Acanthamoeba castellanii causes amoebic keratitis which is a painful sight-threatening disease of the eyes. Its eradication is difficult because the amoebas encyst making it highly resistant to anti-amoebic drugs, but several medicinal plants have proven to be more effective than the usual therapy. This study aimed to evaluate the in vitro amoebicidal activity of ethanol extracts of Arachis hypogaea L. (peanut), Curcuma longa L. (turmeric), and Pancratium maritimum L. (sea daffodil) on A. castellanii cysts. Acanthamoeba were isolated from keratitic patients, cultivated on 1.5% non-nutrient agar, and then incubated with different concentrations of plant extracts which were further evaluated for their cysticidal activity. The results showed that all extracts had significant inhibitory effect on the multiplication of Acanthamoeba cysts as compared to the drug control (chlorhexidine) and non-treated control, and the inhibition was time and dose dependent. The ethanol extract of A. hypogaea had a remarkable cysticidal effect with minimal inhibitory concentration (MIC) of 100 mg/ml in all incubation periods, while the concentrations of 10 and 1 mg/ml were able to completely inhibit growth after 48 and 72 h, respectively. The concentrations 0.1 and 0.01 mg/ml failed to completely inhibit the cyst growth, but showed growth reduction by 64.4-82.6% in all incubation periods. C. longa had a MIC of 1 g and 100 mg/ml after 48 and 72 h, respectively, while the concentrations 10, 1, and 0.1 mg/ml caused growth reduction by 60-90.3% in all incubation periods. P. maritimum had a MIC of 200 mg/ml after 72 h, while the 20-, 2-, 0.2-, and 0.02-mg/ml concentrations showed growth reduction by 34-94.3% in all incubation periods. All extracts seemed to be more effective than chlorhexidine which caused only growth reduction by 55.3-80.2% in all incubation periods and failed to completely inhibit the cyst growth. In conclusion, ethanol extracts of A. hypogaea, C. longa, and P. maritimum could be considered a new natural agent against the Acanthamoeba cyst.

Acanthamoeba is one of the most common free-living amoebae. It is widespread in the environment and can infect humans causing keratitis. Delayed diagnosis or misdiagnosis leads to extensive corneal inflammation and profound visual loss. Therefore, accurate and rapid diagnosis of Acanthamoeba keratitis is essential for successful treatment and good prognosis. This study was designed to use different staining techniques to facilitate the identification of Acanthamoeba cysts. Acanthamoeba cysts were isolated by cultivation of either corneal scraping specimens or tap water samples onto non-nutrient agar plates seeded with Escherichia coli. Subcultures were done from positive cultures until unique cysts were isolated. Acanthamoeba cysts were stained temporarily using iodine, eosin, methylene blue, and calcofluor white (CFW) stains and as permanent slides after processing for mounting using modified trichrome, Gimenez and Giemsa staining. These stains were compared on the basis of staining quality including clarity of morphological details, differentiation between cytoplasm and nuclei, color and contrast, and also other characteristics of the staining techniques, including ease of handling, time taken for the procedure, and cost effectiveness. The cysts of Acanthamoeba were recognized in the form of double-walled cysts: the outer wall (ectocyst) that was being differentiated from the variably stained surrounding background and the inner wall (endocyst) that was sometimes stellated, polygonal, round, or oval and visualized as separate from the spherical, sometimes irregular, outline of the ectocyst. Regarding the temporary stains, it was found that they were efficient for visualizing the morphological details of Acanthamoeba cysts. In CFW staining, Acanthamoeba cysts appeared as bluish-white or turquoise oval halos although the internal detail was not evident. On the other hand, the results of permanent-stained slides showed the most consistent stain for identification of Acanthamoeba cysts was modified trichrome followed by Gimenez stain and lastly Giemsa stain that gave poor visibility of Acanthamoeba cysts due to the intense staining background and monochrome staining of parasite. In the present study, multi-attribute ranking of the used staining techniques showed the highest rank for iodine stain (92 %) followed by eosin stain (84 %), Gimenez stain (76 %), methylene blue (72 %), CFW (64 %), modified trichrome (56 %), and the least was Giemsa stain (44 %). In conclusion, the staining techniques enhance the overall visibility of Acanthamoeba cysts.

Toxoplasmosis may present as a severe disease among some Egyptian patients with chronic liver disease (CLD) due to their impaired immune system, changing the course of the disease. The classical diagnosis of toxoplasmosis by serological tests is inadequate for such patients. This study was performed to highlight the role of real-time quantitative PCR (qrtPCR) test in the accurate diagnosis of toxoplasmosis among Egyptian patients with CLD. Seventy patients with CLD and 50 healthy controls were enrolled in this study. All were subjected to full clinical examinations, abdominal ultrasonography, and biochemical analysis of liver enzymes and they were investigated for markers of hepatitis B virus (HBV) and hepatitis C virus (HCV). In addition, Toxoplasma gondii (T. gondii) parasitemia was determined using qrtPCR. The results showed that T. gondii parasitemia was positive in 30% of CLD patients with highly statistically significant (p < 0.001) compared with the control group (6%). Co-infection in both T. gondii/HBV and T. gondii/HCV was 33.3% and 31.4%, respectively, with a highly significant association between T. gondii parasitemia and HCV viral load. Moreover, the results showed a significant increase of liver enzymes in the serum of patients positive for T. gondii compared with negative patients. An association between T. gondii infection and CLD was observed, and further studies will be needed to define the mechanism of this association.

Patients with chronic liver diseases (CLD) are often highly susceptible to parasitic infection due to a depressed immune system. The objective of this study was to detect the most commonly intestinal parasites found among Egyptian children with CLD. The present study was conducted on 50 children with CLD of different etiology (25 were having different intestinal symptoms, 25 without intestinal symptoms) and 50 non-CLD children with gastrointestinal complaints served as controls. All cases were subjected to stool examination and investigated by liver function tests. Also, anthropometric measurements were taken for all children including weight and height. It was found that the most commonly intestinal protozoa identified in the patients with CLD in order of frequency were: Entamoeba histolytica/Entamoeba dispar (16 %), Giardia lamblia (14 %), Blastocystis hominis (14 %), Cryptosporidium parvum (10 %), E. histolytica and G. lamblia (2 %), E. histolytica and B. hominis (2 %), G. lamblia and B. hominis (2 %), B. hominis and Entamoeba coli (2 %), Microsporidium (2 %) and no cases were found infected with Strongyloides stercoralis. As compared to the controls, the observed incidence of these organisms in CLD patients was significantly higher (p \ 0.045) as regards stool examination by unstained techniques while, there was no significant difference between both groups as regards stool examination by stained techniques (p \ 0.478). In addition, this study showed that the weight and height of studied patients were affected by parasitic infection while, there was no significant correlation between parasitic infection and liver function tests. In conclusion, chronic liver diseases affect the immunity of the patients as shown in significant increase in the incidence of intestinal parasites in cases compared to controls.