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Aspergillus flavus , the primary mold that causes food spoilage, poses significant health and economic problems worldwide. Eliminating A. flavus growth is essential to ensure the safety of agricultural products, and extracellular compounds (ECCs) produced by Bacillus spp. have been demonstrated to inhibit the growth of this pathogen. In this study, we aimed to identify microorganisms efficient at inhibiting A. flavus growth and degrading aflatoxin B 1 . We isolated microorganisms from soil samples using a culture medium containing coumarin (CM medium) as the sole carbon source. Of the 498 isolates grown on CM medium, only 132 bacterial strains were capable of inhibiting A. flavus growth. Isolate 3BS12-4, identified as Bacillus siamensis , exhibited the highest antifungal activity with an inhibition ratio of 43.10%, and was therefore selected for further studies. The inhibition of A. flavus by isolate 3BS12-4 was predominantly attributed to ECCs, with a minimum inhibitory concentration and minimum fungicidal concentration of 0.512 g/ml. SEM analysis revealed that the ECCs disrupted the mycelium of A. flavus . The hydrolytic enzyme activity of the ECCs was assessed by protease, β-1,3-glucanase, and chitinase activity. Our results demonstrate a remarkable 96.11% aflatoxin B 1 degradation mediated by ECCs produced by isolate 3BS12-4. Furthermore, treatment with these compounds resulted in a significant 97.93% inhibition of A. flavus growth on peanut seeds. These findings collectively present B. siamensis 3BS12-4 as a promising tool for developing environmentally friendly products to manage aflatoxin-producing fungi and contribute to the enhancement of agricultural product safety and food security.

In tropical regions, the viability of outdoor photo-fermentative biohydrogen production faces challenges arising from elevated temperatures and varying light intensity. This research aimed to explore how high temperatures and outdoor environments impact both biohydrogen production and the growth of purple non-sulfur bacteria. Our findings revealed the potential of Rhodopseudomonas spp. as a robust outdoor hydrogen-producing bacteria, demonstrating its capacity to thrive and generate biohydrogen even at 40 °C and under fluctuating outdoor conditions. Rhodopseudomonas harwoodiae NM3/1-2 produced the highest cumulative biohydrogen of 223 mL/L under anaerobic light conditions at 40 °C, while Rhodopseudomonas harwoodiae 2M had the highest dry cell weight of 2.93 g/L. However, R. harwoodiae NM3/1-2 demonstrated the highest dry cell weight of 3.99 g/L and Rhodopseudomonas pentothenatexigens KKU-SN1/1 exhibited the highest cumulative biohydrogen production of 400 mL/L when grown outdoors. In addition, the outdoor enhancement of biohydrogen production was achieved through the utilization of a cluster of ten bioreactors system. The outcomes demonstrated a notable improvement in biohydrogen production efficiency, marked by the highest daily biohydrogen production of 493 mL/L d by R. pentothenatexigens KKU-SN1/1. Significantly, the highest biohydrogen production rate was noted to be 17 times greater than that observed in conventional batch production methods. This study is the first to utilize R. pentothenatexigens and R. harwoodiae for sustained biohydrogen production at high temperatures and in outdoor conditions over an extended operational period. The successful utilization of a clustered system of ten bioreactors demonstrates potential to scale-up for industrial biohydrogen production., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Protease is a widely used enzyme particularly in the detergent industry. In this research, we aimed to isolate alkaline protease-producing bacteria for characterization as a laundry detergent additive. The screening of alkaline protease production was investigated on basal medium agar plus 1% skim milk at pH 11, with incubation at 30°C. The highest alkaline protease-producing bacterium was 6BS15-4 strain, identified as Bacillus gibsonii by 16S rRNA gene sequencing. While the optimum pH was 12.0, the strain was stable at pH range 7.0-12.0 when incubated at 45°C for 60 min. The alkaline protease produced by B. gibsonii 6BS15-4 using dairy effluent was characterized. The optimum temperature was 60°C and the enzyme was stable at 55°C when incubated at pH 11.0 for 60 min. Metal ions K + , Mg 2+ , Cu 2+ , Na + , and Zn 2+ exhibited a slightly stimulatory effect on enzyme activity. The enzyme retained over 80% of its activity in the presence of Ca 2+ , Ba 2+ , and Mn 2+ . Thiol reagent and ethylenediaminetetraacetic acid did not inhibit the enzyme activity, whereas phenylmethylsulfonyl fluoride significantly inhibited the protease activity. The alkaline protease from B. gibsonii 6BS15-4 demonstrated efficiency in blood stain removal and could therefore be used as a detergent additive, with potential for various other industrial applications.

This research aimed to evaluate the diversity of yeasts recovered from fermented foods gathered from some areas of Northeastern Thailand. The fermented food items included Pla-som, Nham-pla, Kem-buknud, Isan-sausage, Pla-ra, Mhum-neu, Mhum-Khai-pla, Nham-neu, Nham-mu, Kung-joom, Som-pla-noi, and Poo-dong. Their probiotic characteristics were also investigated. A total of 103 yeast isolates of nine genera were identified using 28S rDNA sequencing. The yeast genera were Candida (20.3%), Diutina (2.9%), Filobasidium (1.0%), Kazachstania (33.0%), Pichia (3.9%), Saccharomyces (1.0%), Starmerella (28.2%), Torulaspora (2.9%), and Yarrowia (6.8%). Based on probiotic characteristic analysis of ten selected yeast strains, Kazachstania bulderi KKKS4-1 showed the strongest probiotic characteristics in terms of hemolytic activity, antimicrobial activity against pathogenic bacteria, tolerance to low pH and bile salt and hydrophobicity. Isolated yeasts with probiotic characteristics may be useful in fermented food and animal feed production to improve their nutritional values., Competing Interests: Conflict of interest: The authors declare no conflict of interest., (© 2022 the Author(s), licensee AIMS Press.)

Saccharomyces cerevisiae is a yeast strain often used to improve the feed quality of ruminants. However, S. cerevisiae has limited capacity to provide biomass when inoculated with carbon sources and a low ability to produce cellulase enzymes. Here, we hypothesized that yeast in the rumen produces a large amount of biomass and could release cellulase enzymes to break down fiber content. Therefore, the aim of this study was to screen, isolate and identify yeast from the rumen fluids of Holstein Friesian steers and measure the efficiency of biomass production and cellulase activity. A fermentation medium containing sugarcane molasses as a carbon source and urea as a nitrogen source was optimized. Two fistulated-crossbred Holstein Friesian steers averaging 350 ± 20 kg body weight were used to screen and isolate the ruminal yeast. Two experiments were designed: First, a 12 × 3 × 3 factorial was used in a completely randomized design to determine biomass and carboxymethyl cellulase activity. Factor A was the isolated yeast and S. cerevisiae . Factor B was sugarcane molasses (M) concentration. Factor C was urea (U) concentration. In the second experiment, potential yeasts were selected, identified, and analyzed for 7 × 4 factorial use in a completely randomized design. Factor A was the incubation times. Factor B was the isolated yeast strains, including codes H-Khon Kaen University (KKU) 20 (as P. kudriavzevii -KKU20), I-KKU20 ( C. tropicalis -KKU20), and C-KKU20 (as Galactomyces sp.-KKU20). Isolation was imposed under aerobic conditions, resulting in a total of 11 different colonies. Two appearances of colonies including asymmetric colonies of isolated yeast (indicated as A, B, C, E, and J) and ovoid colonies (coded as D, F, G, H, I, and K) were noted. Isolated yeast from the rumen capable of providing a high amount of biomass when inoculant consisted of the molasses 15% + urea 3% (M15 + U3), molasses 25% + urea 1% (M25 + U1), molasses 25% + urea 3% (M25 + U3), and molasses 25% + urea 5% (M25 + U5) when compared to the other media solution ( p < 0.01). In addition, 11 isolated biomass-producing yeasts were found in the media solution of M25 + U1. There were 4 isolates cellulase producing yeasts discovered in the media solution of M25 + U1 and M25 + U5 whereas molasses 5% + urea 1% (M5 + U1), molasses 5% + urea 3% (M5 + U3), molasses 5% + urea 5% (M5 + U5), molasses 15% + urea 1% (M15 + U1), molasses 15% + urea 3% (M5 + U3), and M25 + U3 were found with 2, 3, 1, 2, 1, and 2 isolates, respectively. Ruminal yeast strains H-KKU20, I-KKU20, and C-KKU20 were selected for their ability to produce biomass. Identification of isolates H-KKU20 and I-KKU20 revealed that those isolates belonged to Pichia kudriavzevii -KKU20 and Candida tropicalis -KKU20 while C-KKU20 was identified as Galactomyces sp.-KKU20. Two strains provided maximum cell growth: P. kudriavzevii -KKU20 (9.78 and 10.02 Log cell/mL) and C. tropicalis -KKU20 (9.53 and 9.6 Log cells/mL) at 60 and 72 h of incubation time, respectively. The highest ethanol production was observed in S. cerevisiae at 76.4, 77.8, 78.5, and 78.6 g/L at 36, 48, 60, and 72 h of incubation time, respectively ( p < 0.01). The P. kudriavzevii -KKU20 yielded the least reducing sugar at about 30.6 and 29.8 g/L at 60 and 72 h of incubation time, respectively. The screening and isolation of yeasts from rumen fluids resulted in 11 different yeasts being obtained. The potential yeasts discovered in the rumen fluid of cattle were Pichia kudriavzevii -KKU20, Candida tropicalis -KKU20, and Galactomyces sp.-KKU20. P. kudriavzevii -KKU20 had higher results than the other yeasts in terms of biomass production, cellulase enzyme activity, and cell number.

A new yeast species (KKU-FW10) belonging to the Candida genus was isolated from Jasminum adenophyllum in the Plant Genetic Conservation Project under The Royal Initiative of Her Royal Highness Princess Maha Chakri Sirindhorn area, Chulabhorn Dam, Konsan district within Chaiyaphum province in Thailand. The strain was identified via analysis of nucleotide sequences from the D1/D2 domain of 26S ribosomal DNA and based on its morphological, physiological and biochemical characteristics. The sequence obtained from yeast isolate KKU-FW10 was 97 percent identical to that of Candida chanthaburiensis (GenBank accession number AB500861.1), with 506/517 (nucleotides identity/total nucleotides) matching nucleotides, nine substitutions and two gaps being detected. This species belonged to the Candida clade. Regarding morphological characteristics, isolate KKU-FW10 presents cream-colored butyrous colonies, vegetative reproduction through budding and, round cells without filaments or ascospores. The major ubiquinone detected was Q-9. The above results suggest that isolate KKU-FW10 is a new member of the genus Candida, and the name Candida konsanensis is proposed for this yeast. The type strain of the new species is KKU-FW10(T) (= BCC 52588(T), = NBRC 109082(T), = CBS 12666(T)). In addition, this KKU-FW10 could potentially produce 58.24 Units/ml of carboxymethyl cellulase when it was cultured in YP broth containing 1.0 % carboxymethyl cellulose for 24 h.