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Giardia Presentation

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Evaluation of the Therapeutic Effect of Medium Chain Saturated Fatty Acid (Lauric Acid) against Giardia lamblia in Infected Hamsters Mona M. Aly 2 , Maisa A. Shalaby 1 , Samar S. Attia 2 , Shaimaa H. El Sayed 1 , Amira Helmy 1 , Olfat Hammam 1 , Soheir S. Mahmoud 1 , Soad I. Hassan 1 1 Theodor Bilhars Research Institute, Imbaba, Giza, Egypt 2 Medical Parasitology department, Kasr Al-Ainy Faculty of Medicine, Cairo University, Cairo, Egypt
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Page 1: Giardia Presentation

Evaluation of the Therapeutic Effect of Medium Chain Saturated Fatty Acid

(Lauric Acid) against Giardia lamblia in Infected Hamsters

Mona M. Aly2, Maisa A. Shalaby1, Samar S. Attia2, Shaimaa H. El Sayed1, Amira Helmy1, Olfat Hammam1, Soheir S.

Mahmoud1, Soad I. Hassan1

1Theodor Bilhars Research Institute, Imbaba, Giza, Egypt2Medical Parasitology department, Kasr Al-Ainy Faculty of

Medicine, Cairo University, Cairo, Egypt

Page 2: Giardia Presentation

Giardia duodenalis continues to be one of the most common intestinal parasitic protozoa reported in humans worldwide.

Recently, giardiasis has become a significant public health concern due to its association with multiple waterborne outbreaks.

WHO reported that 3 billion people live in developing countries and the rate of giardiasis among them approaches 30%, contributing to 2.5 million deaths annually from diarrheal diseases.

Introduction.

Page 3: Giardia Presentation

Introduction

There are many problems associated with the currently used chemotherapeutic agents in treatment of giardiasis including treatment failure, unpleasant side effects, activity against normal intestinal flora, possible carcinogenicity and parasite resistance.

These adverse reactions had led to search for other suitable natural agents from different origins which are readily accepted, free of toxicity and may augment or even replace some of the standard chemotherapeutic agents presently employed in the treatment of giardiasis.

Page 4: Giardia Presentation

Lauric acid is a naturally derived fatty acid belonging to the medium chain saturated fatty acids (MCSFAs) which is reported to has anti-giardial effect through accumulation within the parasite cytoplasm resulting in rupture of cell membrane and finally trophozoite death.

Introduction

The best and richest natural sources of lauric acid are coconuts and coconut oil.

Page 5: Giardia Presentation

• To evaluate the efficacy of lauric acid in treatment of giardiasis.

• To compare between metronidazole and lauric acid in treatment of giardiasis.

• To evaluate the combination of metronidazole and lauric acid as a line of treatment of giardiasis hopping to reduce the dose and long duration of treatment with metronidazole to avoid the resulting side effects.

Aim of work

Page 6: Giardia Presentation

MATERIAL & METHODS

Experimental Animals, Parasite & Drugs

Animals grouping

Plan of work

Assessment of Anti-giardial Activity

Page 7: Giardia Presentation

Experimental Animals, Parasite & Drugs

Experimental Animals:

• Laboratory bred Syrian hamsters with a weight range of 100-110 gm, provided by the Schistosome biological supply program (SBSP) in Theodor Bilharz Research Institute (TBRI).

Parasite:• Giardia lamblia cysts used for infection of hamsters were

obtained from diarrheic patients attending the outpatient clinic of TBRI.

Drugs:• Metronidazole (Flagyl) was given orally in a suspension form

in a dose of 7 mg/hamster/day for seven consecutive days• Lauric acid (Monolaurin) in a dose of 20 µg/hamster/day for

seven consecutive daysFlagyl

Lauric

acid

Page 8: Giardia Presentation

Group A

(Normal

Control)

Group B

( Drug control

)

Group C(Infecte

d control)

Group D(Metronidazole

treated)

Group E(Lauric

Acid treated

)

Group F(Combined treated)

Number of Hamster

10 15 10 10 10 10

Infection No No Yes Yes Yes Yes

Treatment No Yes No Yes Yes Yes

Dose ( for 7 consecutive days

--- Differs accordin

g to subgrou

p

---- 7 mg per Hamster/

day

20μg per Hamster/

day

3.5 mg (Metronidaz

ole + 10μgLauric

acid)per Hamster/da

y

Animal Groups

Page 9: Giardia Presentation

Intestinal sections were dissected & subjected for histopathological & E. M. examination

duodenal contents were subjected to parasitological examination in order to count the number of Giardia lamblia trophozoites in five successive fields/animal

Scarification of Hamsters

Two weeks later, stool samples were collected and subjected to parasitological examination to count the number of Giardia lamblia cysts per each gram of stool.

Hamsters were given the corresponding medication for 7 consecutive days

Three weeks later, Stool samples were collected and subjected to parasitological examination to detect Giardia lamblia cysts and to ensure that hamsters have

been infected.

Oral Infection of hamsters with Giardia lamblia cysts in a dose of 10.000±10 cysts/hamster.

Plan of Work

Page 10: Giardia Presentation

Parasitological examination

I. Microscopic examination of stool samples

• Direct wet smear

• Merthiolate iodine formaldehyde concentration method (MIFC).

II. Examination of the duodenal contents in order to count the number of Giardia lamblia trophozoites in five successive fields/animal

Assessment of Anti-giardialActivity

Page 11: Giardia Presentation

• Sections of small intestines of sacrified hamsters were done to detect the histopathological changes that occurred due to giardiasis and to assess the cure rates and the degree of healing of intestinal mucosa after drug administration.

Histopathological examination:

Assessment of Anti-giardial Activity

Page 12: Giardia Presentation

Electron micrroscopic examination

• Sections of small intestines of sacrified hamsters were subjected to TEM for higher and detailed magnification of the detected histopathological changes in the small intestine due to giardiasis.

Assessment of Anti-giardial Activity

leika ulta-microtome Phillips 208S Transmission Electron Microscope

Page 13: Giardia Presentation

Results

Results

Parasitological results

stool examination

contents of small

intestine examination

Histopathological results

Electron microscopic

results

Page 14: Giardia Presentation

Parasitological Results

Page 15: Giardia Presentation

70.00%75.00%80.00%85.00%90.00%95.00%

100.00%93.77%

82.03%

98.83%

Pe

rce

nta

ge

of

Re

du

cto

n

Effect of treatment with metronidazole, lauric acid and combination of both drugs on the percentage of cysts reduction in hamsters infected with Giardia lamblia two weeks after administration of drugs.

Parasitological resultsResults of stool

examination

Page 16: Giardia Presentation

metronidazole (D)

lauricacid (E) lauric acid + metronidazole (F)

60.00%

65.00%

70.00%

75.00%

80.00%

85.00%

90.00%

95.00%

100.00%95.50%

78.76%

96.95%

Perc

en

tag

e o

f

Red

uct

ion

Effect of treatment with metronidazole, lauric acid and combination of both drugs on the percentage of reduction in the number of

Giardia trophozoites in the contents of the small intestines of infected hamsters two weeks after administration of drugs.

Parasitological resultsResults of examination of the contents of

small intestine of hamsters

Page 17: Giardia Presentation

Histopathological Results

Page 18: Giardia Presentation

Ulceration of the lining intestinal epithelium (blue

arrow) with Giardia trophozoites (red arrow) .

 Histopathological results

G C

G D GE G F

Partial healing of the intestinal villi

Partial healing of villi with presence of few Giardia

trophozoites in the intestinal lumen

Complete healing with no Giardia

trophozoites in the lumen

Page 19: Giardia Presentation

Electron microscopic results

Page 20: Giardia Presentation

Electron micrograph showing destructed

intestinal cell projection (red

arrow) with marked degeneration of

intestinal submucosa (blue arrow)

Electron microscopic results

GC

GD GE GF

partial repair of the intestinal cell projection &complete repair of the

submucosa

partial repair of the intestinal cell projection

with degenerated trophozoite (black arrow)

complete repair of the intestinal cell projection

and submucosa with absence of Giardia

trophozoites

Page 21: Giardia Presentation

• It was concluded that lauric acid (monolaurin) or coconut offers an alternative therapy to be used in place of, or concurrently with conventional antimicrobials used for treatment of giardiasis.

• The combination of lauric acid and metronidazole, the drug most currently used for the treatment of Giardia lamblia, revealed the highest percentage of reduction in cyst count in stool, vegetative forms in the small intestine, marked improvement of the histopathological and ultrastructural changes of the small intestine caused by Giardia lamblia infecton.

• This study may open fresh avenues for development of natural drug therapy in which food supplementation may augment or even replace some of the standard chemotherapeutic agents currently employed in the treatment of giardiasis and possibly other infectious intestinal diseases.

Conclusion

Page 22: Giardia Presentation

AcknowledgmentTheodor Bilhars Research Institute, Imbaba, Giza, Egypt.• Soad I. Hassan.• Maisa A. Shalaby.• Soheir S. Mahmoud.• Amira M. Helmy.• Olfat A. Hammam.

Medical Parasitology department, Kasr Al-Ainy Faculty of Medicine, Cairo University, Cairo, Egypt• Mona M. Aly• Samar S. Attia

Page 23: Giardia Presentation

THANK YOU


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