Gene migration of giardiasis in Iran; a microevolutionary scale for reflecting transmission patterns of Giardia lamblia assemblages in symptomatic patients.

In the microevolutionary scale of Giardia lamblia, the gene migration indicates how G. lamblia assemblages have transmitted between adjacent counties. 33 positive fecal samples were taken from patients suffering gastrointestinal disorders (nausea, bloating, burping constipation and fatty diarrhea) at Tabriz and Ardabil cities, where located in the cold regions of northwest Iran. Following parasitological examinations, DNA samples were extracted, amplified and digested by single-step PCR-RFLP assay, targeting the glutamate dehydrogenase (gdh) locus to distinguish within and between assemblages A and B. PCR products were directly sequenced to reconfirm their heterogeneity traits and phylogenetic analysis. Of the 33 isolates, 81.9% (n: 27), 9% (n: 3) and 9% (n: 3) were successfully identified as assemblages A (genotype AII), B (genotype BIII) and the mixed of genotypes AII and B, respectively. Despite the presence of heterogeneous clinical backgrounds, a low genetic diversity of sub-assemblage AII was identified among symptomatic cases. A low value of pairwise fixation index showed that G. lamblia sub-assemblage AII is not genetically differentiated among northwest regions of Iran. The occurrence of haplotypes TAB-1/ARD-1 between two regional populations indicates that there is a dawn of G. lamblia gene flow due to transfer of alleles through host mobility and/or ecological alterations. To assess the hypothetical evolutionary scenario, further studies are essential for multilocus genotyping of G. lamblia in tropical regions of Iran and neighboring countries.

Genetic variability and transcontinental sharing of Giardia duodenalis infrapopulations determined by glutamate dehydrogenase gene.

Microevolutionary data of Giardia duodenalis sub-assemblages is a prerequisite for determining the invasion zoonotic patterns of the parasite. To infer transmission patterns that could not be differentiated by the phenotypic features, a population genetic investigation is crucial for the elucidation of the genetic structure of G. duodenalis among the continents. Forty G. duodenalis positive fecal samples were collected from different foci of Northwest Iran. The specimens were subjected to Trichrome staining and sucrose gradient flotation. DNA samples were extracted, amplified, and sequenced by targeting glutamate dehydrogenase (gdh) gene. The global gdh sequences of sub-assemblages AII and BIV retrieved from NCBI GenBank were analyzed to estimate diversity indices, neutrality indices, and gene migration tests. Sequencing analyses indicated various levels of genetic variability of sub-assemblages AII and BIV among the five continents. Sub-assemblage BIV had greater genetic variability (haplotype diversity: 0.975; nucleotide diversity: 0.04246) than sub-assemblage AII. The statistical Fst value demonstrated that the genetic structure of sub-assemblages AII and BIV are moderately differentiated between European-American populations (Fst: 0.05352-0.15182), whereas a significant differentiation was not seen among other geographical population pairs. We conclude that a high gene flow of G. duodenalis sub-assemblages AII and BIV is unequivocally sharing among the continents. The current findings strengthen our knowledge to assess the evolutionary patterns of G. duodenalis in endemic foci of the world and it will become the basis of public health policy to control human giardiasis.

Introducing nitazoxanide as a promising alternative treatment for symptomatic to metronidazole-resistant giardiasis in clinical isolates.

OBJECTIVE: To identify the frequencies (F) of ferredoxin and nitroreductase mutations on Iranian clinical isolates of Giardia lamblia in order to predict whether the nitazoxanide can be prescribed as suitable drug for symptomatic to metronidazole-resistant giardiasis. METHODS: Forty Giardia lamblia isolates as of 38 symptomatic and two metronidazole-resistant patients were collected from Iran. DNAs were extracted and amplified by targeting ferredoxin and GlNR genes. The amplicons were directly sequenced to determine gene mutations. RESULTS: The various amino acid substitutions (F: 20%, Haplotype diversity: 0.891, Tajima's D: -0.44013) were identified by analyzing ferredoxin gene in four symptomatic and two resistant isolates. Only two haplotypes (F: 5%, HD: 0.345; Tajima's D: 0.77815) characterized in metronidazole-resistant isolates of GlNR, however, no point mutations was found in symptomatic isolates. CONCLUSIONS: Non-synonymous mutations of ferredoxin oxidoreductase gene reduce translational regulatory protein's binding affinity which concludes reduction of ferredoxin expression and its activity. This leads to decrease in metronidazole drug delivery into the cells. Mutations in these isolates may lead to their resistance to metronidazole. No to low synonymous mutations of GlNR demonstrates that nitazoxanide can be prescribed as promising alternative treatment for symptomatic to metronidazole-resistant giardiasis in Iranian clinical isolates.