Is GRA6 Gene a Suitable Marker for Molecular Typing of Toxoplasma Gondii? A Scoping Systematic Review.

Toxoplasmosis is a zoonotic disease with a worldwide prevalence that is caused by Toxoplasma gondii. This study aimed to summarize available data on genotyping T. gondii strains based on the GRA6 gene marker in different hosts around the world. We conducted a comprehensive literature search using five international databases (PubMed, Scopus, Science Direct, Web of Science, and Google Scholar) from inception until December 2021. We identified 32 papers eligible for inclusion in this systematic review. The majority of studies (50%) were carried out in Iran (n = 16) to identify T. gondii genotypes based on the GRA6 gene. Other countries with reported studies include China, Japan, Sweden, and Italy (n = 2 each). Out of 3,434 samples collected from various hosts, most studies (n = 11) focused on human samples (34.4%), followed by ovine (n = 7), pig (n = 4), goat (n = 3) and soil and cattle (n = 2).Using various molecular methods such as conventional PCR, nested-PCR, real-time PCR, microsatellite analysis, and Restriction Fragment Length Polymorphism (RFLP), we found DNA positive results in 805 out of 3,434 samples. Of these, 285 (35.40%), 207 (25.71%), 182 (22.60%), 65 (8.07%), and 18 (2.23%) were infected with types I, II, III, mix I, II, III, and mix II, III, respectively. Our data demonstrate that the GRA6 gene marker has sufficient polymorphism to detect three types of T. gondii genotypes in various hosts. Identifying the specific genotype could be valuable in developing new strategies for treatment, vaccination, diagnosis, control, and prevention of T. gondii infection.

Anti-leishmanial effects of Eryngium planum and Ecbilliun elaterum methanolic extract against Leishmania major.

Leishmaniasis is a vector-borne disease, one of the most important neglected tropical diseases. Existing anti-leishmanial treatments are not effective for a long time and associated with toxic side effects so searching for a new, effective and safe alternative treatments against infectious diseases is greatly needed. This study is aimed to assess the leishmaniacidal effects of methanolic extracts of Eryngium planum (E. planum) and Ecbilliun elaterum (E. elaterum) on Leishmania major (L. major), In vitro. The selected plants were collected from northern areas of Iran. The methanolic extract from the aerial parts of plants were prepared using maceration methods. GC- Mass analysis was used to determine the compounds of the plants. Promastigotes of L. major was cultured in RPMI-1640 medium and the anti-leishmanial and cytotoxicity effects of extracts at concentrations of 100, 200, 400 and 800 µg/ml were assessed using MTT assay. The data obtained from gas chromatography revealed that α-Pinene, Caryophyllene oxide, ß-Caryophyllene, Bicyclogermacrene and α-Bisabolol are the main compounds extracted from E. planum and α-Pinene, Germacrene D, Caryophyllene oxide, γ-Eudesmol and α-Bisabolol are the main components of E. elaterum. The results of MTT Assay revealed that E. planum at concentrations of 800 µg/ml after 24 h at 400 µg/ml after 48 h and the E. elaterium at concentrations of 800 µg/ml after 48 h at 400 µg/ml after 72 h had similar anti-leishmanial effects to the positive control. These results indicated that E. planum and E. elaterum are the potential sources for the discovery of novel anti-leishmanial treatments.

Promising potent in vitro activity of curcumin and quercetin nano-niosomes against Trichomonas vaginalis.

Trichomonosis, caused by infection with a motile protozoan parasite called Trichomonas vaginalis, is the most common non-viral sexually transmitted disease worldwide. Since the 1960s, metronidazole has been used as a drug of choice. Considering increased resistance to anti-trichomonial drugs, alternative treatments are urgently needed. In this study, the standard strain of T. vaginalis was cultured in TYM medium. Curcumin and quercetin loaded with hyaluronic acid niosomes were prepared by the thin film hydration method. The mean vesicle size, polydispersity index, and zeta potential of each prepared formulation were characterized, and its anti-Trichomonas activity was assessed by concentrations of 0.01, 0.1, 1, 10 and 100 mg/ml. The cytotoxicity effects of the mentioned drugs were determined using a MTT assay on L929 fibroblast cell viability. The particle sizes of curcumin, quercetin, and curcumin-quercetin entrapped modified nano-niosomes were characterised as 243 ± 5.28, 223 ± 7.21 and 266 ± 4.81 nm. The results showed that quercetin and curcumin at a concentration of 100 mg/ml after 24 h had anti-T. vaginalis activity. However, curcumin at a concentration of 100 at time 3h with 97% growth inhibition had better performance than positive control (metronidazole). According to the results of the MTT assay, all drugs, even at the highest concentration (400 mg/ml), had no toxic effect on the fibroblast cell line. According to potent in vitro activity of curcumin and quercetin nanoniosomes against T. vaginalis in comparison with metronidazole, it can be concluded these compounds could be promising therapeutic candidates for trichomonosis in future.

Peganum harmala and Nigella sativa: anti-leishmanial activity against Leishmania major promastigotes and amastigotes: in vitro and ex vivo experiment.

Leishmaniosis is one of the most important vectors borne disease that is endemic in tropical and subtropical areas. There are many approved treatment for different types of leishmaniosis but all are with some adverse side effects that limited its uses. Here, we attempt to evaluate in vitro and ex vivo anti-leishmanial activities of Peganum harmala (P. harmala) and N. sativa (Nigella sativa) on promastigotes and amastigotes of L. major. The plants were extracted by maceration method and prepared in concentrations of 7.8, 3.9, 1.9, and 0.9 µg. L. major were cultured in RPMI-1640 medium alone and in J774 cell line separately. The extracts at different concentrations were assessed against promastigote (in vitro assay) and amastigotes (ex vivo assay) of L. major for 72 h at 22 and 37°C, respectively. In current work, N. sativa at highest concentration (7.8 µg/ml) showed 54.4 and 60% anti-leishmanial activity with IC50 of 5.3 and 3.278 µg/ml, respectively. Also, P. harmala at highest concentration (7.8 µg/ml) showed 68.9 and 58.6% antileishmanial activity with IC50 of 2.4 µg/ml for both of them, respectively. The SI value was 38.22 for N. sativa, 25.9 for P. harmala, 19.4 for Amphotericin B, and 16.33 for Glucantime. The results of our study indicated that N. sativa and P. harmala are effective against L. major promastigotes and amastigotes and could be consider as an alternative treatments for leishmaniosis. Therefore, it is recommended that further studies be performed to confirm the efficacy and evaluate the toxicity of the herbal extracts.

Prevalence of Intestinal Parasites among Patients with Chronic Urticaria in Northern Iran.

BACKGROUND: Chronic urticaria (CU) has a range of clinical demonstrations and causes. Parasitic infections are mentioned as one of the main causes of the CU. OBJECTIVES: The aim of this study was to investigate the prevalence of intestinal parasites in patients with CU (with duration time of more than 6 weeks) compared healthy subjects. METHODS: A total of 169 cases and 210 controls were included in this study. Ages ranged from 1 to 77 years old. Three samples were gathered from each patients and direct wet mount, formol-ether concentration, Ziehl-Neelsen and trichrome staining were used. RESULTS: Out of the 379 individuals examined here, 208 were from urban areas and 171 from rural areas. Three stool samples were taken from each patient in three consecutive days. Based on parasitological tests, 7 (4.1%) cases from 169 patients with CU and 6 (2.9%) cases from 210 non- CU group individuals were positive for intestinal parasites. CONCLUSION: The prevalence of various parasites between case and control groups was not significant.

Promising In Vitro Anti-Toxoplasma gondii Effects of Commercial Chitosan.

CDATA[Background: Toxoplasmosis is a disease that results from infection with an obligate intracellular T. gondii parasite, one of the world's most common parasites. Considering the complications of chemical drugs and the need for an appropriate drug combination for treatment of toxoplasmosis and considering the antimicrobial potential of chitosan, as a natural source, this study was aimed to evaluate in vitro activity of commercial chitosan (CC) on T. gondii. METHODS: In this experimental study, the tachyzoites of T. gondii were collected from the peritoneal exudates from infected Balb/c mice. The tachyzoites were diluted in phosphate buffer saline (PBS) solution. Chitosan with low molecular weight was commercially purchased. Then, at concentrations of 10, 50, 100, and 200 µg/mL and after 30, 60, 120, and 180 minutes, the viability of tachyzoites was determined by using trypan blue 0.1%. Anti-T.gondii activity of CC in all concentrations was significantly higher than pyrimethamine as the control group (P=0.05). RESULTS: The concentration of 200 µg/mL of CC had the highest effects and killed 30.5, 52, 59, and 81.5% of tachyzoites after 30, 60, 120, and 180 minutes. Moreover, IC50 values of CC were 515, 171, 12.5, and <10 µg/mL in comparison with pyrimethamine as 58.82 µg/mL for 30, 60, 120, and 180 min of exposure time. CONCLUSION: Our results indicate that chitosan in low molecular weight had potent activity against T. gondii tachyzoites and could be an appropriate candidate for the treatment of at least acute toxoplasmosis, certainly, after complementary in vivo experiments.

Status of Anaplasma spp. infection in domestic ruminants from Iran: A systematic review with meta-analysis.

Anaplasma species are tick-borne pathogens that are obligatory intracellular of ruminants and other mammalians. In this investigation, we systematically reviewed the distribution of anaplasmosis among domestic ruminants in Iran. Five and four English and Persian databases were studied, respectively, based on keywords and throughout 17 years (2001-2017). Thirty-eight articles were included in this systematic review and meta-analysis. Totally, 5093 cattle, 1958 sheep, and 1232 goats corresponding to prevalence of Anaplasma infection from different areas of Iran were examined. The total prevalence of Anaplasma infection was estimated to be 34% (95% CI 27%, 41%) in domestic ruminants. Based on our data, Khozestan (54%) and Khorasan Razavi (46%) provinces were the most prevalent areas in Iran and Kerman (3%) and Hamedan (1%) provinces are the lowest. The highest prevalence of Anaplasma spp. infection was belonged to A. ovis (44%) and the lowest to A. phagocytophilum (1%) with a significant difference among them (p < .001). In addition, the most common diagnostic tests were PCR (54%), microscopy (35%) and ELISA (7%) assays. The high prevalence of ovine and bovine anaplasmosis in Iran, confirms the stability situations of animal anaplasmosis in the studied regions particularly northeastern and southwestern parts of the country. Our data offer valuable and encouraging information as regards the current situation of anaplasmosis in domestic livestock in Iran, which might be useful for active and passive surveillance and preventing plans.

Genotyping and phylogenic study of Acanthamoeba isolates from human keratitis and swimming pool water samples in Iran.

OBJECTIVE: Acanthamoeba keratitis cause severe corneal infection and lead to poor vision and blindness. This disease is caused by a unicellular amphizoic protozoon called Acanthamoeba spp. that present in different environments. This study aimed to represent the existence and genotyping of Acanthamoeba spp. in patients with keratitis and swimming pool water (SPW) in Tehran Province, Central Iran. METHODS: In this descriptive study, 56 clinical samples were collected from patients with keratitis and 30 water samples were collected from different swimming pools in Tehran Province. All samples were examined based on the morphological and molecular techniques. The genotypes were determined by sequencing the partial of 18S rRNA gene. RESULTS: Of 56 clinical (corneal) and 30 environmental (SPW) samples, 30.3% and 40.0% were positive for Acanthamoeba spp., respectively. According to sequencing analysis, 94.1% of amoebic keratitis isolates were belonged to T4 genotype and only one (5.8%) isolate was belonged to T11 genotype. All genotypes were detected from SPW samples were identified as T4 genotype. CONCLUSION: According to our results, use of contact lens and swimming in pool poses the major risk factor for amoebic keratitis in the studied area (Tehran). Moreover, T4 genotype was the predominant genotype of human keratitis and swimming pool samples there. Consequently, essential and practical measures are urgently needed to prevent subjects against this ocular seriously disease.

A case of fatal disseminated strongyloidiasis accompanied with intestinal obstruction.

Strongyloides stercoralis is an endemic parasite in some regions including the tropical and subtropical areas with high humidity. Most infections are asymptomatic with nonspecific signs and symptoms, making the final diagnosis complicated. Here, we report a patient referred to our hospital with signs consistent with sepsis, intestine obstruction, which finally died with the diagnosis of strongyloidiasis. The patient was from northern parts of Iran which are considered as endemic areas for S. stercoralis. In conclusion, there is an important message in this history, i.e. physicians should be aware of specific and non-specific signs of strongyloidiasis especially in people living in endemic areas to make an accurate final diagnosis by proper clinical and paraclinical examinations.

Potent antileishmanial activity of chitosan against Iranian strain of Leishmania major (MRHO/IR/75/ER): In vitro and in vivo assay.

BACKGROUND & OBJECTIVES: Leishmaniasis is one of the major neglected zoonotic parasitic diseases whose treatment and control is very complex. Pentavalent antimonials remain the primary drugs against different forms of leishmaniasis, however, resistance to antimony and its toxic effects has necessitated the development of alternative medications such as use of medicinal plants and natural compounds. The aim of the current study was to assess the in vitro and in vivo activities of chitosan as a natural resource against Leishmania major. METHODS: Low molecular weight chitosan, with 95% degree of deacetylation was melted in normal saline to a final concentration of 50, 100, 200 and 400 µg/ml. Then, the promastigotes of L. major (Iranian strain) were added to the wells of 96-well plate and 20 µl of each concentration was added to the RPMI 1640 medium. Live and dead promastigotes were counted after adding 0.1% eosin stain. The efficacy of the chitosan was also examined in BALB/c mice infected with Iranian strain of L. major. All in vitro experiments were performed in triplicate and the results of in vitro and in vivo tests were compared to the acetic acid and NaOH as negative control and glucantime as positive control. RESULTS: The low molecular weight chitosan was completely effective at concentrations of 100, 200 and 400 µg/ml on promastigotes of L. major after 180 min of application. Moreover, in the in vivo study, the mean size of dermal lesions significantly decreased in the groups treated with the chitosan compared to the control group. INTERPRETATION & CONCLUSION: According to the results of the study, it can be concluded that chitosan is a potent active compound against L. major and could be evaluated as a new antileishmanial drug in the future.

Anti-Toxoplasma activities of methanolic extract of Sambucus nigra (Caprifoliaceae) fruits and leaves / Actividad anti-Toxoplasma de extractos metanólicos de frutos y hojas de Sambucus nigra (Caprifoliaceae)

Toxoplasma gondii is an obligatory intracellular parasite that infects a wide range of warm-blooded animals and humans. Considering the severity of toxoplasmosis, side effects of current treatments, and the contribution of the ethnopharmacological knowledge for the treatment of parasitic infections, the aim of the present study was to investigate the efficacy of methanolic extracts from the fruits and leaves of Sambucus nigra against tachyzoite of T. gondii. For this, fruits and leaves of S. nigra were collected from Mazandaran province, Iran, were dried under the shade, and powdered using a commercial electrical blender. For extractions, methanol was used as solvent. Virulent RH strain of T. gondii was maintained in mice and macrophages containing tachyzoites were aspirated from the peritoneal cavity. Four concentrations (5, 10, 25 and 50mg/mL) of S. nigra extract were incubated with infected macrophages for 30, 60, 120 and 180 minutes and the viability of the tachyzoites were evaluated by trypan blue staining. Results showed that S. nigra fruit extracts at the concentrations of 5 and 10mg/mL killed 100% of T. gondii tachyzoites after 60 and 120 minutes, respectively; and concentrations of 25 and 50mg/mL killed 100% of the tachyzoites after 30 minutes. Additionally, extract of S. nigra leaves, at the concentrations of 5, 10 and 25mg/mL after 180 minutes, and concentration of 50mg/mL after 60 minutes, resulted with the highest efficacy. Our results showed that S. nigra has acceptable efficacy in vitro and the parasiticidal effect of fruit extract was significantly better than leaf extract. However, in vivo efficacy of this extract needs further investigation. Rev. Biol. Trop. 63 (1): 7-12. Epub 2015 March 01.

Toxoplasma gondii es un parásito intracelular obligatorio que infecta a una gran variedad de animales y seres humanos. Teniendo en cuenta la gravedad de la toxoplasmosis, los efectos secundarios de los tratamientos actuales, y la contribución de los conocimientos etnofarmacológicos para el tratamiento de infecciones parasitarias, el objetivo del presente estudio fue investigar la eficacia de los extractos metanólicos de los frutos y hojas de Sambucus nigra contra el taquizoito de T. gondii. Para esto, se recogieron frutos y hojas de S. nigra en la provincia de Mazandaran, Irán, se secaron a la sombra, y se pulverizaron con una batidora eléctrica comercial. Para las extracciones, se empleó metanol como disolvente. La cepa virulenta RH de T. gondii se mantuvo en ratones y los macrófagos con taquizoitos se aspiraron de la cavidad peritoneal. Cuatro concentraciones (5, 10, 25 y 50mg/ mL) de extracto de S. nigra se encubaron con los macrófagos infectados durante 30, 60, 120 y 180 minutos y la viabilidad de los taquizoitos se evaluó mediante tinción con azul de tripano. Los resultados mostraron que los extractos de frutos de S. nigra en las concentraciones de 5 y 10mg/mL mataron al 100% de los taquizoitos de T. gondii después de 60 y 120 minutos, respectivamente; y las concentraciones de 25 y 50mg/mL mataron al 100% de los taquizoitos después de 30 minutos. Además, el extracto de hojas de S. nigra, en concentraciones de 5, 10 y 25mg/mL después de 180 minutos, y una concentración de 50mg/mL después de 60 minutos, resultaron más eficientes. Nuestros resultados mostraron que S. nigra tiene una eficacia aceptable in vitro y el efecto parasiticida del extracto de frutos fue significativamente mejor que el del extracto de hoja. Sin embargo, la eficacia in vivo de este extracto necesita más investigación.