First molecular subtyping and zoonotic significance of Blastocystis sp. in Dromedary (C. dromedarius) and Bactrian (C. bactrianus) camels in Iran: A molecular epidemiology and review of available literature.

BACKGROUND: Blastocystis sp. is a zoonotic protozoan parasite, and there is limited information about its molecular prevalence and subtypes (STs) distribution in camels globally, especially in Iran. OBJECTIVES: This study aimed to examine the prevalence, STs distribution, and zoonotic potential of Blastocystis sp. in one-humped and two-humped camels in Ardabil province, northwestern Iran. METHODS: A PCR-sequencing tool using the SSU rRNA gene was employed to examine the occurrence and genetic variation of Blastocystis sp. in 150 faecal samples from Bactrian (Camelus bactrianus, 50 samples) and Dromedary (Camelus dromedarius, 100 samples) camels in Ardabil province. RESULTS: The overall prevalence of Blastocystis sp. in camels was determined to be 12% (18/150) through microscopy and PCR analyses. Phylogenetically, this study identified three distinct zoonotic STs: ST7, ST10, and ST14. ST10 was the most prevalent, comprising 50% (9/18) of the isolated STs from camels. ST14 closely followed with 38.9% (7/18), while ST7 made up 11.1% (2/18) of the total STs. In brief, ST10, ST14, and ST7 represented 50% (7/14), 35.7% (5/14), and 14.3% (2/14) of the Blastocystis-positive cases in one-humped camels, respectively. Further, each of the ST10 and ST14 accounted for 50% (2/4) of the Blastocystis-positive samples in two-humped camels. An analysis of the available data reveals that out of the 37-44 identified Blastocystis STs, 15 (ST1-ST7, ST10, ST14, ST15, ST21, ST24, ST25, ST26, and ST30) have been reported in camels. The predominant STs observed are ST10 and ST14. Furthermore, among the 15 zoonotic STs (ST1-ST10, ST12-ST14, ST16, and ST23) of Blastocystis reported thus far, nine zoonotic STs (ST1-ST7, ST10, and ST14) have been found in camels. CONCLUSIONS: These findings indicate that camels serve as a proper reservoir for a diverse array of Blastocystis STs and thereby can play a significant role in the transmission of this protozoan infection to humans, animals, and water reservoirs.

Development of New PCR Protocols to Detect Genetic Diversity in the Metronidazole Metabolism Genes in Susceptible and Refractory Clinical Samples of Giardia duodenalis.

PURPOSE: Investigating the genetic variation in thioredoxin reductase (TrxR) and nitroreductase (NR) genes in both treatment-resistant and -sensitive Giardia duodenalis isolates can provide valuable information in identifying potential markers of resistance to metronidazole. The rapid increase in metronidazole treatment failures suggests the presence of genetic resistance mechanisms. By analyzing these genes, researchers can gain insights into the efficacy of metronidazole against G. duodenalis and potentially develop alternative treatment strategies. In this regard, four G. duodenalis isolates (two clinically sensitive and two clinically resistant to metronidazole) were collected from various hospitals of Shiraz, southwestern Iran. METHODS: Parasitological methods including sucrose flotation and microscopy were employed for the primary confirmation of G. duodenalis cysts in stool samples. Microscopy-positive samples were approved by SSU-PCR amplification of the parasite DNA. All four positive G. duodenalis specimens at SSU-PCR were afterward analyzed utilizing designed primers based on important metronidazole metabolism genes including TrxR, NR1, and NR2. RESULTS: Unlike TrxR gene, the results of NR1 and NR2 genes showed that there are non-synonymous variations between sequences of treatment-sensitive and -resistant samples compared to reference sequences. Furthermore, the outcomes of molecular docking revealed that there is an interaction between the protein sequence and spatial shape of treatment-resistant samples and metronidazole in the position of serine amino acid based on the NR1 gene. CONCLUSION: This issue can be one of the possible factors involved in the resistance of Giardia parasites to metronidazole. To reach more accurate results, a large sample size along with simulation and advanced molecular dynamics investigations are needed.

Comparative molecular epidemiology, subtype distribution, and zoonotic potential of Blastocystis sp. in Equus animals (horses, donkeys, and mules) in northwestern Iran.

A total of 500 fecal samples were collected from Equus animals in six different cities (Ardabil, Namin, Nir, Meshginshahr, Germi, and Khalkhal) of Ardabil Province, northwestern Iran, with 200 samples from horses, 200 from donkeys, and 100 from mules. Of the horse samples, 100 were from racing horses under special monitoring and care, while the remaining 100 were from non-racing horses, including those used for herding or in rural areas. All fecal samples were examined for the presence of Blastocystis sp. using PCR amplification of the SSU rRNA gene's barcode region after DNA extraction. The molecular prevalence of Blastocystis infection in Equus animals was 7.6% (38/500). Blastocystis was more common in horses [11.5% (23/200)] than in donkeys [5.5% (11/200)] and mules [4% (4/100)] (P > 0.05). Compared to racing horses [3% (3/100)], non-racing/rural horses [20% (20/100)] exhibited a substantially higher prevalence of Blastocystis (P < 0.05). The prevalence of Blastocystis in diarrheal samples and younger animals was remarkably higher than in formed samples and older animals, respectively (P < 0.05). No significant difference in Blastocystis infection prevalence was found between the genders of examined animals (P > 0.05). In Equus animals, 38 Blastocystis isolates included eight STs: ST10 [31.6% (12/38)], ST1 [21.1% (8/38)], ST2 [15.8% (6/38)], ST3 [10.5% (4/38)], ST4 [7.9% (3/38)], ST7 [5.2% (2/38)], ST14 [5.2% (2/38)], and ST6 [2.6% (1/38)]. These results suggest that Equus animals act as a proper reservoir for numerous Blastocystis STs, consequently playing a crucial part in the spread of this protozoan infection to humans, animals, and water reservoirs.

Global epidemiology of Giardia duodenalis infection in cancer patients: a systematic review and meta-analysis.

BACKGROUND: Application of chemotherapeutics in cancer patients may provide an immunosuppressive milieu, favourable for parasitic infections. Giardia duodenalis is an important zoonotic intestinal parasite responsible for diarrhoea in humans worldwide. METHODS: The present systematic review and meta-analysis was conducted to estimate the prevalence of G. duodenalis and respective odds ratios (ORs) in cancer patients around the globe. Four online databases-PubMed, Scopus, Web of Science and Google Scholar-were carefully explored for relevant literature without time limitation until 28 November 2020. Meta-analysis was done based on a random effects model to pool the estimations and define 95% confidence intervals (CIs). RESULTS: The overall weighted prevalence of G. duodenalis infection in cancer patients was calculated to be 6.9% (95% CI 0.5 to 9.3) globally, based on data from 32 studies. Although not statistically significant, eight case-control studies revealed that cancer patients were 1.24 times (95% CI 0.66 to 2.31; p=0.501) more exposed to G. duodenalis infection than healthy controls. Moreover, the prevalence of infection was not significantly associated with quantitative variables, including publication year (regression coefficient -0.0135, p=0.578), sample size (regression coefficient -0.0007, p=0.074) and human development index (regression coefficient -1.6263, p=0.419). Also, subgroup analysis of the pooled G. duodenalis infection was performed for publication year, World Health Organization regions, countries, continents, cancer types and country income. CONCLUSIONS: Altogether, the epidemiology of G. duodenalis infection and its associated risk factors in immunocompromised individuals, especially cancer patients, is still open to question and deserves comprehensive investigations.

A systematic review and meta-analysis on the global prevalence and subtypes distribution of Blastocystis sp. infection in cattle: A zoonotic concern.

Blastocystosis is an interesting parasitosis, since the parasitic infection is still seriously neglected and a considerable zoonotic evidence is emerging. Blastocystis sp. infects the intestinal lumen of humans and a wide range of animals, while there is a lack of comprehensive information on Blastocystis epidemiology in cattle worldwide. Thus, the present systematic review and meta-analysis were performed by exploring four electronic databases (PubMed, Web of Science, Scopus, and Google scholar) for relevant published papers up to 7th November 2020, utilizing a random-effects model to pool estimations and assign 95 % confidence intervals (CIs). Results of 28 studies (29 datasets) on cattle showed a 24.4 % (95 % CI: 16.9-33.9 %) prevalence for Blastocystis infection. Also, 16 out of 26 reported subtypes (STs) were isolated from cattle, with ST10 (18 datasets) as the highest-reported [32.3 % (95 % CI: 21.6-45.3)] as well as ST24 and ST25 (one study each) as the lowest-reported STs [1.4 % (95 % CI: 0.2-9.1)]. Additionally, among nine well-known zoonotic STs (ST1-ST8 and ST12), all STs except for ST8 were reported from cattle worldwide, demonstrating this animal species as a potential reservoir for human infections. Meanwhile, the overall prevalence of Blastocystis in various subgroups (publication year, WHO regions, countries, continents, and age groups) was analyzed separately. The finding of the present review article highlights the cattle as a significant source of zoonotic transmission of Blastocystis infection to humans, which must be considered for preventive measures.