Therapeutic effects of silver nanoparticle and L-carnitine on aerobic vaginitis in mice: an experimental study.

Introduction: Aerobic vaginitis (AV) is a type of vaginal infection that occurs at the reproductive age of women. In this study, we aimed to study the possible anti-AV therapeutic effects of silver nanoparticles (AgNPs) and L-carnitine (LC) in the mouse model. Methods: AV model was established by intra-vaginal inoculation of 108 CFU/mL Staphylococcus aureus and Escherichia coli (1:1) in adult NMRI mice. Susceptibilities of the bacteria were examined against AgNPs by inhibitory concentration (IC-50 and IC-90) and minimum biofilm inhibitory concentration (MBIC- 90) methods. The regimens therapy was intra-vaginal inoculation of AgNPs at MBIC- 90 and a daily injection of 250 mg/kg LC for two weeks. Mice were classified into healthy (control) and AV groups and then treated by LC, AgNPs, and AgNPs + LC. The vaginal smears were taken daily and tissue sections were prepared using the hematoxylin and eosin (H & E) method. Results: Minimum inhibitory concentrations (MICs) of AgNPs for E. coli, S. aureus, and their mixture were 250, 125, and 500 ppm, and their MBIC-90% were 500, 250, and 1000 ppm, respectively. The estrus cycle of mice treated with co-administration of AgNPs and LC was similar to the control group (P < 0.05). The results of histology also showed that infected mice were treated with AgNPs and LC, simultaneously. Conclusion: Single bacteria are more sensitive than their mixed model to these NPs. Co-administration of AgNPs as an antibacterial agent and LC as an antioxidant agent can treat AV in the infected mice.

Effect of Silver Nanoparticles and L-Carnitine Supplement on Mixed Vaginitis Caused by Candida albicans/ Staphylococcus aureus in Mouse Models: An Experimental Study.

The evolution of antimicrobial-resistant pathogens is a global health and development threat. Nanomedicine is rapidly becoming the main driving force behind ongoing changes in antimicrobial studies. Among nanoparticles, silver (AgNPs) have attracted attention due to their versatile properties. The study aimed to investigate the effects of AgNPs and L-carnitine (LC) on mixed Candida albicans and Staphylococcus aureus in the mice vaginitis model. Study of antimicrobial activity of AgNPs evaluated by Minimum Inhibitory Concentration (MIC) and Minimum Biocidal Concentration (MBC) assays. AgNPs inhibited biofilm formation of microbial strains, which was tested by using crystal violet staining. In the current study, we evaluated the effects of AgNPs and LC in NMRI mice infected intravaginally with C. albicans/ S. aureus for two weeks. The proportion of mice in each stage of the estrous cycle (proestrus, estrus, metestrus, and diestrus) was examined. Histological properties were assessed by hematoxylin/ eosin (H&E) staining of formalin-fixed, paraffin-embedded vaginal tissue sections. Based on the results, MICs of AgNPs against S. aureus, C. albicans, and their combination were 252.3, 124.8, and 501.8 ppm, and their minimum biofilm inhibitory concentration (MBIC) was 500, 250, and 1000 ppm, respectively. The estrous cycle in the treated group was similar to the control. Vaginal histology and cytology showed that LC can improve tissue damages caused by vaginitis and AgNPs. This study demonstrates the promising use of AgNPs as antimicrobial agents and the combination of AgNPs/ LC could be a great future alternative in the control of vaginitis.

Computational insight into the protective mechanism of Allium iranicum Wendelbo. Alliaceae in a mouse model of Staphylococcosis: focus on dietary phytocannabinoid trans-caryophyllene.

This study was aimed to investigate the prophylactic effects of hydro-alcoholic extract derived from bulbs of Allium iranicum Wendelbo. (Alliaceae; AI) on mouse model of Staphylococcosis, and to decipher which phytochemicals of AI may involve in its anti-staphylococcal property. Male mice were allocated into four groups, i.e. normal control (NC) and three other groups received AI at 0.192, 0.384 and 0.768 mg/ml in drinking water for 9 days. Thereafter, mice were intravenously injected 106 colony forming unites (CFUs/ml) of Staphylococcus aureus suspension at 10th day and tissue homogenates were colony counted for S. aureus 9 days post-inoculation. Molecular docking among cardinal proteins involved in Staphylococcosis and phytochemicals of AI has been performed using PyRx software and the best ligand submitted to compute molecular and biological attributes. Induction of murine Staphylococcosis and inclusion of AI did not adversely alter bodyweights of mice while colony counts in selected tissues of mice infected with S. aureus were different among groups (PANOVA = 0.016). Generally, the colony counts tended to decrease in AI 0.192 (P = 0.099) and AI 0.768 (P = 0.818) groups as compared to NC, however AI 0.384 (P = 0.005) group showed lesser colony counts than NC. In addition, colony counts decreased in AI 0.384 as compared to AI 0.768 (P = 0.009). The colony counts in kidneys of AI 0.0384 group were lesser than those of NC (P = 0.051) and AI 0.768 (P = 0.048). Among target proteins, trans-caryophyllene (TCP) showed reliable binding affinities (kcal/mol) to three hydrolase enzymes [beta toxin (sphingomyelin phosphodiesterase - 8.1), sortase B (protease - 8.1), and FtsZ (GTPase - 8.7) of S. aureus]. The ADMET properties of TCP showed that it followed the Lipinski's rule of five with one violation with respect to its miLogP 5.17. In addition, Molinspiration bioactivity score indicated ion channel modulatory and enzyme inhibitory effect for TCP. Computational findings of admetSAR software revealed that TCP leads to carcinogenicity, Tetrahymena pyriformis, fish, rat, and honey bee toxicities, weak inhibition of human ether-a-go-go-related gene, and cytochromes inhibitory promiscuity. The TCP showed promising in human intestinal absorption, blood-brain barrier permeability, Caco-2 penetration, and solubility. The results of Toxtree software showed that TCP is not an endogenous molecule of the body and contains no functional groups associated with enhanced toxicity and considered as class I toxic compound close to terpenes. In conclusion, we found the hydro-alcoholic extract derived from of bulbs AI has a significant protective effect against Staphylococcosis in mouse model. In silico findings demonstrated that TCP has acceptable ADMET score to be considered as a bioactive compound for designing phytobiotics. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s40203-021-00078-x.

First comprehensive report of bacteria spp. associated with cloaca of Laudakia nupta (Sauria: Agamidae) in Iran using molecular studies.

Iran bears a remarkable variety of reptiles. One of the lizard families occurring in Iran is the Family Agamidae which is widely are distributed throughout the old world. The large-scaled rock agamid, Laudakia nupta, is one of the well-known agamid. There are few reports of cloacal microbial on reptiles hence their function in cloacae remains unknown. Laudakia nupta usually live in rural and urban areas and close vicinity to man, they are likely to play an important role in the spread of disease that may be caused by these microorganisms and their transmission to man. Therefore, the aim of this study was to identify the bacterial flora colonizing the cloacal region of Laudakia nupta using molecular studies. The cloacal fluids were directly placed on nutrient agar (NA) plates and incubated at 25 ± 2 °C for 48 h. The resulting bacterial colonies were transferred to fresh nutrient agar (NA) plates for molecular studies. Twelve isolates were obtained from 17 specimens of Laudakia nupta. All bacteria isolates were identified as Bacillus subtillis (5), Bacillus cereus (4), Bacillus sp. (1), Pseudomonas putida (1), and Pseudomonas sp. (1) based on partial sequences of the 16 s rRNA gene. This is the first comprehensive report of bacteria spp. associated with cloaca of Laudakia nupta using molecular studies. In this research, we found that Laudakia nupta can be a carrier of bacteria which can transfer microorganisms to hosts.

Molecular identification of entomopathogenic Fusarium species associated with Tribolium species in stored grains.

Fusarium species are common pathogens of plants, animals and insects worldwide, including Iran. The occurrence of entomopathogenic Fusarium species isolated from Tribolium species as one of the most important insect pests of stored grains were sampled from various provinces in western Iran. In total, 15 Tribolium species belonging to T. castaneum (Herbst) and T. confusum (Du Val) (Col: Tenebrionidae) were detected and 8 isolates from Fusarium spp. were collected from them. Based on morphological features, the Fusarium isolates were classified into F. keratoplasticum and F. proliferatum. The phylogenetic trees based on tef1 dataset clearly separated all morphological taxa. DNA sequences of ITS regions and ß-tubulin gene were also confirmed morphological taxa. All of the Fusarium isolates were evaluated for their pathogenicity on T. confusum. Maximum mortality rate was observed for F. keratoplasticum (isolate FSSCker2) and this isolate may be considered as a good candidate for biological control in the ecosystem of stored grains. This is the first report on molecular identification of Fusarium species isolated from insects in Iran and F. keratoplasticum and F. proliferatum were isolated for the first time from Tribolium species as two entomopathogenic fungi.

Computational approaches to the in vitro antibacterial activity of Allium hirtifolium Boiss against gentamicin-resistant Escherichia coli: focus on ribosome recycling factor.

Persian shallot, Allium hirtifolium Boiss. (AH), is an Iranian native medicinal plant belongs to Alliaceae family. Here, we investigated in vitro antibacterial activity of hydro-alcoholic extract derived from bulbs of AH. We also employed in silico molecular docking to decipher mechanisms of its antibacterial effects. Minimum inhibitory concentrations (MIC) and minimum bactericidal concentration (MBC) against E. coli ATCC 25922 were determined. Molecular docking was performed for major phytochemicals of AH against ribosome recycling factor (RRF). E. coli ATCC 25922 was gentamicin-resistant while AH showed MIC (42 ± 18 µg/ml) and MBC (106 ± 36 µg/ml) against E. coli. In silico results reported all phytochemicals of AH shown acceptable negative binding affinity (kcal/mol) with RRF. In essence, the binding affinities of alliogenin (-11.6), gitogenin (-11.6), kaempferol (-10.2), linoleic acid (-8.4), oleic acid (-8.0), palmitic acid (-7.4), palmitoleic acid (-8.4), quercetin (-10.8), and shallomin (-13.4) with RRF were comparable to that of gentamicin (-12.6). In sum, hydro-alcoholic extract of bulbs of AH could be considered as a commercial phytobiotics if in-depth antibacterial assays employed in future studies. More interestingly, shallomin showed more promising binding affinity with RRF and can be considered as lead molecule for future drug discovery.

Dermatitis in the Fringe-Toed Lizard, Acanthodactylus nilsoni Rastegar-Pouyani, 1998 (Sauria: Lacertidae) Associated with Fusarium proliferatum.

From July to October 2013, nine out of 40 Acanthodactylus nilsoni collected from Western Iran, showed clinical signs of dermatitis in the dorsal and ventral surface of neck and fingers. Therefore, the aim of this survey was to identify the fungal flora colonizing the skin of A. nilsoni using morphological and molecular studies. Nine isolates of Fusarium were obtained from infected lizard samples and identified as Fusarium proliferatum through study of morphological characters. In the present study, selected F. proliferatum isolates (USMGFSC 230-112, USMGFSC 186-113, and USMGFSC 33-114) were examined and phylogenetically analysed on the basis of partial sequences of the tef1 and tub2 genes. Sequence analysis supported the morphological data, and all isolates were placed within F. proliferatum species. This is the first report on morphological and molecular identification of F. proliferatum isolated from lizards' dermatitis in Iran.

Morphological and phylogenetic analysis of Fusarium solani species complex in Malaysia.

Members of Fusarium solani species complex (FSSC) have been known as plant, animal, and human pathogens. Nevertheless, the taxonomic status of such an important group of fungi is still very confusing and many new species as well as lineages have been elucidated recently. Unfortunately, most of the new taxa came from temperate and subtropical regions. Therefore, the objectives of the present study were to identify strains of FSSC recovered from different sources in Malaysia. In the present study, 55 strains belonging to the FSSC were examined and phylogenetically analyzed on the basis of internal transcribed spacer (ITS) regions and partial translation elongation factor-1 (TEF-1α) sequences. Based on morphological features, a total of 55 strains were selected for molecular studies. Based on morphological features, the strains were classified into four described Fusarium species, namely Fusarium keratoplasticum, Fusarium falciforme, FSSC 5, and Fusarium cf. ensiforme, and one unknown phylogenetic species was introduced. Although the data obtained from morphological and molecular studies sufficiently supported each other, the phylogenetic trees based on ITS and TEF-1α dataset clearly distinguished closely related species and distinctly separated all morphological taxa. All members of FSSC in this research were reported for the first time for Malaysian mycoflora.

Molecular phylogenetic and pathogenetic characterization of Fusarium solani species complex (FSSC), the cause of dry rot on potato in Iran.

Members of Fusarium solani species complex (FSSC) are common pathogens of potato, causing dry rot in the west of Iran which involved Hamedan, Kermanshah, Eilam and Kurdistan provinces. Therefore, the objectives in this study were to isolate and identify disease-causing FSSC from infected potato tubers based on the morphological and molecular characteristics. Forty-five isolates of Fusarium were obtained from potato tubers collected from the wet market in different regions of the west of Iran and identified as FSSC through morphological characters. All of the isolates were evaluated for their pathogenicity on healthy potato tubers in the planthouse. The tubers rot symptoms were observed on the 21st day after inoculation of Fusarium isolates on the tubers tested. In the tubers inoculation tests, lesion sizes were quite variable; therefore, the measurement was done to compare the depth and width of lesion expansion among the isolates. Based on the sequence data from translation elongation factor (EF-lα) gene and internal transcript spacer (ITS) regions analysis, all of the selected FSSC isolates were divided into two major groups. This is the first report on molecular identification of FSSC strains isolated from potato tubers in Iran and Fusarium falciforme was reported for the first time in Iran.

Occurrence of Fusarium spp. and fumonisins in stored wheat grains marketed in Iran.

Wheat grains are well known to be invaded by Fusarium spp. under field and storage conditions and contaminated with fumonisins. Therefore, determining Fusarium spp. and fumonisins in wheat grains is of prime importance to develop suitable management strategies and to minimize risk. Eighty-two stored wheat samples produced in Iran were collected from various supermarkets and tested for the presence of Fusarium spp. by agar plate assay and fumonisins by HPLC. A total of 386 Fusarium strains were isolated and identified through morphological characteristics. All these strains belonged to F. culmorum, F. graminearum, F. proliferatum and F.verticillioides. Of the Fusarium species, F. graminearum was the most prevalent species, followed by F. verticillioides, F. proliferatum and then F. culmorum. Natural occurrence of fumonisin B1 (FB1) could be detected in 56 (68.2%) samples ranging from 15-155 µg/kg, fumonisin B2 (FB2) in 35 (42.6%) samples ranging from 12-86 µg/kg and fumonisin B3 (FB3) in 26 (31.7%) samples ranging from 13-64 µg/kg. The highest FB1 levels were detected in samples from Eilam (up to 155 µg/kg) and FB2 and FB3 in samples from Gilan Gharb (up to 86 µg/kg and 64 µg/kg).