فهرست مطالب منصور بیات

Identifying new antimicrobial compounds is a way to deal with spread of bacterial resistance, which bacterial pigments are one of the antimicrobial compounds. The most common causes of treatment-resistant infections in humans and animals are bacteria such as Pseudomonas aeruginosa and Acinetobacter. The purpose of this research is to compare the antibacterial properties of prodigiosin pigment from Serratia marcescens and carotenoid pigment from Rhodotroella glutinis on Pseudomonas aeruginosa and Acinetobacter. After separating the bacteria and identifying them, by using differential tests, Serratia marcescens (PTCC: 1111) and Rhodotrola (PTCC: 5256) pigments were extracted through chromatography. In order to evaluate the effect of different dilutions of pigments on bacteria, sterile 96-well microplates were used and the minimum inhibitory concentration (MIC) and minimum lethal concentration (MBC) were determined by broth microdilution method. Welling was done with different volumes of pigments. Out of the 50 burn wound samples, 26 samples were positive for the presence of two bacteria, all of which were sensitive to colistin and resistant to gentamicin, imipenem, ceftazidime, amikacin and ciprofloxacin. After the effect of both pigments on Pseudomonas aeruginosa, the highest MBC observed was 25 microliters, and after the effect of prodigiosin pigment on Acinetobacter bacteria, the highest MBC was 6.25 microliters, and this value for carotenoid pigment was 25 microliters. In welling, the largest average diameter of the zone of inhibition related to two pigments against these bacteria was observed in the volume of 200 microliters

In general, animals are an important source of alopecia for humans, both in urban and rural areas. Therefore, the issue of controlling dermatophytosis in animals is important for public health. The presence of infected epidermal shells in public places can be a potential source of infection. Given the high prevalence of dermatophytosis and considering the treatment of problems in some forms of the disease and the side effects of antifungal drugs, the need for appropriate replacement with current drugs seems essential.

Standard Trichophyton mentagrophytes ATCC18748 isolate and clinical isolates of Trichophyton mentagrophytes were used in the present study. 1% terbinafine in nano-form of terbinafine). Inoculation in guinea pigs was performed by the method described by Tagami et al. The site of daily lesion was followed for 20 days in terms of clinical symptoms.

In studies performed on lesion score average in the nano-form group, terbinafine was combined on the first day of treatment, showing a score of 3 (significant redness with large crust). And for the normal form of the drug terbinafine (group 4) shows the number 4 (ulcers and scars along with those in grade 3), the process of reducing the degree of lesions in the treatment groups of nano-drugs, occurred between 10 and 15 days. Which is accompanied by an increase in the next 5 days. And continued with a logical process until the 40th day. On day 40, all groups had a zero score except for positive control.

This study shows that nano-drugs are suitable for the treatment of dermatophytosis caused by Trichophyton mentagrophytes and in terms of the effectiveness of treatment with the usual form of drugs have a more comparable intensity and speed of recovery.

Background of the study:

 In the cultured and ornamental fish industry, fungal organisms threaten young and adult cultured populations and lead to the decay of eggs and larvae. Fungal Infections Secondary infections in fish are caused by primary parasitic, viral and bacterial organisms.

In this research, the number of fungal infections and their type of infection in ornamental fish should be determined.

This study was conducted on ornamental fish with clinical symptoms. The samples were obtained from different places. After identifying the fungi, their abundance was checked by culture and PCR.

Out of the total of investigated cases, the skin and gills had lesions in cases, and the result of mushroom culture was positive. In the PCR analysis of these samples, the fungal species identified by gene sequence were Cladosporium, Candida, Penicillium, and Aureobasidium pullulans and Rhodotorula yeast. Pathological lesions in the gills were dark and hyperemic; under the microscope, epithelial cells were shed, and leukocytes were present. It was seen in the skin as macroscopic and microscopic wounds. Finally, these fungi are responsible for forming lesions seen under certain conditions, such as environmental instability and immunosuppressive factors.

Cryptococcosis is usually a subacute or chronic infection in nature and Cryptococcus neoformans is the cause of this infection.This yeast is found in bird droppings and can be transmitted through dust and inhalation. People with immunodeficiency disorders are at high risk for this infection. In this study, samples were collected from nasal mucosa of domestic cats. The suspension was prepared samples cultured in S, Sc and BHI media and incubated at and °C. After hours, we assessed sample by using Chinese Cotton Blue stain solution. Afterwards, samples were identified by using sugar fermentation tests, urea hydrolysis and germ tube formation. We examined suspected colonies of Cryptococcus neoformans by PCR. The results shown a variety of filamentous fungi such as Penicillium, Aspergillus, Alternaria, Scopulariopsis, Mucor, Rhizopus and yeasts such as Rhodotorula, Candida albicans, Candida Sp, Pachia were isolated and Cryptococcus neoformans were not isolated. Molecular tests of suspected samples of Cryptococcus were negative, The reason of this results could be personal hygiene in the case of domestic cats, keeping them indoor and not have contact with other animals and outdoor environments.

Mastitis, as a most costly disease of dairy cows, causes a lot of damage to the livestock industry in the world every year. One of the most important bacteria which causes clinical mastitis in cattle is S. aureus, which has become resistant to most common antibiotics in veterinary medicine. Therefore, researchers are looking for new methods in the treatment of such infections, in which the pigments of microorganisms fall into this category. The aim of this recent study was to optimize the production of carotenoid pigments by Rhododorula yeast and to investigate its antimicrobial effect on S. aureus isolates To isolate S. aureus, samples were taken from the milk of 100 cows with mastitis and the femA gene was identified for molecular confirmation among suspicious isolates of Staphylococcus by PCR method. Also, the isolate of R. glutinis obtained from one of the samples of mastitis was further examined and used as a sample of yeast producing pigment. The results showed that in addition to S. aureus as one of the causative agents of mastitis in dairy cattle, but also the yeast R. glutinis can be a causative agent of mastitis. The inhibitory effect on the growth of Staphylococcus using the pigment extracted from R. glutinis was also quite evident and more than 80% of S. aureus isolates were sensitive to the pigment at a concentration of 200 μg. R. glutenis inhibits excellent growth in S. aureus as one of the leading causes of mastitis in cows. The cause is the appearance of synthetic antibiotics and the reduction of microbial resistance.

Candida albicans is a common yeast in opportunistic fungal diseases world wide, which is frequently colonized on the surface of the skin and mucous membrane. The aim of this study was to investigate the effect of all nanocomposite copper-silver-chitosan on the mentioned yeast. To synthesis of the nanocomposite, first the chitosan was dissolved in water using ultrasonic device, the binding of chitosan and glutaraldehyde was determined by FT-IR technique and the size and morphology of the synthesized nanocomposite were determined by SEM microscopy. In the animal study section, 43 male Wistar rats weighing 200-250 gin 5 groups of 7 were examined. The immune system of mice was weakened by 30 mg/kg cyclophosphamide injection through peritoneal injection and was given candidiasis by oral inoculation of candida suspension. After treatment with the nanocomposite and nystatin, it was found that the number of inflammatory cells in other groups was higher than the group treated with nystatin. Also, the number of cells in nanocomposite coper- silver- chitosan- treated group was much lower than in the other group of mice with weakened immune system and untreated candidiasis. The findings of the present study showed that the therapeutic effect of the nanocomposite against candidiasis was relatively high and had a positive effect on reducing inflammation. Although the anti-yeast effect of copper and chitosan nanoparticles have not been proven separately, but based on the current results, it was found that their combination has a slight synergistic effect and to some extent, slightly effective in their antifungal properties.

Treatment of breast cancer has many problems due to the systemic effects of chemotherapy. The use of certain molecules with cell killing effects, such as aflatoxin B1, can have fatal effects on breast cancer cells. In this study, we investigated the inhibitory effect of topical aflatoxin B1 use on breast cancer. Class 4T1 cancer cells were inoculated subcutaneously in female mice. Doses of 50 and 500 μg / ml of aflatoxin B1 were then injected around the tumors in comparison with doxorubicin. The ratio of tumor size to animal weight in different groups was evaluated along with histopathology by immunofluorescence on Ki-67 and VEGF indices. Aflatoxin B1 significantly reduces tumor volume, necrosis and suppresses angiogenesis in the tumor. However, liver metastasis occurred in all groups. Aflatoxin B1 in topical injections of 50 and 500 μg / ml around the tumor has anti-cancer effects on the breast. But it can not prevent metastasis.Aflatoxin-induced hepatotoxicity was also observed.

Aspergillosis is an opportunistic fungal infection in animals. The disease is mainly respiratory, but other disease manifestations also occur in poultry. Researchers have shown that, one of the reasons for the increase in drug resistance in Aspergillus species is the overexpression of the CYP51A gene. This study compared CYP51A gene expression of Aspergillus fumigatus isolated from poultry against fluconazole and nano-fluconazole. Fluconazole liposomal-nanoparticles were prepared by the thin layer hydration method. We dissolved 5.12 mg of fluconazole powder in 1 ml of sterile distilled water and 6 ml of chloroform-methanol organic solvent. We added 51 mg of lecithin and 5 mg of cholesterol to it. The size of nanoparticles was 88.9±12.1 nm and the surface charge of these nanoparticles was -20.12±1.88 mv. We also used a Scanning-Electron Microscope to study the structure of nanoparticles. Thirty samples of A. fumigatus were collected from poultry lung nodules. Minimum inhibitory concentration (MIC) was performed by standard Broth Microdilution method according to NCCLS-M38A2 to evaluate the MIC of fluconazole and nano-fluconazole against isolates. We selected two high-resistance isolates to fluconazole and used them to determine the CYP51A gene expression level by real-time PCR. The results showed that nano-fluconazole had a lower MIC than fluconazole and in lower concentrations of the drug inhibited the growth of Aspergillus fumigatus isolates. CYP51A gene expression was increased in fluconazole and nano-fluconazole-treated isolates compared to the untreated state. Conversely, a decrease in CYP51A gene expression was observed in the exposure to nano-drug compared with the normal drug.

Recent advancements in drug delivery system through development of nanomedicine has significantly improved the delivery of the medicine to the target tissue, consequently reduced the required therapeutic dose, toxicity and side effects. The aim of this study was test antifungal susceptibility of trimethylchitosan nanoparticles in combination with amphotericin B (NPs-AmB) through broth micro-dilution method described in CLSA-A3 guidelines on resistant (R) and susceptible (S) Candida albicans isolates and the cell viability and proliferation assays. Minimum inhibitory concentrations (MICs) 90 % of NPs-AmB and AmB determined to be 0.25 and 0.75 µg/mL, and minimum fungicidal concentration (MFC) were 0.5 and 1 µg/mL for S-isolate. MIC 90 for R-isolate were 8 and 32 µg/mL and MFC were 32 and 64 µg/mL respectively demonstrating a significant improvement in antifungal activity (p

Ascomycota contains the most numerous groups of fungi and playing important roles in most of ecosystems. Diverse aspects of their activities, ascomycetes are known as fascinating subjects of biological studies, including exploration of new strategies for alteration of fungal lifespan. The gene los1, directly involving in nucleus-cytoplasm tRNA trafficking, plays a critical role in fungal growth rate, as well as its drug resistance. Here, we investigated an upstream region of the gene los1; among 10 species belong to Ascomycota. Furthermore, its homologue was knocked out from Aspergillus fumigatus, and its subsequent influences on any changes in expression of some related genes and drug resistance were evaluated. Our results suggest that in Ascomycota, the upstream region of los1 contains recognition sites for transcription factors SFP1 and FKH1. There is also a low probability that the transcription factor YAP6 binds to the upstream region of this gene. Besides, its deletion may lead to increase of fungal lifespan, as well as its resistance to bleomycin, while it presents more sensitivity to cycloheximide. The results of this study can be used in more targeted and intelligent selection of fungal antibiotics. In addition, information about the role of los1 gene in the growth and lifespan of ascomycetes, as well as their drug resistance and sensitivity, can be used in the design and manufacture of industrial strains.

Cancer is the second factor of death in world on the basis of WHO reports on 2018. Application of fungal polysaccharides is one of the cheaper, less dangerous, less side effects, and newer in the treatment and prophylaxis of cancer. Object of this research is isolation and identification of endemic fusarium fungus, its glucan extraction, determination of chemical characteristics by HPLCand FT-IR and anticancer effects on cell lines of LCL, Hela.In this study fusarium fungus was isolated from soil of karaj district and identified on the basis of microscopic and macroscopic and genetical characteristics as a fusarium genus .Then fungus was grown into selective broth medium for obtaining of the most biomass for extraction of glucan .Glucan of fungus was extracted by boiling water method.Different amounts of extracted glucan were treated to the cell cultures of lcl and Hela and its cytotoxicity effects were surveyed.Results showed that glucan polysaccharide had anticancer effects against cell lines of LCL and no anticancer effects against Hela cell lines. Cytotoxicity effects of glucan showed by colorimetry MTT method.

Epidermophyton floccosum is one of the worldwide anthropophilic dermatophytes that invade keratinized structures such as hair, skin, and nail, causing dermatophytosis by secreting important proteases such as subtilisin. This study aimed to evaluate the presence of SUB3 and SUB6 encoding serine proteases in the isolate, which received from the fungi culture collection of Tehran University of Medical Sciences, Faculty of Public Health. Special primers designed according to the highly conserved regions of similar genes in other dermatophytes. Genomic DNA and designed primers used in PCR, then PCR products sequenced with ABI PRISM®3730XL automated Sanger sequencer, and presence of 2 new subtilisin genes (SUB3 and SUB6) were confirmed and recorded in NCBI(with the accession numbers MN206114, MN177931 respectively). The coding sequence of SUB3 found to contain 861 nucleotides, which encodes a polypeptide with 287 amino acids. The coding sequence of SUB6 found to contain 699 nucleotides that encode a polypeptide with 233amino acids. Comparing the sequences with Gene Bank database information, revealed significant homology with other dermatophytes. Achieving a better understanding of the molecular characteristics of virulence genes may help develop effective therapies and prevention strategies.

Shigella is a gram-negative intestinal bacillus whose infections have caused serious problems in developed and developing countries. The prevalence of Shigella infections is very low due to the low dose of pathogenicity of this bacterium as well as its easy transmission from one person to another and also the indirect contamination of people through the consumption of contaminated food and water. Objective of isolation of Shigella sonnei from children's feces, Effect and isolation of spring grass extract, Molecular isolation of tetracycline genes by multiplex PCR, Effect of extract on tetracycline gene expression by multiplex PCR.

Out of 60 Shigella sonnei strains isolated from children with diarrhea after isolation of strains on different media and MacConkey Agar (MAC) medium in terms of shape and color of colonies were examined and recorded. First, the DNA of each sample was extracted by a DNA extraction kit, and after extraction, the DNA of the template was amplified by PCR. To investigate the effect of antibiotic resistance of spring grass to the extracted gene resistant to tetracycline, MIC of plant extract on different concentrations of Shigella sonnei was investigated. Strains with tetracycline-resistant genes were treated with MIC, using both treatment and non-treatment groups, and after incubation for 18 hours, stationary lag log was performed using RNA extraction kit x plus RNA extraction. Then, using RT-PCR ABI plus device, the expression of tet A and tet B genes in the treated and untreated groups under the influence of spring grass extract was read quantitatively (numerically) CT and the expression of the gene under the influence of treatment was interpreted.

Shigella sonnei isolated results showed a high percentage of the highest resistance to streptomycin, Nalidixic acid, tetracycline and ampicillin and chloramphenicol with 100%, 100%, 86.7%, 91.8% and 50%. Also, the highest sensitivity to gentamicin and ciprofloxacin was observed with 85% and 86.6%, respectively. Multiple PCR results showed that out of all 60 Shigella sonnei isolates, 3 isolates had only tetB gene (5%) and 55 isolates had tetA gene (91.6%).

The results of Fisher test indicate that there was a significant relationship between tetA genes and antibiotic resistance at 5% error level (p-value <0.05). The results showed that at the 5% error level there was no significant relationship between tetB genes and antibiotic resistance (p-value = 0.05). The results showed that at the 5% error level, there was no significant relationship between positive and negative genes of all genes with antibiotic resistance (p-value <0.05).

Due to the increasing problems and side effects of the use of chemical antibacterial agents as well as antibiotic resistance, this study aimed to evaluate the effects of aloe vera gel on biofilm gene expression of sulfate-reducing bacteria (SRB) isolated from patients with periodontal infection by Real time-PCR method.

For this study, 100 individuals including 50 patients and 50 healthy individuals were recruited and examined by a periodontist. After identifying sulfate-reducing bacteria by biochemical tests and specific media, the effect of aloe vera extract on them was investigated and the expression of BFR gene against housekeeping gene (16srDNA) was determined by Real time- PCR test via T-test analysis method.

The data showed that 12 strains of sulfate-reducing bacteria were isolated from the samples, 5 of which had BFR gene and their gene expression was significantly reduced by aloe vera gel (P<0.05).

The data of this study in proving the anti-biofilm and antibacterial effects of aloe vera extract showed that the expression of the target gene is reduced. It seems that this substance can be used as an alternative to oral hygiene chemicals.

The ABC family of transporters, in addition to their role in cell division and storage of cell volume in fungi, they are reported to play a role in natural metabolism and drug resistance. The purpose of this study is to investigate the molecular separation and cloning of the ATP-binding Cassette (ABC) from Trichophyton rubrum.

Sixty samples of dermatophytosis were isolated from the patients. Isolates were identified by chemical and molecular tests. Afterward, PCR was performed using specific primers for the ABC gene. PCR product was cloned in the PTG19 vector. Real-Time PCR was performed to evaluate the expression level of ABC gene.

A total of 12 isolates of Trichophyton Rubrum were identified. Six isolates were positive for ABC gene. The expression of the ABC gene cloned in E. coli XL1blue was confirmed by Real time PCR.

Expression of efflux pumps considered a way of resistance to antibiotics in microorganisms. In this regard, the researchers have developed the efflux pump inhibitor compounds and used them in combination with antibiotics to combat infections. However, because of the toxicity of most of these compounds in humans, their extensive use is prohibited. Therefore, finding new inhibitory compounds with little or no toxicity is challenging.

For centuries Herbal medicines have been used to treat various diseases in many societies. Among of them essences have well known properties. The essence of Origanum vulgare have been shown to possess effects like as antioxidant, antibacterial, antifungal, carminative, antispasmodic and analgesic activities. In this study, the Ketoconazole 2% antifungal cream drug and Origanum vulgare essence were used to treat dermatophytosis induced by Microsporum canis in an experiment on cats. The 36 healthy domestic short hair male cats were divided in 3 groups randomly for clinical evaluation. As the following, Minimum inhibitory concentration (MIC) of Origanum vulgare essence was detected at 0/19 μl/ml. The treatment started 5 days after infection as 12 hours’ regimen and was considered until 40 days’ post infection. Patients in both Ketoconazole 2% cream and Origanum vulgare essence groups had completed cure at day 40. Results discussions show that the Origanum vulgare essence similar to Ketoconazole 2% cream is an effective drug to treat Microsporum canis induced dermatophytosis.

Subtilisin genes, which encoding serine protease, have an important role in the pathogenicity of dermatophytes. The aim of this study was considered the presence of Subtilisin gene (SUB) and its similarity in 25 clinical and nonclinical samples of two dermatophytes, Trichophyton verrucosum (T. verrucosum) and Microsporum gypseum (M. gypseum), by using molecular method and designing dendrogram.64% (16/25) strains showed the active presence of the gene. Statistical analysis of data showed the correlation among the presence of SUB gene, the host (human/ animal/ soil) and the type of dermatophytes (p0.05).By using NTSYS software, UPGMA method and cut off number 70%, the strains were classified in 10 different genotypes according to their similarities to SUB gene (the Simpson coefficient of primers was 0/88).Due to different reports of presence/absence of Subtilisin gene in clinical and nonclinical strains of Trichophyton verrucosum and Microsporum gypsum,in this study, new primers were designed and synthesized to confirm the presence of Subtilisin gene in strains 64% of the, which, of course, requires more studies for further confirmation.

Dermatophytosis is one of the most common fungal infection in human and animals. Today in some cases drug resistance were reported, so designing the new antifungal agents were necessary. In this study we evaluated the antifungal susceptibility of nano fluconazole compare with fluconazole on zoophilic dermatophyte isolates by CLSI M38-A2 method. Dermatophyte species were isolated from patients and animals with dermatophytosis and identified with PCR sequencing of ITS1-5.8S-ITS2 region. 47 isolates of zoophilic dermatophytes were identified, including 23 cases of T. mentagrophytes, 12 cases of T. verrocosum and 12 cases of M. canis. The MIC50  of fluconazole for three species including T. mentagrophytes, T. verrocosum, and M. canisisolates were 32, 32, and 8 μg/ml, respectively. These value for nano fluconazole were 8, 16 and 4 μg/ml for three mentioned species. Also MIC90 for these three mentioned species were 64, 64, and 16 μg/ml for fluconazole and 16, 32, and 8 μg/ml for nano fluconazole. The MIC value for nano fluconazole was lower than fluconazole in all dermatophytes species, so nano fluconazole could inhibit the growth of dermatophytes better than fluconazole in lower concentration of drug.

Trichophyton rubrum is one of the most common causes of dermatophytosis. Increasing the expression of the TruMDR2 gene, which codes efflux pumps, results in resistance to T., rubrum versus terbinafine. Researching on new antifungal agents that can reduce the expression of this gene leading to decreases resistance to terbinafine is undeniable. The purpose of this study was to evaluate the expression of TruMDR2 gene on clinical isolates of T., rubrum against terbinafine and nanotrobinifin. In this study, two isolates of T. rubrum isolated from the onychomycosis lesions were identified by phenotyping and genotyping, and the anti-fungal susceptibility testing of terbinafine and nano-terbinafine was done by Broth microdilution according to the CLSI method. Then gene expression of TruMDR2 in two distinct isolates were investigated in sub MIC concentrations of terbinafine and nano-terbinafine. The results showed that in the presence of sub-MIC concentrations of terbinafine, the expression of the gene was 16/222 and 18/636, while the expression of the gene was in the sub MIC concentrations of nano-terbinafine 7/913 and 9/190. According to the findings of this study, the expression of TruMDR2 gene expression in both two isolates was lower increasing in present of sub MIC concentration of nano terbinafine in compared to terbinafine.

In this research, using FRET (fluorescence resonance energy transfer) from Cd/Te quantum dot (anti Ochratoxin A antibody immobilized on the external surface of quantum dot or QDs) to (Ochratoxin A labeled with Rhodamine 123 bound to albumin), a sensitive competitive immunoassay was developed for measuring Ochratoxin A. The highly specific immune-reaction between anti-Ochratoxin A antibody on the QDs and labeled Ochratoxin A brings the Rho fluorophore and the QDs in close spatial proximity, and following photo-excitation of the QDs, causes FRET to occur between the Rho fluorophore (acting as the acceptor) and the QDs (acting as the donor). In the absence of free Ochratoxin A, the immune reaction between labeled Ochratoxin and the anti-Ochratoxin antibody on the QDs induces emission and causes FRET to occur. In the presence of free Ochratoxin A, it competes with the labeled Ochratoxin A-albumin complex for binding to the antibody-QDs conjugate in the Nanobiosensor, leading to reduction in Rhodamine emission after FRET. The reduction in the fluorescence intensity of the Rhodamine acceptor directly correlates with the concentration of free Ochratoxin A in the sample. This method has a detection limit of 220pg per ml. It has also been used to measure Ochratoxin A in human serum samples. A linear relationship is found between the increase in the fluorescence intensity of Rho 123 at580 nm and the concentration of OTA in spiked samples over the 100-800 pg⋅mL−1 concentration range. This highly sensitive homogeneous competitive detection scheme is simple, rapid and efficient. It does not require multiple separation steps and excessive washing.