الفهرس 
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Abstract
Trichinella spiralis (T. spiralis), is a nematode parasite that is present in a wide range of hosts including man (Ashour and Elbakary, 2011). It is widely distributed all over the world in most climates except for deserts with a burden of approximately 10,000 people per year and 0.2% mortality rate (Garcia et al., 2014).
Trichinellosis infection is initiated by the ingestion of raw or poorly cooked meat containing encysted T. spiralis larvae. Due to the presence of digestive fluids in the stomach, larvae are released. They enter the mucous membrane of duodenum and jejunum where they grow to attain sexual maturity. The males die after fertilizing the females and the females start producing larvae 6–7 days after infection. Juveniles enter the lymph and small intestinal capillaries. They are distributed by blood throughout the body. They reach many organs, but are able to survive only in striated muscles. The juveniles grow within muscles and the muscle tissue starts forming cysts. Trichinella spiralis causes specific humoral and cellular responses. Antibodies IgG, IgM and IgA together with eosinophil’s are a dead end for the larvae (Alva et al., 2015).
Oral administration of dosage forms offers significant advantages over other routes. It is a straightforward and comfortable route, providing many different options to control drug release and patient compliance during a chemotherapeutic treatment (Priotti et al., 2017).
The current experimental study was carried out on one hundred male albino mice which were classified into six groups. Each group was subdivided into a and b subgroups. Two hundred T. spiralis larvae were given orally to each mouse of all groups except group I (control negative). Four types of drugs were used, albendazole orally, chitosan nanoparticles orally, chitosan nanoparticles loaded with full dose of albendazole orally and chitosan nanoparticles loaded with half dose of albendazole orally. Drugs were taken after three days of infection for all subgroups a except subgroup I a for the intestinal phase of infection and were administrated after one month post infection for the muscular phase of the infection. For drugs evaluation, parasitological, histopathological, immunohistochemical and ultrastructural studies were done.
One hundred male albino mice were used in this study and were classified into six experimental groups:-
group I (n = 10): Control group, non-infected and non-treated mice (control negative)
group II (n = 10): Infected non treated mice (control positive).
group III (n = 20): Infected mice treated with albendazole
group IV (n = 20): Infected mice and treated by chitosan nanoparticles.
group V (n = 20): Infected mice and treated by chitosan nanoparticles loaded with full dose of albendazole.
group VI (n = 20): Infected mice and treated by chitosan nanoparticles loaded with half dose of albendazole.
Then all groups were divided into two subgroups:
Subgroup a: Mice were sacrificed after seven days p.i. to evaluate the effect of the drugs on the intestinal phase of the infection.
Subgroup b: Mice were sacrificed after 49 days p.i. to evaluate the effect of the drugs on the muscular phase of the infection.
The adult T. spiralis were isolated from the small intestine of the mice that were sacrificed in subgroups a in all groups for counting and preserved in glutraldehyde for SEM study. Parts from small intestinal tissue in the same mice were preserved in formalin for histopathological and immunohistochemical studies. The larvae of T. spiralis were isolated from the mice that were sacrificed in subgroups b in all groups for counting and preserved in glutraldehyde for SEM study. Parts from hind limb muscular tissue in the same mice were preserved in formalin for histopathological and immunohistochemical studies.
The present study showed that:
Concerning the effect of drugs administration on the adults and larval counts, the highest percentage of T. spiralis adult count reduction (99.1%) and larval count (97.3 %) were detected in the subgroups GVa and GVb respectively, which were treated with chitosan nanoparticles loaded with full dose of albendazole. Followed by the infected subgroups that were treated with chitosan nanoparticles loaded with half dose of albendazole (98.8 %) GVIa and 96.7% (GVIb). Followed by the infected subgroups that were treated with albendazole 90.2 % (GIIIa) and 71.3% (GVIb). While, the subgroups treated with chitosan nanoparticles alone showed the lowest percent of reduction 63.3% (IVa) and 60.9% (IVb).
In the present study, the lowest intensity of intestinal inflammation and iNOS expresion were noticed in subgroup GVa that was treated with chitosan nanoparticles loaded with full dose of albendazole, where marked decrease in histopathological changes and iNOS expression including the inflammatory cellular infiltrate in the small intestine as compared to the infected control group appeared. Followed by subgroup treated by chitosan nanoparticles loaded with half dose of albendazole (GVIa) with no significant difference between the two subgroups (GVa& GVIa). Followed by subgroup that with albendazole that showed moderate degree of inflammation and moderate degree of iNOS expression with significant differences between this subgroup (GIIIa) and all treated subgroups (GIVa, Va and VIa). While subgroup treated with chitosan nanoparticles alone (GIVa) showed the highest degree of inflammation and the strongest iNOS expression between the treated subgroups.
Concerning the muscular phase of the infection in this study the lowest intensity of larval deposition, inflammation and iNOS expression were noticed in subgroups treated with chitosan nanoparticles loaded with full dose of albendazole (GVb) as compared to the infected control group. Followed by subgroup treated with chitosan nanoparticles loaded with half dose of albendazole (GVIb) with no significant difference between the two subgroups (GVb& GVIb). Most of the infected mice that were treated with albendazole showed moderate degree of larval deposition, inflammation and moderate degree of iNOS expression with significant differences between this subgroup (GIIIb) and all treated subgroups (GIVb, Vb and VIb). While subgroup treated with chitosan nanoparticles alone (GIVb) showed the highest degree of larval deposition, inflammation and the strongest iNOS expression between the treated subgroups.
In addition, regarding the muscular phase in the current study, the subgroup treated with chitosan nanoparticles loaded with full dose of albendazole (GVb) showed marked decrease in histopathological changes including the inflammatory cellular infiltrate as compared to the infected control group.. In addition, degeneration of the larvae in the form of fragmentation, vacuolation and invasion by inflammatory cellular infiltrate was noticed. Most of the larvae showed homogenization (replaced by eosinophilic material) and invasion by inflammatory cellular infiltrate. Moreover, the capsules around most of the larvae showed thinning in some areas, splitting, vacuolation and areas of breakdown.
Here in all treated groups in the intestinal phase of the infection, showed changes in the structures of T. spiralis adults and larvae regarding SEM. In albendazole treated group (GIIIa& b) treated with albendazole showed flattening of the worm, rupture of the cuticle with complete disappearance of internal content and multiple fissures with loss of the normal annulations of the cuticle. While, the group treated by chitosan nanoparticles (GIVa, b) showed sloughing of some areas of the cuticle with loss of the normal annulations of the cuticle. Moreover, chitosan loaded with full dose of albendazole treated group (GVa, b) showed sloughing and destruction of the cuticle, multiple fissures, loss of the normal annulations of the cuticle, widening of the hypodermal glands, melting and dissolving of the worm with multiple large cauliflower masses and loss of the normal annulations of the cuticle. In addition, group treated with chitosan loaded with half dose of albendazole (GVIa, b) showed multiple vesicles along the cuticle with loss of normal annulations and widening of hypodermal glands.