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Transcriptomic responses of catalase, peroxidase and laccase encoding genes and enzymatic activities of oil spill inhabiting rhizospheric fungal strains
2018
Asemoloye, Michael Dare | Ahmad, Rafiq | Jonathan, Segun Gbolagade
Fungi are well associated with the degradation of hydrocarbons by the production of different enzymes, among which catalases (CBH), laccases (LCC) and peroxidases (LiP and MnP) are of immense importance. In this study, crude oil tolerance and enzyme secretions were demonstrated by rhizospheric fungal strains. Four most abundant strains were isolated from the rhizosphere of grasses growing in aged oil spill sites and identified through morphological characterization and molecular PCR-amplification of 5.8–28S ribosomal rRNA using ITS1 and ITS4 primers. These strains were subjected to crude oil tolerance test at 0–20% concentrations. Presence and transcriptase responses of putative genes lig (1–6), mnp, cbh (1.1, 1.1 and 11), and lcc encoding lignin peroxidase, manganese peroxidase, catalase, and laccase enzymes respectively were also studied in these strains using RT-PCR. In addition, activities of secreted enzymes by each strain were studied in aliquots. The strains were identified as Aspergillus niger asemoA (KY473958), Talaromyces purpurogenus asemoF (KY488463), Trichoderma harzianum asemoJ (KY488466), and Aspergillus flavus asemoM (KY488467) through sequencing and comparing the sequences’ data at NCBI BLAST search software. All the isolated strains showed tolerance to crude oil at 20% concentration, but the growth rate reduced with increasing in oil concentrations. All the isolated strains possess the tested genes and lig 1–6 gene was overexpressed in A. niger and T. harzianum while lcc and mnp genes were moderately expressed in all the four strains. Almost 145 U.mL⁻¹ of lignin and manganese peroxidase, 87 U.mL⁻¹ of catalase, and 180 U.mL⁻¹ of laccase enzymes were produced by these strains and it was also observed that these strain mostly produced studied enzymes in response to increasing crude oil concentrations. Considering the robust nature and diverse production of these catalytic enzymes by these strains, they can be exploited for various bioremediation technologies as well as other biotechnological applications.
Afficher plus [+] Moins [-]Biodegradation of HDPE by Aspergillus spp. from marine ecosystem of Gulf of Mannar, India
2015
Sangeetha Devi, Rajendran | Rajesh Kannan, Velu | Nivas, Duraisamy | Kannan, Kanthaiah | Chandru, Sekar | Robert Antony, Arokiaswamy
High density polyethylene (HDPE) is the most commonly found non-degradable solid waste among the polyethylene. In this present study, HDPE degrading various fungal strains were isolated from the polyethylene waste dumped marine coastal area and screened under in vitro condition. Based on weight loss and FT-IR Spectrophotometric analysis, two fungal strains designated as VRKPT1 and VRKPT2 were found to be efficient in HDPE degradation. Through the sequence analysis of ITS region homology, the isolated fungi were identified as Aspergillus tubingensis VRKPT1 and Aspergillus flavus VRKPT2. The biofilm formation observed under epifluorescent microscope had shown the viability of fungal strains even after one month of incubation. The biodegradation of HDPE film nature was further investigated through SEM analysis. HDPE poses severe environmental threats and hence the ability of fungal isolates was proved to utilize virgin polyethylene as the carbon source without any pre-treatment and pro-oxidant additives.
Afficher plus [+] Moins [-]Determination of Mycotoxigenic Fungi and Total Aflatoxins in Stored Corn from Sites of Puebla and Tlaxcala, Mexico
2024
K. Saez-Gomez, R. Avila-Sosa, M. Huerta-Lara, F. Avelino-Flores and R. Munguia-Pérez
This paper aimed to evaluate the contamination with mycotoxigenic fungi and total aflatoxins in stored corn from different sites in Puebla and Tlaxcala, Mexico. Methodology. The study was conducted at two sites in Puebla (San Salvador El Seco and Junta Auxiliar La Resurrección) and two sites in Tlaxcala (Tlaltepango and Nativitas). A total of 80 samples of stored corn were collected. Identification of Aspergillus flavus was performed by microculture techniques and specific taxonomic keys (macromorphological and micromorphological). Then, samples of contaminated corn were selected, and aflatoxin production was confirmed using a direct solid-phase ELISA kit. A total of 25 A. flavus strains were identified. Other possible mycotoxin-producing fungi were Penicillium (n=52) and Fusarium (n=19). Regarding total aflatoxin contamination, all samples were contaminated within a range of 1.589 to 11.854 μg/kg, and the average concentration was 6.3 μg/kg corn. Implications. The detection of mycotoxigenic fungi in the samples tested and of aflatoxins in corn highlights the importance of monitoring these fungi. Since food safety is at risk, it shows the need for methods to control these fungi and their metabolites.
Afficher plus [+] Moins [-]Evaluation of bio-aerosols type, density, and modeling of dispersion in inside and outside of different wards of educational hospital
2022
Yousefzadeh, Ameneh | Maleki, Afshin | Athar, Saeed Dehestani | Darvishi, Ebrahim | Ahmadi, Manochehr | Mohammadi, Ebrahim | Tang, Van Tai | Kalmarzi, Rasoul Nassiri | Kashefi, Hajar
Exposure to bioaerosols in the air of hospitals is associated with a wide range of adverse health effects due to the presence of airborne microorganisms. Intensity and type of health effects depend on many factors such as the type, density, and diversity of bioaerosols in hospital environments. Therefore, identifying and determining their distribution in hospital environment contribute to reduce their adverse effects and maintain the physical health of patients and staff, as well as find the source of infections and possible allergies due to the presence of bioaerosols. Therefore, the present study was conducted to determine the type and concentration of the bacterial and fungal bioaerosols, and their distribution in the indoor and outdoor air of a teaching hospital to establish a reference for future studies or measures. The air samples were collected with a one-stage Anderson sampler and particle mass counter for a period of four months in the fall and winter of 2019. In total, 262 bacterial and fungal samples were collected from the air of the wards of Tohid Hospital, Sanandaj, Iran. Antibiotic resistance test, bacterial identification by PCR method, and modeling the dispersion of concentrations of bio-aerosols were also conducted. In order to identify bacteria and fungi, some biochemical and molecular tests and microscopic and macroscopic characteristic methods were applied, respectively. The results showed that the highest and lowest densities of the bioaerosols were observed in lung and operating wards (336.67 and 15.25 CFU/m³). Moreover, the highest and least concentrations of particles were seen in the emergency and operating wards, respectively. The most common fungi isolated from the hospital air were Penicillium (24.7%), Cladosporium (23. 4%), Aspergillus niger (13.3%), and Aspergillus Flavus (11.4%). Furthermore, the highest concentration of the isolated bacterium was Staphylococcus hemolyticus (31.84%). Most bacteria showed the highest resistance to gentamicin. The overall average hospital air pollution to bioaerosols was slightly higher than the standards proposed by international organizations. Due to the high concentration of bioaerosols and particles in the studied hospital, providing suitable conditions such as temperature, humidity, proper ventilation, and intelligent air conditioning system using efficient ventilation systems, and restricting the entrance of wards can reduce airborne particles in hospital environment.
Afficher plus [+] Moins [-]Chemically characterized nanoencapsulated Homalomena aromatica Schott. essential oil as green preservative against fungal and aflatoxin B1 contamination of stored spices based on in vitro and in situ efficacy and favorable safety profile on mice
2022
Tiwari, Shikha | Upadhyay, Neha | Singh, Bijendra Kumar | Singh, Vipin Kumar | Dubey, N. K.
Present study deals with the efficacy of nanoencapsulated Homalomena aromatica essential oil (HAEO) as a potent green preservative against toxigenic Aspergillus flavus strain (AF-LHP-NS 7), storage fungi, AFB₁, and free radical-mediated deterioration of stored spices. GC–MS analysis revealed linalool (68.51%) as the major component of HAEO. HAEO was encapsulated into chitosan nanomatrix (CS-HAEO-Ne) and characterized through SEM, FTIR, and XRD. CS-HAEO-Ne completely inhibited A. flavus growth and AFB₁ biosynthesis at 1.25 μL/mL and 1.0 μL/mL, respectively in comparison to unencapsulated HAEO (1.75 μL/mL and 1.25 μL/mL, respectively). CS-HAEO-Ne caused significant reduction in ergosterol content in treated A. flavus and provoked leakage of cellular ions (Ca⁺², Mg⁺², and K⁺) as well as 260 nm and 280 nm absorbing materials. Depletion of methylglyoxal level in treated A. flavus cells illustrated the novel antiaflatoxigenic efficacy of CS-HAEO-Ne. CS-HAEO-Ne exhibited superior antioxidant efficacy (IC₅₀ ₍DPPH₎ = 4.5 μL/mL) over unencapsulated HAEO (IC₅₀ ₍DPPH₎ = 15.9 μL/mL) and phenolic content. CS-HAEO-Ne depicted excellent in situ efficacy by inhibiting fungal infestation, AFB₁ contamination, lipid peroxidation, and mineral loss with acceptable sensorial profile. Moreover, broad safety paradigm (LD₅₀ value = 7150.11 mg/kg) of CS-HAEO-Ne also suggests its application as novel green preservative to enhance shelf life of stored spices.
Afficher plus [+] Moins [-]Chemical diversity and biological activities of essential oils from native populations of Clinopodium menthifolium subsp. ascendens (Jord.) Govaerts
2021
Debbabi, Haïfa | El Mokni, Ridha | Nardoni, Simona | Chaieb, Ikbal | Maggi, Filippo | Nzekoue, Franks Kamgang | Caprioli, Giovanni | Hammami, Saoussen
This study is focused on the analysis of regional variation of the chemical compositions of three Clinopodium menthifolium subsp. ascendens (Jord.) Govaerts Tunisian accession, as well as their inhibition toward fungi and insect pests. The diversity of the chemical constituents and biological activities in front of the aforementioned variations was found to be remarkable. Essential oils were obtained by hydrodistillation of the aerial parts and analyzed by gas chromatography-mass spectrometry (GC-MS). A total of 41, 42, and 30 compounds were identified respectively from Clinopodium menthifolium essential oils harvested from three Tunisian regions, namely Ain-Draham (ADEO), Babouch (BEO), and Tabarka (TEO). All analyzed oils were rich in oxygenated monoterpenes with different major constituents. Piperitenone (34.5%), cis-piperitone oxide (26.1%), and pulegone (47.9%) were the dominant compounds in the three volatile oils, respectively. The antifungal activity was investigated in vitro using six targeted fungal strains (Aspergillus flavus, Aspergillus terreus, Candida albicans, Microsporum canis, Microsporum gypseum, and Trichophyton mentagrophytes). The toxicity and repellency of essential oils were evaluated against the stored product pest Tribolium confusum. The tested samples were differently effective toward the target fungi and the pest depending on the variability of their chemical compositions. BEO exhibited the highest fungitoxic properties toward A. terreus mold, M. canis dermatophyte, and C. albicans yeast (the MIC values ranged from 40 to 400 μg mL⁻¹). In addition, the data showed that TEO repelled T. confusum moderately (PR = 42.5% at 2 h after exposure). Concerning the contact treatment, both ADEO and BEO were proved to possess slightly toxic effects toward T. confusum pest (% of mortality 27.5–32.5% at 5% concentration). The results showed that the geographic origin greatly influenced the chemical composition and the associated bioactivities of Clinopodium menthifolium subsp. ascendens.
Afficher plus [+] Moins [-]Assessing the Levisticum officinale Koch. essential oil as a novel preservative for stored chia seeds (Salvia hispanica L.) with emphasis on probable mechanism of action
2021
Deepika, | Chaudhari, Anand Kumar | Das, Somenath | Singh, Vipin Kumar | Prasad, Jitendra | Dubey, N. K.
The present study was undertaken to explore the inhibitory effect of Levisticum officinale Koch. essential oil (LₒffEO) on the growth and aflatoxin B₁ secretion by Aspergillus flavus (AF-LHP-SH1, aflatoxigenic strain) causing deterioration of stored chia seeds (Salvia hispanica). The chemical profile analysis of LₒffEO by GC-MS analysis revealed the presence of α-terpinyl acetate (26.03 %) as a major component followed by terpineol <1- > (24.03 %) and citronellal (24.03 %). Results on antifungal and antiaflatoxigenic activity indicated that LₒffEO at 2.0 and 1.75 μL/mL caused complete inhibition of growth and aflatoxin B₁ production, respectively. Antifungal toxicity of LₒffEO was strongly correlated with the inhibition of ergosterol content, leakage of cellular ions, and disintegration of membrane permeability. Reduction in cellular methylglyoxal by LₒffEO indicated a novel antiaflatoxigenic mechanism of action. The LₒffEO showed moderate free radical quenching activity in DPPH assay (IC₅₀ = 26.10 μL/mL) and exhibited remarkable inhibitory efficacy against lipid peroxidation of chia seeds. In addition, LₒffEO presented strong in situ antiaflatoxigenic efficacy, and exhibited non-phytotoxic nature, acceptable sensory characteristics, and favorable safety profile (LD₅₀ = 19786.59 μL/kg), which recommends its practical utilization as a novel and safe preservative to improve the shelf life of stored chia seeds from fungal infestation and aflatoxin B₁ contamination.
Afficher plus [+] Moins [-]Bioremediation of a Benzo[a]Pyrene-Contaminated Soil Using a Microbial Consortium with Pseudomonas aeruginosa, Candida albicans, Aspergillus flavus, and Fusarium sp
2015
Waszak, Dafne Q. | da Cunha, Ana Cristina B. | Agarrallua, Marcio R. A. | Goebel, Cristine S. | Sampaio, Carlos H.
Many studies have been conducted regarding the degradation of PAHs. One of the technologies that has been widely used is bioremediation due to its relatively low cost and greater efficiency for those compounds with structural complexity. Biotechnology has been used in several countries for many years and consists in the use of microorganisms (bacteria and fungi) to transform contaminants into inert substances, which is a result of the microbial activity from biochemical processes. This study aimed to develop a bioremediation methodology for the pollutant benzo[a]pyrene (B[a]P), which belongs to the group of PAHs. The potential use of a microbial consortium with Pseudomonas aeruginosa, Candida albicans, Aspergillus flavus, and Fusarium sp. for bioremediation was assessed. To confirm the pollutant reduction, quantifications of the samples were performed via gas chromatography–mass spectrometry (GC-MS). The contamination was prepared with a soil previously contaminated with B[a]P at the concentration of 3.74 mg kg⁻¹. The microbial consortium was added (16 μL g⁻¹), and samples were incubated for 42 days in an oven at 35 °C. The microbial growth curves showed representative differences between the samples in the presence and absence of the pollutant, demonstrating the possibility of bioremediation process. The final quantification of soil showed a mean concentration of 1.29 mg kg⁻¹, showed that 65.51 ± 0.95 % of the pollutant was degraded, which is an important and representative performance.
Afficher plus [+] Moins [-]Determination of Mycotoxigenic Fungi and Total Aflatoxins in Stored Corn from Sites of Puebla and Tlaxcala, Mexico
2024
K. Saez-Gomez, R. Avila-Sosa, M. Huerta-Lara, F. Avelino-Flores and R. Munguia-Pérez
This paper aimed to evaluate the contamination with mycotoxigenic fungi and total aflatoxins in stored corn from different sites in Puebla and Tlaxcala, Mexico. Methodology. The study was conducted at two sites in Puebla (San Salvador El Seco and Junta Auxiliar La Resurrección) and two sites in Tlaxcala (Tlaltepango and Nativitas). A total of 80 samples of stored corn were collected. Identification of Aspergillus flavus was performed by microculture techniques and specific taxonomic keys (macromorphological and micromorphological). Then, samples of contaminated corn were selected, and aflatoxin production was confirmed using a direct solid-phase ELISA kit. A total of 25 A. flavus strains were identified. Other possible mycotoxin-producing fungi were Penicillium (n=52) and Fusarium (n=19). Regarding total aflatoxin contamination, all samples were contaminated within a range of 1.589 to 11.854 μg/kg, and the average concentration was 6.3 μg/kg corn. Implications. The detection of mycotoxigenic fungi in the samples tested and of aflatoxins in corn highlights the importance of monitoring these fungi. Since food safety is at risk, it shows the need for methods to control these fungi and their metabolites.
Afficher plus [+] Moins [-]Elucidating Mycotoxin-Producing Aspergillus Species in River Water: An Advanced Molecular Diagnostic Study for the Assessment of Ecological Health and Contamination Risk
2024
R. Ravikiran, G. Raghu and B. Praveen
The primary goal of this research is to isolate mycotoxin-producing fungus from the Nagavali River. Examining isolated fungi involved analyzing their mycelium growth on culture media and detailed microscopic inspection. We employed PCR analysis utilizing universal primers ITS1 and ITS4 to accurately identify the species. Furthermore, we sequenced the amplified ITS region and rigorously analyzed the sequences using NCBI-BLASTn and the ITS2 database. The analysis found a high 96.38% genetic similarity to the Aspergillus flavus strain, resulting in a 600-base pair fragment size. The sequence was given the accession number OR536222 in the NCBI GenBank database. Phylogenetic analysis was performed to ascertain the particular strain of A. flavus and its source. Remarkably, this analysis led to the identification of a single new strain gene, which represents a novel discovery in the field of fungal research. These results underscore the vital significance of molecular techniques in promptly and precisely identifying organisms. This research enhances our understanding of mycotoxin contamination in water, providing valuable insights to improve detection and prevention strategies. It accentuates the overarching importance of conserving our water resources and upholding ecological equilibrium, ultimately safeguarding the well-being of both humanity and the environment.
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