MiR-770-5p, miR-661 and miR-571 expression level in serum and tissue samples of foot ulcer caused by diabetes mellitus type II in Iranian population.

BACKGROUND: Microvascular complications related to diabetes mellitus type II such as foot ulcers are the reason of many mortalities among T2DM patients. The role of microRNAs (miRNAs, miRs) as potent regulators of gene expression is studied in different diseases such as diabetes mellitus and primary studies revealed their importance as early detecting biomarkers. Therefore, in this study it is tried to evaluate the expression level of some miRNAs (miR-770-5p, miR-661 and miR-571) in serum and tissue samples of T2DM related foot ulcer among Iranian patients. METHODS: 30 samples of blood and 30 muscle tissue were collected from T2DM patients suffering foot ulcer (T2DM + FU), 30 blood samples collected from T2DM patients without foot ulcer (T2DM-FU). 30 tissue samples collected from patients with trauma and 30 blood samples were selected as healthy controls. RESULTS: The three studied miRNAs were statistically significant in all groups in comparison to control blood group. Also, comparison between other groups showed a significant increase of all studied miRNAs especially in the blood and tissues of T2DM + FU patients. The only significant correlation detected between the FBS level and miR-571 expression pattern in blood samples of T2DM + FU group. Finally, the results showed that miR-571, -661, and -770 has a statistically significant discriminative character for differentiating T2DM + FU patients from T2DM-FU both in tissue and blood samples. CONCLUSION: Although more studies are essential for certifying these findings, our results showed that miR-770-5p, miR-661 and miR-571 are correlated with the microvascular complications related with T2DM such as foot ulcer.

Molecular Characterization of Fasciola spp. from a Donkey (Equus asinus) Using Partial Sequencing of cox1 and nad1.

BACKGROUND: Fasciola hepatica as an important parasite affects health of humans and animals in some tropical and subtropical areas of the world, including Iran. Little is known about the molecular diversity of Fasciola in Equidae. Therefore, this study aimed to characterize the genetic polymorphisms among parasites. METHODS: Eight adult Fasciola spp. isolates were collected from a working donkey after necropsy in Shiraz, southwestern Iran, in 2018. Primarily, various parameters were measured morphologically. Subsequently, DNA was extracted from each fluke and molecular markers of cytochrome C oxidase (cox1) and NADH dehydrogenase 1(nad1) from individual Fasciola isolates were amplified using PCR assay and sequence data were employed for molecular and phylogenetic analysis. Genetic diversity between isolates was evaluated by comparing the sequences of these two mitochondrial regions. RESULTS: Based on the morphological and analyzed mitochondrial sequences, all of eight donkey isolates (100%) were identified as F. hepatica. Moreover, nine and five nucleotide polymorphisms were identified in the cox1and nad1 region sequences, respectively. CONCLUSION: Accordingly, phylogenetic data revealed five and four haplotypes among donkey isolates based on the cox1and nad1 markers. Similarly, some of these haplotypes have been previously reported from different host species in Iran as well as all around the world.

Molecular characterization of ß-tubulin gene associated with benzimidazole resistance in larvae of field isolates of Parascaris (Nematoda: Ascarididae).

Since the past 2 decades, an increasing number of resistance to the Benzimidazoles (BZs) have been reported in nematode parasites of livestock. More recently, detection of single-nucleotide polymorphisms (SNPs) at codons of 167, 198 or 200 of the ß-tubulin gene has been attributed to the occurrence of resistance. In the present study, we investigated the presence of those SNPs in the ß-tubulin isotype-1 gene in different isolates of Parascaris in horse. Also, the mitochondrial (mt) and ribosomal genes were sequenced for species confirmation of the isolates. The analysis of sequences inferred from COII gene confirmed that those isolates were P. equorum. The distance between mt genes obtained here and several ascarid species in equids and other hosts suggests the need for the combination of more genetic data with morphologic and other diagnostic measures. The analysis on ß-tubulin isotype-1 gene revealed no resistance-related SNPs or substitutions at the expected codon positions and selection pressure with BZs has not occurred for Parascaris worms. Although the molecular data showed the susceptibility of Parascaris isolates against BZs, other mechanisms of resistance should be also investigated to confirm the validity of molecular results.

Influence of hydatid disease on the pregnancy outcomes: An experimental study.

AIM: Hydatid disease during pregnancy may lead to various complications in both mother and child. Accordingly, enough knowledge is required to manage these complications. This study was done to evaluate the interaction between hydatid disease and pregnancy in laboratory mice. METHODS: Twenty female BALB/c mice (Mus musculus) were divided into test and control groups, 10 animal in each. The mice of the test group were infected intraperitoneally by injection of 1000 protoescolices. Six months after infection, the mice of test and control groups were mated with male mice for a week. After parturition, the size and weight of babies were measured and compared between two groups. RESULTS: While all the mice of control group delivered healthy babies, 3 out of 10 mice of test group showed no pregnancy. The infected mice delivered lower number of babies than the mice of control group. The mean size of fetuses or babies obtained from the infected mice were statistically lower than those obtained from the mice of control group (P = 0.000). The weight of babies delivered by the infected mice not only at the time of birth (P = 0.005) but also 10 days after birth (P = 0.021), were significantly lower than those delivered by the mice of control group. The level of progesterone and estradiol in the mice of test group were significantly lower (P = 0.009) and higher (P = 0.043), respectively, in comparison to the mice of control group. CONCLUSION: Hydatid disease during pregnancy may considerably affect the pregnancy outcomes.

The field efficacy of Nigella sativa and Berberis vulgaris methanolic extracts against Haemoproteus columbae.

OBJECTIVE: The methanolic extracts of Nigella sativaL. seeds (MENS) and Berberis vulgaris L.(MEBV) were investigated for treatment of Haemoproteus columbae-infected pigeons (Columba livia domestica). MATERIALS AND METHODS: One hundred twenty naturally-infected pigeons were randomly divided into four groups of thirty each. Two groups were treated separately with the extracts, while the positive and negative control groups were given buparvaquone (Butalex®) and distilled water, respectively. The parasitaemia rate was calculated in all groups before and after the experiment at four-day intervals for16 days. RESULTS: The results showed a high therapeutic effect for MENS with a progressive decrease in average parasitaemia rate from 18.17% before treatment to 0.73% at the end of treatment (p<0.05), while Butalex® was able to suppress the parasitemia rate from 18.90% before treatment to 0.23% at the end of experiment (p<0.05). However, no significant changes in parasitemia rate were evident in groups treated with MEBV (p>0.05). CONCLUSION: Methanolic extracts of N. sativa showed therapeutic effects against H. columbae and may be regarded as a suitable choice for further studies to develop new drugs against blood parasites, in both animals and human beings.

The field efficacy of Lepidium latifolium and Zataria multiflora methanolic extracts against Varroa destructor.

Varroa destructor is the most serious pest of honeybee (Apis mellifera), causing high economic losses in the beekeeping industry worldwide. The intensive utilization of many chemical substances against the mites resulted in resistance development. One of the applicable and alternative treatments being used for their control is plant-derived products (PDSS). The aim of this study was to evaluate the acaricidal activity of Lepidium latifolium and Zataria multiflora leaf extracts on V. destructor in field conditions. Four different concentrations (100, 200, 400, and 500 ppm) of the methanolic extracts were sprayed to treat each colony. The efficacy and side effects on adult bees were compared to Apistan chemical strips (ACSS). The acaricidal activity was the highest (100%) for L. latifolium extract at 500 ppm after 12 days and 86.26% for Z. multiflora. The infestation rate was decreased to 0.0% with L. latifolium and to 13.74% with Z. multiflora. The highest reduction was observed with L. latifolium followed by Z. multiflora extract at 500 ppm concentration. Both of the extracts showed negligible effect on bees, and it can be concluded that these PDSS as biodegradable agents could be used for V. destructor control in honeybee colonies.