Hymenolepiasis is the most prevalent tapeworm infection worldwide, it infects mainly children owing to the lack of faecal hygiene together with bad feeding habits and poverty. H.nana is being considered an opportunistic parasite with a life-threatening effect among immunosuppressed patients thus hymenolepiasis should be eliminated before initiating immunosuppressive therapy for fear of cysticercoids dissemination and persistence of infection.
Praziquantel is the drug of choice for the treatment of H.nana but because of the tapeworm’s relative resistance to chemotherapy and the potential tendency for autoinfection; the patient’s stool should be checked for eggs one and three months after treatment. Although praziquantel has a wide margin of safety it was found hepatotoxic, genotoxic and carcinogenic in albino rats.
There is an increased demand for using plants in therapy "back to nature" instead of using synthetic drugs, which may have adverse effects that may be more dangerous than the disease itself. Rosemarinus officinalis oil is one of the most common folk medicine remedies which is known to have immune modulator action, antioxidant, anti- inflammatory and antimicrobial.
The aim of the present work was to evaluate the parasitological, histopathological, changes in the small intestines of H.nana experimentally infected mice before and after treatment with Rosemarinus officinalis in comparison with praziquantel, the drug of choice for treatment of H.nana infection.
Each of the albino mice used in this study received an oral dose of about 100 H.nana eggs that were freshly obtained and prepared.
The study was conducted on infected albino mice, divided into 2 groups as follows:
A. Group A for studying effect of the drugs on the immature stage of Hymenolepis nana:
Animals in this group classified into 5 subgroups of 10 mice each:
Subgroup A1 (GA1): Non-infected, non- treated (normal control).
Subgroup A2 (GA2): Infected non-treated mice (infected control).
Subgroup A3 (GA3): Infected mice treated with Rosemarinus officinal oil.
Subgroup A4 (GA4): Infected mice treated with Praziquantel.
Subgroup A5 (GA5): Infected mice treated with Praziquantel and Rosemarinus officinal oil.
B. Group B for studying effect of the drugs on the mature stage of Hymenolepis nana:
Animals in this group classified into 5 subgroups of 15 mice each:
Subgroup B1 (GB1): Non-infected, non- treated (normal control).
Subgroup B2 (GB2): Infected non-treated mice (infected control).
Subgroup B3 (GB3): Infected mice treated with Rosemarinus officinal oil and a second dose given after one week.
Subgroup B4 (GB4): Infected mice treated with Praziquantel
Subgroup B5 (GB5): Infected mice treated with Praziquantel and Rosemarinus officinal oil.
C. Dosage schedules:
Group A:
Mice received the treatment from the first day after infection, to investigate the effect of PZQ and Rosemarinus officinalis oil on cysticercoid stage and were sacrificed on the 4thday after infection.
Group B:
Mice harboring adult stage, confirmed by egg detection in stool 14 days after infection, received the treatment to investigate the action of PZQ and Rosemarinus officinalis oil on the fully matured worms already laying eggs and were sacrificed on the 1st, 3rd and 6th day after treatment. The mice group which took Rosemarinus officinalis oil received first course of treatment (400UL/K.g for 5days) and were sacrificed during the course on the 1st, 3rd and 6th day from the start of treatment, the remaining mice received another course of treatment one week apart from the first course and sacrificed on the 5th day from the start of treatment
Mice of the infected control group were sacrificed in accordance, for comparison of the normal development of the parasites to those affected by the treatments.
Studies of the parasitological parameters were concerned with:
I-Effects on total worm burden in the intestine at different sacrifaction times.
II-Effects on the adult worms' morphology using conventional microscopy.
III- Effects on the Total Egg Output (TEO) at different sacrifaction times.
IV-Effects on egg viability.
V-Histopathological study of cysticercoids in the intestinal mucosa if infected mice (size and number).
Concerning the effect of drugs on the number and size of csticercoids, there was remarkable effect of combined PZQ and Rosemarinus officinalis oil treatment on the reduction the number of cysticercoids ,the number of cysticercoids was zero and the percentage of reduction was 100% .There was a statistically significant reduction in the number of cysticercoids in Rosemarinus officinalis treated group, the percentage of reduction in number was 54.35% .In Praziquantel treated group, the percentage or reduction in number was 83.49% which is significant.
In combined PZQ and Rosemarinus officinalis oil treatment, the percentage of reduction in the mean diameter of cysticercoids was 100% .There was a statistically significant reduction in the mean diameter of cysticercoids in Rosemarinus officinalis treated group, the percentage of reduction in mean diameter was 43.44% . In Praziquantel treated group, the percentage of reduction in mean diameter was 64, 79% which is significant.
Concerning histopathological study of cysticercoids, there was few inflammatory cell infiltration in the lamina propria in PZQ treated group, cysticercoid was dark and shrunked.in Rosemarinus officinalis treated few inflammatory cell infiltration in lamina propria but cysticercoid size less affected.In combined treatment ,there was few inflammation and there was restoration of normal architecture of intestinal villi ,no cysticercoids could be dectected in intestinal villi.
Concerning total worm burden ,there was significant difference between PZQ treated group and infected control group as it showed complete eradication of the worms from the 1st day post treatment.As regard Rosemarinus officinalis treated group, there was non-significant difference in the mean TWB of mice sacrificed one day after treatment(Z=0.88,P=0.37),while upon time progression,the difference in the TWB was statistically significant in mice sacrificed on the 3rd,6th day after treatment(Z=1.96,P=0.05, Z=1.48, P=0.04) for 3rd and 6th day respectively, also there was statistically difference on the TWB in mice received a second course of treatment(Z=1.98, P=0.047).
There was obvious effect of combined praziquantel and Rosemarinus officinalis on the TWB of H.nana infected mice along all experimental time points as it killed all worms when administrated at 1st, 3rd, 6th day time points.
Besides, gravid segments were slightly empty in Rosemarinus officinalis treated group.
As regards the mean total egg output, There was a statistically significant difference between the TEO of PZQ treated mice and those of control infected non treated (Z test =2.09, P value =0.036) at all-time points of the experiment.
The efficacy of Rosemarinus officinalis is influenced by the time point, there was non-significant difference in the mean TEO of mice sacrificed one day after treatment(Z=1.52,P=0.12),while upon time progression,the difference in the TEO was statistically significant in mice sacrificed on the 3rd,6th day after treatment(Z=1.96,P=0.05,Z=1.48,P=0.04) for 3rd and 6th day respectively,also there was statistically difference on the TEO in mice received a second course of treatment(Z=1.99, P=0.046).
There was obvious effect of combined praziquantel and Rosemarinus officinalis on the TEO of H.nana infected mice along all experimental time points as the TEO was zero when administrated at 1st, 3rd, 6th day time points.
Detecting the egg viability using Chausov’s method, it gave a yellowish colour of viable eggs in red stained background, while dead eggs appeared totally red and mostly deformed. Moreover, some eggs showed only red stained shells, maintaining a viable yellow embryo inside, indicating a continuous effect.
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