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Sarcocystis is an intracellular parasite of the apicomplexa phylum. More than one hundred species of Sarcocystis can infect wildlife and livestock animals. Samples of the liver, heart, muscle and diaphragm were collected from sheep, goats and cattle in Gonabad, northeast Iran and subjected to macroscopic, microscopic, tissue digestion, sequencing and phylogenetic analyses of 18S-rRNA region. Tissue sections stained with hematoxylin and eosin were also provided for surveying sarcocysts in the samples. Tissue digestion showed that all animal samples (100%) were infected with Sarcocystis bradyzoites. Phylogenetic analysis indicated that out of 50% of sheep genotypes belonged to S. tenella, and 20% to S. moulei and S. arieticanis. Moreover, three samples of macroscopic specimens of sheep were identified as S. gigantea, 100% of cattle isolates infected with S. cruzi, 80% of goat isolates belonged to S. capracanis and one macroscopic specimen of goat (20%) identified as S. moulei. Sarcocystis infection is highly prevalent in livestock in northeast Iran, where a variety of Sarcocystis species are unequivocally circulating in the region.

Unawareness of canine parasitic diseases among at-risk hosts and an uncontrolled program of stray dog population have caused that zoonotic parasites received great attention in endemic regions of the Middle East. A total of 552 faecal samples were collected between December 2016 to January 2018 from stray (n = 408) and domestic (n = 144) dogs of Iran. All specimens were coproscopically observed following concentration and flotation techniques. Subsequently, the DNAs of taeniid eggs were extracted, amplified, and sequenced by targeting of mitochondrial cytochrome oxidase subunit 1 and small-subunit ribosomal DNA markers. The overall prevalence of canine intestinal parasites found 53.6%. The following parasites and their total frequencies were identified: taeniid (10.5%), Dicrocoelium dendriticum (0.7%), Trichuris vulpis (1.2%), Capillaria spp. (2.3%), Blastocystis spp. (5.2%), Ancylostoma spp. (2%), Eimeria spp. (13.2%), Dipylidium caninum (2.3%), Toxocara canis (3.8%), Giardia spp. (8.5%), and Toxascaris leonina (3.6%). Stray dogs were characterized more likely to be poliparasitized and indicated a higher prevalence of taeniid (10.9%), T. canis (4.4%) Giardia spp. (10.1%) than domestic dogs (P > 0.05). Phylogenetic and sequence analysis of Cox1 and SSU-rDNA indicated a low genetic diversity (Haplotype diversity; 0 to 0.495) in E. granulosus sensu lato G1, G3, G7 genotypes, and Taenia hydatigena. The pairwise sequence distances between G7 isolates showed an intra-diversity of 0.7%–1.5% and identity of 98.5%–100%. The first occurrence of pig strain (G7) from Iranian dogs might have substantial implications in the drug treatment of infected dogs due to the shorter maturation time of G7 compared with G1 genotype. Thus, the preventive strategies should be noticed to determine the risk factors, the importance of applying the hygienic practices, and well adjusting deworming programs for the Iranian dogs and at-risk individuals.

In this study, we investigated the possible association between TB and Toxoplasma gondii infection. One hundred confirmed TB individuals living in northwest Iran were classified into three subgroups; newly diagnosed patients (NTB), old diagnosed patients (OTB) and multidrug resistance patients (MDR-TB). One hundred healthy subjects in the same age and sex distribution were ethnically matched. Sera samples were screened for anti-Toxoplasma antibodies. Nested-PCR was performed by targeting the B1 and GRA6 genes. The frequency of Toxoplasma infection based on IgG titer was 71.1% in the OTB subgroup and 33% in the control group, indicating significant association between Toxoplasma seropositivity and OTB (P = 0.001). According to phylogenetic network, the type I strain of Toxoplasma was identified in the OTB subgroup (10.1%). We concluded that patients with OTB subgroup are at high risk for acquisition of Toxoplasma infection which could reactivate the latent toxoplasmosis.

This study reports development of a novel point of care assay, namely an enhanced immuno-dot blot assay, for discrimination of anti-Toxoplasma IgG and anti-Toxoplasma IgM antibodies. This method has been designed based on formation of a sandwich complex between a gold nanoprobe (chitosan gold nanoparticle-anti-human IgG or anti-IgM) and anti- Toxoplasma lysate antigen (TLA) which holds anti-TLA antibodies, either IgG or IgM. Briefly, anti-human IgG or anti-IgM antibody was conjugated to chitosan gold nanoparticles via glutaraldehyde chemistry. Then, lysate antigen was immobilized on the surface of nitrocellulose membrane, which followed by addition of the sera sample and gold nanoprobes. The positive signals were readily detectable via observation with naked eye. This positive color change was further intensified via gold enhancement chemistry. The intensity of biosensor signal was proportional to the concentration of active antibodies on the surface of nanoparticles, titer of T. gondii antibodies in the sera samples and concentration of Toxoplasma lysate antigen coated on the nitrocellulose membrane. A minimum concentration to use the antibodies for conjugation, to detect titer of Toxoplasma IgG and IgM antibodies, and the concentration of TLA coated in nitrocellulose membrane were 0.5 mg/mL, 2 IU/mL, 10 IU/mL, and 20 μg/mL, respectively. This enhanced immuno-dot blot assay offers a simple diagnostic technique without expensive equipment requirement for distinguishing of anti- T. gondii IgM and IgG antibodies in field conditions, pregnant women, and immunocompromised patients.

Cryptosporidium, as a small protozoan parasite, is a leading cause of persistent diarrhea in children in developing countries and has both a short and long-term impact on the growth of children. In the present study, Cryptosporidium infection was compared in malnourished and well-nourished children by modified acid-fast staining, nested-polymerase chain reaction (nested-PCR) and loop-mediated isothermal amplification (LAMP) methods. As a case-control study, Cryptosporidium infection in 94 malnourished children was evaluated and compared with those of 188 age and gender-matched well-nourished children. Oocysts of Cryptosporidium were detected by modified acid-fast staining method. The extracted DNA was amplified by nested-PCR and LAMP techniques. In addition, positive amplicons were directly sequenced for phylogenetic analysis. Cryptosporidium oocysts were found in the stools of two (2.12 %) children who were hospitalized and had diarrhea by nested-PCR while three isolates (3.2 %) were found by LAMP. Cryptosporidium-positive children were more malnourished compared to those who were negative for Cryptosporidium infection but this important finding was not statistically significant. C. parvum was the main species of Cryptosporidium detected in malnourished children in northwest Iran. LAMP can be considered as a sensitive field monitoring assay in patients with low parasite burden. Nutritional status and socio-demographic factors may have interactive effects on the incidence and severity of parasitic diseases.

Cattle are intermediate host for several species of Sarcocystis, including S. cruzi, S. hirsuta, and S. hominis with high prevalence worldwide. The present study aimed to determine the prevalence of Sarcocystis infection, species identification, and phylogenetic analysis of the parasite in cattle in Northwest Iran. The samples of diaphragm and esophagus from 290 cattle were collected from slaughterhouses in Northwest Iran and subjected to macroscopic, microscopic, and histopathology examinations, PCR-RFLP, sequencing and phylogenetic analyses. Tissue cysts of Sarcocystis spp. were detected in 92% of cattle by digestion and microscopic tests. Based on the PCR–RFLP and specific PCR, 87.9% and 1.03% of isolates were identified as S. cruzi, and S. hominis, respectively. Macrocyst was seen in a single sample that was identified as S. gigantea. The haplotype network exhibited the extension of the various haplotypes of S. cruzi between neighboring provinces in Northwest Iran. Heterogeneity analysis of S. cruzi 18S-rRNA sequences indicated genetic diversity among S. cruzi isolates (Haplotype diversity: 0.733-0.854) consisting 16 haplotypes; however, the nucleotide differences showed low diversity (0.01481 to 0.03351). Pair wise sequence distance matrix amongst S. cruzi sequences indicated an intra-species divergence of 0%-7.8% and identity of 92.6%-100%. Sarcocystis infection is highly prevalent in cattle in Northwest Iran, with the predominance of S. cruzi, and genetic variants of this species are unequivocally distributing in Northwest provinces. First global detection of S. gigantea in cattle reflects new insights of transmission dynamic and biology of this parasite in Iran.

Recurrence of hydatid cysts in cystectomy patients has dramatically remained a serious concern within the surgical community. Predisposing factors for recurrence of hydatid cysts remained to be identified. Toll-like receptor (TLR) plays a pivotal role in bridging between acquired and innate immunity in cystic echinococcosis (CE) infection. 117 CE patients including 66 acute hydatidosis (AH; primary infection) and 51 recurrent hydatidosis (RH; chronic infection), and 117 ethnically matched healthy control (HC) were investigated from endemic regions of Iran in the period of 2015–2018. CE patients were definitely confirmed using histopathological and immunological assays. Genotyping of TLR2 Arg753Gln was carried out by restriction fragment length polymorphism and sequencing. The homozygous mutant-type TLR2 Gln/Gln (A/A) was represented to be associated with the occurrence of RH (P = 0.04) and conferred a 9 fold risk for susceptibility, while the heterozygous mutant-type TLR2 Arg/Gln (G/A) indicated a tendency to be associated with the occurrence of RH (P = 0.07). There was no discrepancy in the frequency of TLR2 Arg753Gln haplotypes between AH patients and HC individuals (P = 0.09). The mutant allele A was observed to be a risk factor for susceptibility to RH patients. Our results point to a clinical association between TLR2 Arg753Gln haplotypes with RH in postoperative patients. It can be inferred that allele G may lead to protection against the CE, while mutant allele A may be a diagnostic hallmark in the screening of RH susceptibility. Nevertheless, further studies with a larger sample size of different ethnic populations are required to authenticate this association.

Toxoplasmosis is an infectious disease caused by the intracellular parasite Toxoplasma gondii. Although almost 1/3 of the world's population are seropositive, there is no effective vaccine against toxoplasmosis. Therefore, the development of an effective vaccine for control of toxoplasmosis is one of major concerns in parasitology. The aim of this study was to evaluate the efficacy of nano-liposomal excretory-secretory antigens (NLESA) in BALB/c mice.Excretory-secretory antigens (ESA) was obtained from tachyzoites, encapsulated in the liposome and studied by scanning electron microscope. BALB/c mice were immunized with NLESA and ESA, sterile phosphate-buffered saline (PBS). Immunization was performed three times at 14-day intervals and challenged with 1 × 10The immunization with NLESA and ESA induced a significant increase of anti-Toxoplasma IgG antibody compared with PBS group (P 0.05). After challenge with tachyzoites, qPCR analyses showed significant reduction of parasite load in NLESA and ESA immunized mice compared with control group (P 0.05). Also, NLESAs were more effective than ESAs and showed significantly reduced parasite load in blood (P = 0.001) and brain tissue (P = 0.01).The vaccination with NLESA showed more promising results comparing to ESA. Further studies are recommended in order to achieve effectiveness of the vaccine against T. gondii.