Anti-Trichomonas vaginalis Activity of Marine Ascidians (Tunicates; Ascidiacea) from the Bushehr Province, Iran

Objective: The aim of the current research is to evaluate the antiparasite effects of compounds isolated from marine ascidian tunicates on Trichomonas vaginalis. Methods: Ascidian tunicates after collection were cut into small pieces, freeze-dried, and powdered. The resulting material was subjected to extraction in double-distilled water, ethanol, n-hexane, and dichloromethane. To fractionate the extracts and identify the most bioactive compound, silica gel column chromatography and GC-M/S analysis were used. Results: Fraction 18 of silica gel column chromatography of ethanol extract was the most effective against T. vaginalis. The respective IC50, CC50, and SI values for fraction 18 were 28.62 µg/mL, ˃800 µg/mL, and ˃27.95. GC-M/S analysis of this fraction identified a major phenolic compound (2, 4-bis (1, 1-dimethyl ethyl), whose toxicity against vero cells was only 10.15%. Conclusion: The ethanolic fraction containing phenol-2,4-bis (1,1-dimethylethyl), which has a potent lethality effect on T. vaginalis, may be considered as an antiparasite drug candidate.

Detection of plasmid-mediated AmpC ß-lactamases in Klebsiella pneumoniae clinical isolates from Bushehr province, Iran.

Background and Objectives: Plasmid-mediated AmpC producers are considered an increasing concern. The aim of this study was to investigate the prevalence of plasmid-mediated AmpC ß-lactamases (pAmpCs) in Klebsiella pneumoniae isolates. Materials and Methods: A total of 228 clinical isolates of K. pneumoniae were collected in Bushehr province, Iran, from December 2017 to February 2019. Cefoxitin disks were applied for screening AmpC-producing isolates. Furthermore, 3 phenotypic confirmatory tests including combine disk test (CDT), double disk synergy test (DDST) and modified three dimensional test (M3DT) were used. Finally, the presence of pAmpC genes was tested by multiplex PCR. Results: We identified 18 pAmpC-KP isolates among the 228 isolates (7.9%): 12 DHA (66.6%) and 6 CMY (33.3%). In the present study only 47% of cefoxitin-resistant isolates were pAmpC producers. The sensitivity of CDT, DDST, and M3DT was 89%, 67% and 100% and the specificity was 90%, 90% and 85%, respectively. In addition, M3DT displayed a higher rate of efficiency (92%) than CDT (89%) and DDST (79%) in detecting plasmid-meditated AmpC producers. Conclusion: DHA was the most prevalent pAmpC beta-lactamase in this study. DDST and CDT tests proved inefficient to detect two and six pAmpC producers, respectively, while M3DT represented the best overall performance.

Investigating the Prevalence of Intestinal Parasites in Immunocompromised Patients in Bushehr Province, Southwest Iran: A Conventional and Molecular Study

Objective: The aim of this study was to determine the status of intestinal parasitic infections in immunocompromised patients in Bushehr province, southwest Iran by conventional and molecular methods. Methods: A total of 201 stool samples were collected from kidney transplant recipients, AIDS patients and patients under chemotherapy. Samples were collected from healthy people as the control group. The specimens were tested using various conventional methods. Polymerase chain reaction (PCR) testing was performed on samples identified as positive for Coccidia by direct microscopic examination. Results: Approximately 32.45% were infected with at least one type of intestinal parasite. The highest (46.8%) and lowest rates of infection (24%) were observed in AIDS and chemotherapy patients, respectively, while the infection rate of the control group was 16%. Isospora spp. and Cryptosporidium spp. were observed in all patient groups, and Sarcocystis spp. sporocysts were detected in one of the transplant recipients. All identified coccidia were confirmed by PCR. There was a significant relationship between the rate of intestinal parasite infection and certain variables. Conclusion: Given the potential risk of certain intestinal parasites in people with immune deficiency, it is recommended that diagnosis of parasitic infections in such patients be based on specific parasitological methods. Thus, it is advisable that physicians refer them to a parasitology laboratory prior to drug administration.

Genotyping and phylogenetic analysis of hydatid cysts isolated from livestock in Bushehr province, Iran.

Hydatid cyst is one of the parasitic zoonoses caused by infection with the larval stage of Echinococcus granulosus tapeworm. The spread of this parasite is global and is of great importance in terms of public health. To date, ten different species of this parasite have been identified that differ in characteristics such as life cycle, epidemiology and pathogenesis. The purpose of this study was to determine the genotype and phylogenetic relationship of hydatid cysts isolated from livestock of Bushehr province, Iran. About 62 samples of hepatic and pulmonary hydatid cysts were collected from slaughtered animals. DNA extracted by phenol-chloroform method was amplified by PCR using primers specific for the cox1 gene. The PCR products of 50 samples were sequenced and analyzed using BioEdit software and compared with sequences in the GenBank. The phylogenetic tree was drawn using Neighbor Joining tree-NJ method, and its reliability was evaluated. Sequencing results showed that out of 50 sequenced samples, 43 samples had the genotype of Echinococcus granulosus and 7 samples had the genotype of Taenia hydatigena. By drawing a phylogenetic tree, all 43 hydatid cyst samples belonged to G1 strain. The predominance of G1 strain of hydatid cyst in livestock of Bushehr province shows the main role of this genotype in establishing the life cycle of parasite in this region and if the genotype of the parasite in dogs and humans is determined, then these findings can be used to disrupt the life cycle of the parasite and reduce the human infections.

Evaluation of a new multi-epitope sequence of eight known Leishmania infantum antigens for HVL diagnosis by ELISA and Western blot.

BACKGROUND & OBJECTIVES: Leishmaniasis, known as a disease with high prevalence proportion throughout the world, is caused by protozoan parasites. Visceral leishmaniasis is the most severe form of this condition reported sporadically from all regions in Iran. Between different diagnostic tests, serodiagnosis of this infection is of utmost importance in both humans and dogs. Although rK39 ELISA test has been extensively validated in endemic areas, there are currently challenges regarding a more appropriate serodiagnostic test. METHODS: A novel multi-epitope construct was designed consisting of highexposedB cell epitopes using eight important antigens of Leishmania infantum (Gp63, KMP-11, HSP70, CPA, H2A, H3, LACK and TRYP). Our artificial sequence, a Multi-epitope Recombinant Protein (MRP), was consequently produced and purified. Then, immunoreactivity was investigated by ELISA test and western blotting as well. RESULTS: In the present study, the cutoff value (1.052) for the new MRP-ELISA was determined by receiver operator characteristic (ROC) curve analysis using 35 known positive and 20 known negative HVL sera previously tested for antibodies to L. infantum by DAT, showing a sensitivity of 93.1% and a specificity of 77.4%. The blotting test also showed a favorable band to detect visceral leishmaniasis. INTERPRETATION & CONCLUSION: According to the results, this new antigen had acceptable potential in detecting VL positive cases once western blotting was utilized, but the ELISA test did not proceed as expected for detecting true negative cases, probably due to some optimization issues.The present study is a promising start.

Impact of exposure to ambient air pollutants on the admission rate of hospitals for asthma disease in Shiraz, southern Iran.

Asthma is a common chronic respiratory disease in the world. Short-term exposure to ambient air pollutants is closely related to acute respiratory diseases and asthmatic symptoms. The purpose of this research was to estimate the correlation between exposure to three air pollutants (O3, NO2, and SO2) and hospital admission because of asthmatic disease (HAAD) in the city of Shiraz, southern Iran. The data were collected from the two real-time monitoring stations located in this city. The acquired information was used for developing predictive models by the AirQ software. The findings of this study were reported for two age groups (<15 and 15-64 years old). The highest levels of O3, NO2, and SO2 were obtained 187.33 µg/m3, 34.1 µg/m3, and 491.2 µg/m3 in 2016, respectively, and 227.75 µg/m3, 92.26 µg/m3, and 190.21 µg/m3, respectively, in 2017. Among the mentioned pollutants, the yearly average concentration of SO2 was 8.62 times more than the WHO guideline, during the studied times. The number of extra cases of HAAD for <15 years and 15-64 years caused by the air pollutants in Shiraz were estimated to be 273 and 36, respectively, in 2016, and 243 and 30 for 2017, respectively. The results of this work displayed that air pollutants have caused respiratory problems in Shiraz city. The AirQ model is a facile and potential tool for the prediction of asthma disease to reduce the health risk of atmospheric pollutants in the worldwide.

Toxic heavy metals and nutrient concentration in the milk of goat herds in two Iranian industrial and non-industrial zones.

This work aimed to explore the concentration of nickel, manganese, iron, copper, chromium, and lead in the milk of goat herds in the industrial area of Asaluyeh (southern Iran) and the non-industrial area of Kaki. The milk of 16 goat herds (each herd had at least ten goats) was collected in several villages in each area, and at the same time, the drinking water and forage of goats were sampled. The concentration of elements in the samples was determined by ICP-OES. The mean concentrations of chromium, copper, iron, manganese, lead, and nickel in milk samples of the Asaluyeh area were 16.423 ± 0.349, 0.146 ± 0.118, 6.111 ± 0.501, 0.239 ± 0.016, 0.141 ± 0.030, and 1.447 ± 0.101 mg/kg, respectively. Concentrations of heavy metals (except for copper) in the milk of goats in the industrialized area of Asaluyeh were significantly higher than that of Kaki (P < 0.05). Also, the content of heavy metals was significantly correlated with lactose levels (P < 0.05). The hazard index for drinking the goat milk was computed to be 0.444 and 0.386 for the Asaluyeh and Kaki area, respectively, which shows a minimal effect of this exposure pathway.

Comparison of various methods for DNA extraction from human isolated paraffin-embedded hydatid cyst samples.

Successful molecular research with reliable results depends on achieving significant and uniform amounts of genomic DNA from the parasite as the first and most basic step. Therefore, selection of an appropriate method that minimizes damage to the DNA of the parasite, is very important. In this study, we are going to describe a method that can extract DNA from human isolated paraffin-embedded hydatid cysts with a high quality and quantity. Formalin fixed and Paraffin-embedded hydatid cyst samples isolated from human lung and archived in the pathology laboratory were used for this purpose. Several sections of the paraffin blocks were prepared with 5 micron thickness and DNA were extracted by three different methods including; modified boiling, commercial kit and the method described by Larissa A. Pikor et al. The obtained DNA were evaluated by Nanodrop in terms of the yield of DNA and possible contaminations. To compare the quality of DNA prepared, cox1 region was amplified using specific primers. It was found that the DNA extracted by modified boiling had the lowest rate of contamination and the best electrophoretic band on the gel, compared to other two performed methods. Considering the findings of this study, this simple and high throughput DNA extraction method with high yield and quality can be recommended for extraction of DNA from formalin fixed and paraffin-embedded hydatid cysts.

Rational Design of Hypoallergenic Vaccines: Blocking IgE-binding to Polcalcin Using Allergen-specific IgG Antibodies.

Chenopodium album polcalcin (Che a 3) is characterized as a major cause of cross-reactivity inallergic patients to the Chenopodiaceae family. Therefore, the present study was conducted to develop a hypoallergenic Che a 3 derivatives as the candidate vaccine for type 1 allergy. Four derivatives were generated from Che a 3. The first was a mosaic peptide derivative computationally identified in Che a 3 which was coupled to keyhole limpet hemocyanin (KLH). The second one was a mutant Che a 3, and the other two derivatives included N- and C-terminal halves of Che a 3 that both coupled to KLH. The IgE-binding capacity of Che a 3 and its derivatives and also their ability to induce there combinant Che a 3 (rChe a 3)-specific IgG antibody, were determined using the enzyme-linked immune sorbent assay (ELISA). Moreover, the lymphopro liferative capacity of rChe a 3 or its derivatives and their pro-inflammatory cytokine response interleukin (IL)-5 and IL-13 were measured in the human peripheral blood mononuclear cells (PBMCs). Among all derivatives, the N-terminal half peptide and mosaic peptide exhibited the lowest IgE-binding capacity. In addition, in comparison to other antigens, KLH-coupled mosaic peptide induced the highest level of the recombinant Che a 3 (rChe a 3)-specific IgG antibody and ther Che a 3 specific-blocking IgG antibody in mice. Moreover, the mosaic peptide lacked lymphopro liferative capacity and down-regulated expression of pro-allergic IL-5 and IL-13 cytokines. Therefore, a peptide-carrier fusion vaccine, composed of the B-cell epitope coupled to the carrier, could be considered as one of the promising hypoallergenic vaccines to treat patients with allergy to low molecular weight allergens such as Che a 3.

Molecular Identification of Species Caused Cutaneous Leishmaniasis in Southern Zone of Iran.

BACKGROUND: Leishmania major and Leishmania tropica are two main species causing cutaneous leishmaniasis (CL) in Iran. Recently, Crithidia spp. has also been reported in the wound of patients with CL. In this study, we determined the species causing CL in the southern of Iran and the role of Crithidia spp. in creating skin ulcers. METHODS: In this cross-sectional study from Apr to Sep 2016, 66 patients with CL referred to Diagnostic Lab of Leishmaniasis, Valfajr Health Center, Shiraz, Iran, were selected. After DNA extraction from the Giemsa stained smears, all samples were amplified in two separate steps using specific primers, firstly, to differentiate Leishmania species and then to identify Crithidia spp. RESULTS: Two species L. major and L. tropica were responsible for 60 and 6 cases, respectively. Moreover, in two patients, mixed infection with Crithidia was confirmed. In mix infection cases, the morphology of the cutaneous ulcers was not different from the wounds of other patients. CONCLUSION: Leishmania major is responsible for the most common CL in southern Iran. In addition, in two patients with L. major and L. tropica, mix infection with Crithidia was confirmed. The potential role of Crithidia as the main factor for CL and the probability of this parasite to have synergistic effects on Leishmania, as a hypothesis, requires more comprehensive researches on the ambiguity of this protozoon.

Molecular and parasitological study of cutaneous leishmaniasis in Bushehr province, southwest of the Islamic Republic of Iran: a cross-sectional study during 2009-2012.

Cutaneous leishmaniasis (CL) is one of the most important parasitic disease in Iran. CL is distributed among more than half of 31 provinces of Iran. Studies on epidemiological aspects of the disease and Leishmania species identification among infected humans are necessary for providing a comprehensive prevention and control program thus; this descriptive cross-sectional study was conducted on all CL suspected patients who referred to Health Centers of Bushehr province from 2009 to 2012. Physical examinations were carried out in suspected individuals and CL cases were confirmed by microscopical examinations. Prepared slides from suspicious cases of CL were fixed with absolute methanol and stained by Giemsa 10 %. All the Giemsa-stained slides examined under a light microscope with high magnification (1,000×) and classified them based on grading of Leishmania parasites. DNA from each slide was extracted, separately. The ribosomal internal transcribed spacer 1 was amplified with specific primers and PCR products were digested by restrict enzymes (HaeIII), run them in 3 % gel agarose for electrophoresis and visualized on a UV transilluminator after staining with ethidium bromide. SPSS version 21 was used for data analyses. A total of 726 suspected CL cases were referred to Health Centers of Bushehr province from 2009 to 2012 and samples were only prepared from 188 of the patients whereas 43 (5.9 %) of them were microscopy positive. The most frequent of CL was observed in November (14 %) and December (12 %). The most distribution of CL lesions were observed on hands (32 %), feet (26 %), and face (21 %), respectively. The highest frequency of CL was observed in 1-9 years old (30 %). Altogether, 50 % of the patients showed one skin lesion and 2-10 skin lesions were occurred in the remained CL patients. Totally, 27 out of 43 (63 %) of the Giemsa stained slides were positive by PCR-RFLP assay because all the PCR-RFLP negative slides were prepared 3-4 years ago and kept without cover slip, and also observed scarce amastigotes during microscopy observations. Leishmania species were identified in 21 desirable slides which 14 of them were L. major and 7 of the remained isolates were identified L. tropica using PCR-RFLP.

The prevalence of intestinal parasites in hemodialysis patients in Bushehr, Iran.

Hemodialysis patients, due to a dysfunction of the immune response, are prone to a variety of opportunistic infections. Studies of intestinal parasitic infections in these patients are limited. Therefore, the present study was performed to determine the prevalence of these infections in patients on hemodialysis in Bushehr. In this cross-sectional study, fecal samples have been collected from all hemodialysis patients who were continuously referred from September 2011 to September 2012 to the dialysis center at Bushehr and tested using routine parasitological methods. From a total of 88 patients studied, 25 patients (28.4%) were infected with one or more intestinal parasites. Blastocystis hominis and Entamoeba coli with 13.6% and 6.7% prevalence had the highest prevalence among the patients, respectively. The age group 51-70 years had the highest rates of infection. Statistical analysis showed no relationship between sex and the risk of intestinal parasites. Seventeen percent of infected patients showed up with diarrhea and this relationship was statistically significant. Considering the high prevalence of intestinal parasitic infection among hemodialysis patients in Bushehr and also the high probability of infection in these patients, it is recommended that periodic examinations and screening patients during dialysis and before kidney transplantation should be a part of routine medical care.

Evaluation of antiviral activities of curcumin derivatives against HSV-1 in Vero cell line.

Antiviral drug resistance is one of the most common problems in medicine, and, therefore, finding new antiviral agents, especially from natural resources, seems to be necessary. This study was designed to assay the antiviral activity of curcumin and its new derivatives like gallium-curcumin and Cu-curcumin on replication of HSV-1 in cell culture. The research was performed as an in vitro study in which the antiviral activity of different concentrations of three substances including curcumin, Gallium-curcumin and Cu-curcumin were tested on HSV-1. The cytotoxicity of the tested compounds was also evaluated on the Vero cell line. The CC50 values for curcumin, gallium-curcumin and Cu-curcumin were 484.2 microg/mL, 255.8 microg/mL and 326.6 microg/mL, respectively, and the respective IC50 values 33.0 microg/mL, 13.9 microg/mL and 23.1 microg/mL. The calculated SI values were 14.6, 18.4 and 14.1, respectively. The results showed that curcumin and its new derivatives have remarkable antiviral effects on HSV-1 in cell culture.

High sensitivity C-reactive protein is associated with the metabolic syndrome independent to viral and bacterial pathogen burden.

AIM: To investigate the influences of bacterial or viral pathogen burden in the relationship of high sensitivity C-reactive protein (hs-CRP) and the metabolic syndrome in a population-based study. METHODS: Data from 1754 men and women aged >or=25 years, from the Persian Gulf Healthy Heart Study were analyzed. The definition of the metabolic syndrome according to the Adult Treatment Panel III was used. Sera were analyzed for IgG antibodies to Chlamydia pneumoniae, Herpes simplex virus type 1, Helicobacter pylori and cytomegalovirus using ELISA. Measurement of CRP by a high-sensitivity CRP assay was done. RESULTS: The subjects with the metabolic syndrome had a higher geometric mean of CRP levels than the normal persons (p<0.0001). A linear relationship between an increase in the number of metabolic syndrome components and CRP concentrations was observed (p for trend<0.0001). In multiple logistic regression models, hs-CRP showed significant associations with the metabolic syndrome after controlling for cardiovascular risk factors and infectious burden divided into 2, 3 and 4 pathogens [OR=2.06, CI (1.32-3.21), p=0.001; OR=1.75, CI (1.26-2.42), p=0. 001; OR=2.12, CI (1.46-3.08), p<0.0001; respectively]. CONCLUSION: There was a strong association between inflammation and the metabolic syndrome independent to viral and bacterial infectious burden.