جستجوی مقالات مرتبط با کلیدواژه « زنده مانی » در نشریات گروه « زیست شناسی »

In recent years, significant efforts have been focused on advancements of novel biomaterials based on natural polymers and utilization of efficient methods such as skin tissue engineering for wound treatment. In this study, a 3D printed polycaprolactone (PCL) scaffold coated via immersion in a 1:4 blend of 40% silk fibroin from Bombyx mori cocoons and TEMPO-oxidized was developed. The pore size and the porosity were 180 µm and 85%, respectively. The results demonstrated an enhancement in exudate absorption (swelling and water uptake of 1342% and 80%, respectively), improvement in storage modulus (G’) from 500 to 4000 Pa, as well as viscoelasticity up to 60%, which all are favorable for wound dressing applications. Moreover, the wettability and biodegradability studies revealed an overall increase in contact angle and degradation rate of 19.9°±3, and 95%, respectively. Cell viability and migration studies on fibroblastic cells (L929) using MTT assay, DAPI/ Phalloidin staining, and scratch test showed over 90% viability up to 7 days and complete scratch repair within 24 hours. These findings show that 3D printed PCL scaffolds coated with silk fibroin and oxidized nanocellulose are promising for wound healing applications and might pave the way to natural polymer-based wound dressings.

Bone is the hardest and one of the most important tissues in the body. In case of bone damage, the current treatments do not completely repair and regenerate the bone. For this reason, cell-based tissue engineering strategies, especially Mesenchymal Stem Cells (MSCs), have received attention. MSCs have the ability to self-renew and differentiate into different cell types, including bone cells, cartilage cells, and fat cells, among others. They are found in various tissues throughout the body, including bone marrow, adipose tissue, and umbilical cord tissue. Today, MSCs are a valuable resource for regenerative medicine and tissue engineering applications. In addition to the cells, scaffolds are another essential element of tissue engineering. One of these scaffolds is decellularized tissue-derived hydrogels, which are three-dimensional network of hydrophilic polymer chains that can absorb and retain a significant amount of water. In tissue engineering, they mimic the natural extracellular matrix of tissues, providing a suitable environment for cells to attach, proliferate, and differentiate. In the current study, we aimed to investigate the effects of decellularized skeletal muscle-derived hydrogel, known as Myogel, on bone marrow-derived MSCs biological behaviors, including proliferation, viability and migration. In this study, MSCs were isolated from tibia and femur of adult Wistar rats. MSCs were cultured in a complete medium (a-MEM containing 15% fetal bovine serum (FBS) and 1% penicillin/streptomycin (Pen/Strep)). The identity of cells was determined by morphology (using inverted microscope) and expression of specific CD markers (using Flowcytometry). Skeletal muscle was decellularized and accuracy of decellularization was evaluated using special staining. Then Myogel was prepared from digested decellularized skeletal muscle. Here, Myogel substrate was used as the control group, gelatin substrate as the positive control, and un-coated plates as the negative control. The effect of Myogel on survival (MTT method), proliferation (drawing the growth curve and calculating the doubling time of the cell population), cell cycle profile (flow cytometry method), and cell migration (scratch method) were investigated. The MTT test showed that the survival of MSCs in Myogel substrate with a concentration of 0.2 mg/ml was higher than the survival of MSCs in gelatin substrate with a concentration of 0.1 mg/ml and the survival of MSCs in gelatin was higher than the survival of control MSCs. Myogel substrate increased the proliferation and migration of cells and decreased the doubling time of MSCs population. Examining the cell cycle profile showed that a high percentage of cells cultured on Myogel were in the G1 and S phase of the cell cycle, indicating an increase in cell division speed by gelatin and, in the next degree, by Myogel. Therefore, Myogel can be used as a suitable substrate to increase the proliferative and migratory potential of MSCs, which are an important factor in tissue engineering.

Nowadays, skin grafts use to replace lost or damaged skin around the world. Natural contaminants can exist on the skin and this contamination can be a threat to graft recipients. In this study, after obtaining human full thickness skin graft sample and culturing it in-vitro condition, skin samples were evaluated for cell viability and the presence of bacterial contamination during 72 hr. The amount of microbial contamination in the presence and absence of antibiotic gentamicin 0.02% was also investigated by standard methods. High cell viability during 72 hr in normal skin tissue samples was confirmed by the acridine orange staining. Electron scanning microscope observations, hematoxylin-eosin, and Masson's trichrome staining images clearly showed the difference between the two experimental groups. Skin graft samples lost the layers of dermis and epidermis (except the stratum corneum layer), in the absence of gentamicin due to contamination with bacteria which can be seen in the obtained images. Finally, the obtained results clearly showed the effect of high microbial contamination on the skin tissue structure, which can be caused by various factors.

One of the environmentally friendly synthesis methods is the synthesis of nanoparticles using plants. Silver nanoparticles are metal nanoparticles with anticancer activity. This study aims  to investigate the effect of silver nanoparticles synthesized from Digitalis nervosa hydroalcoholic extract on MCF-7 breast cancer cell line.

In this experimental study, silver nanoparticles were synthesized from the hydro alcoholic extract of the Digitalis nervosa.MCF-7 cells were then treated with different concentrations of silver(3.125, 6.25, 12.5,25,50 and 100 μg /ml).The toxicity effects of silver nanoparticles were analyzed by MTT method and after determining the concentration of 50% lethality of silver nanoparticles, the expression changes of P53,Bax, Bcl2 and CDH1 genes were investigated by Real - time PCR method. The obtained data were analyzed using SPSS software and one-way variance statistical test and Tukey's test.

The results of MTT showed that the effect of silver nanoparticles on the viability of MCF-7 cell line is dose-dependent and the highest cytotoxicity is related to the concentration of 100 and the lowest is related to 6.25 μg /ml in a significant way. (P<0.001) Also, the effect of silver nanoparticles with a concentration of 50% lethality (19.55 micrograms/ml) on the expression of P53, Bax, Bcl2 and CDH1 genes showed significant changes compared to the control gene.

Silver nanoparticles synthesized by green method from Digitalis nervosa  induce apoptosis in MCF-7 cell line and the use of these nanoparticles can be considered as a promising choice in the treatment of breast cancer.

Abstract Chlorine is an effective disinfection agent to kill pathogenic microorganisms in the municipal water treatment system. However, despite treatment, studies have shown that Acanthamoeba is isolated from different water sources in Iran. In this study, the effect of standard concentrations of chlorine used in urban water treatment systems was evaluated on the survival of Acanthamoeba castellanii and its ultrastructure. Acanthamoeba trophozoites and cysts were exposed to different concentrations (1-10 ppm) of calcium hypochlorite at different times (30 minutes, 1 and 2 hours). Field emission scanning electron microscopy (FESEM) and transmission electron microscopy (TEM) were used to study the ultrastructural changes of amoebic trophozoite. This study showed that conventional chlorine concentrations could not completely eliminate A. castellanii trophozoites and cysts. Cysts were more resistant to different chlorine concentrations and compared to trophozoites, fewer cysts were killed at the same chlorine concentration and exposure time. Alteration of the cell membrane permeability, decrease in the number of pseudopodia, increase in mitochondria, vacuolation of the cytoplasm, and changes in the endoplasmic reticulum were the main ultrastructural changes in the chlorine-treated amoeba. This study showed that standard chlorine concentrations used as a disinfectant could not eliminate the trophozoites and cysts of A. castellanii. Due to the pathogenicity of the amoeba and its role as the reservoir and transmission of microbial agents, revising the guidelines for using disinfectants such as chlorine in the treatment of urban water systems is highlighted by this study.

The prevalence of cancer and the side effects of treatment methods have provided a growing interest in the use of plants as a promising source of treatment.The use of medicinal plants in traditional medicine has been important for a long time and many of these medicinal plants have antioxidant and anticancer effects. The purpose of this study is to investigate the anticancer effects of Artemisia turcomanica extract on breast cancer cell line (MCF-7)and its effect on Bcl2 and Bax gene expression.In this experimental study, the effect of the hydroalcoholic extract of Artemisia turcomanica on viability and expression of Bax and Bcl2 genes in MCF-7 breast cancer cells was examined.For this purpose,MCF-7breast cancer cells were cultured and treated with the hydroalcoholic extract of Artemisia turcomanica(with different concentrations)for 24,48 and72 hours.The effect of the extract on cell viability was evaluated by MTT method.Next,RNA extraction was performed and after cDNA synthesis,the expression level of Bax and Bcl2 genes was measured by Real-time PCR method.This research showed that with the increase in concentration and time,the viability of the cells decreased significantly compared to the control samples.Also, the results of real-time PCR showed that the expression of Bax and Bcl2 genes changed significantly compared to the control sample at 48 and72 hours.The results showed that the extract of Artemisia turcomanica has cytotoxic and apoptotic effects on the MCF-7 cell line, so that by conducting more studies, the extract of this plant can be used as an anti-cancer biological product in the treatment of cancer

The aim of present study was to evaluation the effect of different levels of soybean lecithin in tris extender replaced with egg yolk on semen quality parameters of Arabic ram after freeze-thawing process. For this purpose, the experiment with numbers of 8 adult and healthy Arabic rams with an average weight of 75 ± 5 kg were used in a completely randomized design. Semen samples were collected using electro ejaculator. Semen samples were mixed and after dilution, were divided in to 5 experimental groups. Treatments were increasing levels of soybean lecithin (0.5, 1, 1.5 and 2 percent) and egg yolk (15 percent) in tris base extender and 5 percent of glycerol. The 5 numbers of 0.5 ml straws from each treatment were filled. The straws were equivalence for 2 hours in 5˚C and then freezed. After 2 weeks, straws were thawed at 37˚C. Qualitative parameters of spermatozoa including the motility, viability, morphological abnormalities, membrane integrity and the pH of semen were evaluated. The resulted of this experiment showed that extender containing 1.5 percent of soybean lecithin significantly improved total motility and progressive motility, viability, morphological abnormalities and membrane integrity of Arabic ram spermatozoa compared to the other levels of lecithin as well as egg yolk (P<0.05). The most improvement have been observed at the 1.5 % soybean lecithin. Therefore, this level can be used in tris extender for maintaining of Arabic ram spermatozoa in freeze condition.

Stem cells with high proliferative capacity can be isolated from different tissues of the body. These cells originate from the fetal mesoderm and are found in tissues such as bone marrow, fat tissue, amniotic fluid, and Wharton's jelly. In this study, the survival of Wharton's jelly human mesenchymal stem cells inside alginate capsules after 7, 14 and 21 days has been investigated. In this experimental study, 10 umbilical cord samples were obtained from pregnant mothers during caesarean section, and the vessels of the umbilical cord samples were isolated. Then it was cultured in DMEM-HG medium containing 10% FBS serum for 5 days. To show the stemness of these cells, CD73, CD34 and CD45 markers were evaluated by flow cytometry technique. After confirmation, the cells were encapsulated in alginate hydrogels. The viability of encapsulated cells was evaluated by trypan blue and MTT. The results showed that the capsules are spherical and have a uniform border and are homogeneously dispersed throughout the capsule. Wharton jelly encapsulation of mesenchymal stem cells did not change their morphology and viability. After 21 days, the survival of the encapsulated cells was maintained. Alginate as a three-dimensional biodegradable scaffold with suitable cell viability can be used as a suitable option for cell therapy and tissue engineering with the property of non-graft rejection.

The main cause of death in human societies is cancer, and medicinal plants can play an important role in cancer treatment due to their effective compounds. The purpose of this study is to investigate the anticancer effects of Artemisia turcomanica extract on the viability of cervical cancer cell line (Hela) and its effect on bcl and Bax gene expression. In this experimental study, first, the hydroalcoholic extract of the Artemisia turcomanica was prepared and Hela cancer cells were cultured in RPMI culture medium containing fetal bovine serum and penicillin/streptomycin and treated with different concentrations of the hydroalcoholic extract of the Artemisia turcomanica ( and μg /ml) for , , and hours. The anticancer effect of the extract was evaluated by MTT method and finally, at lethal concentration, the expression of Bax, Bcl genes was evaluated by Real-Time PCR method. Data were analyzed with SPSS software, one-way variance, Tukey test and p less than . This research showed that the anti-cancer effect of the extract on the cells increased significantly with increasing concentration and time compared to the control samples, also, Real-Time PCR results showed that the expression of Bax and Bcl genes changed significantly compared to the control sample at and hours. The results showed that the Artemisia turcomanica extract is an effective anti-cancer compound on cervical cancer cells, and probably with further investigation and identification of the effective compounds in the extract, a step can be taken in the direction of finding and designing new and effective drugs in the treatment of cancer.

The Digitalis nevrosa has anti-cancer properties due to its glycosidic compounds The aim of this study was to investigate the anticancer effects of hydroalcoholic extract of digitalis nevrosa on MCF-7 breast cancer cell line. In this experimental study, hydroalcoholic extract of thistle was produced. MCF-7 cells were then treated with different concentrations (3.125, 6.25, 12.5, 25, 50 and 100 μg / ml). The effects of cytotoxicity of the extract were analyzed by MTT method and after determining the concentration of 50% lethality of the extract, the expression changes of P53, Bax, Bcl2 and CDH1 genes were analyzed by real-time PCR. The data were evaluated using SPSS software and one-way ANOVA and Tukey test. The results of MTT showed that the effect of the extract on viability was dosedependent and the highest cytotoxicity was related to the concentration of the extract at 39/45μg / ml. Which were statistically significant were shown to be significant.(P<0.001). Also, the effect of silver nanoparticles with a concentration of 50% lethality of 39/45 μg / ml on the expression of P53, Bax, Bcl2 and CDH1 genes in comparison with the control gene showed significant changes. Digitalis nevrosa extract induces apoptosis in MCF-7 cell line and the use of this plant can be considered as a promising choice in the treatment of breast cancer.

Silver nanoparticles (AgNPs) have been considered by researchers as a very strong inhibitor due to their biological properties and have many applications in prevention and treatment. However, there is little information about the effect of nanoparticles on healthy cells.The aim of this study was to investigate the effect of silver nanoparticles on the viability percentage of normal skin cell line MHFB-1.

In this study, during 24, 48 and 72 hours, different concentrations of silver nanoparticles (1.56, 3.125, 6.25, 12.5, 25, 50 and 100 μg / ml on MHFB-1 cell line was evaluated by MTT method.Data obtained from the test were statistically analysed using SPSS software, one-way ANOVA, Tukey test and significance level of P <0.05.

Treatment of normal MHFB-1 cells with different concentrations of silver nanoparticles after 24, 48 and 72 hours by  MTT method showed a significant reduction  in the viability of cells at  concentrations of 25, 50 and 100 μg /ml.

The results of this study showed a dose-dependent inhibitory effect of silver nanoparticles on normal skin cell line (MHFB-1).

NIR Laser application in bacteria is often focused on mortality and antibiotic efficacy. The literature records on this point are absolutely diverse from mortality in different degrees to immortality and even viability enhancement. The aim of this study is to investigate 808 nm laser effects on E.coli-DH5α viability and Growth with CFU, MTT and FCM assays. To obtain the purpose, bacteria in LB media put on with 808nm laser on 100 and 200 J/cm2 dosages and were investigated and compared by CFU, MTT and FCM assay. CFU assay results after 24 hours incubation were not significantly different between laser treatments and control. (P=0.06). In contrast, MTT assay results after 1 hours from laser treatment indicated significant deleterious effects in 200 J/cm2 laser treatment compared with control(P=0.006). On the other hand, FCM assay results of laser treatments with using of PI and Triton X100 not only approved MTT assay results but also revealed some dose dependent changes on bacteria ranging from increase membrane permeability to lethal damages. As a conclusion of the results in these method assays, we can state that these different laser doses produce diverse effects on viability and growth in E.coli-DH5α. Consequently the laser treatments could be planned for antibiotic purposes or enhancing gene transformation process.

Fibroblastoma is a common skin malignancy and a common cause of morbidity worldwide. Studies have shown that sex steroid hormones including progesterone have cytotoxic effects on cancer cells. According to this, the present study aims to determine the effects of progesterone cytotoxic concentration on iNOs expression in fibroblastoma (L929) cells.

Cell viability was measured using MTT assay in cells exposed to 0.001, 0.01, 0.1, 1, and 10 mg/ml of progesterone, and IC50 dose was determined. NO concentration levels were also measured using the Griess test. The expression level of iNOS was measured by real-time RT-PCR. Data were analyzed using SPSS by one-way ANOVA.

Viability significantly decreased in fibroblastoma cells exposed to 1 and 10 mg/ml of progesterone (P<0.001). The relative expression level of iNOS significantly increased in cells exposed to IC50 dose of progesterone (P<0.001). The relative concentration of NO significantly increased in fibroblastoma cells exposed to 0.01, 0.1, and 1 mg/ml of progesterone (P<0.001, P<0.001, and P<0.05, respectively).

Progesterone has cytotoxic effects on fibroblastoma cancer cells due partly to the effects of the hormone on iNOS expression level and increased NO that probably induces apoptosis in fibroblastoma cancer cells.

In this study, the effect of Artemisia herba alba essential oil on the survival rate and expression of ODC1 and ADA genes in human breast cancer cell line MCF-7 was studied.

The essential oil (EO) of A. herba alba was extracted by Clevenger. Essential oil composition is analyzed by GC-Mass. MCF-7 cell line was treated by increasing concentrations of EO from 0.01 to 0.03 µg/ml. Then cytotoxicity of EO was investigated by MTT assay and the expression of ODC1 and ADA genes were measured by the Real-time PCR method.

Most essential oils included Camphor, Chrysanthenyl Acetate,  and Davanone. Results showed that by increasing the EO concentration, the cytotoxicity was significantly increased and the calculated IC50 for 24, 48, and 72 h were 0.02104, 0.01858, 0.011751 µg/ml. Moreover, at the concentration of 0.02 µg/ml of EO the expression of the ODC1 and ADA gene decreased by 2 and 1.2-fold against the control, respectively.

The cytotoxicity of A. herba alba EO, which was observed in this study, seems to be the result of decreased expression of ODC1 and ADA genes in the MCF-7 breast cancer cell line.

The studies indicated that sex steroid hormones affect the digestive system function and proliferation of the cancer cells in the cell and molecular level. This study examines the anti-proliferative effects of Nandrolone on colon cancer (HCT) cell line and evaluates the activity of caspases of 3, 8, and 9. In this experimental study, HCT cells were divided into the control group (without treatment) and groups treated with 0.625, 1.25, 2.5, 5, and 10 mg/ml of Nandrolone. The cytotoxic effect of hormone was measured using MTT. The activity level of caspases 3, 8, and 9 were evaluated by the standard kit. The data were compared among the groups using one-way analysis of variance and t-test. HCT cell viability significantly decreased in the groups treated with 2.5, 5, and 10 mg/ml of Nandrolone compared to the control group (p < 0.001); However, caspase 3 and 9 (p < 0.001) and caspase 8 activity (p < 0.01) significantly increased. This study's findings showed that Nandrolone has a cytotoxic effect on colon cancer cells, and this effect is partly mediated by increasing the caspases of 3, 8, and 9 activities.

Although several studies have been carried to investigate the effects of dandelion extracts on the viability of cervical cancer cells, the results on the effects of dandelion on cell viability, particularly in cervical cancer cells, are still challenging. The aim of this study was to investigate the cytotoxic effects of dandelion (Taraxacum officinale) extract on cervical cancer cells compared to non-cancerous cells.

In this laboratory experimental study, HeLa cancer cell line and non-cancerous embryonic kidney cells (Hek293) were purchased from Pasteur Institute, Tehran, Iran. Cells were divided into control and treatment groups. In the treatment group, cells were exposed to 0.002, 0.02, 0.2, and 2 mg/ml of dandelion flower extract. MTT assay was used to evaluate cell viability. Data were analyzed using one-way analysis of variance

Hela cancer cells viability was significantly reduced in the groups exposed to 0.2 and 2 mg/ml of extract compared to the control group (p<0.01 and P<0.001, respectively). None of the concentrations used had a significant effect on the viability of Hek293 cells.

Dandelion flower extract in appropriate concentrations can reduce the viability of cervical cancer cells without side effects on healthy non-cancerous cells. The findings of this study support previous research indicating the anticancer effects of dandelion on cancer cells.

Recently, polymer-based nanofibrous scaffolds have attracted great attention due to their significant antibacterial properties in the field of dermatological applications. In this study, a polycaprolactone-based nanofibrous scaffold has been fabricated using the electrospinning method. The aim of this study was to evaluate the antibacterial effect of electrospun nanofibrous structures.

In this experimental study, the structure and bacterial attachment on polymeric nanofibrous scaffolds were studied by Scanning Electron Microscopy (SEM). In addition, antibacterial properties of nanofibrous scaffolds were studied on two gram-negative bacteria of Escherichia coli and Pseudomonas aeruginosa and two gram-positive bacteria of Staphylococcus aureus and Streptococcus mutans, using microdilution method and biofilm assay. Moreover, MTT assay was performed on HeLa and human fibrosarcoma cell line (HT1080) cancerous cell lines to evaluate the cell viability.

The results of this study showed that nanofibrous scaffold revealed a significant antimicrobial and anti-biofilm formation effect on all of the studied bacterial strains, but in microscopic observations and microdilution assay was observed on Pseudomonas aeruginosa in 1mg/ml of nanofibrous scaffold extract concentration, while the major effect in biofilm assay was observed in 8µg/ml of extract concentration. Moreover, the cell viability studies showed that the most significant effect was shown on HT1080 cell line which has drastically decreased by 40% after 48 hours in comparison with the control.

These results show that electrospun nanofibrous PCL-based scaffolds are potentially promising for dermal tissue engineering applications, due to anti-biofilm effects and capability of reducing the number of cancerous cells in the wound site.

The aim of this study was to evaluate the effects of Lactobacillus plantarum supplementation as probiotic on body composition, growth performance, nutrient efficiency indices, and survival in Astacus leptodactylus. For this purpose, 96 crayfishes (27.88 ± 0.27) were randomly distributed into twelve 100-liter fiberglass tanks, (eight crayfish per tank). Crayfishes were fed with different levels of L. plantarum inclusion comprising 0 (control), 107 (T1), 108 (T2) and 109 (T3) CFU g-1 diet for97 days. Feeding was conducted at a rate 1.5 % of body weight on a daily basis at 9 am and 4 pm. At the end of the feeding trials, crayfish in the T3 showed significantly higher protein and lipid levels compared to the control (P < 0.05). Moreover, protein and lipid efficiency ratio (PPV and LPV respectively) were enhanced in the T3 compared to the control (P < 0.05). In the present study, none of the probiotic supplemented groups showed significant differences in the growth performance parameters and survival rate compared to the control (P > 0.05). These results revealed efficiency of L. plantarum as probiotic on body composition and nutrient efficiency indices in A. leptodactylus.

To evaluate the effects of using various levels of superabsorbent and irrigation regimes on yields of Capparis spinosa plant an experiment was performed based on a randomized complete block design with three replications in the Research Field of Emamzadeh Jaafar Gachsaran in 2015. The main factor consisted of three irrigation regimes (once in a month, once every two months, and no irrigation) and a sub-factor at four levels of superabsorbent by Tarawat A200 applications (no superabsorbent, 75, 150, and 225 g/plant). Results revealed that the highest number of branches, collar diameter, height, and chlorophyll belonged to the plants irrigated every two months with 150 g superabsorbent and the highest value of carotenoids was observed in control plants. The highest plant survival rate was observed in 225 g/plant superabsorbent treatment alone. Irrigation levels had positive effects on the morphological characteristics of the plant so that the irrigation level of once every two months had more yield than the other levels. Findings suggest that the application of superabsorbent material in dry land condition could increase the yield by mitigating the effects of the drought stress.

Khorasanicum currant (Ribes khorasanicum) is one of the endemic species of monogenus Grossulariaceae family which its natural regeneration has been encountered with severe reduction due to livestock grazing, low seeds regeneration potential. It is mainly regenerated by vegetative method via rhizome. Since, the successfulness of regeneration using seed culture and cutting method was not acceptable. So, one of the recommended possible method for conservation of endangered species is multiplying and preservation them using in vitro tissue cultures techniques. But, one of the most probable problems of micropropagated plantlets is the acclimatization to in vivo conditions and high mortality of tissue cultured plantlet during this process. Thus, this research was conducted to find out the most suitable acclimatization medium for this valuable medicinal endemic species. For this purpose the effect of different substrate compounds including peat moss, cocopeat, perlit and soil was evaluated on survival, height and root characteristics during acclimatization process. Results showed that, significant differences were observed in plantlets height, survival percentage, vigourity and leaf numbers between different substrates. But different substrates, has no significant effects on plantlets crown area. Results of analysis of variance on rooting characteristics showed that, significant differences were observed in root numbers, root length and plantlet root length. Totally, according to plantlets characteristics such as: survival percentage, vigourity, plantlet height, leaf numbers and plantlets root length, can be concluded that the best substrate for acclimatization of this species was peat moss substrates.