Secondary Metabolites of Entomopathogenic Fungi, Biological Alternative for the Control of Agricultural Pests and Disease: Present and Perspectives

The use of entomopathogenic fungi (EF) in recent years has been highly effective against the different orders of insects considered pests of agricultural importance and their conidia have been commonly applied, but it has been reported that these are sensitive to the environmental conditions. For this reason, biopesticides products have been formulated based on secondary metabolites, recently. These biomolecules participate as biological control agent, such as: cyclic depsipeptides, amino acids, polyketides, polyphenols and terpenoids, affecting their morphology, life cycle and insect behavior. The use of secondary metabolites of entomopathogenic fungi opens the possibility of application in a more efficient way for the control of agricultural pests in a compatible with the environment and human health;therefore, it is important to know, analyzing the type of molecules, their effects, and their different methods of application.

Study of the Constraints of Millet Production (Pennisetum glaucum (L.) R. Br.) and the Peasant Perception of Biological Control in the Tahoua Region

Millet (Pennisetum glaucum (L.) R. Br.) is the Sahelian crop par excellence due to its adaptation to the particular production conditions in this region. Unfortunately, in recent years this crop has been threatened by very strong parasitic pressure and drought during the production period. The objective of this study is to analyze the main constraints of millet production and the solutions known to producers. A survey was carried out in November 2022 with a sample of 298 producers in five municipalities in the Tahoua region. The main constraints are drought and pressure from crop pests (locust, millet ear miner, floricultural insects) according to 57.9% of respondents. The millet ear miner is the most formidable pest according to 55% of respondents. Thus, the average yield obtained in a year of good production without the leafminer is 194 kg/ha and that obtained in a year of millet ear leafminer is around 27 kg to 43 kg/ha depending on the municipality. The yield obtained this last campaign after the attack of this leafminer varies from 64 to 77 kg/ha depending on the municipalities compared to a potential yield of over 1000 kg/ha. More than half of producers (58.1%) are unaware of the existence of biological control compared to only 12.5% who are aware of this alternative method. Work to popularize this technology is necessary in the five municipalities and the entire region in general.

Coffee Leaf Rust(Hemileia vastatrix)Disease in Coffee Plants and Perspectives by the Disease Control

Coffee Leaf Rust(CLR)is caused by Hemileia vastatrix in Coffea spp.It is one of the most dangerous phytopathogens for coffee plantations in terms of coffee productivity and coffee cup quality.In this review,we resume the problem of CLR in Mexico and the pathogenesis of H.vastatrix.The review abord plant-pathogen interactions which lead a compatible or incompatible interactions and result in CLR disease or resistance,respectively.The review abord Coffea spp.defense response pathways involved in H.vastatrix pathogenicity.Additionally,current measures to control H.vastatrix proliferation and germination were aborded focused on phytosanitary actions,and biological and chemical control.Finally,new trendlines to reduce the impact of CLR as nanoparticles and nanotechnology were analyzed.

Investigation of Kiwifruit Bacterial Canker Disease(Pseudomonas syringae pv.Actinidiae Takikawa) in Guanzhong Area of Shanxi Province and Its Biological Control

[ Objective ] The paper was to grasp the incidence characteristic of kiwifruit bacterial canker disease （Pseudomonas syrirtgate pv. actinidiae Takikawa） in Guanzhong area of Shanxi province, and to screen the endophytic actinomycetes with control effect against kiwifruit bacterial canker. [ Method] The incidence characteristics of kiwifruit bacterial canker disease in 65 orchards of 13 villages in guanzhong area of Shanxi province （Yangling, Zhouzhi, Meixian） were investiga- ted and analyzed. The antagonistic strains against Pseudomonas syringae were simultaneously screened from 242 endophytic actinomycetes, and carried out field dis- ease control test. [ Result] The average incidence rate of kiwifruit bacterial canker disease in guan^ong area of Shanxi province was 7.95% ; the variety of kiwifruit had dose relationship with incidence rate. 76 strains bad inhibition activity agmnst P. syringae, while the inhibition zone diameters of fermentation broths of 35 strains against P. syr/ngae were greater than 20 nun. TIASA5 strain had inhibition activity against four target bacteria; gCLA4 strain not only had inhibition activity against four target bacteria, but also had strong inhibition activity against 10 species of plant pathogenic fungi. Field test showed that the fermentation broth of gCLA4 strain had obvious control effect against kiwifruit bacterial canker disease, and the relative control effect at 21 d could reach 66.7%. [ Conclusion] The an- tibacterial spectrum of fermentation broth of gCLA4 strain is broad, so gCLA4 strain has the potential to be developed into a broad-spectrum agricultural fungicide. Key words Kiwifruit bacterial canker disease; Endophytic actinomycetes; Biological control; Field test

Identification and Characterization of Lysobacter enzymogenes as a Biological Control Agent Against Some Fungal Pathogens

Strain OH11, a Gram-negative, nonspore forming, rod-shaped bacterium with powerful antagonistic activity, was isolated from rhizosphere of green pepper in Jiangsu Academy of Agricultural Sciences of China and characterized to determine its taxonomic position. 16S rRNA gene sequence analysis revealed that strain OH11 belongs to the Gammaproteobacteria and had the highest degree of sequence similarity to Lysobacter enzymogenes strain C3 （AY074793） （99%）, Lysobacter enzymogenes strain N4-7 （U89965） （99%）, Lysobacter antibioticus strain （AB019582） （97%）, and Lysobacter gummosus strain （AB16136） （97%）. Chemotaxonomic data revealed that strain OHI 1 possesses a quinine system with Q-8 as the predominant compound and C15：0 iso, C17：1 iso ω9c as the predominant iso-branched fatty acids, all of which corroborated the assignment of strain OH11 to the genus Lysobacter. Results of DNA-DNA hybridization and physiological and biochemical tests clearly showed that strain OH11 was classified as Lysobacter enzymogenes. Strain OH11 could produce protease, chitinase, and β-1,3-glucanase. It showed strong in vitro antifungal activity against Rhizoctonia solani, Sclerotinia scletotiorum, and several other phytopathogenic fungi. This is the first report of identification and characterization of Lysobacter enzymogenes as a biological control agent of plant diseases in China.

Control of Lymantria dispar L. by biological agents

The experiment on control of Lymantria dispar L by using different kinds ofbiological measures, including nuclear polyhedrosis virus (NPV) of Lymantria dispar L., BtMP-342,sex-attractant as well as botanical insecticide, was carried out in the forest regions of InnerMongolia in 2003. Two concentrations (2.632x10^6 PIB ? ml^(-1) and 2.632x10^7 PIB ?ml^(-1)) of Lymantria dispar L. NPV were sprayed on the 2rd-instar-larvae of L. dispar and 70% and77.8% control effect were obtained respectively. BtMP0-342 was applied to the 3rd- and 4th-instarlarvae and the control effect was around 80%. The sex-attractant provided by Canada Pacific ForestryResearch Center also showed a good result in trapping L. dispar adults. The self-produced botanicalinsecticide, which was extracted from a kind of poisonous plant distributed in Daxing'an Mountains,China, exhibited a good control result in controlling the larvae of L. dispar, and 82% mortalitywas observed when spraying primary liquid of the botanical insecticide on the 3rd-5th-instar-larvaein lab.

Potential of Steinernema carpocapsae(Weiser) as a biological control agent against potato tuber moth, Phthorimaea operculella(Zeller)(Lepidoptera: Gelechiidae)

The entomopathogenic nematode,Steinernema carpocapsae,was evaluated for control of the potato tuber moth,Phthorimaea operculella,under laboratory conditions.We evaluated different concentrations of S.carpocapsae for control of 2 nd,3 rd,and 4 th instar P.operculella.The median lethal concentration(LC50)of S.carpocapsae infective juveniles(IJs)to 2 nd,3 rd and 4 th instar larvae of P.operculella was 200,363,181 IJs mL–1,respectively.With the extension of treatment time,the cumulative mortality increased for 2 nd,3 rd,and 4 th instar larvae and pupae of P.operculella.Fourth instars were the most susceptible for all observation periods.Therefore,our results suggest that S.carpocapsae could be an effective biological control agent for P.operculella.

Biological Control of Tortricidae in Tea Fields in Japan Using Insect Viruses and Parasitoids

Tea is a perennial and evergreen plant. Cultivated tea trees provide a habitat for insect pests and their natural enemies. In Japan, granuloviruses (GVs) have successfully controlled two of the most important pests of tea, Adoxophyes honmai and Homona magnanima (Tortricidae: Lepidoptera). The GVs are produced in vivo and a single application sustains pesticidal efficacy throughout a year, which encompasses 4 to 5 discrete generations of both species. A. honmai and H. magnanima also have various natural enemies, especially hymenopteran parasitoids. Such resident natural enemies also play a role in reducing the pest density in virus-controlled fields, but the effect of virus infection on parasitoids sharing the same host larva has not been well studied. Survival of one of the major parasitoids of A. honmai, Ascogaster reticulata (Braconidae: Hymenoptera), is reduced by virus infection of the host. Viruses, including GV and entomopoxvirus (EPV), and certain koinobiont endoparasitoids, including A. reticulata, are both known to regulate host endocrinology. However, the GV and EPV have distinct host regulation mechanisms, and consequently have different impacts on the survival of A. retuculata, when A. reticulata parasitizes a host that is infected with either GV or EPV. These additional effects on host regulation displayed by both viruses and parasitoids affect the outcome of virus-parasitoid interactions.

Fire Blight Disease Caused by Erwinia amylovora on Rosaceae Plants in Kyrgyzstan and Biological Agents to Control This Disease

Erwinia amylovora species were isolated from the blossoms, exudates, infected fruits, leaves and bent branches of diseased apple, pear and hawthorn trees, selected in the Chy, Osh and Jalal Abad regions. Biochemical and pathogenicity tests, alongside PCR analyses, were conducted to identify the local isolates of Erwinia amylovora. The alternative antagonistic microorganisms which combat bacterium E. amylovora were tested within in vitro and in vivo conditions. The results revealed the ability of Streptomyces antagonistic bacteria to decrease fire blight severity on pear and apple trees during the first stage of the fire blight disease in leaf tissues. Streptomyces strain C1-4 suppressed E. amylovora disease symptoms in the leaf tissues and excised apple and pear shoots. The incidence of fire blight on leaves was reduced by about 70% with two applications of bacterial antagonists. Further studies at different locations in Kyrgyzstan, using large scale application, would allow for stronger recommendations to be made, including studies and recommendations on their ability to prevent disease and to use them as main components in an integrated pest management program.

Integrated Biological Control of Tobacco Bacterial Wilt (<i>Ralstonia solanacearum</i>) and Its Effect on Rhizosphere Microbial Community

Biological control agents and soil amendments have been applied to control tobacco bacterial wilt, but the mechanism is not well-known. In the present study, a field experiment was performed to investigate the soil physicochemical properties, disease index (DI) and disease incidence of tobacco bacterial wilt, and rhizosphere microbial community. The results show that the control efficacy of single application of antagonistic bacteria and calcium cyanamide was 46.43% and 51.92%, respectively. While the combined control efficacy of antagonistic bacteria and calcium cyanamide was 65.79%. Besides, the combined application of antagonistic bacteria and calcium cyanamide could increase soil pH, total N alkaline N, and exchangeable Ca, which were negatively associated with the microbial diversity, soil-borne pathogenic microorganisms, and incidence of tobacco bacterial wilt. Additionally, the combination of antagonistic bacteria and calcium cyanamide can improve the proportion of some antagonistic microbial species, and these antagonistic microbial species were inversely associated with the DI of tobacco bacterial wilt. In conclusion: The integrated measure may influence soil microbial communities through enhancing soil physicochemical properties and rebuild healthy soil microbial community structure to mitigate tobacco bacterial wilt. The current study presented valuable insights into the mechanisms enhancing soil health in the integrated measure.

Integrated biological control of water hyacinths, Eichhornia crassipes by a novel combination of grass carp, Ctenopharyngodon idella (Valenciennes, 1844), and the weevil, Neochetina spp.

The efficacy of grass carp Ctenopharyngodon idella （Cyprinidae） and weevils Neochetina spp. （Curculionidae） to control the aquatic weed, water hyacinth, is investigated in a square net cage （happas） setting at a farm in Cuddalore District, South India. This novel combination of insects and fish is found to be superior to individual treatments for controlling the weed growth within ll0 d. The biomass of the weed, number of plants, percentage of flowered plants and chlorophyll contents were studied. The weed biomass is reduced from 5 kg （day 1） to 0.33 kg （day 110） when exposed to grass carp and weevils. The number of plants is reduced to 0.75 in grass carp and weevil exposed happas, while it is 741.5 in the control. The mean number of leaves per plant is also reduced. In addition, the chlorophyll a and b are significantly reduced in happas exposed to the combination of fish and insects when compared to the other treatments. Based on the results of this study, we consider the combined use of grass carp and weevils to be more efficient and sustainable for managing water hyacinths than the use of these organisms individually.

The Main Biological Hazards in Animal Biosafety Level 2 Facilities and Strategies for Control

Concern about the biological hazards involved in microbiological research, especially research involving laboratory animals, has increased in recent years. Working in an animal biosafety level 2 facility （ABSL-2）, commonly used for research on infectious diseases, poses various biological hazards. Here, the regulations and standards related to laboratory biosafety in China are introduced, the potential biological hazards present in ABSL-2 facilities are analyzed, and a series of strategies to control the hazards are presented.

Controlling Dengue: Effectiveness of Biological Control and Vaccine in Reducing the Prevalence of Dengue Infection in Endemic Areas

With the increased prevalence of dengue infection in tropical countries, concerned members of the public are now pressing their local health ministries to act immediately and effectively in managing the rising numbers of reported cases. This includes reviews of the methodologies and the effectiveness of current combative systems to find other possible novel approaches that might yield better results. One of those novel approaches is the integration of a parasite into mosquito vector, manipulating the parasite-host interaction to reduce the transmission of dengue in endemic hotspots. Another alternative is by Sanofi-Pasteur’s dengue vaccine that showed over 60.8% success rate in reducing severe dengue infection in children aged 9 - 16 during its final clinical implementation phase. This report will compare and contrast these two novel ideas to determine which of the approaches are more likely to be effective in the long run. The aspects covered will include the application, effectiveness, functionality, and problems with these approaches. The results could then be utilised by governments or organizations to select precise and effective methods in reducing the prevalence of dengue infections in their countries.

A Mechanism for Biological Control-Tempo-Spatial Synchronization of Natural Enemy and Insect Pest Population Dynamics at the Peak by Increase of Biodiversity

To reveal the mechanism of control of the Woolly Apple Aphid （Eriosoma lanigerum Hausm, WAA） by increase of biodiversity in the apple orchard, a consecutive two-year field trials were conducted in Yurman province, China. The results showed that the peak of natural enemy density curve in the cover cropping orchard was well temporally synchronized with that of the WAA density on trees, but did not synchronize with that of WAA density on trees in clean orchard. In addition, the frequency of natural enemies and WAA within a same sample in cover cropping orchard was obviously higher than that in clean cultivated orchard during the peak of population curves, indicating that natural enemies spatially synchronized with aphids. The density of WAA for the whole growing season in cover cropping orchard was much less than that in clean orchard. Therefore, the temporal-spatial synchrony of natural enemy and aphid populations was proposed as one of the mechanisms for aphid control in cover cropped orchards.

Research Progress on Occurrence and Biological Control of Citrus Blue Mold

Citrus blue mold. is an important disease of Citrus postharvest storage and transportation, and a major factor affecting the long-term storage of citrus. The pathogens, harm symptoms as well as the infection circulation of Citrus blue mold were introduced, followed by a summary of the progress on the biological control of Citrus blue mold in recent years, with the aim to provide references for the prevention and control of Citrus blue mold.

Primary Study on Biological Control Potential of Trichoderma harzianum TL-1

Trichoderma harzianum is a widely used biocontrol fungus. The growth promoting effect of strain Trichoderma harzianum TL-1 on tomato and pepper and its biological control effects against tomato seedling damping-off and pepper blight were investigated through pot experiments. The results showed that the stain TL-1 had significant promotion effect on growth of pepper and tomato in sterilized and natural soils. With the application dose of 3. 0 and 0. 5g/ pot,their dry weight were increased up to 46% and 150% compared with control,respectively. In addition,TL-1 had good control effects against tomato seedling damping-off and pepper blight. Compared with fungicide treatment,TL-1 treatment could control diseases for long term,without repeat occurrence of diseases.

Utilization of Fungi for the Biological Control of Insect Pests and Ganoderma Disease in the Indonesian Oil Palm Industry

Biological control is usually the first choice of control and prevention method for integrated pest management （IPM） strategies and has now been widely implemented by Indonesian oil palm plantations. Entomopathogenic fungus, i.e., Metarhizium anisopliae, Cordyceps militaris and Beauveria bassiana have been demonstrated to control renowned pests of oil palm. Metarhizium has been used to control Oryctes larvae and the mortality has ranged from 91.67% to 100% in laboratory and 7.4% to 88.75% in the field. Metarhizium has been applied in combination with a termite baiting system （TBS） to control termites in the field for preventive and curative action as well. In many oil palm plantations in Indonesia, Cordyceps has been used to reduce the field moth population ofSetothosea asigna. Application of Cordyceps within the oil palm circle was able to infect S. asigna pupae up to 80%. Meanwhile, Beauveria in an effervescent formulation was demonstrated to have better efficacy on Darna trima larvae. A significant finding on the biological control of basal stem rot disease （Ganoderma） was the isolation of Trichoderma sp. and Gliocladium sp.. The efficacy was conducted with promising result and techniques on the application of Triehoderma have been developed, i.e., hole-in-hole system, surgery and a mounding method. However, as the roots developed, Trichoderma was no longer able to protect the palm from Ganoderma. In spite of that, the use of Trichoderma still prolonged the life ofoil palms by up to 2-3 years. Another fungi belonging to vesicular arbuscular mychorrhiza （VAM） has been developed to control Ganoderma. The efficacy in the nursery showed promising results and the Ganoderma incidence remained low compared to the untreated control. Large scale field trials are ongoing. Challenges on the implementation of biological control in oil palm plantations are because of application and availability of biopesticides/natural enemies. Therefore, advances in research on the formulation of biological control agents are still needed.

Potential of the Integrated Control of Cucumber Root Rot Using Natural, Biological and Chemical Methods

Over the few last years, cucumber （Cucumis sativus L） root rot disease became common and inflicted marked losses to yield in Fayoum. Isolation trails from infected cucumber roots revealed that Thielaviopsis basicola and Fusarium moniliforme were the main cucumber root rot pathogens. The isolation trials from the rhizosphere of healthy cucumber plants revealed that two fungal isolates and four bacterial isolates had antagonistic effects against cucumber root rot fungi. All the tested biocontrol agents reduced the radial growth of all the tested root rots fungi in dual cultures. However, all culture filtrates of the tested biocontrol agent significantly reduced radial growth of all the tested pathogenic fungi, except that T. harzianum. Under field conditions, soil treatment with any of T. harzianum and B. subtilis BI and BF, significantly reduced percentages of infected plants and significantly increased percentages of survived plants and fruit yield compared with the control. Application of the commercial product Harpin protein （Messenger）~ product, as a resistance inducer at 0.3, 0.5 and l g/L significantly reduced the percentage of infected plants under greenhouse and field conditions. Field experiments indicated that the average percentage of infected plants after 90 days significantly decreased. The percentage of the survived plants as well as fruit yield increased by using integrated disease management （IDM） package, including the most effective treatments： tolerant cultivar, Trichoderma harzianum granules formula as soil treatment, Purging cassia plant extract, Harpin protein, and a half dose of Vitavax/thiram comparing with the treatment recommended by the Ministry of Agricultural or applied treatments individually.

Evaluation of Selected Chemical,Biological Fungicides,and Induced Resistance to Control White Rot(Sclerotium rolfsii Sacc.)on Tomato

Tomato(Solanum lycopersicum)is one of the most widely grown and consumed as fresh vegetable in the world.Sclerotium rolfsii Sacc.is one of the most destructive diseases and affects more than 500 plant species.This study was conducted for“Evaluation of Selected Chemical,Biological Fungicides,and Induced Resistant to Control White Rot(Sclerotium rolfsii Sacc.)on Tomato”.The experiment was conducted at the Royal University of Agriculture and divided into two sections.The first section conducted under in vitro condition consists of four treatments T1 control treatment,T2 copper hydroxide,T3 azoxystrobin+difenoconazole,T4 metalaxyl+mancozeb in Nagoya Laboratory,and had been starting from 11st May to 15th May 2020.Section two was conducted in a pot in net-house which started from 1st January 2019 to 19th February 2019 and 1st January to 19th February 2020 and arranged in RCBD(Randomized Completely Block Design)with six replications and nine treatments.T0 negative control,T1 inoculate but not treatment,T2 metalaxyl+mancozeb,T3 azoxystrobin+difenoconazole,T4 copper hydroxide,T5 acibenzolar-S-methyl,T6 chicken dung+Trichoderma harzianum,T7 acibenzolar-S-methyl+chicken dung+Trichoderma harzianum,T8 acibenzolar-S-methyl+metalaxyl+mancozeb.Data were collected on colonizing diameter in vitro.For pot experiment collected on disease incidence,incubation period Based on the result under in vitro condition metalaxyl+mancozeb highly inhibited germination of Sclerotium rolfsii when compared with another treatment.However,the application of copper hydroxide seems less effective compared with control.For in pot experiment,when treated after symptom of southern blight appeared to seem less effective on the severity of southern blight.In contrast,when applied as a protectant and curative when symptoms appeared only systemic fungicide azoxystrobin+difenoconazole significantly reduced the severity of Sclerotium rolfsii and delayed incubation period while compared with other treatments(p<0.05)and was followed by metalaxyl+mancozeb,whereas,copper hydroxide,acibenzolar-S-methyl,chicken dung+Trichoderma harzianum,acibenzolar-S-methyl+chicken dung+Trichoderma harzianum,acibenzolar-S-methyl+metalaxyl+mancozeb were less effective(p>0.05).Based on the three experiments we can assume that fungicide is more effective in reducing the growth of the pathogen and delaying the incubation period of fungal colonization when compared with biological control and induced resistance.

An Attenuated Strain of Cucumber Green Mottle Mosaic Virus as a Biological Control Agent against Pathogenic Viral Strains

Cucumber green mottle mosaic virus (CGMMV), a member of the Tobamovirus genus, causes a severe disease of cucurbits. In the Moscow region of Russian Federation, the incidence of infection on cucumber plants in greenhouses is high;however, the virus is poorly studied. In this work, the full-length genomes of two pathogenic MC-1 and MC-2 strains of CGMMV isolated from cucumber plants grown in greenhouses in the Moscow region and the attenuated VIROG-43M strain were sequenced. Comparison of VIROG-43M nucleotide sequence with those of the pathogenic strains revealed three missense mutations. Their role in attenuation is discussed. For the first time, in a number of trials conducted under laboratory conditions and in commercial greenhouses, the efficiency of the attenuated VIROG-43M strain as a biocontrol agent for cucumber plant protection resulting in significant yield gain was demonstrated. Phylogenetic analysis with 83 full-length CGMMV coat protein genes isolated in 16 different countries showed that Russian strains are related to isolates from Spain, Greece, USA and Israel.