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 Table of Contents  
ORIGINAL ARTICLE
Year : 2014  |  Volume : 27  |  Issue : 1  |  Page : 103-114

Parasitological, histopathological, and immunohistochemical assessment of nitric oxide synthase inhibitor: aminoguanidine versus albendazole in the treatment of experimental murine toxocariasis


1 Department of Parasitology, Faculty of Medicine, Menoufiya University, Menufia, Egypt
2 Department of Pathology, Theodor Bilharz Research Institute, Giza, Egypt

Date of Submission07-Sep-2013
Date of Acceptance30-Oct-2011
Date of Web Publication20-May-2014

Correspondence Address:
Salwa A. Shams El-Din
Department of Parasitology, Faculty of Medicine, Shibin El-Kom-YassinAbd El-Ghaffarstreet 32511
Egypt
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/1110-2098.132778

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  Abstract 

Objectives
The study was intended to evaluate the effect of aminoguanidine compared with that of albendazole on mice infected with eggs of Toxocara canis by parasitological, histopathological, and immunohistochemical studies.
Background
T. canis is a widely distributed parasite. The nematode can cause toxocariasis in man. The infection takes place after swallowing fully embryonated eggs with larvae inside.
Materials and methods
The study was conducted on 117 albino mice classified into four groups: GI (the control group with subgroups Ia, Ib, and Ic); GII (infected with T. canis eggs); GIII (infected with T. canis eggs and treated with albendazole); and GIV (infected with T. canis eggs and treated with aminoguanidine). Sera from different groups of mice were collected for nitrite assay. The lung and brain tissues were taken for larval recovery and histopathological and immunohistochemical [inducible nitric oxide synthase (iNOS)] studies.
Results
The mean larval count decreased significantly from 7th to 45th day post infection (d.p.i), with albendazole and aminoguanidine, which was more effective in decreasing larval count. Aminoguanidine treatment caused early improvement in histopathological lesions initiating from the second d.p.i.; however, albendazole improvement was observed on the seventh d.p.i. The grade of iNOs expression in GII and GIII was high as compared with that in GIV. There was significantly positive correlation between serum nitrite and the grade of iNOS expression, especially on early d.p.i only in groups II and III.
Conclusion
Both albendazole and aminoguanidine treatment for T. canis-infected mice caused decrease in larval count and improvement in histopathological lesions. Aminoguanidine was more effective with early response and decreased tissue damage.

Keywords: Albendazole, aminoguanidine, immunohistochemistry, nitric oxide synthase, Toxocara spp


How to cite this article:
Nassef NA, El-Kersh WM, El-Nahas NS, Shams El-Din SA, Oshiba SF, Nosseir MM. Parasitological, histopathological, and immunohistochemical assessment of nitric oxide synthase inhibitor: aminoguanidine versus albendazole in the treatment of experimental murine toxocariasis. Menoufia Med J 2014;27:103-14

How to cite this URL:
Nassef NA, El-Kersh WM, El-Nahas NS, Shams El-Din SA, Oshiba SF, Nosseir MM. Parasitological, histopathological, and immunohistochemical assessment of nitric oxide synthase inhibitor: aminoguanidine versus albendazole in the treatment of experimental murine toxocariasis. Menoufia Med J [serial online] 2014 [cited 2020 Feb 26];27:103-14. Available from: http://www.mmj.eg.net/text.asp?2014/27/1/103/132778


  Introduction Top


Toxocara canis is a widely distributed parasite that can cause toxocariasis/visceral larva migrans (VLM) in man and many other hosts [1],[2]. VLM can be acquired from dog (T. canis) or cat (Toxocara cati) [3].

The seroprevalence of human VLM was found to be 14% in the USA [4], 31.6% in Argentina [5], and 51.6% in Brazil [6]. However, it was about 11.1% in children in Brazil [7] and 35.6% in Peruvian children [8]. The infection rates were 7.6, 25, and 44.6% among children in Swaziland, Turkey, and Iran, respectively [9],[10],[11].

VLM seropositivity was recorded to be 24.2% in different rural areas in Egypt [12]. Recently, T. canis eggs were identified in the hair samples of 26.6% stray dogs and 10.7% domestic dogs in Kafr el-Sheikh [13].

The disease showed hypergammaglobulinemia, anemia, leukocytosis, and marked eosinophilia, together with the positive result of serological tests [14]. The symptoms were found to decrease by antihelminthic drugs [15].

Albendazole was able to decrease the larval count of Toxocara spp. in the liver and lung and to prevent migration of larvae to the mice brain [16]. It acts by binding to the colchicine-sensitive site of tubulin-inducing degenerative alterations in the tegument and intestinal cells of the worm. Degeneration of the endoplasmic reticulum and mitochondria of the germinal layer and the subsequent release of lysosomes result in decreased production of adenosine triphosphate, which is the energy required for the survival of the helminthes. Because of diminished energy production, the parasite is immobilized and eventually dies [16],[17].

It is known that microorganisms increase inducible nitric oxide synthase (iNOs) in humans and animals. Nitric oxide (NO), a free radical important in inflammation and immunity [18], is produced by macrophages, endothelial cells, and neurons. Studies suggested that NO participates in antiparasite defense, for example trypanosomiasis and malaria [19-21], but it is responsible for some pathological disorders [22]. NO can affect the living cells through oxidation of iron-containing proteins such as ribonucleotide reductase and aconitase [23].

Nitric oxide synthases are a family of eukaryotic enzymes (NOs) that act by conversion of O 2 and L-arginine to NO and L-citrulline in the presence of NADPH and dioxygen (O 2 ) [24]. iNOS limits parasite development in the vectors [19]. Phagocytes are stimulated by interferon-gamma or by tumor necrosis factor to form iNOS [21]. In contrast, transforming growth factor-β inhibits iNOs, whereas interleukin-4 (IL-4) and IL-10 provide weak inhibitory signals with resultant decrease in tissue injury [25].

The control of NO synthesis by affecting a number of important biological processes has been implicated in the treatment of a variety of diseases [26]. Expression of iNOS, which synthesizes nitric oxide, leads to elimination of parasites [18]. iNOS can be affected by aminoguanidine, a selective iNOs inhibitor [24]. Increased expression of transforming growth factor β-1 by aminoguanidine improves wound healing and preserves the ultrastructure of collagen [27]. Aminoguanidine can also inhibit osteoarthritis by suppressing the production of iNOs, and it decreases the apoptosis of some cells [28].

The aim of the present study was to evaluate the effect of aminoguanidine as compared with that of albendazole on mice infected with T. canis eggs by parasitological, histopathological, and immunohistochemical studies and to correlate these effects with serum nitric oxide level in different groups on different days.


  Materials and methods Top


Experimental design

In all, 117 albino mice were used and classified into four experimental groups. Group 1 (GI) included 27 mice that were divided into three subgroups: GIa contained noninfected nontreated mice (negative control), GIb contained noninfected mice receiving albendazole (as a drug control subgroup), and GIc contained noninfected mice receiving aminoguanidine (as a drug control subgroup). GII included 30 mice infected with T. canis eggs. GIII included 30 mice infected with T. canis eggs then treated with albendazole. GIV included 30 mice infected with T. canis eggs then treated with aminoguanidine. On 2, 7, and 45 days post infection (d.p.i.), three animals from the control subgroups and 10 animals from each group (GII, GIII, and GIV) were killed [29]. Tissue and serum samples were collected for detection of serum nitrite and parasitological, histopathological, and immunohistochemical studies.

Experimental animals

Male Balb/c albino mice with an age of 3 months and weight of 25-30 g were used in the current study. The mice were kept in the animal house in Faculty of Medicine, Menoufiya University, exposed to 12 h light/12 h dark, and fed ad libitum on standard diet and tap water.

Experimental infection

T. canis eggs were obtained from the intestines of adult female stray dogs from Cairo and Giza Provinces. The eggs were purified by straining through a sieve with 0.5 mm pores, washed with saline several times, and then kept in 0.5% formol saline solution (99.5 ml physiological saline and 0.5 ml formaldehyde 40%) for 4-8 weeks in petridishes at 28-30°C to induce embryonation. Maturation of the eggs was checked every day after aeration and shaking of eggs to enhance maturation and to prevent sticking. Then, mature embryonated eggs were kept at 4°C until used. Each mouse was inoculated with about 1000 viable T. canis eggs counted by hemocytometer through the oral route [30].

Drugs

Albendazole in the form of Alzental (Epico Pharm Co., 10 th Ramadan city, Egypt) as a white suspension of 100 mg/5 ml was given orally at a dose of 100 mg/kg once daily for 5 consecutive days, diluted in 0.1 ml distilled water on the first day of infection [31].

Aminoguanidine in the form of aminoguanidine bicarbonate as a white powder (Sigma Chemicals, Cairo, Egypt) dissolved in distilled water was given intraperitoneally at a dose of 50 mg/kg on 1, 3, 5, 8, 10, 12, 14, 16, 18, 22, 27, 32, 37, 40, and 44 d.p.i. [32].

Parasitological study for T. canis larval recovery

Samples from the lung and brain tissues (0.5 g each) were sliced and digested in 50 ml pepsin-HCl solution (2.5 g pepsin, 3.5 ml HCl, and 500 ml water) and incubated at 37°C for 24 h to examine the larval recovery [33]. The digest in petridish and larvae were counted at × 40 magnifications under an inverted microscope (Olympus, Tokyo, Japan) [29]. Larvae in the brain were counted directly, after squashing small amounts of fresh unstained brain tissue between two slides to calculate parasite load in 1 g [34].

Serum nitrite determination [29]

Serum samples (50 μl) collected from each mouse in different groups on the tested days were incubated in Griess reagent (1% sulphanilamide, 0·1% naphthylethylenediamide dihydrochloride in distilled water, and 2·5% orthophosphoric acid) for 10 min. The absorbance was determined for individual sample with an automated spectrophotometer at 540nm wave length. The nitrite concentration was calculated with reference to a sodium nitrite linear standard curve [29].

Histopathological study

The lung and brain tissues of each mouse were fixed in 10% formalin, embedded in paraffin, and sectioned and stained with hematoxylin and eosin stain [35] [Figure 1],[Figure 2],[Figure 3],[Figure 4] and [Figure 5].
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Detection of inducible nitric oxide synthase by immunohistochemical stain (iNOS enzyme in the tissue)

Briefly, paraffin-embedded tissues were deparaffinized and blocked with 3% H 2 O 2 (Merck, Taufkirchen, Germany). Then, they were washed and incubated in serum-blocking reagent G (mouse CTS002-kits) [Figure 6],[Figure 7],[Figure 8],[Figure 9] and [Figure 10]. Thereafter, the specimens were incubated with avidin-blocking reagent (SP2001; Vector, Burlingame, California, USA), washed, and then incubated with biotin-blocking reagent (SP2001; Vector). The specimens were incubated with primary antibody (X0907; Dako, Carpinteria, California, USA) and washed, and again incubated with biotinylated secondary antibody [NOS2 (N-20) sc-651, H2306, 200 μg/ml rabbit polyclonal IgG (Santa Cruz Biotechnology Inc., Dallas, Texas, USA) and washed. Then, the specimens were incubated with horseradish peroxidase and washed. The specimens were incubated with DAB chromogen solution, washed, and counterstained with Mayer's hematoxylin [36]. The grading of iNOS expression in tissues was carried out on the basis of the number of cells showing immunopositivity according to Othman et al. [37] [Figure 11],[Figure 12],[Figure 13],[Figure 14] and [Figure 15].
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Ethical consideration

Animals were housed and scarified according to the international guides to use of laboratory animals.

Statistical methods

Results were collected, tabulated, and statistically analyzed by IBM personal computer and statistical package for the Social Science program for Windows (version 13; SPSS Inc., Chicago, Illinois, USA). Two types of statistics performed are as follows.

Descriptive statistics

It included percentage (%), mean, and SD.

Analytic statistics

The Student t-test, the paired t-test, the F-test (analysis of variance), the c2 -test, the Z-test (z), and Pearson's correlation test were performed. The data were considered statistically significant when P-value was less than 0.05* [38] [Figure 16],[Figure 17],[Figure 18],[Figure 19] and [Figure 20].
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  Results Top


Data in [Table 1] show that T. canis larvae were detected in the lung tissue of GII, GIII, and GIV on 2 d.p.i. and their number was significantly decreased up to 45 d.p.i. T. canis larvae were not detected in the brain on 2 d.p.i.; however, it increased significantly on the following days in GII. There was a statistically significant difference in the number of T. canis larvae in the brain and lung tissues in GII, GIII, and GIV on 7 d.p.i. The number of T. canis larvae in the lung and brain tissues decreased significantly when compared on 7 and 45 d.p.i. in all groups (data not shown) [Figure 21],[Figure 22] and [Figure 23].
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Table 1: Comparison between mean number of Toxocara canis larvae in the lung and brain tissues on different days in different mice groups

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[Table 2] shows that serum NO decreased significantly from 2 to 45 d.p.i. in GII, GIII, and GIV. NO level was also significantly lower in GIV than in GII and GIII on all tested days.
Table 2: Comparison of serum nitric oxide level between different groups on different days

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Graph 1[Additional file 1] shows that, in the lung tissues, no significant difference was recorded in the grades of iNOS expression between the studied groups on the second d.p.i. However, on the seventh d.p.i., the percentage of grade II iNOS expression was significantly higher among GIV (P < 0.001***), whereas the percentage of grade III iNOS expression was significantly higher among GIII than among other groups (P < 0.001***). On the 45 th d.p.i., the percentage of grade I iNOS expression was significantly higher among GIV (P < 0.001***), whereas the percentage of grade III iNOS expression was significantly higher among GII and GIII than among GIV (P < 0.001***). In Graph 2 [Additional file 2] concerning the brain tissues, the percentage of grade I iNOS expression was significantly higher among GIV than among other groups, whereas the percentage of grade II iNOS expression was significantly higher among GII than among other groups on the seventh d.p.i. (P < 0.001***). On the 45th d.p.i., the percentage of grade 0 iNOS expression was significantly higher among GIV and the percentage of grade I iNOS expression was significantly higher among GIII, whereas the percentage of grade III iNOS expression was significantly higher among GII than among other groups (P < 0.001***).

In [Table 3] and Graph 3 [Additional file 3], there was significant positive correlation between serum NO level in μmol/l and grade of iNOs expression in the lung on 2 d.p.i. (P < 0.05*).
Table 3: Correlation between serum NO level and grade of iNOs expression on different days post infection for group II

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[Table 4] and Graph 4 [Additional file 4] show significant positive correlation between serum NO level in μmol/l and grade of iNOs expression in the lung on 2 d.p.i. (P < 0.001***).
Table 4: Correlation between serum NO level and grade of iNOS expression on different days for group III

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[Table 5] shows no significant positive correlation between serum NO level in μmol/l and grade of iNOS expression in the lung and brain in GIV.

The lung tissues were moderately inflamed on the second d.p.i., but severe inflammation was recorded in the lung tissues on the seventh and 45th d.p.i. The brain tissue was normal on the second and seventh d.p.i., but on the 45th d.p.i. some of the brain tissues were mildly inflamed.
Table 5: Correlation between serum NO level in m mol/l and grade of iNOS expression on different days for group IV

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After albendazole treatment, mild improvement in histopathological lesions initiated on the seventh d.p.i., whereas evident improvement occurred on the 45th d.p.i. After aminoguanidine treatment, improvement in histopathological lesions was observed from the second d.p.i. and increased on the seventh d.p.i., and the maximum improvement was evident on the 45th d.p.i. compared with the infected nontreated group.

Immunohistochemical results of this study indicated that the grade of iNOS expression in different tissues among GII (infected nontreated mice) was high. Albendazole treatment for T. canis-infected mice caused increased expression of iNOS in different tissues especially lung, whereas aminoguanidine treatment caused diminution of iNOS enzyme in all tissues. There was significant positive correlation between serum nitric oxide level and the percentages of grade of iNOS expression in some tissues of different groups, especially on early d.p.i.


  Discussion Top


This study was conducted on 117 albino mice that were classified into four experimental groups: the control group GI [control negative, (a); drug control for albendazole, (b); and drug control for aminoguanidine, (c)], GII that was infected with embryonated T. canis eggs, GIII that was infected with embryonated T. canis eggs and treated with albendazole, and GIV that was infected with embryonated T. canis eggs and treated with aminoguanidine.

In the current study, the recovery of T. canis larvae initiated in the lung tissue on the second d.p.i. and decreased significantly on the following days in GIII and GIV until disappeared. In contrast, T. canis larvae were not detected on the second d.p.i. in the brain tissues but appeared on the seventh and decreased on the 45th d.p.i. Similar results were reported by Othman et al. [37], Taira et al. [39], and Caldera et al. [40] who found that larval recovery showed progressive increase over the course of infection, with special predilection for the central nervous system.

T. canis larvae detected in the lung and brain tissues were significantly lower in the aminoguanidine-treated group (GIV) than in the albendazole-treated group (GIII) and in the infected group (GII) on days 7 and 45 d.p.i., respectively [Table 2]. Albendazole exhibited antihelminthic effect on the brain (0 larva/mouse) and lung [16]. The study by Fernando et al. [41] coincided with the present study and showed that albendazole treatment in patients with toxocariasis caused clinical improvement.

Serum NO was significantly reduced with the chronicity of infection in GII, GIII, and GIV. NO level was significantly reduced also in GIV than in GII and GIII on all tested days. Lin et al. [42] reported decreased NO level on 28 d.p.i. with T. canis and continued decreasing on the following days. Also, NO level was drastically reduced in thegroup treated with iNOs inhibitor: L-NIL then infected with T. canis.

The lung tissues showed infiltration with inflammatory cells and focal peribronchial and intrabronchial inflammation. The blood vessels showed dilatation, congestion, and interstitial hemorrhage. These manifestations progressed from moderate to severe in GII. Oryan et al. [43] showed similar pathological findings in infected lung tissues.

Production of iNOs during T. canis infection causes direct host damages and is strongly related to the oxidative stress. We propose that larval NO can also be effective in larval migration [18]. These pathological manifestations progressed from moderate to mild in GIII after albendazole treatment. In GIV treated with aminoguanidine, the lung pathology ranged from mild to moderate. L-NIL treatment resulted in large, irregularly shaped granulomas with suppressed collagen contents at 4 weeks post infection (w.p.i). but not at 8 w.p.i. The suppressed collagen contents might have been related to decreased serum NO and Th2-type cytokine of IL-4 but not to Th1-type cytokine of interferon-gamma expression [42]. Enhanced iNOs expression seemed to play a certain role in pathological damage. A potential therapeutic strategy for treatment of nematodes is through manipulation of iNOs expression [33].

The brain tissue of GII contained few scattered larvae and showed congestion and inflammation that progressed from normal to mild. In GIII also, the pathological manifestation progressed from normal to mild after albendazole treatment. The brain tissue was nearly normal in GIV in which mice were treated with aminoguanidine. Inhibition of iNOs has protective effect on the brains of infected mice, and T. canis infection could be related to oxidative stress; hence, NO production and iNOs inhibition can protect the tissue from damage in this infection [44].

Expression of iNOs reported in the lung tissues in the current study changed from grade I to grade III in GII and GIII. In the aminoguanidine-treated GIV, iNOs expression ranged from grade I to II. This is in agreement with the study by Othman et al. [37], who reported increased expression of iNOs in T. canis-infected tissues. These changes were maximal in the chronic stage of infection or were increased over time.

Expression of iNOs reported in the brain tissues of GII was found to be increased from grade 0 to grade III then decreased again to grade 0. Othman et al. [37] observed that iNOS was expressed by the glial cells of Toxocara spp.-infected mice at 2 w.p.i but not in the age-matched control group of uninfected mice. Moreover, there was a significant increase in its expression with increasing duration of infection in GIII. Aminoguanidine treatment decreased iNOs expression from grade III to 0 in GIV. Hamilton et al. [45] showed increased expression of iNOs in murine toxocariasis. These results have implications for the role of iNOs in the cerebral establishment of T. canis and in the cerebral pathology reported during infection. Inducible NOs expression was enhanced in infected and L-NIL-treated mice at 4 w.p.i. but declined at 8 w.p.i. as shown by immunohistochemical study [42].

There was a significant correlation between serum NO level and grade of iNOS expression in the lung of GII and GIII on 2 d.p.i. only.


  Conclusion Top


Serum NO level can be a marker for suspected cases of visceral larvae migrans, mainly in early -d.p.i. iNOs inhibition with aminoguanidine resulted in less damaged tissues and diminished the deleterious effects of the parasite. Aminoguanidine can be used alone for the treatment of VLM, as it was effective in decreasing larval count especially in the brain tissue and it improved histopathological changes in the tissues. Finally, combinations of albendazole and aminoguanidine for treatment of VLM should be further studied as a more effective therapy for VLM.


  Acknowledgements Top


Conflicts of interest

None declared.

 
  References Top

1.Deutz A, Fuchs K, Auer H, Kerbl U, Aspock H, Kofer J, et al. Toxocara-infestations in Austria: a study on the risk of infections of farmers, slaughterhouse staff, hunters and veterinarians. Parasitol Res 2005; 97:390-394.  Back to cited text no. 1
    
2. Smith HV, Holland C, Taylor M, Magnaval JF, Schantz P, Maizels RM, et al. How common is human toxocariasis? Towards standardising our knowledge. Trends Parasitol 2009; 25:182-188.  Back to cited text no. 2
    
3. Markell EK, Voge, M. Markell and Voge′s medical parasitology. 9th ed.. St Louis: W.B. Saunders; 2006. 310-317.  Back to cited text no. 3
    
4. Won KY, Moran DK, Schantz PM, Jones, JL. National seroprevalence and risk factors for zoonotic Toxocara spp. infection. Am J Trop Med Hyg 2008; 79:552-557.  Back to cited text no. 4
    
5. Fillaux J, Santillan G, Magnaval J, Jensen O, Larrieu E, Sobrino-Becaria CD, et al. Epidemiology of toxocariasis in a steppe environment: the Patagonia study. Am J Trop Med Hyg 2007; 76:1144-1147.  Back to cited text no. 5
    
6. Colli CM, Elefant RG, Paludo ML, Guilherme EV, Mattia S, et al. Serological, clinical and epidemiological evaluation of toxocariasis in urban areas of South Brazil. Rev Inst Med Trop Sao Paulo2010; 52:69-74.  Back to cited text no. 6
    
7. Santarem VA, Leli FC, Rubinsky-Elefant G, Giuffrid, R. Protective and risk factors for toxocariasis in children from two different social classes of Brazil. Rev Inst Med Trop Sao Paulo 2011; 53:67-72.  Back to cited text no. 7
    
8. Roldan WH, Cavero YA, Espinoza YA, Jimenez S, Gutierrez, CA. Human toxocariasis: a seroepidemiological survey in the Amazonian city of Yurimaguas, Peru. Rev Inst Med Trop Sao Paulo2010; 52:37-42.  Back to cited text no. 8
    
9. Akdemir C. Visceral larva migrans among children in Kütahya (Turkey) and an evaluation of playgrounds for T. canis eggs. Turk J Pediatr 2010; 52:158-162.  Back to cited text no. 9
[PUBMED]    
10.1Sharif M, Daryani A, Barzegar G, Nasrolahei M, Khalilian, A. Seroprevalence of toxocariasis in schoolchildren in Northern Iran. Pak J Biol Sci 2010; 13:180-184.  Back to cited text no. 10
    
11.1Liao CW, Sukati H, D′Lamini P, Chou CM, Liu YH, Huang YC, et al. Seroprevalence of Toxocaracanis infection among children in Swaziland, Southern Africa. Ann Trop Med Parasitol 2010; 104:73-80.  Back to cited text no. 11
    
12.1Antonios SN, Eid MM, Khalifa EA, Othman AA. Seroprevalence study of Toxocaracanis in selected Egyptian patients. J Egypt Soc Parasitol 2008; 38:313-318.  Back to cited text no. 12
    
13.1El-Tras WF, Holt HR, Tayel, AA. Risk of Toxocaracanis eggs in stray and domestic dog hair in Egypt. Vet Parasitol 2011; 178:319-323.  Back to cited text no. 13
    
14.1Carvalho EA, Rocha, RL. Toxocariasis: visceral larva migrans in children. J Pediatr 2011; 87:100-110.  Back to cited text no. 14
    
15.1Bede O, Szenasi Z, Danka J, Gyurkovits K, Nagy D. Toxocariasis associated with chronic cough in childhood: a longitudinal study in Hungary. J Helminthol 2008; 82:357-363.  Back to cited text no. 15
    
16.1Barrera MG, Leonardi D, Bolmaro RE, Echenique CG, Olivieri AC, Salomon CJ, Lamas MC, et al. In vivo evaluation of albendazole microspheres for the treatment of Toxocaracanis larva migrans. Eur J Pharm Biopharm 2010; 75:451-454.  Back to cited text no. 16
    
17.1Musa D, Senocak G, Borazan G, Altas M, Ozgonul A, et al. Effects of Nigella sativa and albendazole alone and in combination in Toxocaracanis infected mice. J Pak Med Assoc 2011; 61:866-870.  Back to cited text no. 17
    
18.1Demirci C, Gargili A, Kandil A, Cetinkaya H, Uyaner I, Boynuegri B, Gumustas MK, et al. Inhibition of inducible nitric oxide synthase in murine visceral larva migrans: effects on lung and liver damage. Chin J Physiol 2006; 49:326-334.  Back to cited text no. 18
    
19.1Ascenzi P, Gradoni, L. Nitric oxide limits parasite development in vectors and in invertebrate intermediate hosts. Life 2002; 53:121-123.  Back to cited text no. 19
    
20.2Espinoza E, Perez-Arellano JL, Carranza C, et al. In vivo inhibition of inducible nitric oxide synthase decreases lung injury induced by Toxocaracanis in experimentally infected rats. Parasite Immunol 2002; 24:511-520.  Back to cited text no. 20
    
21.2Hsiao HY, Mak OT, Yang CS, Liu YP, Fang KM, Tzeng SF, et al. TNF-alpha/IFN-gamma-induced iNOS expression increased by prostaglandin E2 in rat primary astrocytes via EP2-evoked cAMP/PKA and intracellular calcium signaling. Glia 2007; 55:214-223.  Back to cited text no. 21
    
22.2Lawrence CE, Paterson JC, Wei XO, Liew FY, Garside P, Kennedy MW, et al. Nitric oxide mediates intestinal pathology but not immune expulsion during Trichinellaspiralisinfection in mice. J Immunol 2000; 164:4229-4234.  Back to cited text no. 22
    
23.2Prabhu V, Guruvayoorappan, C. Nitric oxide: pros and cons in tumor progression. Immunopharmacol Immunotoxicol 2010; 32:387-392.  Back to cited text no. 23
    
24.2Alderton WK, Cooper CE, Knowles, RG. Nitric oxide synthases: structure, function and inhibition. Biochem J 2001; 357:593-615.  Back to cited text no. 24
    
25.2Sosroseno W, Bird PS, Seymour, GJ. Nitric oxide production by a murine macrophage cell line (RAW264.7 cells) stimulated with Aggregatibacter actinomycetemcomitans surface-associated material. Anaerobe 2011; 17:246-251.  Back to cited text no. 25
    
26.2Bruckdorfer R. The basics about nitric oxide. Mol Aspects Med 2005; 26:3-31.  Back to cited text no. 26
    
27.2Yavuz D, Tugtepe H, Cetinel S, Uyar S, Kaya H, Haklar G, et al. Collagen ultrastructure and TGF-β1 expression preserved with aminoguanidine during wound healing in diabetic rats. Endocr Res 2005; 31:229-243.  Back to cited text no. 27
    
28.2Yimin Y, Min W, Meng L. The effect of aminoguanidineintraperitoneal injection on apoptosis in chondrocyte with osteoarthritis. J Xi′an Jiaotong University (Medical Sciences) 2008; 3-13.  Back to cited text no. 28
    
29.2Fan CK, Lin YH, Hung CC, Chang SF, Su KE. Enhanced inducible nitric oxide synthase expression and nitrotyrosine accumulation in experimental granulomatous hepatitis caused by Toxocaracanis in mice. Parasite Immunol 2004; 26:273-281.  Back to cited text no. 29
    
30.3Fan CK, Lin YH, Du WY, Su, KE. Infectivity and pathogenicity of 14-month-cultured embryonated eggs of Toxocaracanis in mice. Vet Parasitol 2003; 113:145-155.  Back to cited text no. 30
    
31.3Yarsan E, Altinsaat C, Aycicek H, Sahindokuyucu F, Kalkan, F. Effects of albendazole treatment on haematological and biochemical parameters in healthy and Toxocaracanis infected mice. Turk J Vet Anim Sci 2003; 27:1057-1063.  Back to cited text no. 31
    
32.3Zeromski JZ, Boczon K, Nowak EW, Lisewska, IM. Effect of aminoguanidine and albendazole on inducible nitric oxide synthase (iNOS) activity in T. spiralis-infected mice muscles. Folia Histochem Cytobiol 2005; 43:157-159.  Back to cited text no. 32
    
33.3Horiuchi A, Satou T, Akao N, Koike K, Fujita K, Nikaido T, et al. The effect of free and polyethylene glycol-liposome-entrapped albendazole on larval mobility and number in Toxocaracanis infected mice. Vet Parasitol 2005; 129:83-87.  Back to cited text no. 33
    
34.3Chung LY, Fang BH, Chang JH, Chye SM, Yen, CM. The infectivity and antigenicity of Toxocaracanis eggs can be retained after long-term preservation. Ann Trop Med Parasitol 2004; 98:251-260.  Back to cited text no. 34
    
35.3Kessel RG. In: Kessel RG, ed. Techniques for the study of cells, tissues and organs. Medical histology. Madison Avenue, NY: Oxoford University Press Inc.; 1998. 265-266.  Back to cited text no. 35
    
36.3Morikawa A, Kato Y, Sugiyama T, Koide N, Chakravortty D, Yoshida T, Yokochi T, et al. The role of nitric oxide in lipopolysaccharide-induced hepatic injury in d-galactosamine sensitized mice as an experimental endotoxic shock model. Infect Immun 1999; 67:1018-1024.  Back to cited text no. 36
    
37.3Othman AA, Abdel-Aleem GA, Saied ME, Mayah WW, Elatrash AM. Biochemical and immunopathological changes in experimental neurotoxocariasis. Mol Biochem Parasitol 2010; 172:1-8.  Back to cited text no. 37
    
38.3Morton RF, Hebel JR, Mc Carter, RJ. Medical statistics. In: Morton RF, Hebel JR, Mc Carter, RJ. A study guide to epidemiology and biostatistics (5 th edition). Maryland, Gaitherburg publication; 2001. 71-74.  Back to cited text no. 38
    
39.3Taira K, Saitoh Y, Kapel MO. Toxocaracati larvae persist and retain high infectivity in muscles of experimentally infected chickens. Vet Parasitol 2011; 180:287-291.  Back to cited text no. 39
    
40.4Caldera F, Burlone ME, Genchi C, Pirisi M, Bartoli E. Toxocara encephalitis presenting with autonomous nervous system involvement. Infection 2013;41:691-694.  Back to cited text no. 40
    
41.4Fernando SD, Wickramasinghe VP, Dewasurendra RL, Kapilananda GMG. Comparative effect of albendazole and diethylcarbamazine in the treatment of toxocariasis in children from Sri Lanka: a preliminary study. J Clin Med Res 2011; 3:46-51.  Back to cited text no. 41
    
42.4Lin SM, Liao CW, Lin YH, Lee CC, Kao TC, Fan CK, et al. Inducible nitric oxide synthase inhibition influenced granuloma formation with suppressed collagen expression in myositis caused by Toxocaracanis in mice. Parasitol Res 2008; 102:577-585.  Back to cited text no. 42
    
43.4Oryan A, Sadjjadi SM, Azizi S. Longevity of Toxocaracati larvae and pathology in tissues of experimentally infected chickens. Korean J Parasitol 2010; 48:79-80.  Back to cited text no. 43
    
44.4Gargili A, Demirci C, Kandil A, Cetinkaya H, Atukeren P, Gumustas MK, et al. In vivo inhibition of inducible nitric oxide and evaluation of the brain tissue damage induced by Toxocaracanis larvae in experimentally infected mice. Chin J Physiol 2004; 47:189-196.  Back to cited text no. 44
    
45.4Hamilton CM, Brandes S, Holland CV, Pinelli E. Cytokine expression in the brains of Toxocaracanis-infected mice. Parasite Immunol 2008; 30:181-185.  Back to cited text no. 45
    


    Figures

  [Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5], [Figure 6], [Figure 7], [Figure 8], [Figure 9], [Figure 10], [Figure 11], [Figure 12], [Figure 13], [Figure 14], [Figure 15], [Figure 16], [Figure 17], [Figure 18], [Figure 19], [Figure 20], [Figure 21], [Figure 22], [Figure 23]
 
 
    Tables

  [Table 1], [Table 2], [Table 3], [Table 4], [Table 5]



 

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