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The Central Composite Design (CCD) within the Response Surface Methodology (RSM) was applied to optimize the enzymatic hydrolysis process. This process used Alcalase® to hydrolyze Bactronophorus thoracites protein with the goal of maximizing its antimicrobial effects. Four distinct parameters were identified as independent variables: pH (A: 8.5-10.5), temperature (B: 45-65 °C), hydrolysis time (C: 120-360 min), and enzyme-to-substrate ratio (D: 1.45%-2.65% w/v). Meanwhile, the antimicrobial activity was chosen as the response variable, specifically against Pantoea ananatis (Y1) and Pantoea stewartii (Y2). According to the findings, the constructed quadratic polynomial model showed a significant correlation with the experimental data, as evidenced by the coefficient of determination (R²) values for antimicrobial activity: Y1 being 0.9893 (p < 0.0001) and Y2 at 0.9848 (p < 0.0001). Optimal antimicrobial activity for Bactronophorus thoracites protein hydrolysates (BTPH) was recorded at 46.748% against P. ananatis and 40.768% against P. stewartii. This result was observed under the optimal conditions of pH 9.5, temperature 55°C, hydrolysis duration of 240 minutes, and 2.05% w/v enzyme-to-substrate ratio. There was a notable alignment between the actual and predicted values from our models, with the Residual Standard Error (RSE) values falling under 5%. Furthermore, the established Minimum Inhibitory Concentration (MIC) was 250µg/mL, and the Minimum Bactericidal Concentration (MBC) was 500µg/mL for both P. ananatis and P. stewartii. In conclusion, the findings suggest that the refined BTPH has great promise as an effective bioactive component for agricultural use. [ABSTRACT FROM AUTHOR]

Despite black liquor's (BL) renown as a difficult-to-manage contaminant in the pulp and paper industry, BL has been found as a viable alternative material for adhesive formulation due to its high lignin content. Nevertheless, modification is required to enhance lignin's reactivity, and there is currently a lack of study focusing on this aspect for BL-lignin. This study aims to increase the phenolic hydroxyl content of BL-lignin by phenolation. After being phenolated at lignin to phenol ratio of 1:1, at a temperature of 100°C for 110 minutes, and with the addition of 8% sulfuric acid (H2SO4) as a catalyst, the phenolic hydroxyl content improved by 51.5%. The results of Fourier transform infrared spectroscopy (FTIR), UV/Vis spectrophotometry, proton nuclear magnetic resonance (¹H-NMR), thermogravimetrydifferential scanning calorimetry (TG-DSC), and its differential curve showed that the structural change in phenolated lignin opened up more active sites, implying that this lignin could be a good substitute for phenol in phenol-formaldehyde resin manufacturing. [ABSTRACT FROM AUTHOR]

Banana (Musa spp.), an important food crop in many parts of the world, is threatened by a deadly wilt disease caused by Fusarium oxysporum f. sp. cubense Tropical Race 4 (TR4). Increasing evidence indicates that plant actively recruits beneficial microbes in the rhizosphere to suppress soil-borne pathogens. Hence, studies on the composition and diversity of the root-associated microbial communities are important for banana health. Research on beneficial microbial communities has focused on bacteria, although fungi can also influence soil-borne disease. Here, high-throughput sequencing targeting the fungal internal transcribed spacer (ITS) was employed to systematically characterize the difference in the soil fungal community associated with Fusarium wilt (FW) of banana. The community structure of fungi in the healthy and TR4-infected rhizospheres was significantly different compared with that of bulk soil within the same farm. The rhizosphere soils of infected plants exhibited higher richness and diversity compared with healthy plants, with significant abundance of Fusarium genus at 14%. In the healthy rhizosphere soil, Penicillium spp. were more abundant at 7% and positively correlated with magnesium. This study produced a detailed description of fungal community structure in healthy and TR4-infected banana soils in Malaysia and identified candidate biomarker taxa that may be associated with FW disease promotion and suppression. The findings also expand the global inventory of fungal communities associated with the components of asymptomatic and symptomatic banana plants infected by TR4. [ABSTRACT FROM AUTHOR]

Lignin is a natural biopolymer with a complex three-dimensional network and it is rich in phenol, making it a good candidate for the production of bio-based polyphenol material. This study attempts to characterize the properties of green phenol-formaldehyde (PF) resins produced through phenol substitution by the phenolated lignin (PL) and bio-oil (BO), extracted from oil palm empty fruit bunch black liquor. Mixtures of PF with varied substitution rates of PL and BO were prepared by heating a mixture of phenol–phenol substitute with 30 wt.% NaOH and 80% formaldehyde solution at 94 °C for 15 min. After that, the temperature was reduced to 80 °C before the remaining 20% formaldehyde solution was added. The reaction was carried out by heating the mixture to 94 °C once more, holding it for 25 min, and then rapidly lowering the temperature to 60 °C, to produce the PL−PF or BO−PF resins. The modified resins were then tested for pH, viscosity, solid content, FTIR, and TGA. Results revealed that the substitution of 5% PL into PF resins is enough to improve its physical properties. The PL−PF resin production process was also deemed environmentally beneficial, as it met 7 of the 8 Green Chemistry Principle evaluation criteria. [ABSTRACT FROM AUTHOR]

Aims: Leaf blight disease caused by Pantoea spp. reduces rice yields in numerous nations. However, the exact strategy to combat Pantoea spp. has yet to be determined. Bactronophorus thoracites is a promising source of natural antimicrobial agents due to their potential as a substrate to generate peptides with high antimicrobial activity. This study determined the effects of lactic acid fermentation using Lactobacillus casei ATCC334 as a starter culture on antimicrobial activity against rice pathogens, proximate composition, and amino acid profiles from B. thoracites crude extract. Methodology and results: Bactronophorus thoracites was washed and deshelled to collect the flesh and homogenised at 4 °C before freeze-drying. The freeze-dried samples were fermented with L. casei for 4 to 8 days at 37 °C. The antimicrobial activity, MIC and MBC were determined using a spectrometer. The fermented protein was subjected to proximate and amino acid analyses. The antimicrobial activity of fermented B. thoracites protein (FBTP) was significantly (p<0.05) decreased with the increased fermentation days (from 4 to 8 days). The antimicrobial activity was also increased when the glucose concentration increased from 2% to 3%. However, raising the glucose concentration to 4% decreased the antimicrobial activity. The antimicrobial activity was significantly (p<0.05) increased when the substratewater (S/W) ratio increased from 0.84% to 0.96%. The FBTP (4 days, 3% glucose concentration and 0.96% S/W ratio) showed high antimicrobial activity against Pantoea ananatis and P. stewartii. The MIC and MBC values for FBTP were 500 µg/mL and 250 µg/mL against P. ananatis and P. stewartii. The zones of inhibition value for FBTP were 16.0 ± 0.5 mm (1000 µg/mL) and 9.33 ± 0.57 mm (500 µg/mL) for P. ananatis, and 11.7 ± 0.61 mm (1000 µg/mL), 9.33 ± 0.58 mm (500 µg/mL) and 7.17 ± 0.77 mm (250 µg/mL) for P. stewartii. The proximate composition and amino acid profiles of the freeze-dried protein hydrolysate powder were characterised. FBTP produced a higher value of protein (61.56%) and ash (32.38%) and a lower value of total fat (0.273%) and carbohydrates (6.27%) than the B. thoracites crude extract. Total amino acid content was 39.480 g/100 g in B. thoracites crude extract and 155.442 g/100 g in FBTP. The essential amino acid glutamine was the most abundant in B. thoracites crude extract and methionine in FBTP. Conclusion, significance and impact of study: This study showed that lactic acid fermentation could produce FBTP using L. casei with improved functional characteristics and as a source of a natural antimicrobial agent against rice pathogens. [ABSTRACT FROM AUTHOR]

Used cooking oil (UCO) is a hydrocarbon that causes significant environmental pollution globally. Oil is a difficult pollutant to remove from the environment due to its hydrophobic nature. Bioremediation is preferred over chemical and physical remediation approaches as it is more economical and environmentally friendly. Hence, the search for a microbe with outstanding degrading capability is imperative due to the nature of hydrocarbons as pollutants. The current study evaluates the ability of glyphosate degrader isolates, Burkholderia vietnamiensis AQ5-12 and Burkholderia sp. AQ5-13, to degrade UCO as the sole carbon source. The physico-cultural conditions affecting the activities of the isolates were studied using one-factor-at-a-time (OFAT) and response-surface methodology (RSM). The temperature, agitation speed, UCO concentration, nitrogen supply, yeast extract concentration, and inoculum size were optimised during a 7-day incubation period. Based on the OFAT results, the highest degradations were 62.23% and 58.33% for the isolates AQ5-12 and AQ5-13, respectively. The optimisation of RSM showed 83.42% and 87.09% degradation of the UCO by the AQ5-12 and AQ5-13 isolates. Parameters including the temperature, yeast extract, and ammonium sulphate concentration provided significant interaction for the RSM model in the isolate AQ5-12. Meanwhile, AQ5-13 showed significance in all five parameters: pH, UCO concentration, temperature, yeast extract, and ammonium sulphate concentration. The degradation efficiency validated via RSM demonstrated better results than the OFAT approach. The model validation has verified no significant difference between the experimental and predicted values. The ability of these two bacterial isolates to degrade UCO is a worthwhile procedure in the bioremediation of hydrocarbon-rich areas. [ABSTRACT FROM AUTHOR]

This study aims to increase the in vitro ruminal degradability of oil palm fronds (OPFs) through enzymatic pretreatment. The isolated fungi were selected based on their lignocellulosic degrading enzyme activities. Eleven fungi were successfully isolated, and their enzyme activities were evaluated. Three fungi, F1, F2 and F4 were selected, and they were identified as Trichoderma harzianum MK027305, Trichoderma harzianum MK027306 and Fusarium solani MK027309, respectively. The highest total gas and methane production was produced when OPFs were pretreated with an enzyme extract from 15 and 30 days of solid-state fermentation of T. harzianum MK027305 and T. harzianum MK027306, respectively. Meanwhile, OPFs pretreated with an enzyme extract from F. solani MK027309 after 45 days of solid-state fermentation produced the highest amount of volatile fatty acids. The pretreatment using the enzymes extracted from 45 days of solid-state fermentation of F. solani MK027309 increases the apparent rumen degradable carbohydrate (ARDC) by 35.29% compared to unpretreated OPF. This study showed that pretreatment of the OPFs using selected fungi's enzymes increases the volatile fatty acid production and in vitro ruminal degradability of OPF, hence improving livestock production via increased utilization of agricultural by-products with minimal impact on the production cost. [ABSTRACT FROM AUTHOR]

Endophytic lactic acid bacteria (LAB) isolated from papaya seeds, including a consortium of two LAB isolates, Weissella cibaria PPKSD19 and Lactococcus lactis subsp. lactis PPSSD39 could previously inhibit papaya dieback disease causative agent, Erwinia mallotivora BT-MARDI in vitro, indicating their potential as biofertilizer. However, further characterizations on other plant growth-promoting (PGP) properties of the LABs are prerequisite to use in agricultural settings as bio-inoculum. Hence, this study aimed to evaluate PGP potentials further and in vitro antifungal activity of the LABs against various plant pathogens. The LAB isolates were tested positive in indole-3-acetic acid (IAA), siderophore, and ammonia production and could solubilize phosphate. Weissella cibaria PPKSD19 and L. lactis subsp. lactis PPSSD39 exhibited the strongest in vitro antifungal activity against Fusarium oxysporum TKA and Curvularia lunata. Inoculum concentration of 1x108 cfu/ml of W. cibaria PPKSD19 and L. lactis subsp. lactis PPSSD39 mixture showed the highest increment in shoot and root dry weight. In conclusion, W. cibaria PPKSD19 and L. lactis subsp. lactis PPSSD39 consortium displayed promising plant probiotic potential. These results highlighted the possibility of the bacterial consortium to be exploited as bioinoculant to promote plant growth and inhibit phytopathogens causing plant diseases. [ABSTRACT FROM AUTHOR]

Fusarium oxysporum f. sp. cubense tropical race 4 (Foc TR4) is a destructive necrotrophic fungal pathogen afflicting global banana production. Infection process involves the activation of programmed cell death (PCD). In this study, seven Musa acuminata vacuolar processing enzyme (MaVPE1 – MaVPE7) genes associated with PCD were successfully identified. Phylogenetic analysis and tissue-specific expression categorized these MaVPEs into the seed and vegetative types. Foc TR4 infection induced the majority of MaVPE expressions in the susceptible cultivar "Berangan" as compared to the resistant cultivar "Jari Buaya." Consistently, upon Foc TR4 infection, high caspase-1 activity was detected in the susceptible cultivar, while low level of caspase-1 activity was recorded in the resistant cultivar. Furthermore, inhibition of MaVPE activities via caspase-1 inhibitor in the susceptible cultivar reduced tonoplast rupture, decreased lesion formation, and enhanced stress tolerance against Foc TR4 infection. Additionally, the Arabidopsis VPE -null mutant exhibited higher tolerance to Foc TR4 infection, indicated by reduced sporulation rate, low levels of H2O2 content, and high levels of cell viability. Comparative proteomic profiling analysis revealed increase in the abundance of cysteine proteinase in the inoculated susceptible cultivar, as opposed to cysteine proteinase inhibitors in the resistant cultivar. In conclusion, the increase in vacuolar processing enzyme (VPE)-mediated PCD played a crucial role in modulating susceptibility response during compatible interaction, which facilitated Foc TR4 colonization in the host. [ABSTRACT FROM AUTHOR]

Fusarium wilt is a soil borne disease causing severe losses in tomato plants. The pathogen perseverance in the soil makes the disease difficult to control. Therefore, this study was conducted to examine the efficacy of the biocontrol agent Trichoderma asperellum against Fusarium oxysporum f. sp. lycopersici, causal agent of Fusarium wilt disease of tomato based on in vitro and in vivo conditions. Trichoderma asperellum B1092 inhibited mycelial growth of the pathogen under in vitro condition using poison agar method. As in vivo, formulation of Trichoderma asperellum-enriched oil palm empty fruit bunch was significantly enhanced the growth parameters of tomato seedlings as compared to the control and it also significantly reduced the wilt disease severity. As a conclusion, T. asperellum B1902 may offer the potential for biologically controlling Fusarium wilt of tomato, inducing the growth of tomato seedlings and improve its quality. [ABSTRACT FROM AUTHOR]

Jackfruit-bronzing is caused by bacteria Pantoea stewartii subspecies stewartii (P. stewartii subsp. stewartii), showing symptoms of yellowish-orange to reddish discolouration and rusty specks on pulps and rags of jackfruit. Twenty-eight pure bacterial strains were collected from four different jackfruit outbreak collection areas in Peninsular Malaysia (Jenderam, Maran, Muadzam Shah and Ipoh). Positive P. stewartii subsp. stewartii verification obtained in the study was based on the phenotypic, hypersensitivity, pathogenicity and molecular tests. Multilocus sequence analysis (MLSA) was performed using four housekeeping genes (gyrB, rpoB, atpD and infB) on all 28 bacterial strains. Single gyrB, rpoB, atpD and infB phylogenetic trees analyses revealed the bootstrap value of 99–100% between our bacterial strains with P. stewartii subsp. stewartii reference strains and P. stewartii subsp. indologenes reference strains. On the other hand, phylogenetic tree of the concatenated sequences of the four housekeeping genes revealed that our 28 bacterial strains were more closely related to P. stewartii subsp. stewartii (99% similarities) compared to its close relative P. stewartii subsp. indologenes, although sequence similarity between these two subspecies were up to 100%. All the strains collected from the four collection areas clustered together, pointing to no variation among the bacterial strains. This study improves our understanding and provided new insight on the genetic diversity of P. stewartii subsp. stewartii associated with jackfruit-bronzing in Malaysia. [ABSTRACT FROM AUTHOR]

Vibrio infections are common among marine fish and lead to serious problems in the aquaculture sector. This study reports a recent occurrence of Vibrio species (spp.) isolated from cultured groupers in Peninsular Malaysia using the gyrB and pyrH genes. A total of 147 Vibrio strains were successfully isolated from 77 (64%) groupers using culture method and subjected to gyrB and pyrH sequencing for species identification and confirmation. Results showed that 89% of Vibrio strains were identified and clustered to six groups of Vibrio spp., while 11% were not clustered to any Vibrio spp. using the gyrB sequences. Meanwhile, by analysis of the pyrH sequences all the 147 Vibrio strains (100%) were successfully identified and clustered into 11 groups of Vibrio spp., including the gyrB non‐identified strains. The pyrH gene provides a better resolution for identification of Vibrio spp. compared with the gyrB gene. Thus, the pyrH gene was more suitable for a rapid determination of Vibrio spp. distribution in Peninsular Malaysia. Using the pyrH gene, our study found higher prevalence of Vibrio vulnificus (33%), V. alginolyticus (24%) and V. parahaemolyticus (22%), followed by V. rotiferianus (5%), V. harveyi (3%), V. tubiashii (2%), V. campbellii (2%), V. ponticus (1%), V. diabolicus (1%), V. owensii (1%) and others Vibrio sp. (7%). Thus, the results of this study revealed that the occurrence of pathogenic vibrios among grouper fish is still high in Malaysian aquaculture. In addition, the pyrH gene was proved as a suitable marker for rapid identification of Vibrio species compared with the gyrB gene. [ABSTRACT FROM AUTHOR]

Bacteria are present in stingless bee nest products. However, detailed information on their characteristics is scarce. Thus, this study aims to investigate the characteristics of bacterial species isolated from Malaysian stingless bee, Heterotrigona itama, nest products. Honey, bee bread and propolis were collected aseptically from four geographical localities of Malaysia. Total plate count (TPC), bacterial identification, phenotypic profile and enzymatic and antibacterial activities were studied. The results indicated that the number of TPC varies from one location to another. A total of 41 different bacterial isolates from the phyla Firmicutes, Proteobacteria and Actinobacteria were identified. Bacillus species were the major bacteria found. Therein, Bacillus cereus was the most frequently isolated species followed by Bacillus aryabhattai, Bacillus oleronius, Bacillus stratosphericus, Bacillus altitudinis, Bacillus amyloliquefaciens, Bacillus nealsonii, Bacillus toyonensis, Bacillus subtilis, Bacillus safensis, Bacillus pseudomycoides, Enterobacter asburiae, Enterobacter cloacae, Pantoea dispersa and Streptomyces kunmingensis. Phenotypic profile of 15 bacterial isolates using GEN III MicroPlateTM system revealed most of the isolates as capable to utilise carbohydrates as well as amino acids and carboxylic acids and derivatives. Proteolytic, lipolytic and cellulolytic activities as determined by enzymatic assays were detected in Bacillus stratosphericus PD6, Bacillus amyloliquefaciens PD9, Bacillus subtilis BD3 and Bacillus safensis BD9. Bacillus amyloliquefaciens PD9 showed broad-spectrum of antimicrobial activity against Gram-positive and Gram-negative bacteria in vitro. The multienzymes and antimicrobial activities exhibited by the bacterial isolates from H. itama nest products could provide potential sources of enzymes and antimicrobial compounds for biotechnological applications. [ABSTRACT FROM AUTHOR]

Infection with Pantoea stewartii subspecies stewartii, which causes jackfruit-bronzing disease, is a huge problem faced by the jackfruit industry in Malaysia. This study was carried out to assess the disease ratings and aggressiveness performance of 28 Pantoea stewartii subspecies stewartii isolated from jackfruit-bronzing diseased jackfruits from four collection areas (Jenderam in Selangor State, Maran and Muadzam Shah in Pahang State, and Ipoh in Perak State) in Peninsular Malaysia, inoculated into jackfruit varieties (Tekam Yellow J33, Hong J34 and Subang Chap Boy J39), the sweetcorn variety, Mas Madu (two-week-old seedlings and nine-week-old seedlings), the cucumber variety, Rocky, and the pineapple variety, MD2. The results revealed that Pantoea stewartii subspecies stewartii produced symptoms upon all inoculations in the pathogenicity testing, thus fulfilling Koch's postulates, except in the case of J39 and sweetcorn (two-week-old seedlings). No disease symptoms (disease rating 0) were observed in J39 and sweetcorn (two-week-old seedlings) within 14 days post-inoculation (14 dpi). The disease progression (based on disease ratings) proved that the jackfruit variety J39 was the most resistant, whereas J33 and J34 were susceptible to jackfruit-bronzing disease. The disease ratings of the 14-dpi period revealed a variability of disease progression among the 28 bacterial isolates, where the isolate JEN-14 had the fastest and highest disease ratings when inoculated into J33, J34, nine-week-old sweetcorn seedlings, cucumber, and pineapple. Likewise, the AUDPC value, based on disease rating, across the 28 isolates indicated that JEN-14 is the most aggressive and significant of the isolates (J33, J34, nine-week-old sweetcorn seedlings, cucumber, and cucumber pineapple inoculation; p < 0.05). Even though isolates from Jenderam (other than JEN-14) and Maran had better disease ratings and the area under the disease progress curve (AUDPC) values than isolates from Muadzam Shah and Ipoh, no significant differences were found among the isolates (p < 0.05). Based on our findings, we identified the isolate JEN-14 as the best potential candidate to assist in jackfruit-bronzing disease resistance breeding in the future. Last but not least, the methods, disease ratings, and variations of the aggressiveness profiles among the isolates from this study may be beneficial and significant by providing disease-rating references and appropriate screening approaches when selecting the most appropriately aggressive isolates for evaluating the defense response in the disease resistance breeding program. [ABSTRACT FROM AUTHOR]

Simple Summary: Recently, there has been an emergence of a hypervirulent pathotype of Aeromonas hydrophila, vAh strain. The strain was responsible for the acute mass mortalities among catfish in the USA. One of the unique abilities of the vAh strain is to utilize myo-inositol as a sole carbon source and this ability has been linked to contribute to its virulence. Therefore, the present study was carried out to screen and assess the virulence of myo-inositol-utilizing strains among Aeromonas spp. in Malaysia. Out of the 124 Aeromonas isolates screened, only A. dhakensis strain 1P11S3 was able to utilize myo-inositol as a sole carbon source. The only myo-inositol-utilizing strain was compared with five non-myo-inositol-utilizing Aeromonas spp. in an experimental challenge test using red hybrid tilapia (Oreochromis mossambicus × O. niloticus). Our findings demonstrated that the most virulent strains were A. dhakensis strain 4PS2 and A. hydrophila strain 8TK3, followed by A. dhakensis strain 1P11S3 (the only myo-inositol-utilizing strain), A. veronii strain 6TS5, A. caviae strain 7X11 and the least virulent strain was A. jandaei strain 7KL3 under current disease model. Therefore, more data are needed to assess the influence of myo-inositol utilizing ability on the pathogenesis of Aeromonas spp. The genus Aeromonas has been recognised as an important pathogenic species in aquaculture that causes motile Aeromonas septicaemia (MAS) or less severe, chronic infections. This study compares the pathogenicity of the different Aeromonas spp. that were previously isolated from freshwater fish with signs of MAS. A total of 124 isolates of Aeromonas spp. were initially screened for the ability to grow on M9 agar with myo-inositol as a sole carbon source, which is a discriminatory phenotype for the hypervirulent A. hydrophila (vAh) pathotype. Subsequently, LD50 of six selected Aeromonas spp. were determined by intraperitoneal injection of bacterial suspension containing 103, 105, and 107 CFU/mL of the respective Aeromonas sp. to red hybrid tilapias. The kidneys, livers and spleens of infected moribund fish were examined for histopathological changes. The screening revealed that only A. dhakensis 1P11S3 was able to grow using myo-inositol as a sole carbon source, and no vAh strains were identified. The LD50–240h of A. dhakensis 1P11S3 was 107 CFU/mL, while the non-myo-inositol utilizing A. dhakensis 4PS2 and A. hydrophila 8TK3 was lower at 105 CFU/mL. Similarly, tilapia challenged with the myo-inositol A. dhakensis 1P11S3 showed significantly (p < 0.05) less severe signs, gross and histopathological lesions, and a lower mortality rate than the non-myo-inositol A. dhakensis 4PS2 and A. hydrophila 8TK3. These findings suggested that myo-inositol utilizing A. dhakensis 1P11S3 was not a hypervirulent Aeromonas sp. under current experimental disease challenge conditions, and that diverse Aeromonas spp. are of concern in aquaculture farmed freshwater fish. Therefore, future study is warranted on genomic level to further elucidate the influence of myo-inositol utilizing ability on the pathogenesis of Aeromonas spp., since this ability correlates with hypervirulence in A. hydrophila strains. [ABSTRACT FROM AUTHOR]

The usage of plant growth-promoting (PGP) Streptomyces to improve plant growth is an alternative to existing strategies of chemical fertilizers that commonly caused environmental pollution. The aim of this study was to isolate and characterize PGP Streptomyces from Malaysian rice fields with antagonistic activity against Bacterial Panicle Blight disease in rice caused by Burkholderia glumae. A total of 50 bacterial isolates were recovered from the soil, rhizosphere, and endosphere; 22% showed antagonistic activity against B. glumae. Molecular identification using 16S rRNA amplification and phylogenetic tree analyses revealed that the antagonistic isolates belonged to the genus Streptomyces. Among the antagonistic isolates, Streptomyces sp. TBS5 (26.35 ± 0.14 mm) demonstrated the most antagonistic activity (p < 0.05) against B. glumae. Through API® ZYM analysis, the antagonistic isolates were revealed to have phosphoric hydrolase, ester hydrolase, peptide hydrolase, and glycosidase activities that play a crucial role in plant growth promotion. The antagonistic isolates demonstrated the highest (80%) PGP traits including able to fix nitrogen and solubilize phosphate, as well as produce siderophore and indole-3-acetic acid. Plant growth promotion assay under laboratory and greenhouse conditions were analyzed with the treatment of rice, Oryza sativa, seeds with the antagonistic isolates. Seeds treated with Streptomyces sp. TBS5, JAS2, R2-7, and TKR8 showed improvement in vigor index by more than 100% compared to water-treated control plants under both conditions. Augmentation of root length showed an overall increment of more than 101.5% under laboratory condition and 151.9% under greenhouse condition using Streptomyces sp. TBS5, JAS2, R2-7, and TKR8 treatments. Results suggest that Streptomyces sp. TBS5, JAS2, R2-7, and TKR8 are a good candidate to be developed as both biocontrol agent against B. glumae and PGP agent in rice. [ABSTRACT FROM AUTHOR]

As a major food crop, rice (Oryza sativa) is produced and consumed by nearly 90% of the population in Asia with less than 9% produced outside Asia. Hence, reports on large scale grain losses were alarming and resulted in a heightened awareness on the importance of rice plants' health and increased interest against phytopathogens in rice. To serve this interest, this review will provide a summary on bacterial rice pathogens, which can potentially be controlled by plant growth-promoting bacteria (PGPB). Additionally, this review highlights PGPB-mediated functional traits, including biocontrol of bacterial rice pathogens and enhancement of rice plant's growth. Currently, a plethora of recent studies address the use of PGPB to combat bacterial rice pathogens in an attempt to replace existing methods of chemical fertilizers and pesticides that often lead to environmental pollutions. As a tool to combat bacterial rice pathogens, PGPB presented itself as a promising alternative in improving rice plants' health and simultaneously controlling bacterial rice pathogens in vitro and in the field/greenhouse studies. PGPB, such as Bacillus, Pseudomonas, Enterobacter, Streptomyces, are now very well-known. Applications of PGPB as bioformulations are found to be effective in improving rice productivity and provide an eco-friendly alternative to agroecosystems. [ABSTRACT FROM AUTHOR]

Gene therapy research has advanced to clinical trials, but it is hampered by unstable nucleic acids packaged inside carriers and there is a lack of specificity towards targeted sites in the body. This study aims to address gene therapy limitations by encapsidating a plasmid synthesizing a short hairpin RNA (shRNA) that targets the anti-apoptotic Bcl-2 gene using truncated hepatitis B core antigen (tHBcAg) virus-like particle (VLP). A shRNA sequence targeting anti-apoptotic Bcl-2 was synthesized and cloned into the pSilencer 2.0-U6 vector. The recombinant plasmid, namely PshRNA, was encapsidated inside tHBcAg VLP and conjugated with folic acid (FA) to produce FA-tHBcAg-PshRNA VLP. Electron microscopy revealed that the FA-tHBcAg-PshRNA VLP has an icosahedral structure that is similar to the unmodified tHBcAg VLP. Delivery of FA-tHBcAg-PshRNA VLP into HeLa cells overexpressing the folate receptor significantly downregulated the expression of anti-apoptotic Bcl-2 at 48 and 72 h post-transfection. The 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay demonstrated that the cells' viability was significantly reduced from 89.46% at 24 h to 64.52% and 60.63%, respectively, at 48 and 72 h post-transfection. As a conclusion, tHBcAg VLP can be used as a carrier for a receptor-mediated targeted delivery of a therapeutic plasmid encoding shRNA for gene silencing in cancer cells. [ABSTRACT FROM AUTHOR]

Basal stem rot (BSR) disease caused by Ganoderma boninense basidiomycetous fungus is the most economically important disease in oil palms in South East Asia. Unfortunately, there is no single most effective control measure available. Tremendous efforts have been directed in incorporation of environmentally friendly biocontrol approaches in minimizing BSR disease. This study investigated the performance of two potential biocontrol agents (BCAs), AAT0115 and AAB0114 strains recovered from oil palm on suppression of BSR in planta, and also assessed their plant-growth-promoting (PGP) performance. ITS rRNA-sequence phylogeny discriminated the two ascomycetous Talaromyces apiculatus (Ta) AT0115 and Clonostachys rosea (Cr) AAB0114 biocontrol species with PGP characteristics. In vitro studies have demonstrated both Ta and Cr are capable of reducing linear mycelial growth of G. boninense. Inoculation of individual Cr and Ta—as well as Cr+Ta consortium—induced a significant increment in leaf area and bole girth of oil-palm seedlings five months post-inoculation (MPI) under nursery conditions. At five months post-inoculation, shoot and root biomass, and nutrient contents (nitrogen, phosphorus, potassium, calcium, magnesium and boron) were significantly higher in Ta-inoculated seedlings compared to control treated with non-Ta-inoculated maize. Chlorophyll and carotenoids contents in rapidly growing oil-palm seedlings challenged with Cr, Ta or a combination of both were not negatively affected. Cr, Ta and Cr+Ta consortium treated seedlings had 4.9–60% BSR disease reduction compared to the untreated control. Co-inoculation of Cr and Ta resulted in increased BSR control efficiencies by 18–26% (compared with Cr only) and 48–55% (compared with Ta only). Collectively, Cr and Ta, either individually or in consortium showed potential as BSR biocontrol agents while also possess PGP traits in oil palm. [ABSTRACT FROM AUTHOR]

Aeromonas species are ubiquitous in diverse aquatic habitats. High stocking density and improper farm practices can increase the susceptibility of cultured freshwater fishes to motile Aeromonas septicemia (MAS) outbreak. However, polymorphic phenotypes and genotypes can lead to misidentification of Aeromonas isolates from diseased fish. This study characterized a large collection of Aeromonas spp. that were collected from nine important fish farms in Peninsular Malaysia, and to compare these isolates based on their virulence and antibiotic resistance profiles. A total of 124 isolates were obtained from fish with clinical symptoms of MAS that grew on Aeromonas selective agar, were Gram-negative and were oxidase-positive, indicating these as presumptive Aeromonas spp. Based on a phylogenetic analysis of these isolates using gyrB sequences, along with phenotypic analysis based on a series of biochemical tests, five species of Aeromonas were identified among these isolates. Among these isolates, Aeromonas dhakensis was obtained most frequently (43%), followed by A. veronii (22%), A. hydrophila (20%), A. caviae (8%) and A. jandaei (7%). Moreover, 50% of the isolates were found to harbour at least half of the virulence genes tested such as aer, ahp, alt, hly, lip , fla, ela and/or act , while 2% of the isolates were found to harbour none. The Aeromonas isolates displayed resistance to amikacin (5%), ampicillin (100%), cefotaxime (7%), amoxicillin (65%), gentamicin (2%), chloramphenicol (2%) and tetracycline (15%), but not for levofloxacin (0%). The highest multiple antibiotic resistance (MAR) index of 0.63 was found in 1% of the isolates, while the mean MAR index was determined at 0.25. This study reported the prevalence of Aeromonas in cultured freshwater fishes in Peninsular Malaysia, where A. dhakensis was observed as the predominant species. However, the presence of Aeromonas with multiple virulence genes and antibiotic resistance ability should alarm the freshwater fish industry in this country. • The diversity of Aeromonas in cultured freshwater fish in Peninsular Malaysia was described. • Aeromonas dhakensis was the most dominant species, in contrast to A. hydrophila reported in previous study in Malaysia. • Among the five species isolated, only strains of A. dhakensis (2%) and A. hydrophila (20%) were found with all eight virulence genes studied. • A high percentage of Aeromonas isolates (69%) with MAR index of more than 0.2. [ABSTRACT FROM AUTHOR]

Molybdenum is very toxic to ruminants with level as low as 2 parts per million can cause severe scouring. Its contamination of waters and soils in agricultural areas needs novel removal technology. In this work we demonstrated a novel method of molybdenum removal from aqueous solution using the dialysis tubing method coupled with molybdenum-reducing activity of Serratia sp. strain Dry5. The enzymatic reduction of molybdenum is molybdenum blue, a colloid that does not pass through dialysis tubing. The calculated maximal rate of molybdenum blue production (VMoblueMax) was 0.264±0.034 mM (Mo-blue h)-1 and the concentration of molybdate resulting in the half-maximal rate of reduction (KMo) was 21.78±3.89 mM molybdate and the specific maximal rate of Mo-blue production was approximately 80 mM (Mo-blue.hr.mg cells)-1 indicating an efficient system with high tolerance towards molybdenum. © 2014 Friends Science Publishers [ABSTRACT FROM AUTHOR]

The globally vital oil palm, a major oil producer, confronts productivity challenges due to Ganoderma boninense (Gb), causing output decline. Chemical control efforts have proven ineffective, prompting exploration of microbial-based biocontrol. While single fungal biocontrol research exists, the impact of employing multiple biocontrols concurrently to combat Ganoderma and enhance oil palm growth remains uncharted. This study examined four soil-derived fungal isolates for their ability to antagonize Gb PER71 in vitro. Molecular identification categorized them as Talaromyces spp. and Penicillium sp. Moreover, all isolates were revealed to have at least three plant growth-promoting (PGP) traits and were shown to have phosphoric hydrolase, ester hydrolase, peptide hydrolase, and glycosidase activities which are essential for plant growth. Furthermore, the synergistic evaluation of fungal isolates was tested against Gb PER71. One out of six combinations of fungal isolates showed a synergistic effect in vitro, and two showed a synergistic effect in planta. The application of single and combined fungal isolates tested in planta also suppressed Gb PER71 and enhanced oil palm growth compared to control groups. The findings indicate the promising potential of these isolates as biocontrol agents (BCAs) and bioformulations against Gb in oil palm cultivation., (© 2024 Wiley‐VCH GmbH.)

Rice (Oryza sativa) is a staple food for most of the world's populations, particularly in Asia (Gumma et al. 2011). The rice sector provides Malaysians with a food supply, food sufficiency, and income for growers (Man et al. 2009). From January to February 2022, panicle samples showing symptoms of bacterial panicle blight (BPB) disease, including reddish-brown, linear lesions with indistinct margins on flag-leaf sheaths and blighted, upright, grayish straw-colored florets with sterile and aborted grains on panicles were collected in granary areas in Sekinchan, Selangor, Malaysia with 90% disease incidence in fields. Surface-sterilization of infected leaf tissue was performed using 75% ethanol and 1% sodium hypochlorite, followed by rinsing three times in sterilized water. Leaf tissue was macerated in sterilized water and aliquots were spread on King's B agar medium, then cultured for 24 h to 48 h at 35 °C. All isolated bacteria were Gram-negative rods, positive for catalase and gelatinase but negative for indole, oxidase and hydrogen sulfide (H2S), and utilized sucrose, inositol, mannitol, glucose, and citrate. Colonies were circular and smooth-margined, producing a diffusible yellowish-green pigment on King's B agar medium, which are characteristics of Burkholderia species (Keith et al. 2005). Five representative isolates (UPMBG7, UPMBG8, UPMBG9, UPMBG15, UPMBG17) were selected for molecular and pathogenicity tests. PCR using specific primers targeting the gyrB gene for molecular characterization was performed, and the ∼470 bp amplicons were sequenced (Maeda et al. 2006) and deposited in GenBank (OM824438 to OM824442). A BLASTn analysis revealed that the five isolates were 99% identical to the B. gladioli reference strains MAFF 302533, GRBB15041, and LMG19584 in GenBank (AB190628, KX638432, and AB220898). A phylogenetic tree using Maximum-likelihood analysis of the gyrB gene sequences showed that the five isolates were 99% identical to B. gladioli reference strains (AB190628, KX638432, and AB220898). To verify the identification of these isolates, the 16S rDNA gene was amplified using 16SF/16SR primers (Ramachandran et al. 2021), producing ~1,400 bp amplicons. The resulting sequences of the five isolates (OM869953 to OM869957) were 98% identical to the reference strains of B. gladioli (NR113629 and NR117553). To confirm pathogenicity, 10 ml suspensions of the five isolates at of 108 CFU/ml were inoculated into the panicles and crowns of 75-day-old rice seedlings of local rice varieties MR269 and MR219 grown in a glasshouse with temperatures ranging from 37 °C to 41 °C (Nandakumar et al. 2009). Control rice seedlings were inoculated with sterilized water. All isolates produced BPB disease symptoms like those originally found in the rice fields at four weeks after inoculation. Control seedlings remained asymptomatic. To fulfill Koch's postulates, the bacteria were reisolated from symptomatic panicles and were confirmed as B. gladioli by sequence analysis of the gyrB and 16S rDNA genes. To our knowledge, this is the first report of B. gladioli causing BPB disease of rice in Malaysia. Since BPB disease causes a significant threat to the rice industry, it is crucial to investigate the diversity of this destructive pathogen for effective disease management strategies in Malaysia.

The genus Streptomyces demonstrates enormous promise in promoting plant growth and protecting plants against various pathogens. Single and consortium treatments of two selected Streptomyces strains (Streptomyces shenzhenensis TKSC3 and Streptomyces sp. SS8) were evaluated for their growth-promoting potential on rice, and biocontrol efficiency through induced systemic resistance (ISR) mediation against Xanthomonas oryzae pv. oryzicola (Xoc), the causal agent of rice bacterial leaf streak (BLS) disease. Seed bacterization by Streptomyces strains improved seed germination and vigor, relative to the untreated seed. Under greenhouse conditions, seed bacterization with consortium treatment TKSC3 + SS8 increased seed germination, root length, and dry weight by 20%, 23%, and 33%, respectively. Single and consortium Streptomyces treatments also successfully suppressed Xoc infection. The result was consistent with defense-related enzyme quantification wherein single and consortium Streptomyces treatments increased peroxidase (POX), polyphenol oxidase, phenylalanine ammonia-lyase, and β,1-3 glucanase (GLU) accumulation compared to untreated plant. Within all Streptomyces treatments, consortium treatment TKSC3 + SS8 showed the highest disease suppression efficiency (81.02%) and the lowest area under the disease progress curve value (95.79), making it the best to control BLS disease. Consortium treatment TKSC3 + SS8 induced the highest POX and GLU enzyme activities at 114.32 μmol/min/mg protein and 260.32 abs/min/mg protein, respectively, with both enzymes responsible for plant cell wall reinforcement and resistant interaction. Our results revealed that in addition to promoting plant growth, these Streptomyces strains also mediated ISR in rice plants, thereby, ensuring protection from BLS disease.