Seroprevalence and risk factors associated with toxoplasmosis and hydatidosis among the butchers of Tabriz city, the northwest of Iran: a case control study.

INTRODUCTION: Occupation plays an important role in the spread of infectious diseases in humans. Toxoplasmosis and hydatidosis are world-wide diseases with different routes of transmission. This study aimed to investigate the prevalence of toxoplasmosis and hydatidosis and risk factors associated with these diseases among the butchers of Tabriz City, the northwest of Iran. METHODS: In this case-control study conducted in Tabriz city in 2023, 250 serum samples were collected from butchers (n = 125) and outpatients referred to Imam Reza Hospital (n = 125) and. The ELISA test was used to identify IgG and IgM antibodies against toxoplasmosis and IgG antibodies against hydatidosis. The results were analyzed by statistics tests using SPSS v. 16 software. Risk factors' association was tested using Chi square or logistic regression analysis. RESULTS: The results indicated that 66/125 (52.8%) cases and 40/125 controls (32%) were positive for toxoplasmosis IgG antibody. Also, 5/125 (4%) and 1/125 (0.8%) were positive for toxoplasmosis IgM antibody in the case and control groups, respectively. In addition, 10/125 people (8%) were positive for anti-hydatidosis IgG antibody in the case group, while no positive cases were found in the control group. The main risk factors for toxoplasmosis were age (OR: 1.014), education level (OR: 0.638), and work experience(OR: 1.695), these factors for hydatidosis included age and education level (OR: 1.765 and 0.271) respectivily. CONCLUSIONS: Our results suggest the high prevalence of toxoplasmosis and in butchers of Tabriz, which required special attention and basic measures. Moreover, the prevalence of hydatidosis IgG antibodies also requires more attention to be focused on breaking the transmission and reducing the infection.

Cytotoxic and Immunomodulatory Activity of Curcumin and Chitosan on Experimental Toxoplasmosis.

Background: Toxoplasma gondii is a pathogenic parasite with worldwide distribution. We investigated curcumin and chitosan in combination on the viability of T. gondii tachyzoites in silico, in vitro and in vivo. Methods: A 3D model was employed in Urmia University of Medical Sciences, Urmia, Iran in 2021 to study the interaction between curcumin and dihydrofolate reductase (DHFR). Ramachandran root-mean-square deviation and VERIFY3D validated the model. Cytotoxicity of curcumin and chitosan was evaluated by MTT viability assay. BALB/c mice infected with 104 Toxoplasma organisms were treated with curcumin, chitosan, and the combination of curcumin+chitosan. Serum levels of inducible NO synthetase (iNOs), interferon gamma (IFN-γ), interleukin (IL)-5, glutamate oxaloacetic transaminases(SGOT), and glutamic pyruvate transaminase (SGPT) were determined. Result: Curcumin-DHFR and curcumin-DHPS (dihydropteroate synthase) interactions and calculated enzyme energy indicated an excellent affinity for curcumin with DHFR, but not DHPS. MTT results of concurrent treatments demonstrated IC50 rates of 0.1, 0.05, and 0.01 mg/ml at 24, 48, and 72h, respectively. IFN-γ, IL-5 and iNOs levels in curcumin+chitosan treated mice were 1.71, 0.51, and 1.51 IU/L, while those of SGOT and SGPT were 76 and 84 IU/L, respectively. Conclusion: The combination of curcumin and chitosan increased survival time of infected mice by seven days. Curcumin and chitosan in combination regulated the immune system and reduced liver damage, potentially forming the basis of a new treatment for toxoplasmosis.

Modifications in gene expression and phenolic compounds content by methyl jasmonate and fungal elicitors in Ficus carica. Cv. Siah hairy root cultures.

BACKGROUND: One of the most effective strategies to increase phytochemicals production in plant cultures is elicitation. In the present study, we studied the effect of abiotic and biotic elicitors on the growth, key biosynthetic genes expression, antioxidant capacity, and phenolic compounds content in Rhizobium (Agrobacterium) rhizogenes-induced hairy roots cultures of Ficus carica cv. Siah. METHODS: The elicitors included methyl jasmonate (MeJA) as abiotic elicitor, culture filtrate and cell extract of fungus Piriformospora indica as biotic elicitors were prepared to use. The cultures of F. carica hairy roots were exposed to elicitores at different time points. After elicitation treatments, hairy roots were collected, and evaluated for growth index, total phenolic (TPC) and flavonoids (TFC) content, antioxidant activity (2,2-diphenyl-1-picrylhydrazyl, DPPH and ferric ion reducing antioxidant power, FRAP assays), expression level of key phenolic/flavonoid biosynthesis genes, and high-performance liquid chromatography (HPLC) analysis of some main phenolic compounds in comparison to control. RESULTS: Elicitation positively or negatively affected the growth, content of phenolic/flavonoid compounds and DPPH and FRAP antioxidant activities of hairy roots cultures in depending of elicitor concentration and exposure time. The maximum expression level of chalcone synthase (CHS: 55.1), flavonoid 3'-hydroxylase (F3'H: 34.33) genes and transcription factors MYB3 (32.22), Basic helix-loop-helix (bHLH: 45.73) was induced by MeJA elicitation, whereas the maximum expression level of phenylalanine ammonia-lyase (PAL: 26.72) and UDP-glucose flavonoid 3-O-glucosyltransferase (UFGT: 27.57) genes was obtained after P. indica culture filtrate elicitation. The P. indica elicitation also caused greatest increase in the content of gallic acid (5848 µg/g), caffeic acid (508.2 µg/g), rutin (43.5 µg/g), quercetin (341 µg/g), and apigenin (1167 µg/g) phenolic compounds. CONCLUSIONS: This study support that elicitation of F. carica cv. Siah hairy roots can be considered as an effective biotechnological method for improved phenolic/flavonoid compounds production, and of course this approach requires further research.

Seroprevalence of Fasciola sp. and Toxoplasma gondii Infections in Rural and Urban Inhabitants of Jolfa County, Northwest Iran.

Fascioliasis and toxoplasmosis are the two important zoonotic diseases that are endemic in Iran and share some common transmission routes. The present study is aimed at determining the seroprevalence of human fascioliasis and toxoplasmosis in rural and urban areas of Jolfa County, Northwest Iran. In a cross-sectional study, 600 human sera were collected randomly from humans living in Jolfa County including three cities and 13 villages from 2017 to 2018. Anti-Toxoplasma IgG and anti-Fasciola sp. IgG tests have been performed using the enzyme-linked immunosorbent assay. Four (0.7%) out of 600 human sera showed positive levels of anti-Fasciola IgG. Three out of four seropositive humans were from an urban area, and one (25%) was from rural inhabitants. Considering T. gondii infection, 45% of studied human sera were seropositive for anti-T. gondii IgG. In conclusion, this is the first study reporting Fasciola seropositivity in the area. Based on the findings, human fascioliasis is present in the studied area, Northwest Iran, granted in low prevalence. Considering T. gondii seropositivity, the prevalence is high, yet close to the reports from other regions in the province.

Evaluation of In Vitro Cytotoxic and Apoptotic Effects of Miltefosine on the Toxoplasma gondii RH Strain.

Background: We aimed to investigate the cytotoxic and apoptotic effects of miltefosine on Toxoplasma gondii RH strain by various techniques. Methods: The study was conducted at the Department of Parasitology and Mycology, Urmia University of Medical Sciences, Iran in 2020. Four groups of five BALB/c mice were selected. The cytotoxicity test was conducted by adding miltefosine to T. gondii tachyzoites; control tachyzoites received PBS and MTT assay was done on each suspension. For evaluating the Th1-type immune responses, the serum levels of IFN-γ and nitric oxide (NO) were assessed in mice after injecting tachyzoites and miltefosine, respectively. The flow cytometry technique was performed on T. gondii tachyzoites challenged with IC50 and IC90 doses of miltefosine and unchallenged cells. DNA fragments in T. gondii tachyzoites were detected by Terminal dUTPnick-end labeling (TUNEL) method. Results: Overall, 256, 64, 32, and 16 µg concentrations of miltefosine, respectively could kill more than 50% of viable T. gondii tachyzoites. The infected mice group, treated with miltefosine, significantly produced more IFN-γ relative to other groups (P< 0.001). Moreover, a significant difference was found in inducible NO synthase between the experimental and control groups (P<0.05). The flow cytometry results demonstrated a concentration-dependent apoptosis rate in tachyzoites incubated with miltefosine, though the necrosis rate was non-significant. DNA fragmentation analysis indicated oligonucleotides (18-200 bp) in tachyzoites treated with 11µg of miltefosine for 24, 48 and 72 h. However, this pattern was not observed in untreated control microorganisms. Conclusion: Miltefosine could be a favorable candidate for use as a new treatment for toxoplasmosis.

Molecular examination for Coxiella burnetii and Brucella spp. infections in Iranian women experiencing spontaneous miscarriage.

BACKGROUND: Spontaneous miscarriage, a leading health concern globally, often occurs due to various factors, including infections. Among these, Coxiella burnetii and Brucella spp. may have adverse effects on pregnancy outcomes. While previous research has established a link between infections and spontaneous miscarriage, our study aimed specifically to investigate the presence of these two pathogens in abortion samples from women who experienced spontaneous miscarriages in Iran. Our study can add to the existing knowledge by focusing on Iran, a region with a high prevalence of C. burnetii and Brucella spp. As a result, it could provide a better understanding and unique insights into the relationship of these pathogens with spontaneous miscarriages in endemic regions. METHODS: From March 2021 to March 2022, a total of 728 abortion samples (including placenta and cotyledon) were collected from 409 women who had experienced spontaneous miscarriages in the provinces of Tehran, Fars, and West Azerbaijan in Iran. The specimens included 467 Formalin-Fixed Paraffin-Embedded (FFPE) and 261 fresh frozen samples. After DNA extraction from abortion samples, the quantitative real-time PCR (qPCR) assay targeted a specific fragment of the IS1111 and IS711 elements for molecular identification of C. burnetii and Brucella spp., respectively. Furthermore, the qPCR assay employing specific primers for different species was used to determine the species of Brucella. RESULTS: Among the studied women, 1 out of 409 (0.24%) samples tested positive for Brucella spp., specifically Brucella melitensis. There were no positive specimens for C. burnetii. CONCLUSIONS: Our study contributes to understanding the potential involvement of Brucella species in spontaneous infectious abortion within endemic regions. The identification of B. melitensis in this study highlights the need for further research in this area. However, while our results suggest a relatively low or zero identification of these pathogens in our sample population, this does not rule out the possibility of undetected infections. Therefore, it is critical to acknowledge the limitations of the molecular techniques used (qPCR), which may have potential limitations such as sensitivity and specificity. Moreover, because 64.15% of our samples were FFPE, the sensitivity of the qPCR test may be reduced. These raise concerns about the accuracy of the reported prevalence rates and the potential for false positives or negatives.

Anti-Leishmania major activity of Calotropis procera extract by increasing ROS production and upregulating TNF-α, IFN-γ and iNOS mRNA expression under in vitro conditions.

BACKGROUND: Leishmaniasis, caused by protozoan parasites of the genus Leishmania, is a neglected tropical disease with 700,000 to 1,000,000 global new cases annually. Adverse effects associated with expense, long-term treatment and drug resistance have made conventional therapies unfavorable, encouraging the search for alternative drugs based on plant products. In this study, the effect of Calotropis procera (Asclepiadaceae) extract against viability of promastigotes and amastigotes of Leishmania major was evaluated in vitro. METHODS: The extract from the leaves of C. procera seedlings was prepared using a methanol maceration method. The colorimetric cell viability 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to determine the growth-inhibitory effect of the extract on promastigotes. The level of reactive oxygen species (ROS) in promastigote cultures was determined after treatment with the extract using the 2',7'-dichlorofluorescein diacetate (DCFH-DA) method and compared with untreated cultures (control). After exposure to the extract the expression levels of tumor necrosis factor-α (TNF-α), interferon gamma (IFN-γ) and inducible nitric oxide synthase (iNOS) genes were determined and compared to control in peripheral blood mononuclear cells (PBMCs) infected with L. major. RESULTS: Based on the MTT assay, the C. procera extract significantly reduced the proliferation of L. major promastigotes with IC50 values of 377.28 and 222.44 µg/mL for 24 and 72 h, respectively (p < 0.01). After treatment with 222.44 and 377.28 µg/mL of C. procera extract, ROS production in L. major promastigote cultures increased 1.2- to 1.65-fold and 2- to 4-fold compared to the control, respectively (p < 0.05). C. procera extract induced significant increases in gene expression of TNF-α (2.76-14.83 fold), IFN-γ (25.63-threefold) and iNOS (16.32-3.97 fold) in infected PBMCs compared to control (p < 0.01). CONCLUSIONS: On the basis of its anti-leishmanial activity, C. procera can be considered as a promising new plant source for the potential treatment of leishmaniasis.

In vitro and in silico scolicidal effect of sanguinarine on the hydatid cyst protoscoleces.

We aimed to investigate the scolicidal effects of sanguinarine on hydatid cyst protoscoleces (PSCs) in vitro and in silico. Different targets were docked into the active sites of sanguinarine. Molecular docking processes and visualization of interactions were performed using AutoDock Vina and Discovery Studio Visualizer. Binding energy was calculated and compared (kcal/mol). PSCs were aspirated from the hydatid cysts and washed. The sediments of PSCs were then exposed to various concentrations (50, 25, 12, 6, 3, and 1 µg/mL) of sanguinarine. The viability test was finally evaluated by the Trypan blue solution 4%. Levels of malondialdehyde (MDA), superoxide dismutase (SOD), glutathione (GSH), glutathione peroxidase (GPX), and catalase were analyzed to assess the level of oxidative stress-treated PSCs. Caspase-3 activity rate was determined to evaluate cell apoptosis in treated PSCs. Among the receptors, acetylcholinesterase was identified as the excellent target, with Vina score of -11.8. Sanguinarine showed high scolicidal effects after 12, 24, and 48 h. Also, in the first hour of exposure to the drug, caspase-3 activity and MDA level significantly increased, but the levels of GSH and GPx had a significant reduction after 12, 24, and 48 h (P < 0.05). The findings of this study revealed that sanguinarine have potent scolicidal effects in vitro and in silico and could be considered an opportunity for the introduction of a novel and safe therapeutic agent for the treatment of cystic echinococcosis. However, supplementary studies will be desired to prove the current findings by examining sanguinarine in a clinical setting.

Prevalence and risk factors of Toxoplasma gondii infection among women with miscarriage and their aborted fetuses in the northwest of Iran.

Toxoplasmosis is a worldwide disease of various animals and human and results from infection with the Toxoplasma gondii parasite. Abortion and congenital defects are important consequences of the T. gondii infection. The aim of this study was to determine the Toxoplasma-induced abortions among women with miscarriage and the presence of T. gondii in their aborted fetuses in Urmia, the northwest of Iran. This cross-sectional study was conducted with 215 women with abortion and their aborted fetuses, from 2020 to 2021. Seroprevalence of anti-Toxoplasma IgG and IgM were determined using the sera of the aforesaid women. Nested PCR was carried out using RE-529 gene sequences, and sequencing was performed using the T. gondii GRA6 gene on the remnant of pregnancy after abortion. The tissue positive samples were then subjected to another PCR on GRA6 gene and sequenced for genotyping. Among 215 serum samples of women with abortion, 70 (32.6%) were positive for anti-Toxoplasma IgG, and three (1.4%) were positive for IgM. The RE-529 sequence of T. gondii was positive in three (1.4%) of the aborted fetuses. The analysis of GRA6 gene indicated that all three positive samples carried a GRA6 allele (GRA6I) of T. gondii type I genotype. Our findings suggest that T. gondii is one of the causative agents of spontaneous abortion in West Azerbijan Province, the northwest of Iran.

The Effect of Curcumin on the Expression of INFγ, TNF-α, and iNOS Genes in PBMCs Infected with Leishmania major [MRHO/IR/75/ER].

BACKGROUND: Leishmaniasis, caused by the Leishmania parasite, is one of the most important tropical neglected diseases. The urgent search for effective, inexpensive, and preferably herbal anti-leishmanial agents, is needed. OBJECTIVE: Curcumin is a natural polyphenolic compound derived from turmeric that is well known for its antioxidant, anti-inflammatory, anti-tumor, and anti-cancer activity. METHODS: The present work evaluates the anti-leishmanial [Leishmania major] activity of curcumin. The infected PBMCs were treated with curcumin. The ROS level at 6, 12, 24 h and gene expression levels at 24, 48, and 72 h of PBMCs after treatment with curcumin were determined. RESULTS: Based on the results, the curcumin concentrations of 268 µM [24 h] and 181.2 µM [72 h] were defined as IC50 against L. major promastigotes. Treatment of L. major infected-peripheral blood mononuclear cells [PBMCs] with IC50 concentrations of curcumin, depending on exposure time, significantly induced the reactive oxygen species [ROS] generation and increased the expression levels of interferongamma [IFN-γ], tumor necrosis factor-alpha [TNF-α], and nitric oxide synthase [iNOS] genes. CONCLUSION: These findings suggest the potential of curcumin against Leishmaniasis.

Anti-toxoplasma and Cytotoxic Activities of Holothuria leucospilota Extract and TiO2NPs In vitro and In vivo.

BACKGROUND: An impressive treatment for toxoplasmosis is the combinatory use of sulfadiazine and pyrimethamine. However, both the drugs involve significant side effects and toxicity for the host. Therefore, the discovery of new anti-toxoplasma medications with high efficacy and less to no side effects is urgently needed. OBJECTIVE: This study aimed to evaluate the anti-toxoplasmic effects of Holothuria leucospilota (H. leucospilota) extract and TiO2NPs on the cell death of Toxoplasma gondii (T. gondii) tachyzoites in vitro and serum liver enzymes (AST, ALT, and ALP), and also to evaluate the immune response and production of IL-5, IFN-γ, and TNF-α in a mouse model. MATERIALS AND METHODS: The cytotoxicity of TiO2NPs and H. leucospilota extract against the tachyzoite of T. gondii was evaluated by the methyl thiazolyl tetrazolium (MTT) assay. The levels of serum TNF-α, IFN-γ, IL-5, and liver enzymes were measured, as well. All the groups were subjected to T. gondii, and the survival rate of experimental mice was evaluated. RESULTS: Our findings suggested in vivo and in vitro anti-toxoplasmic activity of TiO2NPs and H. leucospilota extract by inhibiting the proliferation and invasion of T. gondii tachyzoite. In addition, a significant increase in IFN-γ and TNF-α production was observed in mice treated with high doses of TiO2NPs and H. leucospilota extract. However, IL-5 levels decreased in TiO2NPs and H. leucospilota extract-treated mice. Our results also showed a highly significant increase (P < 0.05) in the levels of ALT, AST, and ALP in the groups injected with TiO2NPs and H. leucospilota extract, but not the control group. CONCLUSION: TiO2NPs and H. leucospilota extract have greater anti-toxoplasma effects in vitro and in vivo. These two compounds could be considered as a candidate for use against toxoplasmosis, both therapeutically and prophylactically.

Prevalence of strongyloidiasis in the general population of the world: a systematic review and meta-analysis.

Strongyloides stercoralis is a neglected soil-transmitted helminth affects approximately 100-370 million people globally. The life cycle is unusual as only larvae can be found in stool specimens. Thecurrent review and meta-analysis represented the distribution of strongyloidiasis in general population of the world based on published papers. Five English databases (Science Direct, Scopus, PubMed, Web of Science, and Google Scholar(were explored for literature published before October 2019.Altogether 235 studies (862243 participants) was eligible. Regarding diagnostic method, the overall prevalence for studies performed microscopic, culture, immunological and molecular method was 1.47% (95% CI = 1.56%), 10.08% (95% CI = 8.99% - 11.16%), 23.88% (95% CI =  20.82% - 26.94%) and 9.3% (95% CI  =  7.2% - 11.3%), respectively. Based on microscopic methods, the highest prevalence was related to the Western Pacific region [9.47% (95% CI =  8.55% - 10.39%)]. According to the culture method, Western Pacific region [21.36% (95% CI  =  16.32% - 26.39%)] had the highest estimated pooled prevalence. In immunological studies, Eastern Mediterranean Region [40.72% (95% CI  =  36.74% - 44.70%)] had the highest seroprevalence.Also in molecular surveys, the highest prevalence was related to the African region [19.72% (95% CI  =  16.71% - 22.73%)]. The current study indicated that strongyloidiasis is still considered a health problem in many parts of the world. Thus a comprehensive control program and improvement of public health sectors are required.

Piriformospora indica based elicitation for overproduction of phenolic compounds by hairy root cultures of Ficus carica.

Ficus carica L. is an important source of phenolic and flavonoid compounds with valuable pharmaceutical application across various diseases. The current study was carried out to investigate the influence of Piriformospora indica elicitation on growth, production of phenolic compounds, antioxidant capacity, and expression level of flavonoid biosynthetic pathway genes in hairy root (HR) cultures of F. carica. The maximum improvement in accumulation of phenolic compounds was observed when HR culture of Ficus carica L. was exposed to 2% culture filtrate of P. indica for 72 h: gallic acid (80.5- fold), caffeic acid (26.2-fold), coumaric acid (4.5-fold), and cinnamic acid (60.1-fold), apigenin (27.6-fold) and rutin (5.7-fold). While the highest levels of chlorogenic acid (4.9-fold) and quercetin flavonoid (8.8-fold) were obtained after 48 h elicitation with culture filtrate and cell extract of P. indica at 6% (v/v), respectively. The analysis of biosynthetic genes revealed that the exposure to fungal elicitors resulted in up-regulation of phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS), UDP-glucose flavonoid 3-O-glucosyltransferase (UFGT) and MYB3 transcription factor. This study shows the potential of P. indica as an efficacious elicitor for enhancing the secondary metabolites production by F. carica HRs.

Comparison of diagnostic methods (wet mount, trichrome staining, formol-ether, PCR, and xenic in vitro culture) for the detection of Blastocystis in stool samples in Urmia educational hospitals, the Northwest of Iran.

Blastocystis spp. is known as a common intestinal protozoan parasite in human and animals. The parasite has a worldwide distribution and is frequently detected in faecal samples in clinical parasitology laboratories. The goal of the study was to compare the sensitivity and specificity of formol-ether technique (FECT), trichrome staining, xenic in vitro culture (XIVC), microscopy of faecal smears, and polymerase chain reaction (PCR) methods for detecting Blastocystis spp. in human stool samples. The prevalence of the parasite in the stool samples referred to educational hospitals was also determined. A total of 575 cases were assessed to detect the parasite. After collecting from patients referring to Urmia educational hospitals, the samples were examined by microscopy of faecal smears, trichrome staining, FECT, XIVC using Jones' medium, and PCR, to evaluate the presence of Blastocystis spp. Microscopy of faecal smears, trichrome staining, FECT, and PCR technique detected 94, 100, 96, and 44 positive cases, with the sensitivity of 71.3%, 74.4%, 74.4%, and 80.4% and the specificity of 99.6%, 99.1%, 100%, and 93.1%, respectively. XIVC method identified the highest number of positive cases (129 cases) among the other methods. Our findings indicates that XIVC technique is more sensitive method for the detection of Blastocystis spp. in human stool, as compared to direct smear, trichrome staining, FECT, and PCR methods.

Screening of Cystic Echinococcosis and Toxocariasis in Urmia Municipal Workers, Northwest Iran.

BACKGROUND: Echinococcus granulosus and Toxocara spp., the causative agents of hydatid cyst and toxocariasis in humans and animals, are zoonotic infections with medical and veterinary significance respectively. AIM: Herein, we aimed to investigate the seroprevalence of toxocariasis and cystic echinococcosis in Urmia municipal workers (gardeners and sweepers) in 2018 using ELISA assay. OBJECTIVE: There is no precise report on the seroprevalence of toxocariasis and cystic echinococcosis in Urmia municipal workers using ELISA assay. Therefore, this study found an opportunity to investigate this subject. METHODS: A total of 220 blood samples were obtained from municipal workers with an average age of 41.31 ± 9.42. Commercial anti-Toxocara canis and hydatid cyst ELISA kits were used to assess the seroprevalence rates. RESULTS: Approximately, 14.5% and 2.3% of individuals were seropositive for toxocariasis and hydatidosis, respectively. Consuming unboiled milk and/or raw/uncooked meat and rural conditions are risk factors for toxocariasis. Regarding hydatidosis, consuming unwashed vegetables was the only statistically significant risk factor. Toxocara infection (p = 0.01), but not hydatid cyst (p = 0.05), is more prevalent among sweepers than municipal gardeners. CONCLUSION: Due to the occupational condition of municipal workers, they are exposed to many infectious agents; thus, more researches should be done on these populations.

Sero-epidemiology of Hydatidosis Among General Population of Jolfa County, Northwestern Iran Using IHA, ELISA and Western Blot (2017-2018).

BACKGROUND: Human hydatidosis is mostly a latent and neglected disease with known endemicity in Iran. AIMS: Due to the importance of this infection in the country and its latent nature, we aimed to evaluate the serological status of hydatid cyst in northwestern Iran. OBJECTIVES: Herein, we evaluated the serological status of hydatid cyst in urban and rural inhabitants of Jolfa county, northwestern Iran during 2017-2018. METHODS: In total, 1296 blood samples were obtained from human individuals and the presence of anti-E. granulosus antibodies was investigated using IHA, ELISA and WB. RESULTS: Based on results, 25 IHA positive person were detected in the examined population, however ELISA test showed 14 of 25 IHA positive patients as negative. Also, 269 IHA negative fellows were shown as negative by ELISA. WB analysis of sera from 25 IHA positive subjects revealed consistent results with the ELISA test, and the most reactive SHCF Ag was a 37 KDa protein. The age-standardized seroprevalence of hydatidosis among Jolfa's general population was 1.12% with 95%CI: 1.02-1.20. Moreover, there existed a significant association between keeping/- contact with dogs (P = 0.022) as well as vegetable consumption (P < 0.001) with ELISA positive test results. CONCLUSION: Along with such serological evidence in this region, we highly suggest physical examination and applying imaging techniques for suspected cases in the area for a better understanding of CE.

Seroprevalence of Toxocara Infection in Association with Different Risk Factors among Children of 4-12 Years Old Referred to Some Medical Centers in Aras Free Zone, Northwest Iran.

BACKGROUND: Toxocariosis is a parasitic disease caused by the larval stage of Toxocara species from dog and cat. It has a worldwide distribution with higher prevalence in children. This study aimed to determine seroprevalence of Toxocara infection and its association with some risk factors among children of Aras Free Zone (Jolfa City) in Northwest of Iran. METHODS: Sera were collected from 514 children aged 4-12 yr old attending to some medical centers in the study area from May 2018 to Feb 2019. Anti-Toxocara IgG antibodies assay was performed using commercial ELISA kit (Nova Tec, Germany). The seropositivity rate was determined and its association with different demographic criteria and risk factors were statistically analyzed. RESULTS: The overall seroprevalence was 2.3% (12/514). Risk factors of children's age group and contact with either pet animals (dog and cat) and/or soil were significantly associated with seropositivity. However, there was not any relationship between Toxocara infection and gender of children, place of residency (urban or rural) and their mothers' education level. CONCLUSION: Both girls and boys are at risk of Toxocara infection in the study area. Younger age of childhood and contact with sources of infection were important associated factors. More probably, additional criteria are involved in the initiation of infection.

Association of Toxoplasma gondii infection with cardiovascular diseases: a cross-sectional study among patients with heart failure diseases in Urmia, North-West of Iran.

Toxoplasmosis is a disease caused by the protozoan parasite Toxoplasma gondii. Infection with T. gondii is one of the most common parasitic diseases in humans and other warm-blooded animals with global distribution and generally, one-third of human populations are estimated to be contaminated with this parasite. The prevalence of infection varies according to age, geographical location and dietary habits. The socioeconomic losses caused by the disease can be costly for the community. Acquired toxoplasmosis is potentially associated with schizophrenia, Alzheimer's disease, driving accidents, self-injury and suicide. Also, unusual strains of parasite that are genetically different from the rest (atypical strains) are responsible for several cases of lethal acquired parasites in people with safe immunity, which highlights the potential danger of this parasite in public health. As there is no comprehensive study on the association between toxoplasmosis and cardiovascular diseases in Iran, therefore, current study aimed at assessing the relationship between cardiovascular disease and toxoplasmosis among cardiac patients at the Seyyed al-Shohada specialist Cardiology Centre, Urmia, Iran. This study investigated the seropositivity rate for anti-Toxoplasma IgG antibodies by ELISA in patients with cardiovascular diseases. So, 375 patients with cardiovascular diseases and 336 healthy volunteers were selected for this investigation. The seropositivity rate of anti-Toxoplasma IgG antibodies was significantly higher in cardiovascular patients (63.73%) than in healthy volunteers (37.64%) (P<0.001). Also, a positive association was observed between anti-T. gondii IgG antibody seropositivity and cat contact (P≤0.001, OR: 5.178; 95% CI: 1.97-13.57), consumption of raw or undercooked meat (P≤0.001, OR: 0.3; 95% CI: 0.15-0.61), and consumption of not boiled milk (P≤0.001, OR: 0.26, 95% CI: 0.12-0.54). Our results indicate that T. gondii infection is associated with heart disease and suggest that heart disease might be related with a chronic infection. Risk factors associated with T. gondii exposure found in the present study may help design future prevention strategies against T. gondii infection.

Agrobacterium rhizogenes mediated transformation of Ficus carica L. for the efficient production of secondary metabolites.

BACKGROUND: Ficus carica L., an ancient source of food and medicines, is rich in valuable nutritional and secondary compounds with antioxidant, antimicrobial, and anticancer effects. The present study is the first attempt to examine hairy root (HR) induction of F. carica (Sabz and Siah) by inoculating the 3-week-old shoots and leaves with different strains of Agrobacterium rhizogenes and also to investigate methyl jasmonate (MeJA) elicitation of HRs to produce a fast and high-yield production method for secondary metabolites. RESULTS: The maximum transformation rate (100%) was achieved by inoculating the shoots with Agrobacterium rhizogenes strain A7. Siah HRs elicited with 100 and 200 µmol L-1 MeJA and Sabz HRs with 100 µmol L-1 MeJA showed the highest total phenolic content. The highest flavonoid content was 3.935 mg QE g-1 DW in Siah HRs treated with 200 µmol L-1 MeJA and 2.762 mg QE g-1 DW in Sabz HRs treated with 300 µmol L-1 MeJA. The 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging capacity and ferric reducing antioxidant power (FRAP) value of HRs were affected by MeJA treatments. Methyl jasmonate elicitation also significantly enhanced the content of six phenolic acids (gallic acid, caffeic acid, chlorogenic acid, coumaric acid, rosmarinic acid, and cinnamic acid) and three flavonoids (rutin, quercetin, and apigenin). Thymol, a monoterpene phenol, was the main HR compound detected in gas chromatography mass spectrometry (GC-MS) analysis of the essential oils. CONCLUSION: Induction of HRs and elicitation of F. carica HRs by MeJA resulted in a significant increase in the production of important phenolic compounds and a significant increase in antioxidant capacity. © 2020 Society of Chemical Industry.

In Vitro and in Vivo Scolicidal Activities of Holothuria leucospilota Extract and CeO2 Nanoparticles against Hydatid Cyst.

BACKGROUND: We aimed to investigate the scolicidal effects of Holothuria leucospilota extract and CeO2 nanoparticles against protoscoleces of hydatid cysts in-vitro and in-vivo. METHODS: Hydatid cysts were collected from, Urmia slaughterhouses between years 2016-2017 and the hydatid fluid aspirated from the fertile cysts. Various concentration of H. leucospilota extract, CeO2 NPs and combination of CeO2-NPs/H. leucospilota were used for 10-60 min to evaluate the viability of protoscoleces by 0.1% eosin method. CASPASE -3 activity measured for assessment of cell apoptosis in treated protoscoleces. BALB/c mice were infected intraperitoneally with 2000 viable protoscoleces and treated daily for 4 wk by intragastrical inoculation with H. leucospilota, CeO2 NPs, combination of CeO2 NPs/H. leucospilota and Albendazole. Cyst development was macroscopically analyzed. RESULTS: H. leucospilota extract and combination of CeO2 NPs/H. leucospilota have potent scolicidal activity at concentration of 20 mg/ml and 15 mg/ml after 60 min treatment. Maximum caspase-3 activity was observed when protoscoleces expose with H. leucospilota and combination H. leucospilota & CeO2 NPs. After treatment of cyst infected mice with extract and CeO2 NPs, combination of CeO2 NPs/H leucospilota and albendazole, a significant decrease in number of cysts, size and volume of cyst (P<0.05) was observed. CONCLUSION: This result shows an antihydatic and scolicidal effects of H. leucospilota extract and CeO2 NPs.