Eco-physiological response of two marine bivalves to acute exposition to commercial Bt-based pesticide

Microbial products based on the entomopathogenic bacterium Bacillus thuringiensis (Bt) are among the most common biopesticides used worldwide to suppress insect pests in forests, horticulture and agricultural crops. Some of the effects of commercial Bt have been recorded for terrestrial and freshwater non-target organisms but little research is available on marine fauna. Nevertheless, due to the contiguity of agro-ecosystems and coastal habitats, marine fauna may be highly influenced by this control method. We studied the effect of a commercial Bt product on the physiological and ecological responses and the energy budget of two of the most frequent marine intertidal bivalves in the Mediterranean, the native Mytilaster minimus and the invasive Brachidontes pharaonis. To test the effects experimentally, we simulated the worst scenarios possible using the average dose applied to fields and a hypothetical accumulation dose. The results showed the feeding rates of both species were affected detrimentally by the different experimental conditions; higher concentrations led to higher respiration rates, however neither species showed any significant difference in excretion rates. The biopesticide had a significant effect on the energy budget, the values decreasing with doses. In addition, it led to high mortality for the worst treatments and, in both species, induced significantly higher cardiac activity than in the controls. These results indicate a measurable effect of Bt commercial products on marine organisms, and great attention should be paid to biopesticides composed by entomopathogenic bacteria and addictive compounds. In addition, the results highlight the urgent need to study not only the effects of anthropogenic pressures on target organisms but also to extend our view to other ecosystems not expected to be influenced. Gaining data at the organismal level should help increase the sustainability of pest control and reduce the consequences of side-effects

Odorants of Capsicum spp. dried fruits as candidate attractants for Lasioderma serricorne F. (Coleoptera: Anobiidae)

The cigarette beetle, Lasioderma serricorne F. (Coleoptera: Anobiidae) is an important food storage pest affecting the tobacco industry and is increasingly impacting museums and herbaria. Monitoring methods make use of pheromone traps which can be implemented using chili fruit powder. The objective of this study was to assess the response of L. serricorne to the volatile organic compounds (VOCs) from different chili powders in order to identify the main semiochemicals involved in this attraction. Volatiles emitted by Capsicum annuum, C. frutescens, and C. chinense dried fruit powders were tested in an olfactometer and collected and analyzed using SPME and GC-MS. Results indicated that C. annuum and C. frutescens VOCs elicit attraction toward L. serricorne adults in olfactometer, while C. chinense VOCs elicit no attraction. Chemicals analysis showed a higher presence of polar compounds in the VOCs of C. annuum and C. frutescens compared to C. chinense, with α-ionone and β-ionone being more abundant in the attractive species. Further olfactometer bioassays indicated that both α-ionone and β-ionone elicit attraction, suggesting that these compounds are candidates as synergistic attractants in pheromone monitoring traps for L. serricorne

The Green Synthesis of Silver Nanoparticles from Avena fatua Extract: Antifungal Activity against Fusarium oxysporum f.sp. lycopersici

Using plant extracts as eco-friendly reducing and stabilizing agents for the synthesis of nanoparticles has gained significant attention in recent years. The current study explores the green synthesis of silver nanoparticles (AgNPs) using the Avena fatua extract and evaluates their antifungal activity against Fusarium oxysporum f.sp. lycopersici (Fol), a fungal plant pathogen. A green and sustainable approach was adopted to synthesize silver nanoparticles before these nanoparticles were employed for anti-fungal activity. The primary indication that AgNPs had formed was performed using UV-vis spectroscopy, where a strong peak at 425 nm indicated the effective formation of these nanoparticles. The indication of important functional groups acting as reducing and stabilizing agents was conducted using the FTIR study. Additionally, morphological studies were executed via SEM and AFM, which assisted with more effectively analyzing AgNPs. Crystalline behavior and size were estimated using powder XRD, and it was found that AgNPs were highly crystalline, and their size ranged from 5 to 25 nm. Synthesized AgNPs exhibited significant antifungal activity against Fol at a concentration of 40 ppm. Furthermore, the inhibitory index confirmed a positive correlation between increasing AgNPs concentration and exposure duration. This study suggests that the combined phytochemical mycotoxic effect of the plant extract and the smaller size of synthesized AgNPs were responsible for the highest penetrating power to inhibit Fol growth. Moreover, this study highlights the potential of using plant extracts as reducing and capping agents for the green synthesis of AgNPs with antifungal properties. The study concludes that A. fatua extract can synthesize antifungal AgNPs as a sustainable approach with robust antifungal efficacy against Fol, underscoring their promising potential for integration into plant protection strategies

Antimicrobial activity of the red palm weevil Rhynchophorus ferrugineus

The red palm weevil, Rhynchophorus ferrugineus (Olivier) (Coleoptera Dryophthoridae), is an important pest of palms. Knowl- edge of both its natural enemies and its defensive mechanisms against predators and microorganisms is important to develop methods for an integrated pest control. Antimicrobial activity of the cuticular surface of adults and larvae, as well as of eggs, of this invasive species was investigated. This activity was tested against the Gram-positive bacteria Bacillus subtilis (Ehrenberg) Cohn and Bacillus thuringiensis Berliner, the Gram-negative bacterium Escherichia coli Escherich, and the entomopathogenic fungi Beauveria bassiana (Balsamo) Vuillemin and Metarhizium anisopliae (Metchnikoff) Sorokin. A similar analysis was con- ducted with the hemolymph of R. ferrugineus\u2019 larvae infected by Pseudomonas aeruginosa (Schroter) Migula, E. coli and Staphy- lococcus aureus Rosenbach. Polar surface fraction of extracts from adults and large larvae inhibits Gram-positive bacteria and the B. bassiana\u2019s growth, but not the growth of E. coli and M. anisopliae. Similarly, the hemolymph of larvae and the surface extracts of both small larvae and eggs seemed not to show any inhibition. Chemical analyses of the fraction exhibiting antimicrobial activ- ity show the presence of some polar compounds ranging between 1000 and 1500 Dalton. This study improves our knowledge on the biology of R. ferrugineus and helps to suggest strategies for the biocontrol of this pest

General framework for assessing the risks for in-soil organisms exposed to Plant Protection Products (PPP)

The Panel on Plant Protection Products and their Residues (PPR) has been tasked by EFSA to revise the SANCO guidance Document on Terrestrial Ecotoxicology which is currently used for conducting the risk assessment for terrestrial non-target organisms, except for birds and mammals and non-target arthropods. For each group of organisms, the PPR Panel first summarises the science behind the respective risk assessment in a scientific opinion and, in a second step, EFSA will develop practical guidance on how to perform the risk assessment. The main scope of the revision is to take into consideration: (i) the entry into force of the Regulation 1107/2009 replacing the Directive 91/414/EEC; (ii) the need for developing specific protection goals and (iii) the inclusion of new scientific elements in the environmental risk assessment of in-soil organisms. The Opinion only covers chemical plant protection products (PPPs), while microbial PPPs are not specifically addressed in the opinion. The working group of the PPR Panel reviewed the current environmental risk assessment, identified key drivers that sustain important soil ecosystem services in agricultural landscapes and developed proposals for specific protection goal (SPG) options for in-field and off-field areas, according to the procedure developed in the opinion of the PPR Panel of 2010 Based on the overarching ecosystem services concept, which was boosted in the so-called Millennium Ecosystem Assessment, those ecosystem services that could potentially be directly or indirectly (e.g. via trophic interactions) affected by the normal agricultural use of plant protection products were first identified, followed by the groups of in-soil organisms that are key drivers or service providing units (SPUs) for those ecosystem services. Specific protection goal (SPG) options have to be proposed for each combination of a key driver and ecosystem service. During the development of the opinion, several research needs covering different aspects in the context of pesticides risk assessment for in-soil organisms have been identified. Some of those needs, such as standardisation of toxicity tests with additional species than the standard ones, should be prioritised before the development of the guidance document. A public consultation on the draft opinion was launched in May, 2016 aiming at involving all the interested stakeholders

Assessment of genetically modified maize\ua04114 for food and feed uses, under Regulation (EC) No\ua01829/2003 (application EFSA-GMO-NL-2014-123)

Maize\ua04114 was developed through Agrobacterium\ua0tumefaciens-mediated transformation to provide protection against certain lepidopteran and coleopteran pests by expression of the Cry1F, Cry34Ab1 and Cry35Ab1 proteins derived from Bacillus\ua0thuringiensis, and tolerance to the herbicidal active ingredient glufosinate-ammonium by expression of the PAT protein derived from Streptomyces viridochromogenes. The molecular characterisation data did not identify issues requiring assessment for food/feed safety. None of the compositional, agronomic and phenotypic differences identified between maize\ua04114 and the non-genetically modified (GM) comparator(s) required further assessment. There were no concerns regarding the potential toxicity and allergenicity of the newly expressed proteins Cry1F, Cry34Ab1, Cry35Ab1 and PAT, and no evidence that the genetic modification might significantly change the overall allergenicity of maize 4114. The nutritional value of food/feed derived from maize 4114 is not expected to differ from that derived from non-GM maize varieties and no post-market monitoring of food/feed is considered necessary. In the case of accidental release of viable maize\ua04114 grains into the environment, maize\ua04114 would not raise environmental safety concerns. The post-market environmental monitoring plan and reporting intervals are in line with the intended uses of maize\ua04114. The genetically modified organism (GMO) Panel\ua0concludes that maize\ua04114 is as safe as the non-GM comparator(s) and non-GM reference varieties with respect to potential effects on human and animal health and the environment in the context of the scope of this application

Phylogeny in Aid of the Present and Novel Microbial Lineages: Diversity in Bacillus

Bacillus represents microbes of high economic, medical and biodefense importance. Bacillus strain identification based on 16S rRNA sequence analyses is invariably limited to species level. Secondly, certain discrepancies exist in the segregation of Bacillus subtilis strains. In the RDP/NCBI databases, out of a total of 2611 individual 16S rDNA sequences belonging to the 175 different species of the genus Bacillus, only 1586 have been identified up to species level. 16S rRNA sequences of Bacillus anthracis (153 strains), B. cereus (211 strains), B. thuringiensis (108 strains), B. subtilis (271 strains), B. licheniformis (131 strains), B. pumilus (83 strains), B. megaterium (47 strains), B. sphaericus (42 strains), B. clausii (39 strains) and B. halodurans (36 strains) were considered for generating species-specific framework and probes as tools for their rapid identification. Phylogenetic segregation of 1121, 16S rDNA sequences of 10 different Bacillus species in to 89 clusters enabled us to develop a phylogenetic frame work of 34 representative sequences. Using this phylogenetic framework, 305 out of 1025, 16S rDNA sequences presently classified as Bacillus sp. could be identified up to species level. This identification was supported by 20 to 30 nucleotides long signature sequences and in silico restriction enzyme analysis specific to the 10 Bacillus species. This integrated approach resulted in identifying around 30% of Bacillus sp. up to species level and revealed that B. subtilis strains can be segregated into two phylogenetically distinct groups, such that one of them may be renamed

Susceptibility of ostrinia nubilalis to bacillus thuringiensis var. kurstaki

Baseline susceptibility of European corn borer larvae, Ostrinia nubilalis (Hübner), to a commercial formulation of Bacillus thuringiensis var. kurstaki was determined for different strains in 7 geographic locations in the corn area of North Italy (Po Valley). For the bioassay 7 different doses, raised on a meredic diet, were used and the data was analysed with the Probit analysis. Differences in the susceptibility among the northern Italian populations of O. nubilalis were recorded, which can be attributed to natural variability. The ECB collected near Como and Torino was the most susceptible, respectively with LC50 of 0.23 and 0.57 mg/ml. The most tolerant ones were the larvae coming from the area of Vicenza (LC50 = 3.40 mg/ml). Baseline susceptibility was also determined for one population originally from Iowa but reared for more than 20 years in the laboratory and which had never experimented Bt. The LC50 calculated for this last one was compared to the Italian situation