Development and application of a bacteriophage cocktail for Shigella flexneri biofilm inhibition on the stainless steel surface.

Food contamination and biofilm formation by Shigella in food processing facilities are major causes of acute gastrointestinal infection and mortality in humans. Bacteriophages (phages) are promising alternatives to antibiotics in controlling plankton and biofilms in food matrices. This study isolated two novel phages, S2_01 and S2_02, with lytic activity against various Shigella spp. From sewage samples. Transmission electron microscopy revealed that phages S2_01 and S2_02 belonged to the Caudovirales order. On characterizing their lytic ability, phage S2_01 initially exhibited relatively weak antibacterial activity, while phage S2_02 initially displayed rapid antibacterial activity after phage application. A combination of these phages in a 1:9 ratio was selected, as it has been suggested to elicit the most rapid and sustained lysis ability for up to 24 h. It demonstrated lytic activity against various foodborne pathogens, including six Shigella spp. The phage cocktail exhibited biofilm inhibition and disruption abilities of approximately 79.29% and 42.55%, respectively, after 24 h in a 96-well microplate. In addition, inhibition (up to 23.42%) and disruption (up to 19.89%) abilities were also observed on stainless steel surfaces, and plankton growth was also significantly suppressed. Therefore, the phage cocktail formulated in this study displays great potential as a biological control agent in improving food safety against biofilms and plankton.

An atoxigenic Aspergillus flavus PA67 from Shandong province exhibits potential in biocontrol against toxigenic Aspergillus flavus, Sclerotium rolfsii, and Fusarium proliferatum.

Peanuts and corn are susceptible to various soil-borne fungi, leading to significant economic losses. Atoxigenic Aspergillus flavus have been widely used as biocontrol agents for managing aflatoxin contamination because of their minimal environmental impact, strong competitive ability, and sustained inhibition effect. After multiple identifications and cluster amplification pattern (CAP) analysis, three atoxigenic A. flavus PA04, PA10 and PA67 were isolated from peanut samples in Shandong Province, which can reduce aflatoxin levels by up to 90 %. Our study revealed that atoxigenic A. flavus also competed vigorously with Sclerotium rolfsii and Fusarium proliferatum for nutrition and space, achieving notable inhibition rates of up to 90.4 % and 90.6 %, respectively. The supernatants of atoxigenic A. flavus also inhibited the growth of S. rolfsii and F. proliferatum, with PA67 demonstrating the most significant effect. Whole genome sequencing revealed that PA67 contains multiple glycoside hydrolases and metabolites with antifungal activity. The kojic acid production of PA67 was higher than that of PA04 and PA10, reaching 17.48 g/L, which has a significant inhibition on sclerotia germination. PA67 supernatant significantly inhibited the hyphae growth of S. rolfsii and F. proliferatum, and down-regulated genes related to sclerotia and fumonisin formation. This study demonstrates the biocontrol potential of PA67 against three soil-borne fungi and is the first investigation of atoxigenic A. flavus to inhibit S. rolfsii and F. proliferatum.

Characterization of a novel phage SPX1 and biological control for biofilm of Shewanella in shrimp and food contact surfaces.

Shewanella putrefaciens, commonly found in seafood, forms tenacious biofilms on various surfaces, contributing to spoilage and cross-contamination. Bacteriophages, owing to their potent lytic capabilities, have emerged as novel and safe options for preventing and eliminating contaminants across various foods and food processing environments. In this study, a novel phage SPX1 was isolated, characterized by a high burst size (43.81 ± 3.01 PFU/CFU) and a short latent period (10 min). SPX1 belongs to the Caudoviricetes class, exhibits resistance to chloroform, and sensitivity to ultraviolet. It shows stability over a wide range of temperatures (30-50 °C) and pH levels (3-11). The genome of phage SPX1 consists of 53,428 bp with 49.72 % G + C composition, and lacks tRNAs or virulence factors. Genome analysis revealed the presence of two endolysins, confirming its biofilm-removal capacity. Following the treatment of shrimp surface biofilm with the optimal MOI of 0.001 of phage SPX1 for 5 h, the bacterial count decreased by 1.84 ± 0.1 log10 CFU/cm2 (> 98.5 %). Biofilms on the surfaces of the three common materials used in shrimp processing and transportation also showed varying degrees of reduction: glass (1.98 ± 0.01 log10 CFU/cm2), stainless steel (1.93 ± 0.05 log10 CFU/cm2), and polyethylene (1.38 ± 0.1 log10 CFU/cm2). The study will contribute to phage as a novel and potent biocontrol agent for effectively managing S. putrefaciens and its biofilm, ensuring a reduction in spoilage bacteria contamination during the aquaculture, processing, and transportation of seafood products.

Biocontrol potential of Streptomyces sp. N2 against green and blue mold disease in postharvest navel orange and the action mechanism.

The objective of this study was to provide a promising alternative to chemical fungicides for management of postharvest citrus decay, thereby promoting sustainable citrus fruit production. The postharvest decay of citrus fruit caused by Penicillium digitatum and Penicillium italicum results in substantial economic losses in citrus industry worldwide. With growing fungal resistance issues in P. digitatum and P. italicum, there is an urgent need for searching new methods to address above problems in a safe and environmentally friendly way. Streptomyces sp. N2, a new species from Streptomyces genus, exhibits significant antagonistic activity against Rhizoctonia solani. However, its biocontrol efficacy against postharvest decay caused by P. digitatum and P. italicum and its action mechanism remain unknown. In this study, Streptomyces sp. N2 was found to have significant potential in controlling green and blue mold diseases in postharvest navel oranges. Moreover, the action mechanism of Streptomyces sp. N2 against both P. italicum and P. digitatum was elucidated. On the one hand, Streptomyces sp. N2 stimulated fruit resistance to fight against invading fungal pathogens. It significantly reduced ROS content in navel orange upon the infection of mold disease, increased the production of defense-related enzymes including peroxidase (POD), polyphenol oxidase (PPO), and phenylalanine ammonia-lyase (PAL) and pathogenesis-related proteins of chitinase and ß-1,3-glucanase. On the other hand, Streptomyces sp. N2 secreted bioactive substances to inhibit the growth of P. italicum and P. digitatum so as to prevent the development of postharvest decay. The bioactive substances secreted by Streptomyces sp. N2 significantly inhibited the spore germination and mycelial growth and led to microstructural damages to the cell wall and membrane, ROS burst, and mitochondrial dysfunction in both P. italicum and P. digitatum. This study provides a theoretical reference and application potential for the biological control of Streptomyces sp. N2 on green and blue mold diseases.

Strategies for the biocontrol Pseudomonas infections pre-fruit harvest.

The efficiency of global crop production is under threat from microbial pathogens which is likely to be worsened by climate change. Major contributors to plant disease are Pseudomonas syringae (P. syringae) pathovars which affect a variety of important crops. This opinion piece focuses on P. syringae pathovars actinidiae and syringae, which affect kiwifruit and stone fruits, respectively. We discuss some of the current control strategies for these pathogens and highlight recent research developments in combined biocontrol agents such as bacteriophages and combinations of bacteriophages with known anti-microbials such as antibiotics and bacteriocins.

Comprehensive genomic analysis of Brevibacillus brevis BF19 reveals its biocontrol potential against bitter gourd wilt.

Bitter gourd wilt, a severe vascular disease triggered by the soilborne pathogen Fusarium oxysporum f. sp. momordicae (FOM), markedly constrains bitter gourd yield. In this study, a novel strain BF19 of Brevibacillus brevis was isolated and identified, exhibiting strong antimicrobial activity against FOM through in vivo and in vitro experiments. To comprehensively assess the biocontrol potential of strain BF19, we conducted phenotypic, phylogenetic, and comparative genomics analyses. Phenotypic analysis revealed that BF19 exhibited 53.33% biocontrol efficacy and significantly increased the average plant height, root fresh weight, and dry weight. Whole-genome sequencing and comparative genomic analysis revealed numerous potential genes associated with biocontrol mechanisms in BF19. Importantly, the integration of metabolic cluster prediction with liquid chromatography‒tandem mass spectrometry (LC‒MS/MS) revealed the presence of a macrobrevin antibiotic, a product of polyketide synthases (PKSs), predominantly in BF19 fermentation products. The effectiveness of the Br. brevis strain BF19 and its crude extract against bitter gourd wilt has also been confirmed. This study provides a genetic framework for future investigations on PKSs and establishes a scientific basis for optimizing field applications of microbial biopesticides derived from Br. brevis BF19.

Biocontrol mechanisms of poplar leaf blight due to Nigrospora oryzae.

Nigrospora oryzae, a newly identified pathogen, is responsible for poplar leaf blight, causing significant harm to poplar growth. Here, we describe, for the first time, a biological control method for the control of poplar leaf blight via the applications of 3 dominant Trichoderma strains/species. In this study, dominant Trichoderma species/strains with the potential for biocontrol were identified and then further characterised via dual culture assays, volatile organic compounds (VOCs), and culture filtrates. The biocontrol efficacy of these strains against N. oryzae was found to exceed 60%. Furthermore, the reactive oxygen species (ROS) content in Populus davidiana × P. alba var. pyramidalis (PdPap) leaves pretreated with these Trichoderma strains significantly decreased. Furthermore, pretreatment of PdPap with a combination of these Trichoderma (Tcom) resulted in 9.71-fold and 1.95-fold increases in peroxidase (POD) and superoxide dismutase (SOD) activity, respectively, and 3.87-fold decrease in the MDA content compared to controls. Moreover, Tcom pretreatment activated the salicylic acid (SA) and jasmonic acid (JA) pathway-dependent defence responses of poplar, upregulating pathogenesis-related protein (PR) and MYC proto-oncogene (MYC-R) by more than 12-fold and 17.32-fold, respectively. In addition, Trichoderma treatments significantly increased the number of lateral roots, aboveground biomass, and stomata number and density of PdPap, and Tcom was superior to the single pretreatments. The soil pH also became weakly acidic in these pretreatments, which is beneficial for the growth of PdPap seedlings. These findings indicate that these dominant Trichoderma strains can effectively increase biocontrol and poplar growth promotion.

Employing Bacillus and Pseudomonas for phytonematode management in agricultural crops.

Phytonematodes are responsible for causing significant harm and reducing yields in various agricultural crops. To minimize losses caused by phytonematodes and meet the high demand for agricultural production, it is important to develop effective strategies with minimal environmental impact to manage this biotic stress. Due to the adverse environmental effects associated with synthetic pesticides, it is imperative to use beneficial bacteria, such as Bacillus and Pseudomonas spp., for biocontrol purposes to control phytonematode infestation in agricultural settings. This approach has gained considerable attraction, as there is a promising market for eco-friendly biopesticides based on bacteria that can effectively manage phytonematodes. Furthermore, biocontrol strains of Bacillus and Pseudomonas have the potential to enhance crop productivity by producing various substances that promote plant growth and development. This review aims to explore the role of Bacillus and Pseudomonas spp. in phytonematode management, elucidate different mechanisms by which these bacteria suppress nematode populations, and discuss the future prospects of utilizing these bacteria in agriculture.

Applications of Pythium- and Phytophthora-produced volatiles in plant disease control.

Volatile organic compounds (VOCs) mediate biological interactions and are produced by Pythium and Phytophthora species. These VOCs are biotechnologically relevant because the genera include important plant pathogens, whereby VOCs can aid in disease detection, and biological control agents, whereby VOCs contribute to disease control. Studies on VOC production, identification, and characterization of individual VOCs produced by Pythium and Phytophthora species are reviewed. VOCs detected in plants infected with Phytophthora species are also reviewed as potentially oomycete-derived VOCs. The Pythium- and Phytophthora-produced VOCs are compared with other microorganisms, and the main effects of these VOCs on microbial inhibition and plant-mediated effects are reviewed. These effects are summarized from direct demonstration studies and inferences based on the known functions of the identified Pythium- and Phytophthora-produced VOCs. There are two main applications of VOCs to plant disease control: the use of VOCs to detect pathogenic Pythium and Phytophthora species, e.g., e-nose detecting systems, and the use of VOC-producing biological control agents, e.g., Pythium oligandrum. Future research could understand how the VOCs are produced to engineer VOC levels in strains, analyze more oomycete species and strains, accurately quantify the VOCs produced, and exploit recent developments in analytical chemistry technology. KEY POINTS: • Compiled inventory of volatiles produced by Phytophthora and Pythium species • Volatilomes contain microbe-inhibiting and plant growth-promoting compounds • Volatile potential in disease detection and control supports analyzing more species.

White rot fungi as a multifaceted biocontrol agent: Metabolic disruption and algal inhibition in Microcystis aeruginosa.

Microcystis aeruginosa is a prevalent cyanobacterium linked to water eutrophication and harmful algal blooms. While bacterial control strategies are well-studied, the effects of white rot fungi on Microcystis aeruginosa are less understood. This study examines the impact of whole fungal liquid, its centrifuged supernatant, and sterilized solutions on the algae's physiological and biochemical traits. Metabolomics and multivariate analysis identified significant changes in 47 metabolic markers, including carbohydrates, amino acids, and fatty acids, across treatments. The complete fungal liquid exhibited the strongest algicidal effect, likely due to synergistic solubilization mechanisms mediated by extracellular enzymes such as manganese peroxidase, catalase, and laccase. Notably, algicidal activity persisted even after sterilization, suggesting the presence of non-proteinaceous compounds like polysaccharides or lipids. The metabolic disturbances included downregulation of the TCA cycle and reduced fatty acid synthesis, leading to inhibited photosynthesis and compromised nucleic acid integrity in the algal cells. This research enhances our understanding of how white rot fungi disrupt Microcystis aeruginosa metabolism, providing a theoretical basis for their potential use in bioremediation of eutrophic aquatic environments.

Alternative splicing perspective to prey preference of environmentally friendly biological agent Cryptolaemus montrouzieri.

BACKGROUND: Cryptolaemus montrouzieri (Coccinellidae) is widely utilized as biological control agents in modern agriculture. A comprehensive understanding of its food preference can help guide mass rearing and safety management during field application of pest control. Although some studies have paid attentions to the impacts of prey shift on C. montrouzieri, little is known regarding the role of post-transcriptional regulations in its acclimation to unnatural preys. RESULTS: We performed a genome-wide investigation on alternative splicing dynamics in C. montrouzieri in response to the predation transition from natural prey to unnatural ones. When feeding on undesired diets, 402-764 genes were differentially alternative spliced in C. montrouzieri. It is noteworthy that the majority of these genes (> 87%) were not differentially expressed, and these differentially spliced genes regulated distinct biological processes from differentially expressed genes, such as organ development and morphogenesis, locomotory behavior, and homeostasis processes. These suggested the functionally nonredendant role of alternative splicing in modulating physiological and metabolic responses of C. montrouzieri to the shift to undesired preys. In addition, the individuals feeding on aphids were subject to a lower level of changes in splicing than other alternative diets, which might be because of the similar chemical and microbial compositions. Our study further suggested a putative coupling of alternative splicing and nonsense-mediated decay (AS-NMD), which may play an important role in fine-tuning the protein repertoire of C. montrouzieri, and promoting its acclimation to predation changes. CONCLUSION: These findings highlight the key role of alternative splicing in modulating the acclimation of ladybirds to prey shift and provide new genetic clues for the future application of ladybirds in biocontrol.

Biocontrol of Hemileia vastatrix, the Causal Agent of Coffee Leaf Rust, by Paenibacillus sp. NMA1017.

Coffee leaf rust (CLR), caused by Hemileia vastatrix, is considered a highly important phytosanitary problem in Mexico. Currently, there are few microorganisms used as biocontrol alternatives to chemical control of CLR in organic coffee fields in Mexico. This study evaluates the use of Paenibacillus sp. NMA1017 as a biocontrol agent to inhibit the development of H. vastatrix in in vitro and in vivo (greenhouse) experiments. Hemileia vastatrix urediniospores were placed on water agar plates, and then Paenibacillus sp. NMA1017 was inoculated simultaneously or 8 h later. Urediniospores germination rate was reduced by 94% when the NMA1017 strain was inoculated simultaneously with the urediniospores and reduced by 38% when NMA1017 was inoculated 8 h later. Experiments with 8-month-old Bourbon coffee plants that were infected with H. vastatrix showed that disease incidence was reduced by 38, 90, and 50% when NMA1017 was applied 8 days before, simultaneously, or 8 days after the application of H. vastatrix, respectively. Paenibacillus sp. NMA1017 also reduced the severity of CLR on the leaves by up to 62%. The germination urediniospores of other rust pathogens such as Puccinia sorghi (maize leaf rust), Puccinia triticina (wheat leaf rust), Puccinia graminis f. sp. tritici (black stem rust of wheat), Uromyces striatus (alfalfa leaf rust), and Phragmidium sp. (rosebush leaf rust) were also inhibited. Use of the potential biocontrol agent Paenibacillus sp. NMA1017 might help reduce the application of chemical fungicides for the control of CLR, making coffee a more sustainable crop and providing management options for organic coffee growers.

Antibiotic-producing plant-associated bacteria, anti-virulence therapy and microbiome engineering: Integrated approaches in sustainable agriculture.

Plant health is crucial for maintaining the well-being of humans, animals and the environment. Plant pathogens pose significant challenges to agricultural production, global food security and ecosystem biodiversity. This problem is exacerbated by the impact of climate change, which is expected to alter the emergence and evolution of plant pathogens and their interaction with their plant hosts. Traditional approaches to managing phytopathogens involved the use of chemical pesticides, but alternative strategies are needed to address their ongoing decline in performance as well as their negative impact on the environment and public health. Here, we highlight the advancement and effectiveness of biocontrol strategies based on the use of antimicrobial-producing plant-associated bacteria, anti-virulence therapy (e.g. quorum quenching) and microbiome engineering as sustainable biotechnological approaches to promote plant health and foster sustainable agriculture. Notably, Enterobacterales are emerging as important biocontrol agents and as a source of new antimicrobials for potential agricultural use. We analysed here the genomes of over 250 plant-associated enterobacteria to examine their potential to synthesize secondary metabolites. Exploration of the plant microbiome is of major interest in the search for eco-friendly alternatives for reducing the use of chemical pesticides.

Profiling of secondary metabolites produced by Pseudomonas sp. isolate PY-122 and PY-142 as biocontrol agent againts fusarium wilt disease on chili.

Fusarium wilt is an important disease on chili plants caused by the fungus Fusarium oxysporum. This study aims to determine the effectiveness of secondary metabolites from 2 Pseudomonad isolates in controlling Fusarium wilt in chili plants and their effect on chili plant growth. This research was conducted in two stages, namely in vitro, which was carried out in the plant health laboratory and greenhouse, Faculty of Agriculture, Universitas Pembangunan Nasional Veteran Jawa Timur, and in vivo which was carried out in chili plantations known to be endemic to Fusarium wilt in Menganti, Gresik. The research design used in this study was Completely Randomized Design for in vitro testing and Randomized Group Design for in vivo testing by testing 9 treatments repeated 3 times. The treatment tested was control, secondary metabolite application of Pseudomonas isolates PY-122 and PY-142 with concentrations of 20% 30%, 40% and 50%. The variables observed were inhibition of secondary metabolites on the growth of Fusarium fungus, disease incubation period, disease severity index, and plant growth. Sequencing results with 16rRNA gene markers for both Pseudomonas isolates showed similarities to Pseudomonas sp. The results showed that the treatment of secondary metabolite PY-142 with a concentration of 50% showed the highest consistent inhibition. In the greenhouse and field tests, the two isolates slowed down the disease incubation period and development compared to controls. In agronomical observations, the average plant height, number of leaves, root length, and appearance of the first flower on the treated plants were higher and more numerous than the control plants.

Biocontrol Ability of Strain Bacillus amyloliquefaciens SQ-2 against Table Grape Rot Caused by Aspergillus tubingensis.

Bacillus amyloliquefaciens strain SQ-2, isolated from a cured product, has been demonstrated to exhibit a highly efficacious performance against phytopathogens, including Stemphylium solani, Fusarium moniliforme, Fusarium graminearum, and Aspergillus tubingensis. In particular, with regard to A. tubingensis, which causes summer bunch rot, SQ-2 has been observed to suppress the mycelial growth of all tested grape cultivars by over 40%. Especially on Kyoho grapes, it has the highest inhibition rate of 53%. Scanning electron microscopy (SEM) confirms that SQ-2 is an effective agent for suppressing the mycelia proliferation, differentiation, and spore formation of A. tubingensis. Furthermore, an LC/MS analysis revealed that SQ-2 produces two principal lipopeptides, namely, bacillibactin and surfactin, in addition to a polyketide, bacillaene. Further analysis through gas chromatography-mass spectrometry (GC/MS) identified 41 distinct volatile organic compounds secreted by SQ-2. Transcriptomic analysis indicated that exposure to the metabolite of SQ-2 induced substantial gene expression alterations in A. tubingensis. These data suggest that B. amyloliquefaciens strain SQ-2 exhibits promising crop protection potential of inhibiting plant pathogens through the secretion of bacillibactin, surfactin, bacillaene, and VOCs.

Sterol regulatory element-binding proteins mediate intrinsic fungicide tolerance and antagonism in the fungal biocontrol agent Clonostachys rosea IK726.

Sterol regulatory element-binding proteins (SREBPs) are transcription factors governing various biological processes in fungi, including virulence and fungicide tolerance, by regulating ergosterol biosynthesis and homeostasis. While studied in model fungal species, their role in fungal species used for biocontrol remains elusive. This study delves into the biological and regulatory function of SREBPs in the fungal biocontrol agent (BCA) Clonostachys rosea IK726, with a specific focus on fungicide tolerance and antagonism. Clonostachys rosea genome contains two SREBP coding genes (sre1 and sre2) with distinct characteristics. Deletion of sre1 resulted in mutant strains with pleiotropic phenotypes, including reduced C. rosea growth on medium supplemented with prothioconazole and boscalid fungicides, hypoxia mimicking agent CoCl2 and cell wall stressor SDS, and altered antagonistic abilities against Botrytis cinerea and Rhizoctonia solani. However, Δsre2 strains showed no significant effect. Consistent with the gene deletion results, overexpression of sre1 in Saccharomyces cerevisiae enhanced tolerance to prothioconazole. The functional differentiation between SRE1 and SRE2 was elucidated by the yeast-two-hybridization assay, which showed an interaction between SREBP cleavage-activating protein (SCAP) and SRE1 but not between SRE2 and SCAP. Transcriptome analysis of the Δsre1 strain unveiled SRE1-mediated expression regulation of genes involved in lipid metabolism, respiration, and xenobiotic tolerance. Notably, genes coding for antimicrobial compounds chitinases and polyketide synthases were downregulated, aligning with the altered antagonism phenotype. This study uncovers the role of SREBPs in fungal BCAs, providing insights for C. rosea IK726 application into integrated pest management strategies.

Paenibacillus as a Biocontrol Agent for Fungal Phytopathogens: Is P. polymyxa the Only One Worth Attention?

Control of fungal phytopathogens is a significant challenge in modern agriculture. The widespread use of chemical fungicides to control these pathogens often leads to environmental and food contamination. An eco-friendly alternative that can help reduce reliance on these chemicals is plant growth-promoting bacteria (PGPB), particularly those of the genus Paenibacillus, which appear to be highly effective. The review aims to summarize the existing knowledge on the potential of Paenibacillus spp. as fungal biocontrol agents, identify knowledge gaps, and answer whether other species of the genus Paenibacillus, in addition to Paenibacillus polymyxa, can also be effective biocontrol agents. Paenibacillus spp. can combat plant phytopathogens through various mechanisms, including the production of lipopeptides (such as fusaricidin, paenimyxin, and pelgipeptin), the induction of systemic resistance (ISR), hydrolytic enzymes (chitinase, cellulase, and glucanase), and volatile organic compounds. These properties enable Paenibacillus strains to suppress the growth of fungi such as Fusarium oxysporum, F. solani, Rhizoctonia solani, Botrytis cinerea, or Colletotrichum gloeosporioides. Notably, several strains of Paenibacillus, including P. polymyxa, P. illinoisensis KJA-424, P. lentimorbus B-30488, and P. elgii JCK1400, have demonstrated efficacy in controlling fungal diseases in plants. Importantly, many formulations with Paenibacillus strains have already been patented, and some are commercially available, but most of them contain only P. polymyxa. Nevertheless, considering the data presented in this review, we believe that other strains from the Paenibacillus genus (besides P. polymyxa) will also be commercialized and used in plant protection in the future. Importantly, there is still limited information regarding their impact on the native microbiota, particularly from the metataxonomic and metagenomic perspectives. Expanding knowledge in this area could enhance the effectiveness of biocontrol agents containing Paenibacillus spp., ensuring safe and sustainable use of biological fungicides.

Effects of Paecilomyces lilacinus and Bacillus pumilus on stem nematode and rhizosphere bacterial communities of sweet potato.

Stem nematode (Ditylenchus destructor Thorne) is considered one of the most economically devastating species affecting sweet potato production. Biocontrol offers a sustainable strategy for nematode control. This study conducted a pot experiment to evaluate the biocontrol efficacy of Paecilomyces lilacinus CS-Z and Bacillus pumilus Y-26 against the stem nematode, as well as to examine their influence on the bacterial communities in the sweet potato rhizosphere. The findings indicated that B.pumilus Y-26 and P.lilacinus CS-Z exhibited respective suppression rates of 82.9% and 85.1% against the stem nematode, while also stimulating sweet potato plant growth. Both high-throughput sequencing and Biolog analysis revealed distinct impacts of the treatments on the bacterial communities. At the phylum level, B.pumilus Y-26 enhanced the abundance of Actinobacteria but reduced the abundance of Cyanobacteria, with P.lilacinus CS-Z exhibiting similar effects. Additionally, the treatment with B.pumilus Y-26 resulted in increased abundances of Crossiella, Gaiella, Bacillus, and Streptomyces at the genus level, while the treatment with P.lilacinus CS-Z showed increased abundances of Crossiella and Streptomyces. In contrast, the abundance of Pseudarthrobacter was reduced in the treatment with B.pumilus Y-26. Conversely, the application of the nematicide fosthiazate exhibited minor influence on the bacterial community. The findings indicated that the application of P.lilacinus CS-Z and B.pumilus Y-26 led to an increase in the relative abundances of beneficial microorganisms, including Gaiella, Bacillus, and Streptomyces, in the rhizosphere soil. In conclusion, P.lilacinus CS-Z and B.pumilus Y-26 demonstrated their potential as environmentally friendly biocontrol agents for managing stem nematode disease of sweet potato.

Characterization and screening of new Metarhizium isolates to control the coconut rhinoceros beetle in the Pacific islands.

The coconut rhinoceros beetle (CRB; Oryctes rhinoceros) is one of the most destructive insect pests of coconut and oil palms in tropical Asia and the Pacific islands. Members of a new variant, known as CRB-G (clade I), have recently spread into the Pacific islands, causing significant damage. Biopesticides containing Metarhizium spp. are the strongest candidates for inundative biological control against the emerging CRB threat. Selection of the most virulent and robust isolate may determine the impact of this control option on the pest. In this work, CRB specimens with natural fungal infection were collected in Papua New Guinea (PNG) and Solomon Islands (SI). Putative entomopathogenic fungi were isolated and identified. These new isolates and some previously obtained from other Pacific countries were molecularly identified, characterized, and tested for virulence against CRB larval populations in PNG and SI in laboratory bioassays. Of the new isolates collected, four obtained from SI were identified as Metarhizium majus (conidia length ⁓11-15 µm), and four from PNG were identified as Metarhizium pingshaense (conidia length ⁓4-6 µm). The most virulent isolate was M. majus AgR-F717, which caused 100 % mortality in 20-23 days against a CRB variant from the CRB-S grouping (clade II) in laboratory bioassays carried out in PNG. Isolates of M. pingshaense did not show pathogenicity against CRB larvae. M. majus AgR-F717 was also the most virulent in laboratory bioassays using the mixed SI population (from both CRB-S and CRB-G groupings) and was selected for further evaluation using artificial breeding sites. Under field conditions, this isolate demonstrated its ability to infect CRB, dispersal up to 100 m from treated artificial breeding sites, and persistence in soil for at least four months. The new isolate AgR-F717 of M. majus has demonstrated potential as an augmentative biological control agent for CRB in PNG and SI.

Effects of four potent entomopathogenic fungal isolates on the survival and performance of Telenomus remus, an egg parasitoid of fall armyworm.

Fall armyworm (FAW), Spodoptera frugiperda is a generalist pest known to feed on more than 300 plant species, including major staple crops such as rice, maize and sorghum. Biological control of FAW using a combination of a major indigenous egg parasitoid Telenomus remus and entomopathogenic fungi was explored in this study. Metarhizium anisopliae strains (ICIPE 7, ICIPE 41, and ICIPE 78) and Beauveria bassiana ICIPE 621 which demonstrated effectiveness to combat the pest, were evaluated through direct and indirect fungal infection to assess their pathogenicity and virulence against T. remus adults, S. frugiperda eggs and their effects on T. remus parasitism rates. Metarhizium anisopliae ICIPE 7 and ICIPE 78 exhibited the highest virulence against T. remus adults with LT50 values >2 days. ICIPE 7 induced the highest T. remus mortality rate (81.40 ± 4.17%) following direct infection with dry conidia. Direct fungal infection also had a significant impact on parasitoid emergence, with the highest emergence rate recorded in the M. anisopliae ICIPE 7 treatment (42.50 ± 5.55%), compared to the control ± (83.25 ± 5.94%). In the indirect infection, the highest concentration of 1 x 109 conidia ml-1 of ICIPE 78 induced the highest mortality (100 ± 0.00%) of T. remus adults, and the highest mortality (51.25%) of FAW eggs, whereas the least FAW egg mortality (15.25%) was recorded in the lowest concentration 1 x 105 conidia ml-1 of ICIPE 41. The number of parasitoids that emerged and their sex ratios were not affected by the different fungal strain concentrations except in ICIPE 7 at high dose. This study showed that potential combination of both M. anisopliae and B. bassiana with T. remus parasitoid can effectively suppress FAW populations.