African Journal of Microbiology Research Vol. 3(10) pp. 654-657, 0ctober, 2009
Available online http://www.academicjournals.org/ajmr
ISSN 1996-0808 ©2009 Academic Journals

Full Length Research Paper

Adenosine deaminase level in the serum of the patients
Toxoplasma gondii seropositive and Giardia intestinalisÜlkü Karaman1, Leyla Beytur2, Tu ba Raika Kıran3 and Cemil Çolak4*
1

Department of Parasitology Beyda ı State Hospital, Malatya, Turkey.
Department of Gynecology and Obstetrics, Gözde Hospital, Malatya, Turkey.
3
Inonu University Medical Faculty Department of Biochemistry, Malatya, Turkey.
4
Department of Statistics, Firat University, Elazig, Turkey.
2

Accepted 27 August, 2009

Adenosine deaminase (ADA) is an aminohydrolase making adenosine, deoksiadenozini inozin, anddeocsiniozine deaminise irreversibly and plays role in the catabolism of purine nucleotids. Toxoplasma
gondii is a zoonoses intracellular parasite that causes infection in animals and humans. This parasite
encompasses enzymes that produce free radicals such as superoxide and hydrogen peroxide. In
addition, Giardia intestinalis is another parasite that causes irritations in mucosa, over mucus
discharge, aggravating former inflammations, and various absorption defects. In the present study, it
has been aimed to compare ADA levels between T. gondii seropositive (IgG seropositive but
symptomless patients), G. intestinalis positive patients, and control group. Thus, ADA levels between 32
patients being T. gondii seropositive and 29 controls and between 50 patients’ G. intestinalis positiveand 40 controls have been evaluated. The results were analyzed using independent samples t-test at
the level of p < 0.05. According to this, in the statistical comparison between the parameters of patientand control groups, a meaningful decrease could be determined in ADA levels. This situation can becommented in the way that toxoplasmosis infection being inactive does not necessarily cause anincrease in T lymphocytes. In addition, this decrease can be due to increasing oxidative stress inparasitic infections.
Key words: Toxoplasma gondii, Giardia intestinalis, adenosine deaminase.
INTRODUCTION
Toxoplasmosis is an infection developing asymptomaticand when it is passed in pregnancy period, it can cause
abortion, dead birth and births with congenital anomalies(Kuman, 1987; Roman et al., 2006). It is stated that incidence of the disease increases with age and it ispredicted that 40% of the world population is infected(Yazar, 2000).
Giardiasis is a disease caused by G. intestinalis insmall intestine. The infection can be asymptomatic andalso, causes periodic diarrhea, nausea-vomiting, inapetence, pain in epigastrium, malaise and loss of weight. In
addition, sucking disks of parasite can cause irritations in
irritations in mucosa, over mucus discharge and various*Corresponding author. E-mail: cemilcolak@yahoo.com. Tel.:+90 505 887 04 98. Fax: + 90 312 416 65 93.

absorption defects (Kuman and Altintas, 1996; Unat etal., 1995).
The defense of host immune system against parasites(mature and larval form) is made by means cells. Sitotoxic agents, reactive oxygen and nitrogen intermediary
products in the activated cells play important role. These
products are oxidant molecules and affect parasite viabilities in negative way (Akkus, 1995; Amanvermez andCelik, 2004; Kurt et al., 2002). Adenosine deaminase(ADA) is an aminohydrolase playing role in the catabolism of nucleotids, catalyzes adenosine, deoksiadenosine, and known ribozids in mammals (Ates et al.,2005; Ozvaran et al., 2004). Its structural gene is locatedin 20th chromosome and exists in multiple molecularforms having wide distribution (Ates et al., 2005; Alatas etal., 2003). On the other hand, ADA activity can be foundin lymphocytic cells 10 fold more compared to erythrocytes and also, it is higher in T lymphocytes than to B

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655

Table 1. Serum ADA level of T. gondii and G. intestinalis value.

ParasiteT. gondii
G. intestinalis

GroupsPatient
Control
Patient
Control

lymphocytes (Ates et al., 2005; Ormen et al., 2005).
Considering ADA as a cellular immunity indicator, in thisdirection researchers have determined ADA serum levels
for different diseases. According to this, it has been
demonstrated that ADA activity increases in the activation
period of autoimmune diseases such as; typhoid, infectious mononucleus, brucella, acute pneumonia, tuberculosis, sarcoidosis, liver diseases, acute leukemia, various
malignites and rheumatoid arthritis, systemic lupus
erythematous (SLE), and Behcet’s syndrome (Hatipoglu
et al., 2003; Ozgen et al., 2007) As both cellular andhumoral immunity have disrupted in ADA deficiency, it
has been concluded that ADA activity is significant fornormal lymphocyte function (Ozgen et al., 2007; Cenesiz
et al., 2007).
In the present study, it has been aimed to compare
ADA levels between T. gondii seropositive (IgG seropositive but symptomless patients), G. intestinalis positive
patients, and control group.
MATERIALS AND METHODS
Ethical council report has been taken for the research and only
patients desiring to give sample have been assessed. The stool
and serum samples were examined in the Parasitology and Biochemistry Laboratory in Faculty of Medicine at Inonu University,
Malatya, Turkey between January, 2005 and December, 2007. In
the patient group, only persons seropositive to T. gondii infectionthat were determined using ELISA and IFAT tests and the ones
positive to G. intestinalis using fecal inspection have been comprised in test group. In addition, the patients who had different parasites in their fecal samples, taking some illness medicine such as
typhoid, infectious mononucleus, brucella, acute pneumonia,
tuberculosis, sarcoidosis, liver diseases, acute leukemia and egg.
(Hatipoglu et al., 2003; Ozgen et al., 2007), smoking cigarette andusing alcohol had not been concluded in the research, considering
these situations could cause variable differences at ADA levels.
In order to determine antitoxoplasma antibodies, ELISA andIFAT tests were used. Meddens and Zeus brands were used for
ELISA and IFAT tests, respectively. The volunteer persons without
the history of any parasitic infection, smoking cigarette, taking someillness medicine such as typhoid, infectious mononucleus, brucella,acute pneumonia, tuberculosis, sarcoidosis, liver diseases, acuteleukemia and egg. (Hatipoglu et al., 2003; Ozgen et al., 2007), andusing alcohols were taken to assessment in control group.
The patients that were positive using mentioned tests werecalled again and after making necessary explanations, 5 ml whole
blood samples were taken from the volunteer persons. Its serumhas been separated and stored -20 degree until studying.
Serum ADA levels have been examined by Ellis and Goldberg(Ellis and Galdberg, 1970). According to this method, ammonium
ion that was released from adenosine by the effect of adenosine

n
32
29
50
40

Mean ± SD
9.25 ± 5.09
15.75 ± 10.23
10.87 ± 7.91
21.11 ± 14.08

P value
0.002<0.001deaminase enzyme, forms indophenol complex in green and blue
color as a result of boertholet reaction, also, the intensity of color
increased proportional to enzyme concentration. This complex was
evaluated at 632 nm using spectrophotometer.
Data was described with mean ± standard deviation and normality test was made by Shapiro-Wilk test. For statistical analysis,
independent samples t-test was used through SPSS 13.0 package
program and p < 0.05 value was accepted as a significant result.

RESULTSIn the present study, all the sera samples were negativeto IgM titers. Descriptive statistic for ADA level has beenshown in Table 1. Toxoplasmosis ELISA IgG results ofpatient group were in the range of 1012.97 ± 543.81(mean ± SD) and values of control group were also negative. In the toxoplasmosis IFAT results of patient group,
1/16 and higher values were assessed as positive Table
1. Average age of the group showing T. gondii seropositive results was 29.57 ± 5.02; including 3 men and 29
women. Average age of control group was 35.22 ± 6.03;
including 7 men and 22 women.
Average age of the group with G. intestinalis positive
results was 30.5 ± 8.07; including 33 men and 17 women.
Average age of control group was 40.22 ± 7.01; including
41 men and 9 women.
Significant differences between group of patients
showing toxoplasmosis and their control group and between group of patients infected with G. intestinalis andtheir control group were determined (p = 0.002) anddecreasing in ADA levels of both patient groups could beobserved compared to control groups.
DISCUSSION
It has been stated that parasites have enzymes that
produce free oxygen radicals such as super oxide andhydrogen peroxide and the tissue damages that occurs inparasitic infections take place by the effect of toxindischarged by these organisms (Kurt et al., 2002).
The growth of T. gondii as an intracellular parasite andforming its cysts in human body is controlled by T cells ofcellular immune system. In addition, macrophages, fibroblast cells, astrocytes, and microglia cells found in brain
are cells that can restrict reproduction of toxoplas-mosis(Daubener et al., 1999). Conversely, it has been determined that some T. gondii strains show high virulence inpersons with T lymphocyte deficiency and T lymphocytes

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Microbiol. Res.

play significant role in stimulation of immune reply, andthe resistance occur against tachizoites or intermediaryparasite (Denkers et al., 1998).
G. intestinalis reproduce and place in gastrointestinal
mucosa of humans and animals. In general, 6 - 15 days
after contagion, the special symptoms of parasitic infection start to appear. Immune system in intestines is very
complex and this system can distinguish commensal
bacteria from pathogens. Nitric oxide, reactive oxygen
products, lactoferrin, defensin, phagocyte, mast, anddentric cells play significant role in natural immune
response. Regarding to acquire immune response, IgA
antibodies and T cells have undertaken significant roles(Roxström-Lindquist et al., 2006). The role of T cells in
giardiosis infection in humans could not be determined.
Also, it has been confirmed that chronic giardiosis
develops in rats with CD4+ T cell deficiency (Singer andNash, 2006).
The cells that proliferate in tissues have higher ADA
activity and their ADA percentage is higher in cytoplasm
than nucleus. Also, the highest concentration of ADA has
been in lymphoid tissues (Carson and Seegmiller, 1976).
Furthermore, the activity of lymphocytes increase alongwith mitogenic and antigenic response, meanwhile ADA
activity in T lymphocytes becomes higher than B lymphocytes (Tripathi and al., 2008).
The lack of ADA in erythrocytes and lymphocytes
cause extreme immune deficiency. The inhibition of thisenzyme weakens the maturing and the function oflymphocytes and other immune cells; in addition, whenmonocytes and macrophages are infected with intracellular parasites, they show an increasing in ADA activity.
During the lack of ADA in all cells, immune systemdamage occurs and repetitive vital, fungal, protozal, bacterial infections and lymphopenia can be seen (Cenesiz
et al., 2007). Until now, a study directed to the determination of ADA level in giardiosis infection has not beenencountered. However According to Tripathi et al. (2008),
high ADA levels could be determined in patients showing
visceral leishmaniasis. Again it is stated that ADA levels
increase in malaria (Daddona et al., 1984) and Gakis(1996) acute toxoplasmosis for rats infected with Hitoglou
et al. (2004) Trichinella spiralis. In the present study, ameaningful difference between T. gondii seropositive andcontrol group could be determined and a decrease in
ADA level has been observed according to controls,
either too. Also, as mentioned before, only ADA levels ofpatients with T. gondii IgG seropositive were examined.
The low levels in ADA can be commented in the way that
toxoplasmosis infection being inactive does not cause anincrease in T lympdocytes number or the oxidative stressemerging in parasitic infectios.
In our study, a significant difference between G.
intestinalis and control group could be determined and adecrease in ADA levels could be observed according tocontrols. The duty of T lymphocytes in G. intestinalis infections has not been determined completely yet. It can bethought that ADA levels have been decreased due to oxi-

dative stress increasing resulted from parasitic infec-tion.
Since the lymphocytes of patients infected by T. gondii
and G. intestinalis and control group have not been
examined, data concerning lymphocyte levels does notexist. It is considered that this research will bring light tostudies which will be made in the future in regard tocontrolled test animals on this subject.
ACKNOWLEDGEMENTS
The authors would like to thank to Prof. Nilgün Daldal andAssist. Prof. Metin Atambay and Aysun Bay Karabulut for
their contribution in the study.
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