Biocontrol Potential and Catabolic Profile of Endophytic Diaporthe eres Strain 1420S from Prunus domestica L. in Poland—A Preliminary Study

Recently, Diaporthe has been considered the most frequently isolated genera of endophytic fungi, having a broad spectrum of host plants and a worldwide distribution. The endophytic Diaporthe strain used in the present work came from the Fungal Collection of Phytopathology and Mycology Subdepartment, University of Life Sciences in Lublin (Poland), and was isolated from healthy Prunus domestica shoots during previous studies. Due to the possibility of using the Diaporthe endophytes as a promising option for plant disease management, the main goal of the research was to study the antagonistic effect of endophytic Diaporthe strain against six phytopathogens: Verticillium dahliae, Botrytis cinerea, Fusarium avenaceum, F. sprotrichioides, Alternaria alternata, and Trichothecium roseum based on the dual culture assay and to determine the catabolic profile of the endophyte by using Biolog FF Plates. The dual-culture test assay revealed the ability of the endophytic Diaporthe to limit the growth of all tested pathogens. The growth inhibition percentage ranged from 20% (V. dahliae) to 40% (T. roseum). A distinct zone of inhibition occurred between the endophytic Diaporthe and the pathogens T. roseum, V. dahliae, and B. cinerea in the co-growth combinations. As for the catabolic profile results, the most intensive utilization of carbon substrates was observed after 168 h of incubation. The growth of the analyzed strain was observed on 79 media containing carbohydrates, carboxylic acids, amino acids, amines and amides, polymers, and others. The most effective decomposition was observed in the polymers group, the least in amines and amides. Molecular identification indicated that this strain was closely related to the Diaporthe eres species complex.

Green Fabrication of Silver Nanoparticles using Euphorbia serpens Kunth Aqueous Extract, Their Characterization, and Investigation of Its In Vitro Antioxidative, Antimicrobial, Insecticidal, and Cytotoxic Activities

The silver nanoparticles (AgNPs) were synthesized via green synthesis approach using Euporbia serpens Kunth aqueous extract. The synthesized AgNPs were characterized by UV-visible spectroscopy and Furrier Transformer Infra-Red spectroscopy to justify the reduction and stabilization of AgNPs from its precursors. AgNPs characteristic absorption peak was observed at 420 nm in the UV-visible spectrum. The SEM and TEM analysis demonstrated the spherical shape of the synthesized nanoparticles with particle sizes ranging from 30 nm to 80 nm. FTIR transmission bands at 2920 cm-1, 1639 cm-1, 1410 cm-1, 3290 cm-1, and 1085 cm-1 were attributed to C-H, C=O, C-C, N-H, and C-N functional groups, respectively. XRD peaks could be attributed to (111), (200), (220), and (311) crystalline plane of the faced-centered cube (FCC) crystalline structure of the metallic silver nanoparticles. The AgNPs showed good antibacterial activity against all the tested bacteria at each concentration. The particles were found to be more active against Escherichia coli (E. coli) with 20 ± 06 mm and Salmonella typhi (S. typhi) with 18 ± 0.5 mm zone of inhibition in reference to standard antibiotic amoxicillin with 23 ± 0.3 mm and 20 ± 0.4 mm zone of inhibition, respectively. Moderate antifungal activities were observed against Candida albicans (C. albicans) and Alternaria alternata (A. alternata) with zone of inhibitions 16.5 mm and 15 mm, respectively, compared to the standard with 23 mm of inhibition. Insignificant antifungal inhibition of 7.5 mm was observed against Fusarium gramium (F. gramium). All the tested concentrations of AgNPs showed comparable % RSA with the standard reference ascorbic acid in the range sixty percent to seventy five percent. The percent motility at 3 hours postincubation showed quick response and most Tetramorium caespitum were found deceased or paralyzed. Similarly, the percent mortality showed a linear response at concentration and time. It was observed that 1 μg/mL to 2 μg/mL concentration of AgNPs displayed a significant cytotoxic activity against Artemia salina with LD50 of 5.37 and 5.82, respectively.

Effect of Christmas Melon (Laganaria Breviflorus) extract on toxigenic Mycoflora Isolated from Stored Unpolished Rice sold in major Markets in Abeokuta, Nigeria

Study on toxigenic mycoflora and potential mitigation effect of Christmas Melon (Laganaria Breviflorus) extract in unpolished rice sold in Abeokuta Ogun state of Nigeria was carried out. Unpolished rice gotten from markets in Abeokuta were aseptically transported to the laboratory, serial dilution to reduce the fungal load was carried out and were plated on Potato Dextrose Agar (PDA) and Methyl Red Dessicated Coconut Agar (MRDCA) respectively. Microscopy, macroscopy, toxigenicity test and inhibition studies with the peeled and unpeeled fruit of Laganaria breviflorus fermented for seven days was carried out. Results reveal the predominance of Aspergillus as the major genera, specifically, A. niger, A.flavus, A. parasiticus, A. fumigatus, A. terreus, A. nidulans. Other fungi genera isolated include Penicillium, F`usarium, Mucor, Alternaria and Rhizopus . Of the 11 fungi genera isolated, 9 were toxigenic of which the zones of inhibition of unpeeled whole fruit extract of Laganaria breviflorus range from (3 - 28mm) where A. nidulans shows the highest susceptibility to the whole fruit extract of Laganaria breviflorus while the zone of inhibition of peeled fruit extract of Laganaria breviflorus ranges from (3 - 22mm) where A. parasiticus, Fusarium specie and P.chrysogenum showed the highest susceptibility . As the day progresses the zone of inhibition becomes wider. Unpeeled LB extract exhibited more zones of inhibition than the peeled LB extract. Laganaria breviflorus fruit extracts in the study demonstrates a potential in reducing toxigenic fungi, consequently a means to reducing mycotoxins in staple foods in Nigeria.

Orange-Brown Pigment Production from an Endophytic Fungus Aspergillus Sp. N11 and its Potential Pharmaceutical Applications

Endophytic fungi are the main source of natural compounds including pigments having various industrial applications. Present study describes the production of extracellular orange-brown pigment from an endophytic fungal isolate Aspergillus sp. N11from Teucrium stocksianum. The optimum conditions for pigment production from this isolate was investigated and results showed that highest yield was observed in Potato dextrose broth, at pH 5 and 30 ℃ under shaking condition at 150 rpm for 7-10 days. The pigment was extracted in ethyl acetate and purified using column chromatography. Three different pigments were purified (yellow, light brown and orange-brown) and characterized based on Thin layer chromatography and Fourier transform infrared spectroscopy. The antimicrobial activity of purified fragments showed maximum zone of inhibition of 40 mm against S. aureus while for P. aeruginosa maximum zone of 50 mm and maximum antifungal activity of 20 mm against C. albicans. The antioxidant potential of purified pigment obtained from Aspergillus sp. N11 indicates that maximum scavenging activity of 67%. The results showed that purified pigments are astaxanthins belonging to oxygen containing carotenoids. The purified astaxanthins showed antibacterial, antifungal and antioxidant activities indicating its potential to be utilized in pharmaceutical and food industries.

Phytocompounds of Bistorta macrophylla (D. Don) Sojak. as bioavailability enhancers of fluconazole and amphotericin B to better manage Candida species infections

Bistorta macrophylla (D. Don) Sojak. is a medicinal plant of high altitude and so far, not been scientifically explored? Since prehistoric times, B. macrophylla has been used to cure stomach pain, pyretic fever, flu, lungs infections, diarrhea, vomiting. The present research was aimed to examine the phytochemicals, antifungal, and synergistic potential of methanolic extracts of B. macrophylla. Methanolic extract of B. macrophylla was found to have high phenolic (191.18 ± 29.18 mg g−1 GAE) and flavonoid (26.71 ± 3.21 mg g−1 RE) content. Methanolic extract also demonstrate strong antifungal action with diameter of zone of inhibition of 17.5±0.5 mm (fungicidal) against both the strains of C. albicans (MTCC277 and ATCC90028). The minimal inhibitory concentration (MIC) of methanolic extract was found to be 62.5 µg ml−1 against C. albicans (MTCC277 and ATCC90028). In addition, the combination of methanolic extract of B. macrophylla with antifungal antibiotics (fluconazole and amphotericin B) showed synergistic interaction with MIC reduction from 4-128 folds against both candida strains. GC-MS analysis of methanolic extract revealed the presence of 15 major phytocompounds with area more than 1%. Molecular docking showed that sucrose and 9,9-Dimethoxybicyclo [ 3.3.1] nona-2,4-dione has highest binding energy of -6.3 and -5.1 KJ/mol against Cytochrome P450 14 alpha-sterol Demethylase (PDB ID: 1EA1) protein respectively. Combination of methanolic extract of B. macrophylla with antifungal antibiotics (fluconazole, amphotericin B) can be used to treat drug-resistant candida.

Pseudomonas aeruginosa associated pulmonary infections and in vitro amplification virulent rhamnolipid (rhlR) gene

Background Pseudomonas aeruginosa is a common opportunistic pathogenic bacterium with the ability to develop a strong communication pathway by quorum sensing system and different virulent factors. Among the various important secretions of P. aeruginosa rhamnolipid is important biological detergent, believed to be involved in the development of the biofilm and intercellular communication. It readily dissolves the lung surfactants that are then easily catalyzed by the phospholipases and in this way is involved in the acute pulmonary infection. Objective research work was designed to investigate virulence and gene associated with virulence in P. aeruginosa responsible for pulmonary infections. Methods In current study polymerase chain reaction (PCR) was used for the detection of the rhlR (rhamnolipid encoding) gene of isolated strains. A number of assays were performed that ensured its virulent behavior. Disc diffusion method was used to check its antibiotic resistance. Isolated strains were resistant to a number of antibiotics applied. Result It was found that males are more prone to respiratory infections as compared to females. Male members with age of 44-58 and 59-73 are at a higher risk, while females with age of 44-58 are also at a risk of pulmonary infections. Antibiotic resistance was observed by measuring zone of inhibition in strains GCU-SG-M4, GCU-SG-M3, GCU-SG-M5, GCU-SG-M2, GCU-SG-M1 and GCU-SG-M6. GCU-SG-M2 was resistant to fluconazole (FLU), clarithromycin (CLR), cefixime (CFM) and Penicillin (P10). No zone of inhibition was observed. But it showed unusual diffused zone around the Ak and MEM antibiotic discs. rhl R gene and 16s rRNA gene were characterized and analyzed. Conclusion Findings from current study would help in raising awareness about antibiotic resistance of P. aeruginosa, and also the sequence of rhl R gene can be used as the diagnostic marker sequence to identify the virulent rhl R gene sequence from the samples when isolated from sputum of Pneumonia patients.

Synthesis of Silver Nanoparticle Using Bioactive Phenolic Compound Extracts of Leucas aspera and Leucas cephalotes and Evaluation of its Antibacterial Activity

The present study was carried out to investigate the antibacterial activity of the bioactive phenolic extract from Leucas aspera and Leucas cephalotes. The phenolic compounds were extracted using water: ethanol (1:3, v/v) by hydroethanolic extraction method. The hydroethanolic extracts were subjected to qualitative and FTIR analysis as a confirmatory step for the presence of phenolics. Synthesis of silver nanoparticle from both plants was carried out by acid hydrolysis method and subjected to UV-visible spectrophotometry, SEM, TEM and XRD analysis, for confirmation of tagged bioactive compound to AgNO3. The nanoparticle size distribution ranged between 50-94 nm in L. aspera and 40-67 nm in L. cephalotes. The antibacterial study was carried out using both crude phenolic extract and synthesized nanoparticles and tested against 5 pathogens namely Escherichia coli (ATCC® 8739™), Pseudomonas aeruginosa (ATCC® 25619™), Staphylococcus aureus (ATCC® 6538™), Bacillus subtilis (ATCC® 11774™) and Klebsiella pneumonia (ATCC® 13882™) for their antibacterial activity. From present study, the crude extract of L. cephalotes showed good antibacterial effect against test pathogen species wherein highest inhibition was observed in, P. aeruginosa, followed by B. subtilis and S. aureus with an average zone of inhibition of 23, 14 and 12 mm, E. coli and K. pneumonia measured 9 and 7 mm. The crude extract of L. aspera showed the highest inhibition in P. aeruginosa followed by S. aureus and E. coli with an average zone of inhibition of 12,11 and 10 mm B. subtilis and K. pneumonia measured 8 and 7 mm. Statistical analysis was calculated using One way ANOVA and was found to be statistically significant with p < 0.05.

Teknologi Marinasi Daging Ayam Broiler Dengan Ekstrak Buah Nenas (Ananas comosus (L). Merr) Terhadap Kualitas Mikrobiologi

This study aims to determine the effect of the microbiological quality of broiler chicken meat that is marinated using pineapple extracts with different storage times at refrigerator temperature (180C). The stages of this research consisted of 2 stages, namely the first stage of making pineapple extract from fresh pineapples and the second stage was the marination process in which the broiler chicken meat samples were marinated using pineapple extract with a concentration of 30%. The experimental design used in this study was a completely randomized design (CRD) with one treatment factor (0 days, 3 days, 6 days, 9 days and 12 days) with each treatment repeated 4 times, in order to obtain 5 x experimental units. 4 = 20 experimental units. The microbiological analysis observed was the inhibition zone analysis and Total Plate Count (TPC). Giving marination with pineapple extract to the storage time of chicken meat has a significant effect on the inhibition zone. The highest zone of inhibition was 3.23 mm (for 6 days) while the lowest zone of inhibition was 2.21 mm (for 0 days). Provision of pineapple extract marination on the storage time of broiler chicken has a significant effect on the TPC. The highest TPC was 2.29 (for 12 days) while the lowest TPC was 0.30 (for 0 days). Keywords: Broiler chicken; Marination; Microbiological quality; Pineapple extract. Abstrak Penelitian ini bertujuan untuk mengetahui pengaruh kualitas mikrobiologi daging ayam broiler yang dimarinasi menggunakan ekstrak buah nenas dengan lama penyimpanan yang berbeda pada suhu refrigerator (180C). Tahapan penelitian ini terdiri dari 2 tahapan yaitu tahap pertama pembuatan ekstrak buah nenas yang berasal dari buah nenas segar dan tahap kedua adalah proses marinasi dimana sampel daging ayam broiler dimarinasi dengan menggunakan ekstrak buah nenas dengan konsentrasi 30%. Rancangan percobaan yang digunakan dalam penelitian ini adalah Rancangan Acak Lengkap (RAL) dengan satu faktor perlakuan (0 hari, 3 hari, 6 hari, 9 hari dan 12 hari) dengan masing-masing perlakuan diulang sebanyak 4 kali, sehingga diperoleh unit percobaan 5 x 4 = 20 unit percobaan. Analisis mikrobiologi yang diamati adalah analisis zona hambat dan Total Plate Count (TPC). Pemberian marinasi dengan ektrak buah nenas terhadap lama penyimpanan daging ayam berpengaruh nyata terhadap zona hambat. Zona hambat tertinggi 3,23 mm (selama 6 hari) sedangkan zona hambat terendah 2,21 mm (selama 0 hari). Pemberian marinasi ekstrak buah nenas terhadap lama penyimpanan daging ayam broiler berpengaruh nyata terhadap TPC. TPC tertinggi 2,29 (selama 12 hari) sedangkan TPC terendah 0,30 (selama 0 hari). Kata Kunci: Daging broiler; Ekstrak nanas; Kualitas mikrobiologi; Marinasi.

Physicochemical Properties, Antioxidant and Antimicrobial Activities of Fennel (Foeniculum vulgare Mill) Seed and Leaf Oils

Foeniculum vulgare Mill. Commonly known as fennel has been used in traditional medicinal plant belonging to Apiaceae. The aim of this study was to examine quality and biological activities of fennel seed and leaf oils. The oil extraction was done in Soxhlet apparatus using hexane as a solvent. The result for physicochemical properties presented significantly higher oil yield (4.39%) and peroxide value (3.90) was observed for seed oil. Significantly higher antioxidant activities with respect to DPPH (24.45±3.74) and hydrogen peroxide (62.70±0.28) free radical scavenging activities for leaf oil. However, ascorbic acid was found to be significantly higher for seed oil (82.44±4.63). The strongest antibacterial activity with maximum zone of inhibition (14.25mm), minimum inhibitory concentration (MIC, 0.25µl/ml) and corresponding minimum bactericidal concentration (MBC, 0.50 µl/ml) was recorded for leaf oil extract against Staphylococcus aureus. On the other hand, the strongest antifungal activity with maximum zone of inhibition (13.50mm), MIC (0.38µl/ml, the least value) and minimum fungicidal concentration (MFC, 0.75µl/ml) was recorded for leaf oil against Aspergillus Niger. It can be observed from the result in this study that leaf oil extract has demonstrated more effective biological activities including both antioxidant and antimicrobial potentials.

Antibacterial efficacy of spice extracts against Streptococcus mutans and Lactobacillus acidophilus – An in-vitro study

Objectives: To evaluate the antibacterial activity of black pepper, Indian bay leaf, cinnamon, and cumin against Streptococcus mutans and Lactobacillus acidophilus in-vitro and to determine their minimum inhibitory concentration (MIC). Materials and Methods: The spices (cinnamon, cumin, Indian bay leaf, and black pepper) were obtained from local market, were dried and powdered. Solvent extracts were prepared with methanol by maceration, followed by filtration and evaporation. The antimicrobial activity was assessed using cup plate diffusion method, followed by determination of MIC of the extracts. Statistical analysis was performed using one-way analysis of variance and Tukey’s post hoc test was used for pairwise comparison. P < 0.05 was considered statistically significant. Results: All the four extracts showed significant antimicrobial activity. Cinnamon demonstrated maximum activity against S. mutans (zone of inhibition of 18.1 mm ± 0.30) and L. acidophilus (zone of inhibition of 17.9 mm ± 0.44) with the least MIC against the organisms (<0.05 mg/ml). Conclusion: All the spice extracts tested demonstrated significant antibacterial activity against S. mutans and L. acidophilus. On comparison of the antibacterial activities of all the four extracts, cinnamon extract emerged as the potent agent.