High occurrence of Blastocystis sp. subtype 3 in individuals referred to medical laboratories in Kermanshah, Iran.

Aim: The current study investigated the prevalence and genotypes of Blastocystis sp. in individuals who referred to medical laboratories in Kermanshah, Iran. Background: Blastocystis sp. is a common intestinal protozoan found in humans and a wide range of animals, and it is involved in the development of gastrointestinal disorders. Methods: A total of 950 stool samples were examined using the standard formalin-ether concentration technique. All specimens were cultured in Robinson xenic medium. Subsequently, DNA extraction and PCR amplification of subtype specific sequence-tagged site (STS) were conducted. Results: Microscopic examination showed that 86 out of 950 samples (9.05%) were infected with Blastocystis sp. Subsequently, 33 of 86 positive samples were cultured and molecularly confirmed by conventional PCR, indicating six subtypes (ST1-ST6). Of note, ST3 (45.0%) was the predominant subtype, followed by ST1 (15.15%) and ST5 (12%). Conclusion: Based on the current findings, ST3 was the most frequent subtype among all positive samples. Having a better understanding of Blastocystis sp. subtype distribution and risk factors would lead to improved preventive measures.

Towards the First Multiepitope Vaccine Candidate against Neospora caninum in Mouse Model: Immunoinformatic Standpoint.

Neospora caninum is an economically significant parasite among livestock, particularly in dairy cattle herds, causing storm abortions. Vaccination seems necessary to limit the infection and its harsh consequences. This is the first steps towards developing a multiepitope vaccine candidate against N. caninum using in silico approaches. High-ranked mouse MHC-binding and shared linear B-cell epitopes from six proteins (SRS2, MIC3, MIC6, GRA1, IMP-1, and profilin) as well as IFN-γ-inducing epitopes (from SAG1) were predicted, screened, and connected together through appropriate linkers. Finally, RS-09 protein (TLR4 agonist) and histidine tag were added to N- and C-terminal of the vaccine sequence, yielding 486 residues in length. Physicochemical properties showed a stable (instability index: 27.23), highly soluble, antigenic (VaxiJen score: 0.9554), and nonallergenic candidate. Secondary structure of the multiepitope protein included 58.85% random coil, 20.99% extended strand, and 20.16% alpha helix. Also, the tertiary structure was predicted, and further analyses validated a stable interaction between the vaccine model and mouse TLR4 (binding score: -1261.6). Virtual simulation of immune profile demonstrated potently stimulated humoral (IgG+IgM) and cell-mediated (IFN-γ) responses upon multiepitope vaccine injection. Altogether, a potentially immunogenic vaccine candidate was developed using several N. caninum proteins, with the capability to elicit IFN-γ upsurge and other components of cellular immunity, and can be used in prophylactic purposes against neosporosis.

Neospora caninum SRS2 Protein: Essential Vaccination Targets and Biochemical Features for Next-Generation Vaccine Design.

Vaccination is a standout preventive measure to combat neosporosis among cattle herds. The present in silico study was done to evaluate the physicochemical properties and potent immunogenic epitopes of N. caninum SRS2 protein as a possible vaccine candidate. Web-based tools were used to predict physicochemical properties, antigenicity, allergenicity, solubility, posttranslational modification (PTM) sites, transmembrane domains and signal peptide, and secondary and tertiary structures as well as intrinsically disordered regions, followed by identification and screening of potential linear and conformational B-cell epitopes and those peptides having affinity to bind mouse major histocompatibility complex (MHC) and cytotoxic T lymphocyte (CTL). The protein had 401 residues with a molecular weight of 42 kDa, representing aliphatic index of 69.35 (thermotolerant) and GRAVY score of -0.294 (hydrophilic). There were 53 PTM sites without a signal peptide in the sequence. Secondary structure comprised mostly by extended strand, followed by helices and coils. The Ramachandran plot of the refined model showed 90.2%, 8.8%, 0.5%, and 0.5% residues in the favored, additional allowed, generously allowed, and disallowed regions, correspondingly. Additionally, various potential B-cell (linear and conformational), CTL, and MHC-binding epitopes were predicted for N. caninum SRS2. These epitopes could be further utilized in the multiepitope vaccine constructs directed against neosporosis.

Determination of B and T Cell Epitopes in Neospora caninum Immune Mapped Protein-1 (IMP-1): Implications in Vaccine Design against Neosporosis.

Prevention of neosporosis is advantageous for cattle health and productivity. Previously, several vaccine candidates were nominated for vaccination against Neospora caninum. This study was premised on in silico evaluation of N. caninum IMP-1 in order to determine its physicochemical features and immunogenic epitopes. We employed a wide array of network-based tools for the prediction of antigenicity, allergenicity, solubility, posttranslational modification (PTM) sites, physicochemical properties, transmembrane domains and signal peptide, secondary and tertiary structures, and intrinsically disordered regions. Also, prediction and screening of potential continuous B cell peptides and those epitopes having stringent affinity to couple with mouse major histocompatibility complex (MHC) and cytotoxic T lymphocyte (CTL) receptors were accomplished. The protein had 393 residues with a molecular weight of 42.71 kDa, representing aliphatic index of 85.83 (thermotolerant) and GRAVY score of -0.447 (hydrophilic). There were 47 PTM sites without a signal peptide in the sequence. Secondary structure comprised mostly of extended strand and helices, followed by coils. The Ramachandran plot of the refined model showed 90.1%, 9.9%, 0.0%, and 0.0% residues in the favored, additional allowed, generously allowed, and disallowed regions, correspondingly. Additionally, various potential B cell (linear and conformational), CTL, and MHC binding epitopes were predicted for N. caninum IMP-1. The findings of the present study could be further directed for next-generation vaccine design against neosporosis.

Subtype distribution of Blastocystis sp. isolated from humans in Iran: a systematic review and meta-analysis.

Aim: This systematic review and meta-analysis evaluated the subtyped Blastocystis sp. isolated from humans in Iran. Background: Blastocystis sp. is an anaerobic intestinal protozoan that infects humans as well as domestic and wild animals, i.e. mammals, amphibians, reptiles, and arthropods. Methods: A comprehensive search for papers published before April 2022 was undertaken utilizing English and Persian databases. The following MeSH keywords were used in the electronic search: (Blastocystis sp.) AND (molecular OR subtype) AND (prevalence OR epidemiology) AND Iran. The quality of the included studies was evaluated. Thereafter, a random-effects meta-analysis was conducted to estimate the pooled prevalence and odds ratios regarding the included studies. Results: A total of 32 studies comprised of five case-control studies and 27 cross-sectional studies met the eligibility criteria. The overall pooled prevalence of subtyped Blastocystis sp. in Iran was estimated to be 10% (95% confidence interval: 6 to 15%). Eight subtypes of Blastocystis sp. (ST1- ST7 and ST9) were identified in our study, of which ST3 was the most common subtype (0.04); 0.02-0.07). The difference in subtypes between two case and control groups in reported studies was not significant, but the odds ratio of infection by ST3 (0.98; 95% CI, 0.30 to 3.20) was higher in cases. Conclusion: The current systematic review showed that with the exception of ST8 and ST12, all human Blastocystis sp. subtypes reported in the world are found in different parts of Iran.

Toxoplasma oocysts in the soil of public places worldwide: a systematic review and meta-analysis.

Toxoplasmosis is a zoonotic and cosmopolitan infection. Although a few studies have evaluated the prevalence of Toxoplasma oocysts in the soil of public places, the present study was conducted to provide insights into environmental contamination levels and its potential transmission to humans on a global scale. A systematic search was conducted using bibliographic databases through 30 August 2020. A random effects model was utilized to estimate pooled prevalence with 95% confidence intervals (CIs). Subgroup analysis and meta-regressions were also performed on the geographical and environmental parameters. Finally, 22 articles, wherein 15 420 soil samples were examined, met the systematic review and meta-analysis requirements. The mean pooled prevalence of Toxoplasma oocysts was estimated at 16% (95% CI 10 to 26) in public places. The estimated prevalences in Europe, South America, Asia and North America were 23% (95% CI 4 to 65), 22% (95% CI 18 to 26), 15% (95% CI 0.06 to 33) and 8% (95% CI 0.00 to 97), respectively. An increasing trend was observed in the prevalence of Toxoplasma oocysts with increasing latitude (41-56°), decreasing longitude (0-40°) and increasing relative humidity (≥76%). Loop-mediated isothermal amplification and polymerase chain reaction methods revealed the highest and lowest prevalence rates, respectively, in the detection of Toxoplasma oocysts. Awareness of the health authorities and people about Toxoplasma prevalence in the soil of public places and its risk factors is of great importance to developing effective strategies to prevent infection.

Parasitic Infections of Wild Boars (Sus scrofa) in Iran: A Literature Review.

BACKGROUND: Swine species are an important source of meat production worldwide, except in Islamic countries where pig breeding and pork consumption are forbidden. Hence, they are often neglected in these regions. A considerable number of wild boars (Sus scrofa) inhabit Iranian territories, particularly in dense forests of north, west and southwest of the country, but our knowledge regarding their parasites is very limited. OBJECTIVE: The lack of a comprehensive record in this connection encouraged us to review the whole works of literature in the country. METHODS: The current review presents all the information about the parasitic diseases of wild boar in Iran extracted from articles available in both Persian and English databases until June 2017. RESULTS: So far, 8 genera of protozoa (Toxoplasma, Balanthidium, Tritrichomonas, Blastocystis, Entamoeba, Iodamoeba, Chilomastix and Sarcocystis) and 20 helminth species, including four cestode species, two trematode species, thirteen nematode species as well as a single species of Acanthocephala have been described in Iranian wild boars. CONCLUSION: This review sheds light on the veterinary and public health aspects of the parasitic diseases of wild boars in the country and alerts authorities for future preventive measures.

Intra-Species Diversity of Leishmania major and L. tropica from Clinical Isolates of Cutaneous Leishmaniasis in Southwest Iran Inferred by ITS1-rDNA.

BACKGROUND: Cutaneous leishmaniasis (CL) as a public health concern is increasingly circulating by causative agents of Leishmania tropica and L. major in Iran. As regard to recent treatment failure and controlling problems, the accurate elucidation of heterogeneity traits and taxonomic status of Leishmania spp. should be broadly addressed by policymakers. This study was designed to determine the genetic variability and molecular characterization of L. major and L. tropica from Iranian CL patients. METHODS: One hundred positive Giemsa-stained slides were taken from clinical isolates of CL at Pol-e-Dokhtar County, Southwest Iran, from May 2014 to Sep 2016. DNAs were directly extracted and amplified by targeting ribosomal internal transcribed spacer (ITS) gene following microscopic observation. To identify Leishmania spp. amplicons were digested by restriction enzyme HaeIII subsequent PCR-RFLP technique. To reconfirm, the isolates were directly sequenced to conduct diversity indices and phylogenetic analysis. RESULTS: Based upon the RFLP patterns, 84 and 16 isolates were explicitly identified to L. tropica and L. major respectively. No co-infection was found in clinical isolates. The high genetic diversity of L. tropica (Haplotype diversity 0.9) was characterized compared to L. major isolates (Hd 0.476). The intra-species diversity for L. tropica and L. major isolates corresponded to 3%-3.9% and 0%-0.4%, respectively. CONCLUSION: Findings indicate the L. tropica isolates with remarkable heterogeneity than L. major are predominantly circulating at Pol-e-Dokhtar County. Occurrence of high genetic variability of L. tropica may be noticed in probable treatment failure and/or emerging of new haplotypes; however, more studies are warranted from various geographic regions of Southwest Iran, using large sample size.

Prevalence of Pneumocystis jirovecii among immunocompromised patients in hospitals of Tehran city, Iran.

Pneumocystis jirovecii is an opportunistic organism that can cause extreme complications such as Pneumocystis pneumonia in immunocompromised individuals. There is no comprehensive study was conducted Iran to determine the prevalence of this infection in susceptible individuals. In the present study, 160 sera samples were collected from immunocompromised patients, including acquired immunodeficiency syndrome (AIDS) patients, diabetic patients, Hodgkin lymphoma patients and non-Hodgkin lymphoma patients. The specimens were collected from Imam Khomeini and army's 501 hospitals. The specimens were examined using indirect fluorescent antibody test. The results of the study showed that 39.30% specimens were found positive, with different rates in different groups, including 20, 22.50, 37.50, and 77.50% of diabetic patients, non-Hodgkin patients, Hodgkin lymphoma patients, and AIDS patients, respectively. This occurrence is relatively high and can be a potential life-threatening hazard to infected patients in studied groups, on the other hand the organism can be transmitted from infected people to other susceptible individuals.