrade toxic compounds can be overwhelmed by

Garlic, cilantro and chlorella’s effect on liver histoarchitecture changes in Cd-intoxicated Prussian carp (Carassius gibelio)

Romanian Biotechnological Letters, Vol. 22, No. 6, 2016 12065

elevated concentrations of these compounds, and could subsequently result in structural

damage [27]. The microscopic examination of the liver sections of control group revealed that

liver parenchyma consists of liver cells arranged in uniform cordons (figure 1). Hepatocytes

have a polygonal appearance, the membrane is well defined and nucleus is spherical, centrally

or slightly eccentrically located with obvious nucleolus. Binucleated cells are also seen, they

being common among hepatocytes and they can account for as much as 25% of the

hepatocytes polpulation [28]. Sinusoidal capillaries with normal appearance are arranged

between hepatic cell cords (figure 2). Connective tissue from stroma structure is less obvious.

Figure 1. Liver–LC

uniform liver cords;

centered or easily

eccentrically

nucleus; obvious

nucleolus

(trichrome Mallory

staining, 400x)

Figure 2. Liver–LC

sinusoidal

capillaries with

normal appearance

(trichrome Mallory

staining,1000x)

Liver belonging to cadmium-poisoned fish loses its normal architecture and frequently

sinusoidal capillaries hypertrophy and congestive processes are manifested (figures 3 and 4).

Interlobular spaces are wide. Venulare and arteriolar vascular endings are frequently

congested and increased in volume into interportal spaces. The hepatocytes are arranged in

cords, they have a polygonal form, and clear cytoplasm due to the accumulation of numerous

microlipid droplets (fatty degeneration). Intense hypertrophic processes, nuclear chromatin

condensation, karyolysis processes are reported on wider territories, resulting in cell death.

Unlike the control group, in the individuals of this one, intralobular connective stroma

becomes visible. Consequently, interstitial fibrosis processes occur in liver of these specimens

(figure 5).

Figure 3. Liver–E1

sinusoidal capillaries hypertrophy

(trichrome Mallory staining, 400x)

Figure 4. Liver–E1

congestive processes

(trichrome Mallory staining, 400x)

Figure 5. Liver–E1

interstitial fibrosis

(trichrome Mallory staining 200x)

Light microscopic study of the liver sample from fish exposed to Cd and garlic powder

supplemented diet showed several changes: hepatocytes have polygonal form, clear and

central, slightly eccentric nucleus, or eccentric. Frequently, in the liver parenchyma cell

hypertrophy occurs. Cells are increased in volume, cytoplasm is clear, and a large number of

cells have no nucleus. Mild perisinusoidal lymphocytic infiltrates are present in the

connective tissue. Hypertrophic sinusoids capillary, containing erythrocytes, are arranged

between hepatocytes. Interlobular spaces are filled with fine blades of connective tissue,

containing fine capillaries (figure 6) and slightly hypertrophic vascular ramifications also can

be found in interportal spaces. Histological changes in liver specimens exposed for 21 days to

Cd intoxication and feed with an incorporated coriander diet were hepatic parenchyma

consists of dense cords of hepatocyteswith a polygonal appearance, a spherical central,

slightly eccentric or totally eccentric nucleus, with obvious nucleolus; the cytoplasm is clear,

but has fine granulations diffusely spread; hypertrophic hepatocytes with pyknotic nucleus or

without nucleus appear on smaller areas (figure 7). Hypertrophic sinusoidal capillary network

with rich erythrocytes content are distributed among the hepatocyte cords. Hypertrophic

MARIOARA NICULA, GABI DUMITRESCU, NICOLAE PACALA, LAVINIA ŞTEF, CAMELIA TULCAN, MONICA

DRAGOMIRESCU, DOREL DRONCA, ADRIANA GHERBON, LILIANA PETCULESCU CIOCHINA

12066 Romanian Biotechnological Letters, Vol. 22, No. 6, 2016

processes appear in the peri- and interlobular venules and arterioles, vessels in which also is

viewing erythrocyte content. Lymphocytic infiltrates (figure 8) are reported surrounding the

connective tissue of the blood vessels. Histopathological alterations observed after sublethal

Cd exposure in liver of fish fed with a chlorella incorporated diet were: hepatocytes arranged

in cords, have clear cytoplasm with central, slightly eccentric or totally eccentric spherical

nucleus; hepatocytes in various stages of atrophy, increased in volume, with a single block

condensed chromatin, or showing karyolysis are observed on large areas (figure 9).

Figure 6. Liver E2: cell

hypertrophy; cells without a

nucleus;

mild perisinusoidal lymphocytic

infiltrates;

hypertrophic sinusoids capillary

(trichrome Mallory staining, 400x)

Figure 7. Liver E3

dense cords of hepatocytes;

hypertrophic hepatocytes;

hepatocytes with pyknotic nucleus;

hepatocytes without nucleus, spherical

central, slightly eccentric or totally

eccentric nucleus; obvious nucleolus;

clear cytoplasm with fine diffusely

spread granulations (trichrome Mallory

staining, 1000x)

Figure 8. Liver E3

hypertrophic sinusoidal capillary

network;

rich erythrocytes content; lymphocytic

infiltrates

(trichrome Mallory staining,

400x)

Figure 9. Liver E4: atrophied

hepatocytes; condensed chromatin;

karyolysis;

vascular congestions

(trichrome Mallory staining, 1000x)

Sinusoidal capillaries have wide lumen containing abundant red cells. Also, the

terminals veins are hypertrophic and have content. The liver is frequently identified as a target

organ in toxicity studies [29], partly attributable to its anatomical location and functional

complexity. If accumulation of toxins is faster than the liver metabolizing ability, hepatic

damage may occur [30]. Most of the hepatotoxic agents, including cadmium, damage liver

cells mainly by inducing lipid peroxidation and by generation of reactive oxidative

intermediates in liver [30]. Hepatic tissues of the fish exposed to cadmium chloride showed

high degree of congestion (table 1) that may be attributed to the decline in hematological

parameters as H. A. KAOUD & al. [31] observed since the hemoglobin contents are

insufficient for the respiration of the tissue. Congestion of the hepatic central vein and

sinusoids was also reported by H. A. KAOUD & al. [31]; Z. A. EL-GREISY & A. H. A. EL-

GAMAL [32]. In our study, the abundance of intrahepatic lymphocytes (table 1) found in

cadmium treated fish and fed with an incorporated garlic (E2) and cilantro (E3) diet, indicate

that liver is severely affected by the toxic chemical. This action on the immunological defense

of the fish was confirmed in many works as response to heavy metals stress [33], showing

evidence of cell irritability, inflammation and hypersensitivity to Cd. The fibrosis is other

liver damage caused by cadmium poisoning in the present work (table 1) and confirmed in a

lot of studies [34]. Although hepatic parenchymal fibrosis is rare, J. C. WOLF & M. J.

WOLFE [35] affirms that is not unusual to observe a scirrhous reaction in the fish liver that is

located around the bile ducts. The fibrogenic effect of oxidative stress induced by Cd [36] is

characterized by excessive connective tissue deposition in extracellular matrix. Lipocytes

(hepatic stellate cells (HSC)) may be the cell type responsible for periductal fibrosis [37]. In a

healthy liver, HSCs are quiescent cells and contain numerous A vitamin lipid droplets,

constituting the largest reservoir of A vitamin in the rats body [38]. When the liver is injured

due to viral infection or any hepatic toxins, HSCs receive signals secreted by damaged

hepatocytes and immune cells, causing them to trans-differentiate from a resting A vitamin-

rich cell into active, proliferating, fibrogenic, and contractile cell leading to hepatic fibrosis.

Garlic, cilantro and chlorella’s effect on liver histoarchitecture changes in Cd-intoxicated Prussian carp (Carassius gibelio)

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Oxidative stress represents an imbalance between oxidant and antioxidant agents [39].

Cytoplasmic vacuolation and fatty degeneration associated with lipid accumulation was the

pathological features observed by us in all experimental groups (table 1).

Table 1. List of severity of liver lesions in Prussian carps specimens

Liver lesions

Control group

E1

10 ppm CdCl2

E2

10 ppm CdCl2

+2% garlic

E3

10 ppm CdCl2

+2% cilantro

E4

10 ppm CdCl2

+2% chlorella

Congestive processes - +++ - - -

Lymphocytic infiltrates - - + + -

Fibrosis of hepatic tissue - +++ - - -

Vacuolization - +++ ++ + ++

Necrosis

Pyknotic nucleus - +++ ++ + ++

Karyolysis - +++ ++ + ++

(−) none, (+) mild, (++) moderate, (+++) severe occurrence.

Liver fat accumulation in fish may be due to a various causes such as: greater capacity of wild

or captive fish species to store large amounts of lipid (triglyceride) in their livers compared to

mammals (G. J. ROSSOUW (1987), cited by J. C. WOLF & M. J. WOLFE [35]), overfeeding

of captive fish with an excessively energy-rich diet [40], lipid peroxidation, caused by the

feeding with diets high in polyunsaturated fats coupled with a lack of adaptive peroxisomal

proliferation, the suppression of vitamin [41] or toxicant-induced [42]. In our study, this lipid

vacuolization has led to an increase in the size of hepatocytes. But there are authors who

found that beside the toxicological causes of hepatocites’ hypertrophy there are physiological

ones such can be seen in reproductively active female fish [43] or in male fish which have

been exposed to exogenous estrogenic compounds, due to the ability of estrogens to boost

hepatocyte metabolism for vitellogenin (yolk lipoprotein) production [44]. Vacuolation of

hepatocytes is a cellular defense mechanism against injurious substances and it is associated

with inhibited protein synthesis, energy depletion, disaggregation of microtubules, or shifts in

substrate utilization, as described by E. K. AJANI & B. U. J. AKPOILIH [45]. A. ANNABI

& al. [46] previously noted the cytoplasmic vacuolization with lipid droplets and glycogen

accumulation in the liver tissue of Mosquito fish (Gambusia affinis) tissues following acute

and chronic cadmium exposure as well. Nuclear pyknosis observed in the four experimental

groups (table 1) suggests that the manifestation of apoptotic cell death and karyolysis are

followed by hepatocyte necrosis. This pathologic response of the fish liver to Cd is in

agreement with those of S. A. OMER & al. [47]. The results listed above highlights the

ameliorative effect of garlic, cilantro and chlorella powder on the fish liver histopathological

changes. This could be due to their antioxidant properties in combating the free radical-

induced oxidative stress and liver injury resulting from cadmium chloride exposure. In the

same time, these plants produce chelating compound involved in heavy metal detoxification.

Oxidative stress as a crucial factor in liver diseases, (H. CICHOŻ-LACH & A. MICHALAK

[39]), affecting the major cellular components: proteins, lipids and DNA as a result of

excessive production of reactive oxygen species (ROS) [48] such as superoxide ions,

hydroxyl radicals, and hydrogen peroxide. Protective actions against ROS are performed by

several enzymes (e.g., superoxide dismutase-SOD, catalase and glutathione peroxidase), as

well as no-enzymatic compounds (e.g., tocopherol, E vitamin beta-carotene, ascorbate and

glutathione-GSH) [49] many of them can be found in vegetables and microalgae. Thus,

carotenoids play an important role in quenching reactive oxygen species (ROS) generated

during photosynthesis, especially singled oxygen [50]. Green leafy vegetables are rich in

carotenoids, which have several proven and postulated health benefits. Carotenoids mostly

consumed in general diets are α-carotene, β-carotene, lutein, zeaxanthin, neoxanthin,

violaxanthin, and lycopene. Among these carotenoids, β-carotene is the most important

MARIOARA NICULA, GABI DUMITRESCU, NICOLAE PACALA, LAVINIA ŞTEF, CAMELIA TULCAN, MONICA

DRAGOMIRESCU, DOREL DRONCA, ADRIANA GHERBON, LILIANA PETCULESCU CIOCHINA

12068 Romanian Biotechnological Letters, Vol. 22, No. 6, 2016

provitamin [51]. There are studies showing that carotenoids contribute significantly to the

total antioxidant capacity of microalgae [52]. L. C. WU & al. [53], has demonstrated that

antioxidants from spirulina and chlorella water extracts are able to inhibit the proliferation of

HSCs. A potential treatment for liver fibrosis is to inhibit activated hepatic stellate cell (HSC)

proliferation and, subsequently, to induce HSC apoptosis. The phenolic compounds salicylic,

trans cinnamic, synaptic, chlorogenic, chimic and caffeic acids found in the methanolic

chlorella extract may contribute to its higher antioxidant activity [54]. F. J. KAO & al. [55]

analyzed the total carotenoid content (TCC; i.e., lutein, zeaxanthin and β-carotene) in 25 fresh

vegetables commonly consumed in Taiwan, and found cilantro, Thai basil leaves, sweet

potato leaves, and choy sum contain the highest amount of TCC carotenoid-rich vegetables.

Garlic increases antioxidant action by scavenging reactive oxygen species (ROS), enhancing

the cellular antioxidant enzymes and increasing glutathione in the cells [16]. M. ABDEL-

TAWWAB & al. [56] reported that garlic contains numerous sulphur components, including

the most valuable sulphhydryl groups, which oxidize mercury, cupper, cadmium and lead,

make these metals water-soluble, and facilitate the body excretion of these substances. On the

other hand, chelators have the effect of mobilizing metals from tissues and maintaining the

chelate moiety during circulation to the kidneys for excretion in urine, and to the liver for

excretion in the bile [57]. Not only animals, but also plants produce chelating compounds (R.

PAL and J. P. N. RAI, [58] and metallothionein content of foods may affect bioavailability as

well as metabolism of toxic metals such as cadmium [59]. Thus, chlorella is a mild chelator.

Its detoxification capacity is due to its unique cell wall extremely resistant to breakage and the

material associated with it, sporopollenin (a naturally occurring carotene like polymer, which

is resistant to degradation) [60]. Cilantro is also considered as a natural chelator. In animals, it

decreased lead absorption into bone and inhibition of the delta-aminolevulinic acid

dehydratase (ALAD) enzyme [61]. Garlic is a good source of N-Acetyl-L-Cysteine (NAC).

N-acetyl-cysteine (NAC), contained in garlic is a chelator of toxic elements and may

stimulate glutathione synthesis, and production.

4. Conclusions The current result indicates that Cd contamination definitely affects the liver, inducing

pathological lesions, which can be summarized as follow: cytoplasmic vacuolization, blood

congestions, lymphocytic infiltrates, fibrosis and necrosis. Concomitant treatment with

Cd+cilantro, Cd+garlic and Cd+chlorella powder showed evident recovery and an almost

normal liver architecture with mild residual degeneration. Among the three lyophilized

products, cilantro powder proved to have the most chelating and antioxidant potential (liver

samples showed few lipid droplets and little focal necrotic area), while the response at

chlorella treatment was less effective.

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