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Cryptosporidium spp. are opportunistic intestinal protozoans with global distribution and are of great importance as zoonotic protozoans are common to humans and domestic animals, including cattle and calves. Identification and detection of parasite species using precise methods including molecular methods can be an effective step in treating and controlling parasites.This study aimed to investigate the prevalence of Cryptosporidium among breeding calves of Khorramabad city, Lorestan province, Western Iran, using PCR.The faecal samples were taken from 181 healthy and diarrhoeal calves and after the Ziehl Neelsen Acid-fast staining and microscopic evaluation, the genomic DNA was extracted for molecular evaluations. To detect Cryptosporidium species, specific primers targeting the SAM-1 gene of Cryptosporidium and a commercial master mix were used for PCR.Out of 181 faecal samples of breeding calves in Khorramabad city, 9 samples (5%) were positive for Cryptosporidium spp. using the PCR method. Statistical analysis of the data showed that there was no significant statistical relationship between Cryptosporidium infection of the calves and variables of age, breed, type of water consumption, clinical signs of diarrhoea, and sampling location, while parasite infection had a significant relationship with calf gender so that all Cryptosporidium positive samples were from male calves (p ≤ 0.05).Considering the presence of Cryptosporidium infection, the region's traditional grazing system, and the close relationship between livestock and humans, there is a possibility of human infection in the region. So preventive measures such as periodic animal testing with sensitive and accurate diagnostic techniques including PCR, pharmacological treatment of livestock, water hygiene and the use of industrial grazing instead of traditional grazing to improve the hygiene of food consumed by livestock are recommended.

Free-living amoebae (FLA) thrive in diverse environmental conditions. The present study aimed to define the FLA distribution from the Caspian Sea as well as from hospital ward dust from Guilan, Iran. Seawater (20) and hospital ward dust samples (100) were collected from May to June 2018. Seawater samples were vacuum filtered through a 0.45 μm pore-size membrane. Dust was collected using sterile gauze, washed with sterile distilled water, with washings collected thereafter. Washings were similarly filtered as seawater samples. FLA from the filtered material was cultivated in non-nutrient agar. Molecular analysis was performed by PCR and sequencing using specific primers for Acanthamoeba, Naegleria, and Vermamoeba/Hartmanella. Culture and PCR returned 50 and 65% positivity, respectively, for seawater samples where sequencing revealed Acanthamoeba T2, T5 and T6 genotypes and A. palestinensis and A. lenticulata, as well as N. dobsoni and N. clarki. In addition, 30% amoebic growth and 16% PCR detection were observed from hospital ward dust samples where sequencing revealed Acanthamoeba T2, T4 and T11 genotypes and A. castellanii, A. palestinensis and A. stevensoni as well as N. clarki. For both seawater and dust samples, Acanthamoeba was the dominant isolate. The detection of potentially pathogenic FLA from seawater may pose a threat to the public, while the presence of the same in dust spells threats to both hospital staff and patients, in particular, immunocompromised individuals. Public education, awareness, improved sanitation and hygiene, and the crafting of diagnostic strategies for the early detection of FLA in humans are necessary for the mitigation and management of potential human infection cases. HIGHLIGHTS Free-living amoebae (FLA) detected in environmental and infrastructure-derived samples.; Pathogenic genotypes of FLA detected in the Caspian Sea.; Pathogenic FLA detected in hospital wards for immunocompromised patients.; Pathogenic genotypes of FLA detected in hospital ward dust.; Hygiene and policies are necessary to protect humans against FLA infections.

The incidence of metabolic syndrome (MetS) in menopausal women is one of the main health care concerns. MetS clusters are related to an imbalance in pro- and anti-inflammatory adipokines such as secreted frizzled-related protein 5 (SFRP5) and wingless-type mammary tumor virus integration site family, member 5A (WNT5A). WNT5A induces an inflammatory state to induce insulin resistance and further pathologic consequences. Recent strategies to prevent progression of MetS to diabetes have focused on conservative treatments such as exercise and herbal medicine. The aim of this study was to investigate the mechanistic effects of cotreatment with cinnamon extract and 12-wk high-intensity endurance training on MetS components considering the non-canonical WNT5A signaling.Thirty-two female ovariectomized Wistar rats were divided into the following four groups (n = 8/group): exercise (Ova+Exe), cinnamon extract (Ova+Cin), exercise with cinnamon extract (Ova+Exe+Cin) and saline (Ova+Sal). One group of rats undergoing surgery without removal of the ovaries was considered as a sham. After 3 mo of experimental intervention, waist circumference, serum concentrations of glucose, insulin, lipid profile, tumor necrosis factor-α, WNT5A, and SFRP5 were measured.Data showed a significant reduction in serum glucose, low-density lipoprotein, homeostasis model assessment estimate of insulin resistance, and tumor necrosis factor-α, but an increase in SFRP5 level in Ova+Exe, Ova+Cin and Ova +Exe+Cin groups compared with Ova+Sal group (P0.05). Serum WNT5A significantly was reduced only in Ova+Exe+Cin group (P = 0.02).The present study indicated that high-endurance training combined with aqueous cinnamon extract supplementation for 12 wk more efficiently alleviated insulin resistance and metabolic dysfunctions via reduction in noncanonical WNT signaling in ovariectomized rats.

Background: Nowadays a number of methods are used for the extraction of genomic DNA from fecal specimens. Identification and selection of an effective DNA extraction method is considered as one of the most important steps in molecular assays of Giardia duodenalis. Objectives: We compared the effects of 3 different DNA extraction techniques in PCR amplification of a specific area of SSU rRNA gene. Methods: A total of 20 fecal samples containing purified cysts were aliquoted in 3 sub-samples. DNA extraction was performed using the Phenol-Chloroform Isoamyl alcohol (PCI), QIAamp DNA stool mini kit, and YTA Stool DNA Isolation mini Kit. The quantity and purity of extracted DNA were compared. The potency of extracted DNA was tested. Results: The results showed that the most concentrated DNA was obtained from phenol/chloroform/Isoamyl alcohol method and the best purity based on comparing the ratio of A260/230 was obtained from QIAamp DNA stool mini kit. In this study the diagnostic sensitivity of QIAamp DNA stool mini kit, phenol/chloroform/Isoamyl alcohol, and YTA Stool DNA Isolation mini Kit methods was 60%, 70%, and 60%, respectively. Conclusions: The application of a proper DNA extraction method leads to obtaining reliable and reproducible results in molecular assays and supports treatment and control strategies of G. duodenalis. In this study, PCR amplification targeting a 350-bp fragment of SSUrRNA gene demonstrated phenol/chloroform/Isoamyl alcohol as the most efficient method.

More recently, detection of circulating tumor cells (CTCs) has been considered as an appealing prognostic and diagnostic approach for cancer patients. CTCs as a type of tumor-derived cells are secreted by the tumor and released into the blood circulation. Since the migration of CTCs is an early event in cancer progression, patients who still have tumor-free lymph nodes have to be well examined for the CTCs presence in their blood circulation. Nowadays, there is a broad range of detection methods available to identify CTCs. As artificial RNA oligonucleotides or single-stranded DNA with receptor and catalytic characteristics, aptamers have been standing out, owing to their target-induced conformational modifications, elevated stability, and target specificity to be implemented in biosensing techniques. To date, several sensitivity-enhancement methods alongside smart nanomaterials have been used for the creation of new aptasensors to address the limit of detection (LOD), and improve the sensitivity of numerous analyte identification methods. The present review article supports a focused overview of the recent studies in the identification and quantitative determination of CTCs by aptamer-based biosensors and nanobiosensors.

We studied the prevalence of intestinal parasites (IPs), their risk factors and associated symptoms among patients with gastrointestinal disorders. A total of 1,301 participants aged 22 days-90 years were enrolled in this study. We used a structured questionnaire to obtain socio-demographic and stool examination to investigate intestinal parasite infections. Data analysis was performed using SPSS16. The overall prevalence of intestinal parasites (IPs) was 32.2% (419/1,301). Three hundred and fifty nine cases/1,301 (27.6%) were infected with a single parasite and 60/1,301 cases (4.6%) presented polyparasitism. The most common IP was Blastocystis sp. 350/1,301 (26.9%), followed by Entamoeba coli 38/1,301 (2.92%), Giardia lamblia 30/1,301 (2.3%) and Cryptosporidium spp. 17/1,301 (1.3%). Regarding the socio-demographic variables, educational status (p = 0.001), contact with domestic animals and soil (p = 0.02), age above 15 years (p = 0.001) and seasons (p = 0.001) were significantly associated to intestinal parasitic infections. Concerning clinical characteristics, the presence of IPs was significantly associated to diarrhea (OR = 1.57; CI 95% = 1.24-1.98; p < 0.001) and dysentery (OR = 1.94; CI 95% = 1.03-3.66; p < 0.04). Our findings suggest that IPs are one of the main causal agents of gastrointestinal disorders. Improving the knowledge on local risk factors such as poverty, low level of education, poor sanitation, contact with soil and contact with domestic animal is warranted.

Background: Intestinal parasites of humans are one of the most important health problems worldwide, especially those located in tropical and subtropical areas. Objectives: The aim of this study was to determine the frequency of intestinal parasites in patients with gastrointestinal disorders, in different parts of Iran. Patients and Methods: A total of 1520 stool samples were collected from patients with gastrointestinal disorders. The stool specimens were examined by direct wet mount, formalin-ether concentration and a modified version of the Ziehl-Neelsen staining technique. Amoeba-positive samples were cultured for further differentiation of Entamoeba histolytica, E. dispar, E. moshkovskii. DNA-based methods were used to differentiate these amoebas and to detect Cryptosporidium- positive samples. Statistical analysis was carried out by SPSS ver. 16. Results: Out of the 1520 individuals studied, 153 (10.06%) were infected at least with one intestinal parasite. 781 (51.4%) of patients were male and 738 (48.6%) were female. The prevalence of protozoan parasites 148 (9.7%) was significantly higher than helminth parasites 5 (0.3%) (P < 0.001). The frequency of intestinal parasites was as follows: Blastocystis sp., 72 (4.73%); Giardia intestinalis, 35 (2.30%); Entamoeba coli 21 (1.38%); Endolimax nana 10 (0.92%); Cryptosporidium spp., 1 (0.06%); Entamoeba dispar, 1 (0.06%); Dientamoeba fragilis, 1 (0.06%); Hymenolepis nana, 3 (0.19%); Dicrocoelium dendriticum, 2 (0.13%). In five (0.32%) of the positive samples, co-infections with two parasites were found. G. intestinalis was more prevalent in male 22/35 (62.86%) than female 13/35 (37.14%) as well as in 0-9 years old group. In one sample Heterodera ova contained larva were seen. Conclusions: Blastocystis and G. intestinalis were the predominant intestinal parasites detected in patient with gastrointestinal disorders. The results indicated that the intestinal parasites, particularly helminth infections have been significantly declined in recent years.