Protozoan Parasites of Rodents and Their Zoonotic Significance in Boyer-Ahmad District, Southwestern Iran

Backgrounds. Wild rodents are reservoirs of various zoonotic diseases, such as toxoplasmosis, babesiosis, and leishmaniasis. The current study aimed to assess the protozoan infection of rodents in Boyer-Ahmad district, southwestern Iran. Materials and Methods. A total of 52 rodents were collected from different parts of Boyer-Ahmad district, in Kohgiluyeh and Boyer-Ahmad province, using Sherman live traps. Each rodent was anesthetized with ether, according to the ethics of working with animals, and was dissected. Samples were taken from various tissues and stool samples were collected from the contents of the colon and small intestines. Moreover, 2 to 5 mL of blood was taken from each of the rodents and the sera were examined for anti-Leishmania antibodies, by ELISA, or anti-T. gondii antibodies, by modified agglutination test (MAT). DNA was extracted from brain tissue samples of each rodent and PCR was used to identify the DNA of T. gondii. Results. Of the 52 stool samples of rodents studied by parasitological methods, intestinal protozoa infection was seen in 28 cases (53.8%). From 52 rodents, 19 (36.5%) were infected with Trichomonas, 10 (19.2%) with Giardia muris, and 11 (21.2%) with Entamoeba spp. Also, 10 cases (19.2%) were infected with Blastocystis, 3 (5.8%) were infected with Chilomastix, 7 (13.5%) were infected with Endolimax, 1 (1.9%) was infected with Retortamonas, 3 (5.77%) were infected with T. gondii, and 6 (11.54%) were infected with Trypanosoma lewisi. Antibodies to T. gondii were detected in the sera of 5 (9.61%) cases. Results of the molecular study showed T. gondii infection in 3 (5.77%) of the rodents. Findings of this study showed that rodents in Kohgiluyeh and Boyer-Ahmad province, southwestern Iran, are infected with several blood and intestinal parasites; some of them might be potential risks to residents and domestic animals in the region

Expression Analysis of Multiple Genes May Involve in Antimony Resistance among Leishmania major Clinical Isolates from Fars Province, Central Iran

Background: Treatment of Cutaneous Leishmaniasis (CL) is being faced with serious difficulties in Fars Province, due to emerging of resistance against meglumine antimonite (Glucantime®). In this context, determining some biomarkers for drug sensitivity monitoring seems to be highly essential. Different studies have been carried out to decipher the genes might be involved in antimony resistant phenotype in Leishmania spp. Here, we selected three genes: AQP (as drug transporter), TDR-1-1(as drug activator), and γ-GCS (inducing reduction environment) for comparative expression analysis on clinical resistant and sensitive isolates of L. major. Methods: The clinical isolates of L. major were collected from CL patients referred to Valfajr Health Center, Shiraz from Oct 2011 to Feb 2012. The susceptibility test was performed to confirm drug sensitivity of strains in vitro as well. Then, the gene expression analysis was performed by quantitative real-time PCR using SYBR® Green. Results: By comparison of expression level between strains, up regulation of γ-GCS gene and down regulation of AQP gene were observed in resistant strains compared to the sensitive isolates; however, down regulation of AQP was not statistically specific. Analysis of TDR-1-1 gene unexpectedly showed a high level of expression in the non-responsive cases. Conclusion: The γ-GCS, at least, can be considered as a suitable molecular marker for screening antimony sensitivity in clinical isolates, although AQP and TDR-1-1gene seem not to be reliable resistant markers.

Preparation of meglumine antimonate loaded albumin nanoparticles and evaluation of its anti-leishmanial activity: an in vitro assay

Cutaneous leishmaniasis is still a health problem worldwide, especially in tropical and subtropical areas. Currently, pentavalent antimony compounds are used to treat leishmaniasis. These compounds cause various side effects in the body. Therefore, there is a need to discover new drugs with less toxicity and more therapeutic effects. In this study, we encapsulated the meglumine antimonate into the albumin as a drug carrier and evaluated the anti-leishmanial effect of the prepared nanoparticles. The precipitation method was used for this purpose by applying different concentrations of glutaraldehyde and N-(3-Dimethylaminopropyl)-N-ethyl carbodiimide hydro chloride Ethyl (DEC) and then, field emission test was performed using Scanning Electron Microscopy for evaluating the morphology and size particles. The cytotoxicity and inhibitory of drugs were evaluated on J774 macrophages and Leishmania major promastigotes, respectively. Nanodrugs prepared using glutaraldehyde (10 μl/ml) and DEC (13 mg/ml) had the smallest and largest size, respectively. The highest anti-leishmanial activity was observed in the drugs prepared with glutaraldehyde (10 μl/ml). Also this nanodrug had the lowest cytotoxicity against macrophages. Given that meglumine antimonate loaded albumin nanoparticles prepared with glutaraldehyde (10 μg/ml), can improve the anti-leishmanial effects of this old drug, it can be a good option as a drug delivery syste

The prevalence of toxoplasmosis in hemodialysis patients in Bushehr, Iran in 2012

Background and purpose: Toxoplasmosis infection in hemodialysis patients may cause severe complications, and even in some cases the cause of death. It can be impose considerable costs to health systems. Since the high prevalence of this parasite has been reported in various parts of the country, this study was conducted to determine the prevalence of toxoplasmosis in hemodialysis patients in Bushehr, Iran. Materials and methods: In this cross-sectional study, all hemodialysis patients who were continuously referred from September 2011 to September 2012 to the dialysis center at Bushehr, were evaluated by the ELISA method for determine the presence of anti-Toxoplasma IgG & IgM antibodies. Results: From a total of 84 patients studied, 32 (38.1%) and 2 (2.4%) patients were positive for IgG and IgM antibodies, respectively. The results showed that there is no significant relationship between infection and some variables analyzed, such as consuming and washing vegetables and keeping cats. Conclusion: Considering the high prevalence of toxoplasmosis infection among hemodialysis patients in Bushehr in comparison with healthy people in the region and since the fact that toxoplasmosis is a risk factor for life-threatening of hemodialysis patients, it is recommended that these patients should be screen for toxoplasmosis before dialysis as well as kidney transplant to prevent the spread of the infection through the process of dialysis

Efficacy of some metronidazole derivatives against Giardia lamblia, in vivo study

Giardiasis is a protozoal infection of small intestine caused by Giardia lamblia. This disease is usually asymptomatic, though it can present as acute or chronic diarrhea. Giardiasis, which is a major cause of intestinal infection, is endemic in Iran. Despite reports about drug resistance, long course treatment and various side effects of metronidazole, it is the drug of choice for giardiasis. In this study, we investigated the in vivo effects of five new derivatives (a-e) of metronidazole (MTZ) on the Giardia lamblia trophozoite in infected mice. Giardia intestinalis cysts were isolated from a patient and purified by sucrose gradient method. Fifty purified cysts were fed to the mice. After development of infection, the new metronidazole derivatives were given to the mice and the results were compared with metronidazole as the positive control group. Compounds a and b showed desirable antigiardiasis activity and could destroy the cyst and trophozoite of Giardia lamblia in mice after both two and four days, but the activity of the other compounds appeared only after 4 days

Anti-Toxoplasma Activity of 2-(Naphthalene-2-γlthiol)-1H Indole

Background:This study was undertaken to evaluate the viability, infectivity and immunity of Toxoplasma gondii tachyzoites exposed to 2-(naphthalene-2-ylthio)-1H-indole.  Methods:Tachyzoites of RH strain were incubated in various concentrations of 2- (naphthalene-2-ylthio)-1H-indole (25-800μM) for 1.5 hours. Then, they were stained by PI and analyzed by Fluorescence-activated cell sorting (FACS). To eval-uate the infectivity, the tachyzoites exposed to the different concentrations of the compound were inoculated to 10 BALB/c mice groups. For Control, parasites ex-posed to DMSO (0.2% v/v) were also intraperitoneally inoculated into two groups of mice. The immunity of the exposed tachyzoites was evaluated by inoculation of the naïve parasite to the survived mice. Results:The LD50 of 2-(naphthalene-2-ylthio)-1H-indole was 57 μmol. The lon-gevity of mice was dose dependent. Five mice out of group 400μmol and 3 out of group 800μmol showed immunization to the parasite.  Conclusion:Our findings demonstrated the toxoplasmocidal activity of the com-pound. The presence of a well-organized transporter mechanism for indole com-pounds within the parasite in conjunction with several effective mechanisms of these compounds on Toxoplasma viability would open a window for production of new drugs and vaccines

Antigiardiais Effects of New Metronidazole Derivatives on Trophozoites in TYI-S-33: Antigiardiais effects of metronidazole derivatives

Giardia is a flagellate protozoan with worldwide distribution that causes significant gastrointestinal diseases. The life cycle of the organism alternates between the active, proliferating trophozoite and the dormant cyst. 5-Nitroimidazoles have been used extensively in the treatment of amoebiasis, giardiasis, and trichomoniasis. We have previously synthesized some new analogues of metronidazole containing a phenyl or cyclohexanol ring in the side chain of the imidazole ring, and evaluated their antigiardiasis activity on giardia cyst. In the present study, we evaluated their activity against trophozoites of the parasite. For this purpose TYI-S-33 media was used and their MIC were compared with metronidazole and DMSO as positive and negative controls, respectively. The results showed that the new compounds had desirable antigiardiasis activities. Analogues which contain phenyl group in their structure are more active than those which contain cyclohexanol moiety. Although all the new compounds had higher MIC than metronidazole but their antigiardiasis activity were comparable to metronidazole and they may prove good alternatives for metronidazole. Keywords: Giardiasis; Metronidazole; Trophozoite

Zoonotic intestinal protozoan of the wild boars, Sus scrofa, in Persian Gulf’s coastal area (Bushehr province), Southwestern Iran

Aim: Wild boars, Sus scrofa, are potential reservoirs of many zoonotic diseases, and there are a possibility of transmission of the zoonotic diseases from these animals to humans and also domestic animals. This study aimed to evaluate the protozoan contamination of wild boars in the Persian Gulf’s coastal area (Bushehr Province), southwestern Iran. Materials and Methods: A total of 25 crossbred boars were collected during a course of vertebrate pest control in Bushehr province, in 2013. Samples were collected from the gastrointestinal tracts of each boar in 5% formalin, Bouin’s solution, sodium acetate-acetic acid-formalin, and polyvinyl alcohol fixatives. Fixed stool smears examined by trichrome and Ziehl–Neelsen staining. Results: Each of the 25 wild boars was infected with at least one of the intestinal protozoans. The rate of contamination with intestinal protozoan was 64% for Balantidium coli, 76% for Iodamoeba sp., 52% for Entamoeba polecki, 44% for Blastocystis sp. and 8% for Chilomastix sp. No intestinal coccidian was detected in studied boars when the stool samples were evaluated by Ziehl–Neelsen staining method. Conclusion: Findings of this study demonstrated that wild boars in the Persian Gulf coastal area are contaminated by many protozoans, including zoonotic protozoan, which poses a potential risk to locals as well as the domestic animals of the area

Prevalence and risk factors of intestinal protozoan infections: a population-based study in rural areas of Boyer-Ahmad district, Southwestern Iran

Abstract Background Parasitic infections are still a significant health problem in rural areas in developing countries including Iran. There is no recent population-based data about the prevalence of human intestinal parasites in most rural areas of Iran. The current study aimed to determine the prevalence of intestinal protozoan infection in inhabitants of rural areas of Boyer-Ahmad district, Southwestern Iran. Methods A total of 1025 stool samples were collected from the inhabitant of 50 randomly selected villages in Boyer-Ahmad Township. The stool samples were evaluated by parasitological methods including, direct wet-mounting, formalin ethyl acetate concentration, zinc sulfate floatation, and Trichrome permanent stain for detection of protozoan infections. Diarrheic samples were further evaluated with a modified Ziehl–Neelsen staining method for detection of coccidian parasites. Results The prevalence of both pathogenic and nonpathogenic intestinal parasites in the population was 37.5% (385 out of 1025 cases), some individual with multiple infections. Giardia lamblia was detected in 179 (17.46%), Blastocystis hominis in 182 (17.76%), Entamoeba histolytica/dispar in 9 (0.87%), Endolimax nana in 216 (21.07%), Entamoeba coli in 151 (14.73%), Ioedamoeba butschlii in 45 (4.39%), Chillomastix mesnili in 22 (2.14%), Trichomonas hominis in 2 (0.19%) and Dientamoeba fragillis in 2 (0.19%) of cases. Multivariate logistic regression revealed significant associations between protozoan infection (pathogenic protozoa) and contact with animals (OR yes/no = 2.22, p < 0.001) and educational status (OR higher/illiterate = 0.40, P = 0.01). Conclusion Findings of this study demonstrated that protozoan infection rate in rural areas of southwestern Iran is still high and remained as a challenging health problem in these areas

Evaluation of Correlation between the in Vitro Susceptibility of Field Isolates of Leishmania major and Clinical Outcomes of Meglumine Antimoniate Therapy in Fars Province, Iran

Background: This study was designed to detect whether there is a correlation between in vitro susceptibility of field isolates of Leishmania major and the clinical outcomes of meglumine antimoniate (Glucantime®) therapy, the main­stay of cutaneous leishmaniasis treatment in Iran. Methods: Forty-three patients infected with L. major were enrolled in this study from October 2009 to March 2010 and categorized as responsive or unresponsive to Glucantime® treatment after receiving the appropriate therapy. Then, intracellular amastigote approach was conducted on these field strains to investigate in vitro drug susceptibility as well. Results: At clinical level, out of 43 patients, 15 were clinically non-responsive and 28 were responsive to antimony therapy. All those 28 clinically sensitive strains were susceptible to antimony in the in vitro assay, whereas merely 11 isolates from 15 non-healing isolates were resistant in vitro. Finally, a good correlation (78.9%) with high sensi­tivity, specificity (100/73) between clinical outcomes and the in vitro susceptibility test was achieved.  Conclusion: The intracellular amastigote model could be an appropriate assay for evaluation of the in vivo drug sensitivity of field isolates. However, more comprehensive studies with larger sets of isolates are needed to confirm these preliminary data