Molecular Genotyping of Giardia duodenalis in Humans in the Yazd County, Central of Iran.

Background: We aimed to investigate the molecular genotyping of Giardia duodenalis in Humans in the Yazd County, Central, Iran. Methods: Total of 35 fecal samples were collected from patients referred to Yazd Central Laboratory, Yazd, Iran from February to July 2022. All the samples were included in this study after microscopic observation of G. duodenalis. DNA samples were extracted using related kit and were analyzed by Nano Drop. The molecular assessment was carried out using semi-nested PCR using the target gene of gdh. All amplified samples were sequenced using Sanger method. BLAST analyzed the sequences for assemblage identification. Results: Out of 35 samples, 24 (68.57%) and 11 (31.43%) were male and female, respectively. All included samples were amplified using the specific gdh primer pair. The molecular analysis showed 17 isolates (48.57%) as assemblage BIV, 8 isolates (22.86%) as assemblage BIII, 6 isolates (17.14%) as assemblage AII and 4 isolates (11.43%) as assemblage AIII (P<0.05). Conclusion: Assemblages A and B are the most prevalent in Central Iran. The molecular identification of G. duodenalis isolates from animals and implementing control programs.

Expression profile in Leishmania major exposed to Staphylococcus aureus and group A beta-hemolytic Streptococcus.

BACKGROUND & OBJECTIVES: The interaction of Leishmania spp. with microbiota inside the midgut vector has significant output in pathogenesis. This study aimed to identify the profile of Leishmania majorgene expression of LACK, gp63, and hsp70after exposure to Staphylococcus aureusand group A beta-hemolytic Streptococci (GABHS). METHODS: Leishmania major (MRHO/IR/75/ER) promastigotes were exposed with S. aureus, with GABHS, and with both GABHS and S. aureus at 25°C for 72 h. The gene expression analysis of Lmgp63, Lmhsp70,and LmLACKwas assessed using SYBR Green real-time PCR by ΔΔCt. All experiments were repeated in triplicate. Statistical analysis was done using two-way ANOVA. A P-value less than 0.05 was considered significant. RESULTS: Lmgp63 was expressed in the group exposed to GABHS with 1.75-fold lower than the control group (p=0.000). The LmLACK had expression in both groups exposed with GABHS and GABHS with S. aureus with 2.8 and 1.33-fold more than the control group, respectively (p=0.000). The Lmhsp70 gene expression was reported in the group exposed with GABHS with relative quantification of 5.7-fold more than the control group. INTERPRETATION & CONCLUSION: This study showed that the important genes encoding LACK, gp63, and hsp70 changed their expression after exposure to the S. aureus and GABHS.

In vitro evaluation of nanoliposomes berberine chloride against Leishmania major promastigotes.

Leishmaniosis caused by Leishmania major is one of the main infectious diseases that infected populations in developing countries around the world. We assessed the effectiveness of berberine chloride nanoliposomes (BcNLs) against L. major promastigotes in vitro. Nanoliposomal berberine chloride was prepared using thin film hydration method and characterized based on encapsulation efficiency, size and zeta potential. Anti-Leishmania effect of different concentrations (0.05-60 µg/ml) of BcNLs as studied in L. major (MRHO/IR/75/ER) at 24, 48 and 72 h using the hemocytometer technique. Berberine chloride was successfully loaded into nanoliposomes with encapsulation efficiency of 85.54%. The surface charge of nanoparticle is neutral and the morphology of nanoliposomal berbrine chloride is spherical without any agglomeration. Cell viability assay was performed on HFF cell line to show biocompatibility of liposome nanoparticles. IC50 of BcNPs at 24, 48 and 72 h against L. major were found to be 7.6, 5.96 and 3.19 µg/ml, respectively. BcNLs showed a significant anti-Leishmania effect and induced a better and more tangible effect on the survival of L. major promastigotes and could be suitable candidates for further investigation. The results showed that the BcNLs agent is effective against L. major promastigotes and may be a promising alternative to current treatments.

Leishmaniasis of the Feet Sole: A Case Report.

Here we report the case of a patient with cutaneous leishmaniasis, who was referred to our clinic in Yazd, Iran. On examining the patient, who was a housekeeper, we found a small plaque in the palmoplantar region due to cutaneous leishmaniasis. She had not any history from an identical case in this patient. After treatment, the lesions improved.

In vitro anti-leishmanial activity of Satureja hortensis and Artemisia dracunculus extracts on Leishmania major promastigotes.

Cutaneous leishmaniasis is one of the important skin diseases with diverse clinical manifestations. With an incidence of 0.7-1.3 million cases annually, this disease is often reported from six countries, including Iran. Accordingly, the purpose of this study was to evaluate the anti-leishmanial effect of the three plant hydroalcoholic extracts including fleawort (Plantago psyllium L.), savory (Satureja hortensis L.) and tarragon (Artemisia dracunculus L.) on Leishmania major promastigotes. The hydroalcoholic extract from each plant was extracted and its anti-leishmanial effect was evaluated in different concentrations (100-1000 µg/ml) and at various hours (24, 48 and 72 h). Savory herb inhibitory concentration 50 % (IC50) at 24, 48 and 72 h was 790.81, 398.11 and 298.42 µg/ml, respectively. In addition, tarragon herb IC50 at 24, 48 and 72 h was 962.03, 688.36 and 585.51 µg/ml, respectively. Moreover, the fleawort extract was showed the lowest effect, considering that its effect at the concentration of 1000 µg/ml was 48 % after 72 h (P > 0.05). Furthermore, the statistical analysis showed a significant difference for interaction between concentration and time regarding the tarragon and savory extracts with a P value of lower than 0.05. According to the results, the anti-leishmanial effect of the tarragon and savory extracts may make it possible to use them in the treatment of cutaneous leishmaniasis as a complementary or alternative therapy; however, further studies are necessary and should be evaluated in cell culture and in vivo conditions to confirm it.