Co-production of 1,3-propanediol and phage phiKpS2 from the glycerol fermentation by Klebsiella pneumoniae.

As an alternative to antibiotics in response to antimicrobial-resistant infections, bacteriophages (phages) are garnering renewed interest in recent years. However, the massive preparation of phage is restricted using traditional pathogens as host cells, which incurs additional costs and contamination. In this study, an opportunistic pathogen, Klebsiella pneumoniae used to convert glycerol to 1,3-propanediol (1,3-PDO), was reused to prepare phage after fermentation. The phage infection showed that the fed-batch fermentation broth containing 71.6 g/L 1,3-PDO can be directly used for preparation of phage with a titer of 1 × 108 pfu/mL. Then, the two-step salting-out extraction was adopted to remove most impurities, e.g. acetic acid (93.5%), ethanol (91.5%) and cells (99.4%) at the first step, and obtain 1,3-PDO (56.6%) in the top phase as well as phage (97.4%) in the middle phase at the second step. This integrated process provides a cheap and environment-friendly manner for coproduction of 1,3-PDO and phage.

In Vivo and In Vitro Interactions between Exopolysaccharides from Bacillus thuringensis HD270 and Vip3Aa11 Protein.

Bacillus thuringiensis (Bt) secretes the nutritional insecticidal protein Vip3Aa11, which exhibits high toxicity against the fall armyworm (Spodoptera frugiperda). The Bt HD270 extracellular polysaccharide (EPS) enhances the toxicity of Vip3Aa11 protoxin against S. frugiperda by enhancing the attachment of brush border membrane vesicles (BBMVs). However, how EPS-HD270 interacts with Vip3Aa11 protoxin in vivo and the effect of EPS-HD270 on the toxicity of activated Vip3Aa11 toxin are not yet clear. Our results indicated that there is an interaction between mannose, a monosaccharide that composes EPS-HD270, and Vip3Aa11 protoxin, with a dissociation constant of Kd = 16.75 ± 0.95 mmol/L. When EPS-HD270 and Vip3Aa11 protoxin were simultaneously fed to third-instar larvae, laser confocal microscopy observations revealed the co-localization of the two compounds near the midgut wall, which aggravated the damage to BBMVs. EPS-HD270 did not have a synergistic insecticidal effect on the activated Vip3Aa11 protein against S. frugiperda. The activated Vip3Aa11 toxin demonstrated a significantly reduced binding capacity (548.73 ± 82.87 nmol/L) towards EPS-HD270 in comparison to the protoxin (34.96 ± 9.00 nmol/L). Furthermore, this activation diminished the affinity of EPS-HD270 for BBMVs. This study provides important evidence for further elucidating the synergistic insecticidal mechanism between extracellular polysaccharides and Vip3Aa11 protein both in vivo and in vitro.

Plants recruit insecticidal bacteria to defend against herbivore attacks.

Pest feeding affects the rhizobacteria community. The rhizomicrobiota activates salicylic acid and jasmonic acid signaling pathways to help plants deal with pest infestation. However, whether plants can recruit special pesticidal microorganisms to deal with attack from herbivores is unclear. A system composed of peanuts and first-instar larvae of Holotrichia parallela were used to analyze whether peanuts truly enrich the insecticidal bacteria after feeding by larvae, and whether inoculation of the enriched bacteria promotes the resistance of plants to herbivore. In this study, high-throughput sequencing of 16 S rRNA gene amplicons was used to demonstrate that infestation of the subterranean pest H. parallela quickly changed the rhizosphere bacterial community structure within 24 h, and the abundance of Enterobacteriaceae, especially Enterobacter, was manifestly enriched. Root feeding induced rhizobacteria to form a more complex co-occurrence network than the control. Rhizosphere bacteria were isolated, and 4 isolates with high toxicity against H. parallela larvae were obtained by random forest analysis. In a back-inoculation experiment using a split-root system, green fluorescent protein (GFP)-labeled Enterobacter sp. IPPBiotE33 was observed to be enriched in uneaten peanut roots. Additionally, supplementation with IPPBiotE33 alleviated the adverse effects of H. parallela on peanuts. Our findings indicated that herbivore infestation could induce plants to assemble bacteria with specific larvicidal activity to address threats.

CEA cell adhesion molecule 5 enriches functional human hematopoietic stem cells capable of long-term multi-lineage engraftment.

Hematopoietic stem cell (HSC) surface markers improve the understanding of cell identity and function. Here, we report that human HSCs can be distinguished by their expression of the CEA Cell Adhesion Molecule 5 (CEACAM5, CD66e), which serves as a marker and a regulator of HSC function. CD66e+ cells exhibited a 5.5-fold enrichment for functional long term HSCs compared to CD66e- cells. CD66e+CD34+CD90+CD45RA- cells displayed robust multi-lineage repopulation and serial reconstitution ability in immunodeficient mice compared to CD66e-CD34+CD90+CD45RA-cells. CD66e expression also identified almost all repopulating HSCs within the CD34+CD90+CD45RA- population. Together, these results indicated that CEACAM5 is a marker that enriches functional human hematopoietic stem cells capable of long-term multi-lineage engraftment.

Greater wax moth control in apiaries can be improved by combining Bacillus thuringiensis and entrapments.

The greater wax moth (GWM), Galleria mellonella (Lepidoptera: Pyralidae), is a major bee pest that causes significant damage to beehives and results in economic losses. Bacillus thuringiensis (Bt) appears as a potential sustainable solution to control this pest. Here, we develop a novel Bt strain (designated BiotGm) that exhibits insecticidal activity against GWM larvae with a LC50 value lower than 2 µg/g, and low toxicity levels to honey bee with a LC50 = 20598.78 µg/mL for larvae and no observed adverse effect concentration = 100 µg/mL for adults. We design an entrapment method consisting of a lure for GWM larvae, BiotGm, and a trapping device that prevents bees from contacting the lure. We find that this method reduces the population of GWM larvae in both laboratory and field trials. Overall, these results provide a promising direction for the application of Bt-based biological control of GWM in beehives, although further optimization remain necessary.

Analysis of Synergism between Extracellular Polysaccharide from Bacillus thuringensis subsp. kurstaki HD270 and Insecticidal Proteins.

Bacillus thuringiensis (Bt) is the most widely used biopesticide worldwide and can produce several insecticidal crystal proteins and vegetative insecticidal proteins (Vips) at different growth stages. In our previous study, extracellular polysaccharides (EPSs) of Bt strain HD270 were found to enhance the insecticidal activity of Cry1Ac protoxin against Plutella xylostella (L.) and promote the binding of Cry1Ac to the intestinal brush border membrane vesicles (BBMVs). Whether the synergistic activity of Bt EPSs is common to other Cry1-type or Vip proteins is unclear, as is the potential synergistic mechanism. In this study, crude EPS-HD270 was found to increase the toxicity of Cry1-type toxins and Vip3Aa11 against different lepidopteran pests by approximately 2-fold. The purified EPS-HD270 also possessed synergistic activity against the toxicity of Cry1Ac and Vip3Aa11 against Spodoptera frugiperda (J.E. Smith) and Helicoverpa armigera (Hübner). Furthermore, we found that EPS-HD270 had a strong binding ability with Vip3Aa11 and promoted the binding of Vip3Aa11 to the BBMVs of H. armigera and S. frugiperda. Bt EPS-HD270 also protected Vip3Aa11 from proteolytic processing in larval midgut juice. Bt EPSs had universal synergistic effects on Cry1-type or Vip toxins against S. frugiperda and H. armigera. Bt EPS-HD270 exhibited synergistic activity with Vip3Aa through promotion of binding to BBMVs and protection from digestion by midgut protease. The results indicated that synergistic activity with Bt toxins was an important function of Bt EPSs, which was very different from other Bacillus spp.

Enterobacter Strain IPPBiotE33 Displays a Synergistic Effect with Bacillus thuringiensis Bt185.

The discovery and isolation of new non-Bt insecticidal bacteria and genes are significant for the development of new biopesticides against coleopteran pests. In this study, we evaluated the insecticidal activity of non-Bt insecticidal bacteria, PPBiotE33, IPPBiotC41, IPPBiotA42 and IPPBiotC43, isolated from the peanut rhizosphere. All these strains showed insecticidal activity against first- and third-instar larvae of Holotrichia parallela, Holotrichia oblita, Anomala corpulenta and Potosia brevitarsis. IPPBiotE33 showed the highest toxicity among the four strains and exhibited virulence against Colaphellus bowringi. The genome of IPPBiotE33 was sequenced, and a new protein, 03673, with growth inhibition effects on C. bowringi was obtained. In addition, IPPBiotE33 had a synergistic effect with Bacillus thuringiensis Bt185 against H. parallela in bioassays and back-inoculation experiments with peanut seedlings. IPPBiotE33 induced a decrease in hemocytes and an increase in phenol oxidase activity in H. parallela hemolymph, known as the immunosuppressive effect, which mediated synergistic activity with Bt185. This study increased our knowledge of the new insecticidal strain IPPBiotE33 and shed new light on the research on new insecticidal coaction mechanisms and new blended pesticides.

Identification and functional characterization of eight novel tpp family genes from Bacillus thuringiensis.

BACKGROUND: Bacillus thuringiensis (Bt) and its transgenic crops are widely used as biological control agents for agricultural pests. The tpp family is a branch of Bt insecticidal genes and consists of a few members. Research on the Tpp family proteins has focused on the binary toxins Gpp34Ab/Tpp35Ab and Tpp1/Tpp2, which need to function together to achieve insecticidal activity. However, only a few tpp family genes have been reported to exert insecticidal activity independently. This study aimed to identify and characterize tpp family genes that independently perform insecticidal functions. RESULTS: A total of 162 nucleotide sequences homologous to the single component Bt insecticidal gene tpp78Aa were obtained from the genome data of 1368 wild-type Bt strains, and 25 new full-length tpp family genes were identified. Eight new tpp family genes were successfully cloned and expressed, and bioassays of the expressed products were performed against five different pests. Bioassay results showed that these proteins exerted high insecticidal activity only against Laodelphax striatellus, a globally important rice pest, and were named Tpp78Ab1, Tpp78Bb1, Tpp78Ca1, Tpp78Da1, Tpp80Aa3, Tpp80Ac1, Tpp80Ad1, and Tpp80Ae1. The LC50 values of Tpp78Ab1, Tpp78Bb1, Tpp78Ca1, and Tpp80Ae1 against L. striatum were 8.1, 8.6, 10.1, and 9.6 µg mL-1 , respectively. The phylogenetic tree and conserved motifs indicated that the Tpp family had a common evolutionary ancestor. During evolution, the C-terminal pore-forming domain of the Tpp family adopted a similar arrangement; however, the N-terminal conserved motif showed high variability. CONCLUSION: Twenty-five full-length tpp family genes were identified. Eight new tpp family genes were cloned successfully, which could independently achieve insecticidal activity against L. striatellus. This provides abundant genetic resources for the biological control of important rice pests. In this study, we found that the relative conservation of the Tpp family proteins in the lengthy evolutionary process and the diversity generated for adapting to the environment can lay a theoretical foundation for an in-depth analysis of the function and evolution of the Tpp family. © 2023 Society of Chemical Industry.

Microflora for improving the Auricularia auricula spent mushroom substrate for Protaetia brevitarsis production.

Mushroom cultivation is a sustainable agricultural waste utilization method, but the lack of high-value utilization of the produced spent mushroom substrate (SMS) has hindered the development of mushroom cultivation-based circular agricultural systems. Conversion and utilization of SMS via Protaetia brevitarsis larvae (PBL) have proven to be a high-value AASMS utilization strategy. However, Auricularia auricula SMS (AASMS), which contains woodchips, is less palatable and digestible for PBL. To solve this problem, in this investigation, we screened out microflora (MF) for AASMS fermentation by comparing the fermentation performance as well as the effect on PBL feed intake, weight gain, and AASMS phytotoxic compound removal efficiency. In addition, by bacterial community analysis, the genera Luteimonas, Moheibacter, and Pseudoxanthomonas were predicted to be functional bacteria for AASMS fermentation and contribute to palatability and digestibility improvement.

The Influence of Environmental Values on Consumer Intentions to Participate in Agritourism-A Model to Extend TPB.

This study examines the influence of environmental values on consumer intentions to participate in agritourism through the theory of planned behaviour (TPB) and value-belief-norm (VBN) theory. It proposes an integrative model by adding two variables, i.e., environmental benefits and the human-nature coordination concept, to the TPB. The study employs a questionnaire survey method and a sample of 640, which was statistically analysed through structural equation modeling (SEM). The results reveal that the "environmental values-attitudes-behavioural intentions" framework has scientific applicability in agritourism. Environmental values, measured through the variables environmental benefits and the human-nature coordination concept, are positively correlated directly or indirectly with agritourism consumption intentions, while attitudes and subjective norms serve as mediators. However, the mediating effect of perceived behavioural control is not statistically significant, indicating making efforts to influence attitudes and subjective norms is more useful for and effective in stimulating the public's intentions towards agritourism. As this study tests the hypotheses with empirical data, it provides practical implications for policy-makers and programme managers.

One-step salting-out extraction of bacteriophage from its infection broth of Acinetobacter baumannii.

Phages as a potential alternative antibiotic for multidrug-resistant bacterial infections are receiving great attention worldwide. However, the traditional separation and purification of phage are cumbersome, time-consuming, costly and inefficient. In this study, phage phiAB9 for multidrug-resistant Acinetobacter baumannii was separated and purified by a simple and cost-saving one-step salting-out extraction (SOE). Several kinds of salts and organic solvents without effect on phage survival were chosen to form the SOE systems, and the composition of SOE systems were optimized according to the phage recovery rate and impurity removal rate. After one-step SOE by an optimal system composed of 18% (w/w) ammonium citrate and 40% (w/w) ethyl acetate, the recovery rate of phage in the middle phase could reach to 90.82%, and most proteins (99.57%), cells (97.98%) and endotoxin (84.08%) were well removed, with a concentration factor of 210 and the purification factors of phage to proteins, cells and endotoxin were 303.64, 133.55 and 5.36, respectively. Comparing with two-step SOE and traditional aqueous two-phase extraction, one-step SOE may be an available method for the separation and purification of phages.

The efficiency of a clinical pathway to guide combined applications of interventional pulmonology in undiagnosed pleural effusions.

The diagnostic procedure of pleural effusion (PEs) is challenging due to low detection rates and numerous aetiologies. Hence, any attempt to enhance diagnosis is worthwhile. We present a clinical pathway to guide combined application of interventional pulmonology (IP) for tracing causes of undiagnosed PEs. Subjects with undiagnosed PEs were identified in the Hospital Information System of Dalian Municipal Central Hospital from January 1, 2012, to December 31, 2018. Eligible subjects were divided into a group of combined tests and a group of medical thoracoscopy (MT). Optimal and subsequent diagnostic tests were performed depending on the guidance of the clinical pathway by matching profitable chest lesions with the respective adaptation. As the guidance of clinical pathway, common bronchoscopy would be preferentially selected if pulmonary lesions involved or within the central bronchus, EBUS-TBNA was favoured when pulmonary lesions were adjacent to the central bronchus or with the enlarged mediastinal/hilar lymph nodes, guided bronchoscopy would be preferred if pulmonary nodules/masses were larger than 20 mm with discernible bronchus signs, CT-assisted transthoracic core biopsy was preferred if pulmonary nodules were less than 20 mm, image guided cutting needle biopsy was the recommendation if the pleural thickness was larger than 10 mm and pulmonary lesions were miliary. MT was preferred only when undiagnosed PEs was the initial symptom and pulmonary lesions were miliary or absent. A total of 83.57% cases of undiagnosed PEs were eligible for the clinical pathway, and 659 and 216 subjects were included in the combined tests and MT groups, respectively, depending on the optimal recommendation of the clinical pathway. The total diagnostic yields in the combined tests and MT groups were 95.99% and 91.20%, respectively, and the difference in total diagnostic yield was statistically significant (χ2 = 7.510, p = 0.006). Overall, clinical pathway guidance of the combined application of IP is useful for tracing the causes of undiagnosed PEs. The diagnostic yield of undiagnosed PEs is significantly increased compared with that of MT alone.

Lignocellulose degradation in Protaetia brevitarsis larvae digestive tract: refining on a tightly designed microbial fermentation production line.

BACKGROUND: The Scarabaeidae insect Protaetia brevitarsis (PB) has recently gained increasing research interest as a resource insect because its larvae can effectively convert decaying organic matter to plant growth-promoting frass with a high humic acid content and produce healthy, nutritional insect protein sources. Lignocellulose is the main component of PB larvae (PBL) feed, but PB genome annotation shows that PBL carbohydrate-active enzymes are not able to complete the lignocellulose degradation process. Thus, the mechanism by which PBL efficiently degrade lignocellulose is worthy of further study. RESULTS: Herein, we used combined host genomic and gut metagenomic datasets to investigate the lignocellulose degradation activity of PBL, and a comprehensive reference catalog of gut microbial genes and host gut transcriptomic genes was first established. We characterized a gene repertoire comprising highly abundant and diversified lignocellulose-degrading enzymes and demonstrated that there was unique teamwork between PBL and their gut bacterial microbiota for efficient lignocellulose degradation. PBL selectively enriched lignocellulose-degrading microbial species, mainly from Firmicutes and Bacteroidetes, which are capable of producing a broad array of cellulases and hemicellulases, thus playing a major role in lignocellulosic biomass degradation. In addition, most of the lignocellulose degradation-related module sequences in the PBL microbiome were novel. PBL provide organic functional complementarity for lignocellulose degradation via their evolved strong mouthparts, alkaline midgut, and mild stable hindgut microenvironment to facilitate lignocellulosic biomass grinding, dissolving, and symbiotic microbial fermentation, respectively. CONCLUSIONS: This work shows that PBL are a promising model to study lignocellulose degradation, which can provide highly abundant novel enzymes and relevant lignocellulose-degrading bacterial strains for biotechnological biomass conversion industries. The unique teamwork between PBL and their gut symbiotic bacterial microbiota for efficient lignocellulose degradation will expand the knowledge of holobionts and open a new beginning in the theory of holobionts. Video Abstract.

Vip3Aa domain IV and V mutants confer higher insecticidal activity against Spodoptera frugiperda and Helicoverpa armigera.

BACKGROUND: The fall armyworm Spodoptera frugiperda and cotton bollworm Helicoverpa armigera are major insect pests of corn and cotton worldwide. Genetically engineered crops producing Vip3Aa, a potent endotoxin, from the bacterium Bacillus thuringiensis (Bt) are effective in controlling these two harmful pests. However, Vip3Aa efficacy is relatively weak compared to that of other Bt proteins such as Cry1A and Cry1F. This study sought to modify Vip3Aa for increased insecticidal activity and determine the cause of elevated activity. RESULTS: The two triple Vip3Aa mutants in domains IV and V (Vip3Aa-S543N/I544L/E627A and Vip3Aa-S543N/I544L/S686R) exhibited 7.3-fold and 2.8-fold increased toxicity against S. frugiperda, respectively, compared with the wild type while the toxicity of Vip3Aa-S543N/I544L/S686R was 3.2 times that of wild-type protein in H. armigera. The mutants had enhanced stability in midgut juice and 2.6-5.1 times higher binding affinity against S. frugiperda and H. armigera compared with wild type protein. CONCLUSIONS: The enhanced toxicity of Vip3Aa mutants was due to increased stability and binding affinity during infection. The amino acids S543 and I544 combined with E627 or S686 in domains IV and V of Vip3Aa are important for maintaining structural stability and receptor binding. The results match insecticidal activity (LC50 ) with binding activity (Kd ), which provides novel clues for the rational design of Bt insecticidal proteins. © 2022 Society of Chemical Industry.

Characterization of Microorganisms from Protaetia brevitarsis Larva Frass.

Decomposers play an important role in the biogeochemical cycle. Protaetia brevitarsis larvae (PBLs) can transform wastes into frass rich in humic acid (HA) and microorganisms, which may increase the disease resistance of plants and promote plant growth. Beyond HA, the microorganisms may also contribute to the biostimulant activity. To address this hypothesis, we investigated the potential microbial community in the PBL frass samples and elucidated their functions of disease resistance and plant growth promotion. High-throughput sequencing analysis of four PBL-relevant samples showed that their frass can influence the microbial community of the surrounding environment. Further analysis showed that there were many microorganisms beneficial to agriculture, such as Bacillus. Therefore, culturable Bacillus microbes were isolated from frass, and 16S rDNA gene analysis showed that Bacillus subtilis was the dominant species. In addition, some Bacillus microorganisms isolated from the PBL frass had antibacterial activities against pathogenic fungi. The plant growth promotion pot experiment also proved that some strains promote plant growth. In conclusion, this study demonstrated that the microorganisms in the PBL frass are conducive to colonizing the surrounding organic matrix, which will help beneficial microbes to increase the disease resistance of plants and promote plant growth.

Structure characteristics and function of a novel extracellular polysaccharide from Bacillus thuringiensis strain 4D19.

Bacillus thuringiensis (Bt) are entomopathogenic bacteria that produce different kinds of insecticidal proteins. However, studies on Bt exopolysaccharides are lacking. Here, we aimed to explore the characteristics and insecticidal synergism of EPS, an exopolysaccharide from Bt strain 4D19. The molecular weight of EPS-2 was 58.0 kDa, which consisted of mannose (44.2%), GlcN (35.5%), D-GalN (8.0%), glucose (5.5%), arabinose (5.1%), galactose (0.9%), Man-UA (0.3%) and Glc-UA (0.2%). The toxicity of insecticidal proteins against Plutella xylostella was increased by adding EPS. EPS-2 bound to Cry1Ac protoxin and promoted the binding of Cry1Ac protoxin to the gut membrane of P. xylostella, but did not bind to activated toxins. These results suggested that EPS-2 may bind to the protoxin C-terminal region to enhance insecticidal activity. Our findings indicated that Bt strains produce exopolysaccharide to enhance the toxicity of insecticidal crystal proteins, which could be applied in biopesticide research and product development.

Dominant egg surface bacteria of Holotrichia oblita (Coleoptera: Scarabaeidae) inhibit the multiplication of Bacillus thuringiensis and Beauveria bassiana.

Holotrichia oblita (Coleoptera: Scarabaeidae) and some other scarab beetles are the main soil-dwelling pests in China. Bacillus thuringiensis (Bt) and Beauveria bassiana (Bb) are entomopathogens that have been used as biocontrol agents of various pests. However, scarab larvae especially H. oblita exhibited strong adaptability to these pathogens. Compared to other scarabs, H. oblita could form a specific soil egg case (SEC) structure surrounding its eggs, and young larvae complete the initial development process inside this structure. In this study, we investigated the role of SEC structure and microorganisms from SEC and egg surface in pathogen adaptability. 16S rRNA gene analysis revealed low bacterial richness and high community unevenness in egg surface, with Proteobacteria, Firmicutes, Bacteroidetes and Fusobacteria dominating. In terms of OTUs composition analysis, the data show that the egg surface contains a large number of unique bacteria, indicating that the egg bacterial community may be derived from maternal transmission. Furthermore, we found that all culturable bacteria isolated from egg surface possessed antimicrobial activity against both Bt and Bb. The Pseudomonas bacteria with a significantly higher abundance in egg surface showed strong Bt- and Bb antagonistic ability. In conclusion, this study demonstrated a unique and antimicrobial bacterial community of H. oblita egg surface, which may contribute to its adaptability. Furthermore, the specific SEC structure surrounding the H. oblita eggs will provide a stable microenvironment for the eggs and egg surface bacteria, which probably provides more advantages for H. oblita adaptation ability.

Effects of Bt Cry78Ba1 Toxin on Larvae and Adults of Apis mellifera (Hymenoptera: Apidae).

Cry78Ba1 is Bacillus thuringiensis Berliner (Bacillales: Bacillaceae) (Bt) protein found with high insecticidal activity against the piercing-sucking insect Laodelphax striatellus Fallén (Homoptera: Delphacidae) and has broad application prospects for control of the rice planthopper. As honey bees may be exposed to Bt Cry78Ba1 rice pollen by feeding, we evaluated the risk of Bt Cry78Ba1 toxin to Apis mellifera L. workers. A dietary exposure experiment was conducted on worker larvae and adults under controlled laboratory conditions to examine the effects of Cry78Ba1 toxin on honey bees. Worker bee larvae were fed a diet containing Cry78Ba1 toxin (0.01, 0.1, 1, and 10 mg/liter) on day 2 through day 5 after grafting, and adults were exposed to syrup containing Cry78Ba1 for up to 16 d. Negative control (no test substance added), solvent control (1 mM Tris-HCl), and positive control (dimethoate 45 mg/liter for the larva test, 1 and 45 mg/liter for the adult test) groups were established. Compared with the negative control, larvae and adults that consumed food containing Cry78Ba1 toxin exhibited no significant differences in survival, larval weight, or pollen or syrup consumption. This result indicates that chronic oral exposure to Cry78Ba1 toxin has no negative effects on honey bees at the maximum tested concentration.

Characterization of Two Novel Bacillus thuringiensis Cry8 Toxins Reveal Differential Specificity of Protoxins or Activated Toxins against Chrysomeloidea Coleopteran Superfamily.

Scarabaeoidea and Chrysomeloidea insects are agriculture-destructive coleopteran pests. Few effective Bacillus thuringiensis (Bt) insecticidal proteins against these species have been described. Bt isolate BtSU4 was found to be active against coleopteran insects. Genome sequencing revealed two new cry8 genes in BtSU4, designated as cry8Ha1 and cry8Ia1. Both genes expressed a 135 kDa protoxin forming irregular shape crystals. Bioassays performed with Cry8Ha1 protoxin showed that it was toxic to both larvae and adult stages of Holotrichia parallela, also to Holotrichia oblita adults and to Anoplophora glabripennis larvae, but was not toxic to larval stages of H. oblita or Colaphellus bowringi. The Cry8Ia1 protoxin only showed toxicity against H. parallela larvae. After activation with chymotrypsin, the Cry8Ha1 activated toxin lost its insecticidal activity against H. oblita adults and reduced its activity on H. parallela adults, but gained toxicity against C. bowringi larvae, a Chrysomeloidea insect pest that feeds on crucifer crops. The chymotrypsin activated Cry8Ia1 toxin did not show toxicity to any one of these insects. These data show that Cry8Ha1 and Cry8Ia1 protoxin and activated toxin proteins have differential toxicity to diverse coleopteran species, and that protoxin is a more robust protein for the control of coleopteran insects.

Protaetia brevitarsis larvae can feed on and convert spent mushroom substrate from Auricularia auricula and Lentinula edodes cultivation.

The edible mushroom industry produces massive amounts of spent mushroom substrate (SMS). Thus, there is an urgent need for high-value utilization technology to process the SMS, especially SMSs originating from woodchips. Protaetia brevitarsis larvae (PBL) can feed on various types of organic matter and can produce organic fertilizer and insect protein. In this study, we investigated the potential of PBL to utilize and convert SMSs from Auricularia auricula (SMS-AA) and Lentinula edodes (SMS-LE) cultivation. The results showed that the PBL were able to feed on SMS-AA and SMS-LE and form nutrient-enriched organic fertilizer with a low phytotoxicity and high humic acid content. Further analysis of the organic carbon dynamics suggested that PBL can efficiently digest and utilize lignin. This study demonstrates a new strategy for the utilization of SMSs originating from woodchips, and provides a new model for further investigations on the mechanism of lignin decomposition.