S1070: The Working Group on Improving Microbial Control of Arthropod Pests

(Multistate Research Project)

Status: Active

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SAES-422 Reports

Annual/Termination Reports:

[01/14/2023] [01/25/2024]

Date of Annual Report: 01/14/2023

Report Information

Annual Meeting Dates: 11/21/2022 - 11/22/2022
Period the Report Covers: 11/30/2021 - 11/30/2022

Participants

1. Byron Adams Brigham Young University byron_adams@byu.edu
2. Paula Agudelo Clemson University pagudel@clemson.edu
3. Pasco Avery University of Florida pbavery@ufl.edu
4. Robert Behle USDA-ARS, Illinois robert.behle@usda.gov
5. Surendra Dara Oregon State University surendra.dara@oregonstate.edu
6. Emily Duren University of Florida duren.ebliss@ufl.edu
7. Julie Graesch BioWorks Inc. jgraesch@bioworksinc.com
8. Stefan Jaronski Virginia Tech University thebugdoc01@gmail.com
9. Navneet Kaur Oregon State University navneet.kaur@oregonstate.edu
10. Jimmy Klick Driscoll’s, California jimmy.klick@driscolls.com
11. Albrecht Koppenhöfer Rutgers University a.koppenhofer@rutgers.edu
12. Sriyanka Lahiri University of Florida lahiris@ufl.edu
13. Vijay K. Nandula REE-NIFA vijay.nandula@usda.gov
14. David Oi USDA-ARS, Florida david.oi@usda.gov
15. O. P. Perera USDA-ARS, Mississippi op.perera@usda.gov
16. Lorenzo Rossi University of Florida l.rossi@ufl.edu
17. David Shapiro-Ilan USDA-ARS, Georgia david.shapiro@usda.gov
18. Anamika Sharma Florida A & M University anamika.sharma@famu.edu
19. Colin Wong USDA-ARS, Georgia Colin.Wong@usda.gov
20. Shaohui Wu University of Georgia shaohui.wu@uga.edu

Brief Summary of Minutes

BUSINESS MEETING

Introductions: Anamika Sharma (2022 Member-at-large): Welcomed all and began with introductions. Attendees introduced themselves including a short introduction about their affiliation and work.

Minutes of 2021 (prepared by Shaohui Wu and Anamika Sharma): A copy of the 2021 minutes was circulated electronically prior to the meeting. A motion to approve the 2021 minutes was made by Anamika Sharma and was passed unanimously. Minutes of the 2022 meeting are required to be posted within 60 days.

NIFA administrators report (Dr. Vijay K. Nandula):

Dr. Nandula reported the deadlines for NIFA grant applications: https://www.nifa.usda.gov/grants/upcoming-request-applications-calendar

The Crop Protection and Pest Management (CPPM) RFA has been released, and the application deadline is February 13, 2023, 5 pm EST.

https://www.nifa.usda.gov/sites/default/files/2022-11/FY23-CPPM-RFA-508.pdf

A CPPM Applied Research and Development webinar on Thursday, December 01, 2022, 12-1 pm EST is available to guide through the application process (recording available from the web link below).

https://www.nifa.usda.gov/cppm-applied-research-development-rfa-technical-assistance-webinar

Specialty Crop Research Initiative (SCRI): Pre-application deadline on January 12, 2023.

https://www.nifa.usda.gov/sites/default/files/2022-10/FY23-SCRI-Pre-App-RFA-508_0.pdf

Biotechnology Risk Assessment Research Grants Program: Application deadline on January 19, 2023.

https://www.nifa.usda.gov/sites/default/files/2022-10/FY23-BRAG-RFA-508.pdf

Food and Agriculture Service Learning Program: Application deadline on December 08, 2022.

https://www.nifa.usda.gov/sites/default/files/2022-11/FY23-FASLP-RFA-508.pdf

Assistive Technology Program for Farmers with Disabilities (AgrAbility): Application deadline on January 19, 2023.

https://www.nifa.usda.gov/sites/default/files/2022-11/FY23-AgrAbility-RFA-508_0.pdf

From Learning to Leading: Cultivating the Next Generation of Diverse Food and Agriculture Professionals: Application deadline on December 14, 2022.

https://www.nifa.usda.gov/sites/default/files/2022-11/FY22-NG-RFA-508-MOD2.pdf

Food Safety Outreach Competitive Grant Program: Application deadline on February 16, 2023.

https://www.nifa.usda.gov/sites/default/files/2022-11/FY23-FSO-RFA-508.pdf

Methyl Bromide Transition Program: Application deadline on February 13, 2023.

https://www.nifa.usda.gov/sites/default/files/2022-11/FY23-MBT-RFA-508.pdf

AFR webinars are available for many of these programs and can be used to assist with application process.

Dr. Nandula mentioned that NIFA always looks for qualified reviewers. If anyone is interested in serving as a volunteer as a NIFA panelist, the information can be found via: https://prs.nifa.usda.gov/prs/volunteerPrep.do

Accomplishments/Outcomes 2021-2022

Symposium was organized ‘Incorporating Microbials into the Culture of IPM’

Session Date: Sunday, November 13, 2022

Session Time: 1:30 PM - 4:30 PM

Location: Meeting Room 122, Vancouver Convention Centre

Large acreage crops

Shaohui Wu (University of Georgia): Previously, the new strain of entomopathogenic fungus (EPF) Cordyceps javanica wf GA17 was isolated from whitefly epizootics in Georgia and was found to be highly virulent against whiteflies. The new fungal strain was then tested for field persistence and efficacy against whiteflies. However, without formulations the fungus lost over 90% viability within 24 h after application and the field efficacy was not satisfactory in cotton fields as expected. Hence, in collaboration with Robert Behle (USDA-ARS, Peoria, IL) and David Shapiro-Ilan (USDA-ARS), several formulations are being tested for improving the fungal persistence and field efficacy against whiteflies.

In addition, in collaboration with Israeli researchers, Drs. Dana Ment and Guy Mechrez (Volcani Center, Agricultural Research Organization), nanoparticle formulations for entomopathogenic nematodes (EPNs) were tested for their tolerance against ultraviolet (UV) radiation. A TiO2 based pickering emulsions showed excellent protection of EPNs from UV radiation, and the technology has potential to be used for pest management in any crop system. The nanoparticle formulation for EPNs is currently under a pending patent application. A paper has been published based on this research. The work was conducted under a BARD grant in David Shapiro-Ilan’s lab.

Stefan Jaronski (USDA-ARS retired; Virginia Tech): APHIS in Phoenix, Arizona conducted additional Metarhizium bait trials for control of grasshoppers in rangeland and generated much better results than in the past (control mortality was minimal). The technology used a wheat bran carrier and an experimental Metarhizium strain DWR-2009 (ARSEF 10343) with good spore production and shelf-life as bait, which achieved good results and demonstrated the feasibility of using wheat bran as carrier delivering the fungus. The behavior fever of grasshoppers inhibits the activity of most Metarhizium and Beauveria strains to a greater or less extant, except for M. acridum. The grasshoppers were exposed to the fungal bait in the field for 3 days using field cage and then incubated to observe the infection rates so as to avoid effects of behavioral fever, and thus measure extent of infection from the bait.

For Melanotus communis wireworm control in potato, Virginia Tech graduate student, Mika Pagani, presented her mesocosm data from 2022 at this year’s ESA meeting. She tested three granular baits that incorporated the commercial strain, Beauveria bassiana strain ANT-03 – coarse corn meal coated with a methyl carboxymethyl cellulose binder, spent solid substrate of millet, and similar spent rice substrate. In previous assays the strain showed moderate efficacy, best of all the currently commercial strains. In non-sterile soil, indigenous Metarhizium strains infected many wireworms. (Pagani isolated those Metarhizium strains and will deposit them into the ARSEF collection. For years, Kabaluk in Canada has also observed indigenous Metarhiziumthat interfered with wireworm field trials.) Pagani noted that among the granular baits the spent rice substrate was superior. Larval Tenebrio molitor (a more susceptible host) was tested in parallel soil pots with much greater efficacy. The rice spent substrate was also superior to others. Sharma asked if the indigenous Metarhizium strains were from Virginia, which was confirmed by Jaronski. Jaronski added that in a survey conducted two years ago the field soil contained up to a few thousand CFUs/g soil which interferes with field trials. The M. communis, attacking potato in Virginia, was not very susceptible to any commercial strains of EPF including strain PPRI5339 (BASF), ANT-03 (Anatis Bioprotection), GHA (Certis Bio) (no infection), ATCC74040 (Lallemand) (no infection), Met-52 (Lallemand) (very poor infection) when assayed with 10^7 CFUs/g soil. In an immersion bioassay conducted 2 years ago, none of the strains was effective by immersing insects in fungal suspension of 10^7 CFUs/ml; at 10^8 CFUs/ml, BASF strain produced 30% infection after two weeks; ANT-03 strain 50%; remainder 0% (spore viability was >90% for all strains). Pagani is going to look at spore attachment on the cuticle of wireworms, spore germination, penetration through the cuticle, in an effort to understand the overall low susceptibility of this species to the commercial strains.

Sharma brought up that in a previous meeting Jaronski or Graesch mentioned that in Africa it was planned to use airplane or drone to release fungal products for control of locusts. Graesch said the plan was to release the mixture of azadirachtin with Metarhizium acridum using aerial application to large areas, especially breeding grounds with locust nymphs to be more susceptible than adults. Jaronski added that the fungal application in Africa has been made via air spraying with mineral oil formulation (standard Green Muscle type of products), over several hundard thousand hecteres in east Africa. M. acridum successfully controlled the locusts but it is unavailable in the U.S.

Orchard Systems

Collin Wong (USDA-ARS, Byron, GA) (working with Shapiro-Ilan’s group): Entomopathogenic nematodes (EPNs) in Barricade gel formulation were sprayed for controlling the flatheaded apple borer, which attacks a variety of trees in US and Canada. The trial was conducted on maple trees in Tennessee and in walnut orchards in California. Initial tree damage data did not show significant treatment effects for reducing maple tree damage in nurseries; follow-up observations will be made to assess the larval reduction and mortality. In the laboratory tests, the EPNs were able to kill the larvae of this pest, and hence the field trials were pursued to evaluate the efficacy in controlling this pest.

Codling moth granular virus available on the market was sprayed against shuckworms in pecan. Also, Entrust was sprayed in pecan orchards for potential synergistic effects with other organic products.

Pasco Avery (University of Florida): Emily Duren from Avery’s team reported that laboratory bioassays were conducted to screen the products against the citrus red mite. Among them, BotaniGard Maxx was the best product, and BotaniGard ES also showed good efficacy. These two products were then sprayed on citrus trees growing in citrus under protection screen (CUPS) as well as not under protective screens, i.e. in open air. Trees inside and outside CUPS were naturally infested with citrus red mites and biocontrol assessment of the mites was determined by comparing reduction of the population per treatment over time. Leaf samples were taken from trees prior to spraying and on a weekly basis post-spray to check the persistence of fungal spores on leaves over time using leaf presses. In addition, coverslips were pinned onto leaves to check spore deposition on leaves. Spore persistence was assessed both inside and outside CUPS. Samples from inside CUPS showed the best results with longer spore persistence and the mite population was reduced; viable spores were still detected on leaves at 21 days post application. The research team has finished the 1^st trial for both the field (CUPS) and a 2^nd trial is planned in the spring, 2023.

Also, a similar test was conducted using 3 small young cohort lemon trees in mesh cages in the greenhouse per treatment. Destructive samplings of the 3 cohort trees were conducted to count eggs, nymphs and adults to check if the treatments reduce pest densities. The leaves have been collected for the 1^st trial for the greenhouse study; however, they have not been assessed. Another greenhouse trial as before will be conducted in the spring 2023 using mesh cages.

Compatibility studies were conducted on different ag-chemicals, insect growth regulators, insecticides, miticides from different companies with different Beauveria bassiana strains and also Cordyceps javanica. Shapiro-Ilan asked what strains of Isaria (Cordyceps) were tested and was told that it was the Apopka strain from PFR-97. Sharma followed to ask that if all strains tested are commercially available. Avery confirmed it and said that it is important to test the available products and let the growers know whether the products are compatible or not with the agrochemical.

Duren added that problems might occur with rain washing spores off leaves, and they plan to do a field study soon. Avery followed that they were going to expose the treated leaves to rain using a rain simulator to check if spores would adhere to the leaf surface after a rain event. Positive results have been obtained from a preliminary test, and an intern will be expected to join in February to take over the test with the rain simulator.

Behle asked Avery if he has had spore viability issues with commercial products. Avery said spore viability was always checked before use and had not had any problems. Duren added that compatibility was always checked before combining products. Behle asked if viability was checked by germination or CFUs; Avery said both because germination might be more reliable but CFUs reflect the spore coverage on leaf surface; they do leaf press on PDA with dodine or only PDA with antibiotics for washed leaves to count CFUs on underside or top side. Graesch asked if they checked mycosis on mites after infection; Duren confirmed it and said that it was checked individually in bioassays and BotaniGard Maxx worked the best, killed the insects in 2 days and had 90% mycosis. Avery added that the results were un-published but were presented by Duren who gave a presentation in 2022 ESA meeting on the mite project.

Lorenzo Rossi (University of Florida): Rossi collaborated with Pasco Avery on endophyte establishment in citrus rootstocks in the greenhouse with Beauveria bassiana. Three different methods were used to inoculate the fungus: foliar spray, seed coating, and soil drenching. The preliminary results showed that endophyte establishment depended on inoculation methods. The leaf spray was most successful; fungal endophytes were detected in leaves and moved to stem. But there were no good results in soil drench and root; seed treatment was ineffective. The B. bassiana strain used in the study is a commercial strain (BASF strain Velifer) available for growers to use. So far, the tests have been conducted on rootstocks only, and an intern will be recruited to work on grafted plants in the field next year. Behle asked about the seed treatment. Rossi answered that the seeds were soaked in spore suspension for 12 h and allowed to germinate and no CFUs were recovered on leaves, and he suggested that sculping the seed cuticle or increasing the soaking time might help.

In addition, Rossi worked with grower collaborators to increase soil quality with soil amendments, composts, mulch, and cover crop in citrus. Using cover crop for citrus is new because it is usually used for annual crop, while citrus trees and cover crops grow at the same time. Rossi was interested in evaluating if soil quality increases with EPF and other microbes in soil by using cover crops, or what cover crops act on soil, particularly in sandy soil. Shapiro-Ilan asked if cover crops increased the persistence of Beauveria in soil. Rossi answered that a technician in his lab is still looking into that and they also collected soil samples at different seasons (summer and winter) to check seasonality of EPF and other microbes. Shapiro-Ilan added that his lab used cover crops in pecan a while ago and observed enhanced fungal persistence by using cover crops.

Stefan Jaronski (USDA-ARS retired; Virginia Tech): Continued to work with APHIS people in Mission Texas on efficacy testing using whole trees and mesocosm trees of Jasmine or citrus in 40-gallon pots with two EPF strains, No-Fly (Cordyceps strain) and BioCeres (ANT-03 strain of Beauveria). Jaronski also comparing wettable powder and ES formulations of BioCeres products. BioCeres was superior to No-fly so far. Spore viability varied with products. However, neither fungus worked in summer in southern Texas due to heat (leaf temperatures 39-44^o C). Jaronski will continue to work with APHIS for another full season. Both No-Fly and BioCeres are far superior to PFR-97.

David I. Shapiro-Ilan (USDA-ARS, Byron, GA): Shapiro-Ilan’s lab tested fungal endophytes in pecan via seed treatment rolling with fungal spores or soaking in suspension, drenching and foliar spray. Successful establishment was confirmed in all approaches by molecular detection in plant tissues and plating. Wu participated in the study. They are continuing the endophyte project in pecan and interested in looking at the suppression of pecan aphids in endophytic plants.

EPN pheromones enhanced activity of nematodes in infection and dispersal. Previously, in the laboratory and greenhouse pheromones increased EPN efficacy in biocontrol. They recently finished the 2^nd year field trial against pecan weevil and found an increased level of control of the pest. Also, pheromone biomass extracts increased bioactivity of beneficial fungi and other microbial components in a peach system. A patent has been submitted on this microbiome effect (collaborating with Fatma Kaplan, Pheronym, Inc.)

The ambrosia beetle is a major pest problem in pecan and nursery crops. A postdoc Kyle Slusher has found good results in the laboratory showing the efficacy of different EPN species. The EPF will also be tested against the ambrosia beetle. The project is funded by a SCRI grant.

In addition, Shapiro-Ilan’s lab has been working with Tracy Leskey's group on EPNs against spotted lanternfly. Some efficacy was observed against early instars, but the study was still going on; Laura Nixon (postdoc) was leading this study.

Sharma asked about compatibility of biochar with nematodes. Shapiro-Ilan answered that a paper was published several years ago. Behle added the function of biochar, which may act as fertilizer enhancer or soil amendment; it was tested in turf for increasing moisture and nutrients in greens and tees, and a company is producing biochar as a bio-product (contact Steve Vaughn, USDA-ARS-NCAUR, Peoria, Illinois, email: steven.vaughn@usda.gov).

Shaohui Wu (University of Georgia): In collaboration with Drs. Steven Arthurs (BioBee) and Anna Wallingford (University of New Hampshire), and Shapiro-Ilan (USDA-ARS), laboratory and field studies were conducted to evaluate the persistence of a novel capsule formulation of the EPN Steinernema feltiae ENO2 strain (Nemaplus^®) with traditional aqueous applications. Nematode persistence was evaluated by baiting with the larvae of Tenebrio molitor at different times after inoculation. It was consistent in laboratory soil cup studies and field tests in pecan orchards that the capsule formulation persisted longer than the aqueous applications.

Metabolites of the EPN symbiotic bacteria Photorhabdus luminescens and Xenorhabdus bovienii were tested for toxicity to pecan aphids and lady beetles. It was found that both bacterial metabolites were highly toxic to pecan aphids while being safe to lady beetles. The results have been published, and a grant proposal has been submitted based on this project. Shapiro-Ilan added that this is the first time the bacterial metabolites were tested against the beneficial insects.

Ann Hajek (Cornell University): For Lycorma delicatula (spotted lanternfly), studies were conducted and published describing results on bioassays with entomopathogenic fungi against spotted lanternflies of different instars. Analysis was completed documenting the genetic diversity of Beauveria bassiana isolates from L. delicatula and comparing their virulence with a commercially available isolate and this paper has been submitted for publication. Results from bioassays with the poorly known entomophthoralean pathogen Batkoa major were analyzed and published along with a short description of this species. Additional studies with the biology and ecology of B. major were completed and analyzed and a paper is being written. During 2022, we continued epizootiological studies, including evaluating the infection of spotted lanternflies by EPF at field sites from June through November. Many fungal isolates remain to be identified and Koch’s postulates have been evaluated for some.

For Halyomorpha halys (brown marmorated stink bugs), infection of H. halys by the microsporidian Nosema maddoxi from before overwintering through after overwintering were conducted over the 2020-2021 and 2021-2022 winters. During these studies, they discovered another fungal pathogen of H. halys. It has been identified and Koch’s postulates have been conducted to prove pathogenicity and a publication is being prepared. A paper is also being prepared reporting fungal infection from before to after overwintering.

Small Fruits and Vegetables

Albrecht M. Koppenhӧfer (Rutgers University): Study on use of EPNs for control of plum curculio in highbush blueberries was concluded in 2021, and the results have been published. No additional research was conducted in 2022. Further research on EPN use against plum curculio including other components is planned for 2023 onwards.

Graesch asked if the EPNs used were commercial strains. Koppenhӧfer confirmed and the S. riobrave strain 355 was obtained from Shapiro-Ilan’s lab and it is produced by BASF; for quality consistency commercial strains were produced via wax worm larvae for one generation in the laboratory for experimental use.

David Shapiro-Ilan commented that the results that S. riobrave was most effective against plum curculio were consistent with his studies in peaches.

Stefan Jaronski (USDA-ARS retired; Virginia Tech): In grapes, Virginia Tech graduate student Jason Bielski did a 2^nd year test on treating egg masses of the spotted wing lanternfly with Beauveria bassiana. The newly hatched neonates sit on the surface of egg masses for several days while their cuticle hardens, which provides opportunities for the insects to be infected by the fungus previously applied to the surface of the fibrous egg mass. To overcome the challenge of keeping neonates alive long enough to become patently infected, the insects were incubated for only 3 days after exposure and then macerated in acid fuchsin stain to look for B. bassiana blastospores and hyphae, which worked moderately well. Additional nymphs were killed after the three day incubation period, by freezing, surface disinfected and then plated on selective agar to check for outgrowth of B. bassiana. The two methods correlated well in result findings. These may be used for other small insects which are difficult to keep alive long enough for the fungal infection. He tested 0.5x, 1x and 3x of the labeled rate of BotaniGard^® (1 lb/acre); 1 and 3 lb/acre rates achieved >50% infection. Also, ANT-03 strain of B. bassiana was tested in both formulations with similar results. The data were presented in the ESA meeting by Bielski, and a manuscript will be expected after completing another replication this spring.

Jimmy Klick (Driscoll’s): An outbreak of the cyclamen mite in strawberry was observed in organic production in Northern California. An epizootic of EPF in cyclamen mites was noticed. The fungus was slow growing on the media, and mass production has been explored for potential biocontrol. The taxonomy of the fungus is unknown and needs to be identified. Jaronski suggested it might be Hirsutella, a very slow growing fungus, or Isaria. He suggested sending the fungal culture to authority such as the current curator of the ARSEF collection of EPF for morphological and molecular identification.

There have been issues with supplies such as PFR-97^® and Mycotrol for growers. BoteGHA had steady supplies and achieved exciting results. In cages with beans, high mortality (80%) of the mite was induced after 3 or 4 days. Field trials were tested and had good results. Growers in Mexico lost personnel in nematode production, which is important for white grub control. Alternatives are needed, and mustard seed meal has been tested by incorporating in soil for controlling grubs. Different commercial products of EPNs were applied post planting using the appropriate rates (misused inappropriate rates in the past). There was severe problems with chilli thrips in berry production in FL. BotaniGard (B. bassiana) with azadirachtin combinations applied in the field weekly at standard rates caused 70% reduction in chilli thrips, which was better than Spear-T that did not work. The work on chilli thrips was in collaboration with Sriyanka Lahiri. Growers in Mexico have been trying to produce their own fungi for biocontrol.

Sharma asked if BoteGHA was used for mites or thrips. Klick replied that BoteGHA was tested for cyclamen mites and BotaniGard was used for chilli thrips. Graesch commented that ownership of BotaniGard has been switched from Bioworks to Certis Bio and it is no longer available through Bioworks. Jaronski added that BotaniGard and BoteGHA have the same material and formulation. Sharma commented that mites are not listed on BoteGHA label in California, and Graesch explained that is because California registration needs supporting data generated specifically in the region. Klick asked if there is potential to add mites on the label for California, and Graesch answered that mites are listed in the liquid formulation but not in the wettable powder formulation and there is potential to work with industry to generate the data for additional crops, pests and locations. Jaronski suggest contacting Certis if there is an interest for this product.

Graesch added that Certis may come up with new formulations, both liquid and dry, to be launched in 2023, and it is unknown whether they will phase out the old formulations. Jaronski said that the spore concentration in the products will be decreased, confirmed by Graesch that the liquid will drop from 11% to 2% and the wettable powder from 22% to 4%. Jaronski added that a Canadian company has picked up the registration of Naturalis^® and Met-52^®, which will be brought back to the market in Canada and probably also in the U.S. in 2023-2024. Avery asked if Met-52 will be produced in the same formulations; Graesch said from what she heard the formulations will be the same, liquid and granular (in rice). Shapiro-Ilan asked if that will be conidia-based formulations, which was confirmed by Jaronski. Graesch commented that it is easier to keep the same formulations for bringing products back to the market; although formulations will be expected to improve, it takes time and budget.

Sriyanka Lahiri (University of Florida): Lahiri has been working on strawberry pest management, especially the control of chilli thrips, an invasive pest of strawberries, with EPNs. Chilli thrips do not pupate in soil, with the entire life cycle above ground. The challenge was to find an EPN strain that effectively controls the pest in winter strawberry in FL. In a preliminary field study, S. feltiae and H. bacteriophora applied at 100 million infective juveniles (IJs)/acre twice (5 days apart) in December using a backpack sprayer caused significant suppression of the immature stages of chilli thrips. The test will be repeated this year. There is a potential to use EPNs for chilli thrips management in organic strawberry production. A number of growers are concerned that the data were generated from organic strawberry with no fungicide applications, which may be a limitation although they are interested in trying the EPNs in their commercial fields. Lahiri has been working with growers to simply release predators and EPNs recently in a field heavily infested with chilli thrips and will check the results of using EPNs in strawberry.

Graesch asked if any adjuvant was added for foliar application of EPNs, and if any laboratory test was conducted and infection was observed (chilli thrips have very small body size, < 1 mm). Lahiri said they have not done the laboratory test and the study was implemented in the field using the most promising species (S. feltiae and H. bacteriophora). Shapiro-Ilan commented that most EPN products are not desiccation tolerant but there is a chance that the nematodes would infect the thrips if they are close; nematode infection may occur even if the host size is small but with only a few IJs invade the host and produce for only one generation; the efficacy might not be caused by nematode infection directly but rather the function of the symbiotic bacteria. Shapiro-Ilan agreed with Graesch to look for adjuvants to anti-desiccance, such as Barricade, to protect EPNs in above-ground applications and improve the EPN persistence. The fungicides may not affect the activity of EPNs but can affect EPF like Beauveria, but applying fungicides and EPF at different times (e.g., 1 week apart) should minimize the effect. Klick asked if Barricade is organic compatible; Shapiro-Ilan said no, especially in fruit production. Graesch commented that other than Barricade, there are some common options of adjuvants such as capsules and Silwet L-77; Bioworks is also looking at different firegel formulations. BreakThru may be also used as an adjuvant for EPNs. Kaur was concerned about the potential effect of adjuvants for microbial biopesticides on pollinators. Graesch suggest applying them in the early morning or late evening when pollinators are not as active to minize the impact on pollinators, and also the time is most suitable for EPN/EPF applications because UV light (detrimental to EPN/EPF) is weak. Graesch mentioned a study on hibiscus bud weevil control with EPNs and Barricade (conducted by Alexandra Revynthi from University of Florida). Shapiro-Ilan added that the adjuvants sometimes may improve the dispersion of EPNs, but the big issue for applying nematode above-ground is UV and desiccation. Klick mentioned a previous study they conducted in China for using Metarhizium in drenching through irrigation against chilli thrips in blueberry; the fungal application was combined with predatory mites and significant reductions in chilli thrips numbers and damage levels were observed. Lahiri commented that there was no negative effect on predators; Beauveria may affect the predators; Klick said that was probably because the drenching application avoid most contact with predatory mites. Graesch said that could depend on the formulations used; the wettable powder formulation typically does not affect predatory mites, but the oil formulation may reduce the predatory mite population because of the oil component.

Urban and natural landscapes, rangelands, and nurseries

Albrecht M. Koppenhöfer (Rutgers University): Native persistent EPNs may provide longer lasting suppression of various turf insect pests than commercial EPN strains that have been selected for high virulence and effective mass-production at the cost of reduced field persistence. Surveys on golf courses recovered mostly the EPNs Steinernema carpocapsae and Heterorhabditis bacteriophora. Isolates were mixed within species to increase genetic diversity and used to inoculate field plots at 2 golf courses in June 2020 at a rate of 1.25 billion per ha for each species alone and half of that rate for each species in a combination treatment. Plots measured 20 m x 10 m and were half in the fairway and half in the rough. Populations of EPNs, annual bluegrass weevil (ABW), surface-active insects, and soil insects were monitored regularly from June 2020 through October 2022. Overall, EPN numbers were higher in the rough than the fairway and higher in treated than untreated plots for the EPN species the plots were treated with. ABW larval densities in the fairway in mid-June of the three years were 47-89% lower in plots treated with both EPN species than in untreated plots. Among surface-active insects, ABW adult numbers in the fairway were lower in plots treated with S. carpocapsae and the species combination than in untreated plots; black turfgrass ataenius numbers were lower in all EPN treatments than in untreated plots. White grub densities in the rough were lower in plots treated with H. bacteriophora than in untreated plots; similar reduction in the combination treatment were not statistically significant.

EPN, especially S. carpocapsae, can provide adequate control of ABW larvae on golf courses, but due to competition from several insecticides that are even more effective, may only be used on golf courses with insecticide resistant ABW populations. Another EPN species, S. riobrave, has shown superior control of several weevil species but has not been tested against ABW. In greenhouse tests, S. carpocapsae provided significant control even of highly insecticide resistant ABW but was less effective against populations with resistance ratios to the pyrethroid bifenthrin of 100x and 343x than against populations with 1-55x resistance ratios. We tested the field efficacy of S. carpocapsae and S. riobrave at three golf courses with ABW populations with bifenthrin resistance level of 55x, 100x, and 343x, respectively. There was no clear effect of resistance on the performance of the nematodes, albeit S. carpocapsae tended to be somewhat less effective against the 343x population. At the higher rate (2.5 billion nematodes per ha), S. carpocapsae provided 51 to 88% (average 68%) control compared to S. riobrave’s 8 to 57% (average 32%). Hence, S. riobrave does not seem to be an effective control option for ABW larvae whereas S. carpocapsae may be an effective alternative to insecticides against insecticide resistant ABW populations.

Shapiro-Ilan mentioned that several isolates were found in the field and asked if there is a plan to look at them further (hybridization, etc., to check which one is the best). Koppenhöfer said there were different isolates of S. scarabaei found in grubs in different areas; so far he has only looked at the virulence and efficacy in the laboratory and greenhouse, but the new isolates were not better than the original strain (21 years old) and there were no differences in IJ production. Another direction to look at is the persistence, and S. scarabaei persists for long time. Shapiro-Ilan suggest that if one isolate is superior to others in virulence, hybridization may be considered to screen them. Wong asked if the nematodes were passed through resistant pests for several generations to overcome insecticide resistance (e.g. pyrethroid resistance); Koppenhöfer said it was not done due to the difficulties in larval rearing. Kaur asked how the insecticide resistance ratios were determined in ABW, and if there was a susceptible population. Koppenhöfer replied that for laboratory tests ABW adults collected from the field were tested in Petri dishes at different concentrations of insecticides, and the greenhouse tests were conducted in grass pots for both adults and larvae using several different insecticides and ABW populations; there were susceptible population at Rutgers’ Hort Farm and two county golf courses that receive few insecticide applications (2x resistance). In addition, they also did field tests in golf courses with different resistance ratios targeting different life stages, adults, young and old larvae and obtained similar results; the research has been published. Sharma asked if spinosad was tested, and it was confirmed by Koppenhöfer. Avery asked about the synergistic interaction of EPN and EPF; Koppenhöfer said no work was done in his lab but there were some works published including the paper of Wu (mostly additive interactions), and the EPF need to be applied 2-4 weeks ahead of EPN. Klick mentioned his project on grub control (e.g., mustard seed meal with traps) and wondered about the regulatory of getting S. scarabaei across the border to Mexico, and if unlikely the possibility of conducting intensive survey on native EPNs in white grubs in Mexico. Koppenhöfer said a Canadian company is producing S. scarabaei in vivo with wax worms, but in samples he obtained, IJ numbers were much lower than claimed on label; the chance of getting the nematodes across the border depends on the regulation policy, and for surveying it may be worth trying to look for EPN-infected grubs, identify and produce nematodes with the assistance of specialists. Wu and Koppenhöfer further commented on the interaction of EPN and EPF.

Navneet Kaur (Oregon State University): For the project on endophyte‐mediated resistance response, Kaur has one graduate student who is looking at the endophytic fungus (Epichloe sp.) in cool season turfgrass systems. Pear Intasin is investigating a PCR detection method to explore the viability of these endophytes in perennial ryegrass cultivars being evaluated in the National Turfgrass Evaluation Project (NTEP). Once she identifies the cultivars that produce lolines, permines, and other ergot alkaloids, she will perform a series of greenhouse experiments to evaluate the feeding response of a relatively new cutworm species that is a pest of turfgrasses in different settings namely commercial fields of grass grown for seed, golf courses, and lawns for the past decade. They will establish the association of endophytic presence with insect response and evaluate endophyte mediated insect resistance for this project.

For the evaluation of native EPN strains against cutworm and sod webworm pests of grass grown for seed, in 2021, they identified three native EPN strains from commercial grass fields that are being maintained in vitro for infectivity trials against target insect populations. Infectivity trials are now completed using three different cutworm species and one sod webworm species. She did not get a chance for data analyses, but preliminary analyses looked promising. They included some Steinernema and Heterorhabditis strains from Dr. Shapiro's lab as well in these trials. This was the first time they performed insecticide compatibility assays of the native strains in Petri dishes and none of the insecticides that they used seemed to affect the EPN survival and infectivity. They are going to publish these results sometime in 2023.

Kaur presented extension talks on the above research findings to her growers/stakeholder groups on a regular basis. Some of these efforts are steps to identify viable alternatives to chlorpyrifos research. Kaur organized the symposium on "Incorporating Microbials into the Culture of IPM" during the joint Entomological Society of America meeting in Vancouver, BC. Four talks included perspectives of a commercial grower, extension educator, biopesticide industry representative, and Agriculture and Agri Food Canada researcher were presenters. This session was followed by a panel discussion. Challenges regarding pesticide registrations in North America and the need for an improvement of delivery methods were key discussion topics. Audience ~40 attendance taken three times during the session. Kaur and Dr. Surendra Dara were also invited to present the recent trends and history of our subdiscipline, invertebrate microbial control in "Recent Trends and Prospects for the Future in Several Key Plant‐Insect Ecosystems Sub‐Disciplines" symposium. Group effort for writing a review article in Annals of the Entomological Society of America was suggested.

Koppenhöfer commented that there have been different Oscheius species described but not with much potential in biocontrol and commercialization. Shapiro-Ilan said that the limitation has been largely on production and these nematodes do not have the close association with the symbiotic bacteria as other EPNs; they do show some biocontrol potential but not commercializing potential so far.

David Oi (USDA-ARS, Gainesville, FL) (from Steven Valles and David Oi): Solenopsis invicta virus 3 (SINV-3) is a virulent and specific pathogen of red imported fire ant, Solenopsis invicta, colonies. Under laboratory conditions, it consistently killed colonies and was highly transmissible. In 2022, we published the results of field introductions of this virus into fire ant nests where there were significant reductions of 57% in the size of nests and in the number of nests (7-fold decrease compared to controls) after 77 days. SINV-3 also persisted for over 20 months and spread to adjacent uninoculated colonies. A caveat to this study was that it was small, encompassing inoculations of twelve active fire ant nests. Nevertheless, this is the first documentation of a fire ant virus eliminating fire ant colonies under field conditions.

Surveys in June 2022 for the spread of SINV-3 in the Coachella Valley (Palm Desert area) of California from sites where it was introduced into fire ant nests in 2014, indicated the pathogen was detected at least a ¼ mile, or 400 m beyond release sites. This is 4 times further than the 100 m detection made in 2017. Note that introductions were made in a desert climate and fire ants are generally restricted to irrigated urban landscapes such as golf course and parks. Thus, fire ant populations are smaller than populations typically found in the southern U.S. and may be a factor in its limited spread.

Surveys for viral pathogens for the biocontrol of the little fire ant, Wasmannia auropunctata, an invasive, stinging ant were conducted in Florida, Hawaii, Argentina, and Australia. Six new and complete positive sense, single stranded RNA and one negative sense, single stranded RNA virus genomes were identified, sequenced, and characterized from transcriptomes of W. auropunctata collected in Argentina. The virus genome sequences were absent from the transcriptomes of W. auropunctata collected in Hawaii and Florida. Additional field surveys corroborated the absence of viruses in regions where the ant is invasive (USA and Australia). The replicative genome strand of four of the viruses (Electric ant polycipivirus 2, Electric ant solinvivirus, Electric ant virus 1, and Electric ant virus 2) was detected in W. auropunctata from Argentina, indicating that the ant is a host for these viruses. These represent the first viruses from the little fire ant.

Ann Hajek (Cornell University): For Deladenus and Sirex noctilio, Hajek’s research group had collected data on the parasitic nematodes infecting the Eurasian woodwasp Sirex noctilio from 2011-2019. At the same time, they amassed data on parasitoids from these same S. noctilio populations in 204 pine trees. Data were analyzed to evaluate the interactions between and within nematode and parasitoid populations, as well as describing the S. noctilio populations. Results have been analyzed and submitted for publication.

David I. Shapiro-Ilan (USDA-ARS, Byron, GA): The facility at USDA-ARS, SE Fruit and Tree Research Unit (Byron, GA) will get goats and sheep to look at the benefits of animals in pecan orchards and will also study the EPN efficacy against parasites such as ticks that vector diseases (cattle fever). In Texas, using the nematodes in small sprayers onto antelopes at the feeding station worked well against the cattle fever ticks (this work is in collaboration with John Goolsby, USDA-ARS, Texas). APHIS is interested, as there are regulatory concerns that although EPNs do not need registration the use on animals requires additional clearances. This is also part of the goal of this project on goats and sheep, which is to look at the possibility of controlling both internal and external parasites.

Discussions

The theme for the symposium at ESA 2023. The theme of ESA 2023 is ‘Insects and Influence: Advancing Entomology's Impact on People and Policy’. Shapiro-Ilan suggested formulating microbial agents for enhanced efficacy, as there have been new formulations explored for EPNs and EPFs since last discussion years ago and the formulation carrier may prevent the products from organic use; he recommended Wu to give a talk on EPN/EPF formulations for the relative research she has done. Sharma brought up the regulation policy. Graesch said that Bioworks has a regulatory department and may help with that or provide additional resources. Wu suggested inviting someone from EPA; Wong mentioned a retired EPA personnel in Iowa. Klick suggested inviting speakers on genetically modified (possibly a Bt expert). Kaur suggested delivery methods of microbials (bees -entomovectoring). Graesch also mentioned topic on drone release of entomopathogens. Further discussion on the symposium will be made in Janurary, 2023.

Collaboration projects. Sharma mentioned the use of social media, blogs and short extension articles in advocating microbial control. Wu also suggested education programs on entomopathogens as biocontrol agents since this area lacks public recognition. Insecticide resistance with microbial biocontrol was discussed.

Business discussion: The service term for officers (chair, vice-chair, member-at-large, secretary) is for two years. Since elections were made last year, there is no new election this year.

Paula Agudelo mentioned that official memberships for the project need to be registered on NIMSS. The official membership should be approved by the affiliated institutions (USDA or land-grant universities).

1 PM on November 22, 2022 meeting adjourned.

Minutes Attachment:

S1070 meeting report-2022-Jan 14-2023.pdf

Accomplishments

Symposium was organized ‘Incorporating Microbials into the Culture of IPM’ Session Date: Sunday, November 13, 2022 Session Time: 1:30 PM - 4:30 PM Location: Meeting Room 122, Vancouver Convention Centre

Publications

Microbial related publications (research and outreach) from group members (2021-2022): Arnoldi, M., Duren, E. B., Avery, P. B., and L. Rossi. 2022. Assessing the endophytic potential of a commercially available entomopathogenic Beauveria bassiana strain in various citrus rootstocks. Applied Microbiology 2(3), 561-571; <a href="https://doi.org/10.3390/applmicrobiol2030044">https://doi.org/10.3390/applmicrobiol2030044</a>  Avery, P. B., George J., Markle, L., Martini, X., Rowley, A. L., Meagher Jr., R. L., Barger, R. E., Duren E. B., Dawson, J. S., and R. D. Cave. 2022. Choice behavior of a generalist pentatomid predator when offered lepidopteran larvae infected with an entomopathogenic fungus. BioControl. 67: 201-211. doi.org/10.1007/s10526-021-10124-4 Behle, R.W., Wu, S., Toews, M.D., Duffield, K.R., Shapiro-Ilan, D.I. 2022. Comparing production and efficacy of Cordyceps javanica with Cordyceps fumosorosea. Journal of Economic Entomology. 115(2): 455-461. <a href="https://doi.org/10.1093/jee/toac002">https://doi.org/10.1093/jee/toac002</a> Dara, S. K.  2022.  Role of entomopathogenic microorganisms in IPM.  In Advances in integrated pest management technology: Innovative and applied aspects.  Ed. A. S. Tanda, Springer, pp. 145-155.  (Book chapter) Dara, S. K.  2022. Integrated pest management options for the western flower thrips in lettuce. eJournal of Entomology and Biologicals, 1 April, 2022. <a href="https://ucanr.edu/blogs/blogcore/postdetail.cfm?postnum=51928">https://ucanr.edu/blogs/blogcore/postdetail.cfm?postnum=51928</a> Dragone, N. B., J. B. Henley, H. Holland-Moritz, M. Diaz, I. D. Hogg, W. B. Lyons, D. H. Wall, B. J. Adams, and N. Fierer. 2022. Elevational Constraints on the Composition and Genomic Attributes of Microbial Communities in Antarctic Soils. mSystems 7:e01330-01321. Franco, A. L. C., B. J. Adams, M. A. Diaz, N. P. Lemoine, N. B. Dragone, N. Fierer, W. B. Lyons, I. Hogg, and D. H. Wall. 2022. Response of Antarctic soil fauna to climate-driven changes since the Last Glacial Maximum. Global Change Biology 28:644-653. Gaffke, A.M., Shapiro-Ilan, D., Alborn, H.T. 2022. Deadly scents: Exposure to plant volatiles increases mortality of entomopathogenic nematodes during infection of Galleria mellonella. Frontiers in Physiology 13:978359. doi: 10.3389/fphys.2022.978359 Glazer, I., Shapiro-Ilan, D.I., Sternberg, P.W. (Eds). 2022. Nematodes as Model Organisms. CABI, Wallingford, UK, 374 pp. DOI: 10.1079/9781789248814.0000 Glazer, I. and Shapiro-Ilan, D.I. 2022. Genetic improvement of beneficial organisms. In: Nematodes as Model Organisms (Glazer, I., Shapiro-Ilan, D.I., Sternberg, P.W., eds) pp. 346-364. CABI, Wallingford, UK. Gryganskyi, A.P., Golan, J., Hajek, A.E. 2022. Season-long infection of diverse hosts by the entomopathogenic fungus Batkoa major. PLOS ONE 17(5): e0261912. Gutt, J., E. Isla, J. C. Xavier, B. J. Adams, I.-Y. Ahn, C.-H. C. Cheng, C. Colesie, V. J. Cummings, H. Griffiths, I. Hogg, T. McIntyre, K. M. Meiners, D. A. Pearce, L. Peck, D. Piepenburg, R. R. Reisinger, G. K. Saba, I. R. Schloss, C. N. Signori, C. R. Smith, M. Vacchi, C. Verde, and D. H. Wall. 2022. Ten scientific messages on risks and opportunities for life in the Antarctic. Antarctic Environments Portal/SCAR. Hajek, A.E., Clifton, E.H., Stefanik, S.E., Harris, D.C. 2022. Batkoa major infecting the invasive planthopper Lycorma delicatula. J. Invertebr. Pathol. 194: 107821. <a href="https://doi.org/10.1016/j.jip.2022.107821">https://doi.org/10.1016/j.jip.2022.107821</a> Hazir, S., Kaya, H., Touray, M., Cimen, H. and Shapiro-Ilan, D. 2022. Basic laboratory and field manual for conducting research with the entomopathogenic nematodes, Steinernema and Heterorhabditis, and their bacterial symbionts. Turkish Journal of Zoology: Vol. 46(4) Article 1 (Review Article). <a href="https://doi.org/10.55730/1300-0179.3085">https://doi.org/10.55730/1300-0179.3085</a> Jackson, A. C., J. Jorna, J. M. Chaston, and B. J. Adams. 2022. Glacial Legacies: Microbial Communities of Antarctic Refugia. Biology (Basel) 11. Jorna, J., B. Vandebrink, I. D. Hogg, D. H. Wall, and B. J. Adams. 2022. Metabarcoding inventory of an Arctic tundra soil ecosystem reveals highly endemic communities. Polar Biology (in review). Kariuki, E. M., Lovo, E. E., Price T., Parikh, V., Duren, E. B., Avery, P. B., and C. R. Minteer. 2022. The consumption and survival rate of Lilioceris cheni (Coleoptera: Chrysomelidae) on air potato leaves exposed to Cordyceps javanica (Hypocreales: Cordycipitaceae). Florida Entomologist 105: 258-261. doi.org/10.1653/024.105.0313 Koppenhӧfer A.M., Kostromytska O.S., Ebssa L. 2022. Species combinations, split applications, and syringing to optimize the efficacy of entomopathogenic nematodes against Agrotis ipsilon (Lepidoptera: Noctuidae) larvae in turfgrass. Crop Prot. 155 1-8, 105927.  doi.org/10.1016/j.cropro.2022.105927 Kotliarevski, L., Cohen, R., Ramakrishnan, J. Wu, S., Mani, K.A., Amar-Feldbaum, R., Yaakov, N., Zelinger, E., Belausov, E., Shapiro-Ilan, D.  Glazer, I., Ment, D., and Mechrez, G. 2022. Individual coating of entomopathogenic nematodes with titania (TiO2) nanoparticles based on oil-in-water Pickering emulsion: A new formulation for biopesticides. Journal of Agricultural and Food Chemistry. 70, 13518−13527. https://doi.org/10.1021/acs.jafc.2c04424 Li, J., Li, Y., Wei, X. Cui, Y., Gu, X., Li, X., Yoshiga, T., Abd-Elgawad, M.M., Shapiro-Ilan, D., Ruan, W., Rasmann, S. 2022. Direct antagonistic effect of entomopathogenic nematodes and their symbiotic bacteria on root-knot nematodes migration toward tomato roots. Plant and Soil. In Press. l <a href="https://doi.org/10.1007/s11104-022-05808-4">https://doi.org/10.1007/s11104-022-05808-4</a>. Liebhold, A.M., Hajek, A.E., Walter, J.A., Haynes, K.J., Elkinton, J., Muzika, R.-M. 2022. Historical change in the outbreak dynamics of an invading forest insect. Biological Invasions 24: 879-889. Mbata, G.N., Li, Y., and Shapiro-Ilan, D.I. 2022. Evaluation of chemical and microbial control options for Pangaeus bilineatus (Say) (Hemiptera: Cydnidae) infesting peanut crop. Pest Management Science 78, 4719–4727. DOI 10.1002/ps.7092 Morales-Ramos, J., M. G. Rojas, and D. I. Shapiro-Ilan (Eds.). 2023. Mass Production of Beneficial Organisms: Invertebrates and Entomopathogens. Elsevier, 2nd Edition.  620 pp. O'Brien, K. M., E. L. Crockett, B. J. Adams, C. D. Amsler, H. J. Appiah-Madson, A. Collins, T. Desvignes, H. W. Detrich, 3rd, D. L. Distel, S. M. Eppley, B. W. Frable, N. M. Franz, J. M. Grim, K. M. Kocot, A. R. Mahon, T. J. Mayfield-Meyer, J. A. Mikucki, W. E. Moser, M. Schmull, C. A. Seid, C. R. Smith, A. E. Todgham, and G. J. Watkins-Colwell. 2022. The time is right for an Antarctic biorepository network. Proc Natl Acad Sci U S A 119:e2212800119. Olabiyi, D. O., Duren, E. B., Price, T., Avery, P. B., Hahn, P. G., Stelinski, L. L., and L. M. Diepenbrock. 2022. Suitability of formulated entomopathogenic fungi against hibiscus mealybug, Nipaecoccus viridis (Hemiptera: Pseudococcidae), deployed within mesh covers intended to protect citrus from huanglongbing. Journal of Economic Entomology 115: 212-223. doi.org/10.1093/jee/toab243 Oi, D. H., and S. D. Porter. 2022. Parasitoids and pathogens used against imported fire ants in the southern United States, pp. 252-265. In R. G. Van Driesche, R. L. Winston, T. M. Perring and V. M. Lopez (eds.), Contributions of Classical Biological Control to the U.S. Food Security, Forestry, and Biodiversity, vol. FHAAST-2019-05. Forest Health Assessment and Applied Sciences Team, USDA Forest Service, Morgantown, West Virginia, USA. Pick, D. A., Avery, P. B., Qureshi, J. A., Arthurs, S. P., and C. A. Powell. 2022. Field persistence and pathogenicity of Cordyceps fumosorosea for management of Diaphorina citri. Biocontrol Science and Technology 32: 151-162. doi.org/10.1080/09583157.2021.1976727 Pothula, S. K., and B. J. Adams. 2022. Community assembly in the wake of glacial retreat: A meta-analysis. Global Change Biology 28:6973-6991. Robinson, C. M., L. D. Hansen, X. Xue, and B. J. Adams. 2023. Temperature Response of Metabolic Activity of an Antarctic Nematode. Biology. Rodriguez-Saona, C., E. de Lange, and S. K. Dara.  2022.  Editorial: Advances in crop resistance for insect pest control.  Front. Agron. <a href="https://www.frontiersin.org/articles/10.3389/fagro.2022.845961/full">https://www.frontiersin.org/articles/10.3389/fagro.2022.845961/full</a> Schulte, N. O., A. L. Khan, E. W. Smith, A. Zoumplis, D. Kaul, A. E. Allen, B. J. Adams, and D. M. McKnight. 2022. Blowin’ in the wind: Dispersal, structure, and metacommunity dynamics of aeolian diatoms in the McMurdo Sound region, Antarctica. Journal of Phycology 58:36-54. Shapiro-Ilan, Garrigos Leite, L., Han, R. 2023. Production of entomopathogenic nematodes. Pp., 293316 in: Morales-Ramos, J., Rojas, G., and Shapiro-Ilan, D.I (eds.), Mass Production of Beneficial Organisms: Invertebrates and Entomopathogens. Amsterdam (2nd Edition): Academic Press. Shapiro-Ilan, D.I., Hazir, S. and Glazer, I. 2022. Entomopathogenic nematodes as models for inundative biological control. In: Nematodes as Model Organisms (Glazer, I., Shapiro-Ilan, D.I., Sternberg, P.W., eds) pp. 293-308. CABI, Wallingford, UK. Sousa A.L., Rodriguez-Saona C., Holdcraft R., Kyryczenko-Roth V., Koppenhöfer A.M. 2022. Entomopathogenic nematodes for the management of plum curculio in highbush blueberry. Biology 11, 45, 1-14.  doi.org/10.3390/biology11010045 Tian, C., Zhu, F., Li, X., Zhang, J., Puza, V., Shapiro-Ilan, D., Zhao, D., Liu, J., Zhou, J., Ding, Y., Wang, J., Ma, J., Zhu, X., Li, M., Li, J. 2022. Steinernema populi n. sp. (Panagrolaimomorpha, Steinernematidae), a new entomopathogenic nematode species from China. Journal of Helminthology 96, e57, 1–16. <a href="https://doi.org/10.1017/S0022149X22000426">https://doi.org/10.1017/S0022149X22000426</a> Valles, S. M., D. H. Oi, R. D. Weeks, K. M. Addesso, and J. B. Oliver. 2022. Field evaluation of Solenopsis invicta virus 3 against its host Solenopsis invicta. J Invertebr Pathol 191: 107767. Valles, S. M., D. H. Oi, J. B. Oliver, and J. J. Becnel. 2022. Characterization of Solenopsis invicta virus 4, a polycipivirus infecting the red imported fire ant Solenopsis invicta. Archives of Virology: <a href="https://doi.org/10.1007/s00705-00022-05587-00704">https://doi.org/10.1007/s00705-00022-05587-00704</a>. van Nouhuys, S., Harris, D.C., Stephen, F.M., Galligan, L.D., Hajek, A.E. 2022. Association of the native parasitic nematode Deladenus proximus with individuals and populations of the native woodwasp Sirex nigricornis. Agric. For. Entomol. 24(2): 237-246. Wakil, W., Tahir, M.; Ghazanfar, M.U., Qayyum, M.A., Yasin, M.; Maqsood, S., Asrar, M., Shapiro-Ilan, D.I. 2022. Microbes, Dodonaea viscosa and Chlorantraniliprole as components of Helicoverpa armigera IPM Program: A Three Region Open-Field Study. Agronomy, 12, 1928. <a href="https://doi.org/10.3390/agronomy12081928">https://doi.org/10.3390/agronomy12081928</a> Wakil, W., Usman, M., Pinero, J.C., Wu, S., Toews, M.D., and Shapiro-Ilan, D.I., 2022. Combined application of entomopathogenic nematodes and fungi against fruit flies, Bactrocera zonata and B. dorsalis (Diptera: Tephritidae): Laboratory cups to field study. Pest Management Science. 78, 2779–2791. <a href="http://dx.doi.org/10.1002/ps.6899">http://dx.doi.org/10.1002/ps.6899</a> Williams, L., Cherry, R., Shapiro-Ilan, D. 2022. Effect of host size on susceptibility of Melanotus communis (Coleoptera: Elateridae) wireworms to entomopathogens. The Journal of Nematology e2022-1. DOI: 10.2478/jofnem-2022-0033 Wong, C., Oliveira-Hofman, C., Blaauw, B., Chavez, D.J., Jagdale, G., Mizell, R.F., and Shapiro-Ilan, D. Control of peachtree borer (Synanthedon exitiosa) using the nematode Steinernema carpocapsae: optimization of application rates and secondary benefits in control of root-feeding weevils. Agronomy 12, 2689. <a href="https://doi.org/10.3390/agronomy12112689">https://doi.org/10.3390/agronomy12112689</a> Wu, S., Mechrez, G., Ment, M., Toews, M.D., Ananth, K., Amar Feldman, R., and Shapiro-Ilan, D.I., 2022. Tolerance of Steinernema carpocapsae infective juveniles in novel nanoparticle formulations to ultraviolet radiation.  Journal of Invertebrate Pathology 196, 107851. https://doi.org/10.1016/j.jip.2022.107851 Wu, S., Toews, M.D., Cottrell, T.C., Schmidt, J.M., Shapiro-Ilan, D.I. 2022. Toxicity of Photorhabdus luminescens and Xenorhabdus bovienii bacterial metabolites to pecan aphids (Hemiptera: Aphididae) and the lady beetle Harmonia axyridis (Coleoptera: Coccinellidae). Journal of Invertebrate Pathology 194, 107806.  <a href="https://doi.org/10.1016/j.jip.2022.107806">https://doi.org/10.1016/j.jip.2022.107806</a> Xue, X., B. J. Adams, and A. R. Dilman. 2022. A Draft Mitogenome of Plectus murrayi. Journal Of Nematology 54:20220035. Xue, X., B. N. Adhikari, B. A. Ball, J. E. Barrett, J. Miao, A. Perkes, M. Martin, B. L. Simmons, D. H. Wall, and B. J. Adams. 2023. Ecological stoichiometry drives the evolution of soil nematode life history traits. Soil Biology and Biochemistry 177:108891. Yanagawa, A., Krishanti, N.P.R.A., Sugiyma, A., Chrysanti, E., Ragamustari, S. K., Kubo, M., Furumizu, C., Sawa, S., Dara, S. K., Kobayashi, M. 2022. Control of Fusarium and nematodes by entomopathogenic fungi for organic production of Zingiber officinale. Journal of Natural Medicines. 76, 291–297. https://doi.org/10.1007/s11418-021-01572-4

Impact Statements

Date of Annual Report: 01/25/2024

Report Information

Annual Meeting Dates: 11/04/2023 - 11/04/2023
Period the Report Covers: 01/01/2023 - 12/31/2023

Participants

1. Shaohui Wu The Ohio State University wu.6229@osu.edu
2. Albrecht Koppenhöfer Rutgers University a.koppenhofer@rutgers.edu
3. Rogelio Trabanino Zamorano University rtrabanino@zamorano.edu
4. Vijay K. Nandula USDA-NIFA, Missouri vijay.nandula@usda.gov
5. Pasco Avery University of Florida pbavery@ufl.edu
6. Anamika Sharma Florida A&M University anamika.sharma@famu.edu
7. Jimmy Klick Driscoll’s, California jimmy.klick@driscolls.com
8. Marco Toapanta Agrinova, LLC marco.toapanta@hotmail.com
9. Eric Clifton BioWorks, Inc. eclifton@bioworksinc.com
10. Lorenzo Rossi University of Florida l.rossi@ufl.edu
11. Jinbo Wang USDA-APHIS, Maryland jinbo.wang@usda.gov
12. Julie Graesch BioWorks Inc. jgraesch@bioworksinc.com
13. Julien Levy Texas A&M University julienlevy@tamu.edu
14. Brian Lovett USDA-ARS, New York brian.lovett@usda.gov
15. Edwin Lewis University of Idaho eelewis@uidaho.edu
16. Abigail Kropf Iowa State University alkropf@iastate.edu
17. Jermaine Perier USDA-ARS, Georgia jermaine.perier@usda.gov
18. Kyle Slusher USDA-ARS, Georgia eddie.slusher@usda.gov
19. Colin Wong USDA-ARS, Georgia colin.wong@usda.gov
20. M. Eric Benbow Michigan State University benbow@msu.edu
21. Ann Hajek Cornell University aeh4@cornell.edu
22. Suzanne Wainwright Buglady Consulting sw@bugladyconsulting.com

Brief Summary of Minutes

S1070 Regional Research Project Agenda Saturday, November 4, 2023 Gaylord National Resort & Convention Center, Chesapeake B-C National Harbor, Forest Heights, MD 20745 and virtually Stefan Jaronski, Chair Julie Graesch, Vice-chair Anamika Sharma, Member-at-large Shaohui Wu, Secretary Paula Agudelo, Administrative Advisor

BUSINESS MEETING

Introductions: Julia Graesch (2023 Vice Chair): Welcomed all and began with introductions. Attendees introduced themselves including a short introduction about their affiliation and work.

Minutes of 2022 (prepared by Shaohui Wu): A copy of the 2022 minutes was circulated electronically prior to the meeting. A motion to approve the 2022 minutes was made by Shaohui Wu. Revisions were requested by Pasco Avery and the amended minute was passed unanimously. Minutes of the 2023 meeting are required to be posted within 60 days.

NIFA administrators report (Dr. Vijay K. Nandula):

Dr. Nandula provided link for NIFA-funded projects search

https://www.nifa.usda.gov/data/data-gateway

https://www.nifa.usda.gov/data/cris-current-research-information-system

Additional useful links can be found here:

Signing up for NIFA Updates

https://public.govdelivery.com/accounts/USDANIFA/subscriber/new?qsp=USDANIFA_2

Searching for Funding Opportunities

https://www.nifa.usda.gov/grants/funding-opportunities

List of NIFA’s Competitive RFAs

https://www.nifa.usda.gov/grants/request-for-application-list-rfa

Upcoming NIFA RFA Calendar

https://www.nifa.usda.gov/grants/upcoming-request-applications-calendar

Dr. Nandula also mentioned that NIFA always looks for qualified reviewers. If anyone is interested in serving as a volunteer as a NIFA panelist, the information can be found via:

https://prs.nifa.usda.gov/prs/preLogin.do?page=welcome

Dr. Nandula mentioned that grant application deadlines for this year would be similar to those of last year, as indicated in the RFAs. He suggested aligning objectives as closely as possible with the statements in RFAs and recommended discussing research ideas with NIFA project leaders before submitting an application. It was encouraged to participate in grant review panels (with potential involvement of postdocs), as this could help in building networks and acquiring grant-writing skills. Dr. Ann Hajek emphasized the importance of time allocation in grant writing and suggested that studying the structures of successful grants (available on the NIFA website) could serve as excellent models.

NEW PROJECT REVIEW AND PLANNING

Large acreage crops

Shaohui Wu (The Ohio State University): In previous work at the University of Georgia, a new strain of the entomopathogenic fungus (EPF) Cordyceps javanica wf GA17 was isolated from whitefly epizootics in Georgia and found to be highly virulent against whiteflies. The new fungal strain was then tested for field persistence and efficacy against whiteflies. The field efficacy of C. javanica wf GA17 was not different from the commercial strain Apopka97, and both showed low and short-term efficacy in the field. This was attributed to two main factors: (1) the short persistence of fungal spores (both conidia and blastospores), with over 90% of spores losing viability within 24 hours post-application; (2) inadequate contact of spores with the target hosts, as most spores landed on the upper leaf surface while whiteflies were predominantly active on the lower leaf surface. Future improvements will be necessary to enhance formulation and spore delivery (to the lower surface) to increase field persistence and efficacy. This work was conducted in collaboration with Dr. Robert Behle (USDA-ARS, retired) and was published in the Journal of Fungi. Wu mentioned that future research may be directed towards controlled environment applications, where fungal spores would be more protected from adverse environmental conditions, thus likely achieving better control.

Pasco Avery suggested using drone application technology to spray the fungus; Wu responded that drone application would not enhance spore delivery to the lower leaf surface. Pasco also mentioned that chemicals such as gossypol in cotton plants might inhibit fungal activities. Jermaine Perier suggested rotating crops. Eric Clifton asked if the application time would affect fungal persistence. Wu replied that one field trial was conducted in the late afternoon (6-7 pm), but the efficacy only lasted for about one week. In subsequent tests, spores were applied in mid-morning (9-10 am) to test fungal persistence.

Abigail Kropf (Iowa State University): In greenhouse applications with entomopathogenic nematodes (EPN), persistence was evaluated by baiting with wax worm larvae. Julie Graesch brought up indoor drone applications to reduce labor input.

NEW PROJECT REVIEW AND PLANNING

Orchard Systems

Lorenzo Rossi (University of Florida): The diversity of microorganisms, especially entomopathogenic fungi, was assessed in cover crops in different Florida citrus groves at various times of the year. This project is being conducted in collaboration with Pasco Avery.

Additionally, the establishment of endophytic fungi (Beauveria bassiana and Cordyceps javanica) was evaluated in citrus rootstocks. Positive results were observed in the greenhouse but need confirmation in the field. Methods of inoculation included leaf spray, soil drenching, and seed soaking. Among them, leaf spray was the most successful, with a 20% recovery in leaves but localized in leaves and stems only. Soil drenching was not effective, as most spores stayed near the soil surface. Seed treatment did not yield positive results. Future work will be directed towards grafted citrus plants.

Suzanne Wainwright mentioned new products in the market for mixing B. bassiana or Metarhizium brunneum granules in the soil profile to improve conidial distribution. This led to a discussion about black market microbial products. Julie Graesch mentioned that some entomopathogenic fungi (EPF) products from other countries were labeled as biostimulants to be exempted from registration. Jinbo Wang (USDA-APHIS) talked about the Plant Protection Act and emphasized that both the EPA and USDA have regulatory policies on microbial products.

Colin Wong (USDA-ARS, Byron, GA) (working with Shapiro-Ilan’s group): Reports on the control of flat-headed borers with EPN and on the control of hickory sharkworm with viruses.

Shapiro-Ilan’s lab worked on a method of nematode mass production for growers to produce their own nematodes. Waxworm larvae were inoculated by placing a few EPN-infected larvae in a box of healthy larvae, i.e., nematodes emerge from the dead insect and infect healthy hosts. The infected insect cadavers were placed on a mesh above hydrogels, and the emerging nematodes were then harvested from the gel. Suzanne Wainwright mentioned that a bad odor might be produced by dead insects during the nematode production and because of that some growers have stopped rearing their own nematodes.

Kyle Slusher (USDA-ARS, Byron, GA) (working with Shapiro-Ilan’s group): Reports on the control of ambrosia beetles with EPN.

Pasco Avery asked which parts of tree were attacked by ambrosia beetles; Slusher replied it was the tree trunk. The possibility of spraying trunks with EPF was discussed. Julie Graesch mentioned stopab.org and the ambrosia beetle grant. Ann Hajek asked if EPN were applied before or post infestation of ambrosia beetles; Slusher answered that it was post infestation application. Albrecht Koppenhöfer asked what would gel (nematode formulation) do to the ambrosia beetle adults as gels might create physical barriers to prevent adults from entering the tree trunk. Jimmy Klick brought up the ambrosia beetle problems in China, where EPF products were sprayed with water or adjuvants. Avery added that EPF products Met 52 (now formulated as Lalgard M52 OD), PFR-97™ 20% WDG (PFR-97), and BotaniGard® ES (BotaniGard) sprayed with water under the tree canopy persisted for 3 weeks.

Brian Lovett (USDA-ARS): The EPF products (BotaniGard and PFR-97) were evaluated for ambrosia beetle control in apple orchards. In laboratory bioassays, low fungal concentrations were ineffective and high concentrations killed insects. Pasco Avery asked what symbionts the ambrosia beetles carried.

Pasco Avery (University of Florida): Compatibility studies were conducted for PFR-97 mixed with various ag chemicals, no difference at 6 h. Table of compatible agrochemicals with EPF-based products was produced and handed out to interested people and groups at the Citrus Show in Ft. Pierce, FL. Avery also evaluated the potential of PFR-97 incidental drift from 10², 10³ 10⁵ and10⁶ spores ml^-1 affecting the air potato beetle used for biological control of air potato vines, and found that larvae were not affected by exposure to C. javanica spores contained in the product PFR-97. A prior compatibility study using adult air potato beetles also found that they were not affected after exposure to PFR-97 at the various concentrations listed above compared to the control (no spray). Results have been published in the Florida Entomologist journal.

Research has indicated that rainfall is a major deterrent for the persistence and subsequent field efficacy of EPF-based products over time. Therefore, research studies using a rain fast product were conducted to evaluate the persistence of C. javanica spores over time to adhere to the leaf surface of citrus plants using a rain simulator. Rain fast adjuvant product Ampersand® (used as food additive and OMRI labeled) was able to hold spores on the leaf surface even after major rain events. Julie Graesch asked if Ampersand would affect insects picking up spores on the sprayed surface; Avery said it had not been investigated yet.

Ann E. Hajek (Cornell University) An epizootic caused by fungal pathogens occurred among Halyomorpha halys, brown marmorated stink bugs, while they were overwintering, with infections also occurring after overwintering. We report that one of the two pathogens responsible was Colletotrichum fioriniae (Marcelino & Gouli) Pennycook, a species well known as a plant pathogen and endophyte and which has only previously been reported naturally infecting elongate hemlock scales, Fiorinia externa. To prove pathogenicity, H. halys adults challenged with conidia died from infections and the fungus subsequently produced conidia externally on cadavers.

The microsporidian, Nosema maddoxi Becnel, Solter, Hajek, Huang, Sanscrainte & Estep, infects brown marmorated stink bug, Halyomorpha halys (Stål) (Hemiptera: Pentatomidae), populations in North America and Asia and causes decreased fitness in infected insects. This host overwinters as adults, often in aggregations in sheltered locations, and variable levels of mortality occur over the winter. We investigated pathogen prevalence in H. halys adults before, during, and after overwintering. Population level studies resulted in detection of N. maddoxi in H. halys in 6 new US states, but no difference in levels of infection by N. maddoxi in autumn versus the following spring. Halyomorpha halys that self-aggregated for overwintering in shelters deployed in the field were maintained under simulated winter conditions (4°C) for 5 months during the 2021–2022 winter and early spring, resulting in 34.6 ± 4.8% mortality. Over the 2020–2021 and 2021–2022 winters, 13.4 ± 3.5% of surviving H. halys in shelters were infected with N. maddoxi, while N. maddoxi infections were found in 33.4 ± 10.8% of moribund and dead H. halys that accumulated in shelters. A second pathogen, Colletotrichum fioriniae Marcelino & Gouli, not previously reported infecting H. halys, was found among 46.7 ± 7.8% of the H. halys that died while overwintering, but levels of infection decreased after overwintering. These 2 pathogens occurred as co-infections in 11.1 ± 5.9% of the fungal-infected insects that died while overwintering. Increasing levels of N. maddoxi infection caused epizootics among H. halys reared in greenhouse cages after overwintering.

The prevalence of the microsporidium Nosema maddoxi in populations of brown marmorated stink bug (Halyomorpha halys) from three sites in Guria and eight sites in Samegrelo, Georgia was described. Investigations were conducted on adults collected in fall and spring from fall 2020 to spring 2022. The prevalence of infection differed by both region and season. Total infection of N. maddoxi in adults from the Guria region was significantly higher than the Samegrelo region. Infection levels in fall were higher than in spring in the Guria region, but inconsistent over seasons in the Samegrelo region. Infection levels did not differ by sex. We hypothesize that higher population densities and more adult aggregation in Guria compared with Samegrelo helps to explain the higher infection levels in Guria.

Hajek also mentioned the Insect Pathology Short Course has been offered every other year at Cornell University and can only take 24 students each time.

Edwin Lewis (University of Idaho): Larvae of hard pearl (?) were difficult to control with commercial EPN species. EPF was used to target adults. A trapping method was developed by employing fungal pathogens that did not kill insects rapidly in pheromone traps attracting males, so that infected adults were be able to move and transfer the pathogen to their mate. The larval stage lasted 3-4 years, and adults did not feed.

Pasco Avery asked where eggs were laid; Lewis answered that it was around the soil surface and larvae burrow down after hatching. Color had no impact on the trap that took effects by pheromone.

Shaohui Wu (The Ohio State University): In the previous work in Shapiro-Ilan’s lab, Wu explored formulations for EPN and EPF. In searching for novel formulations of EPNs, a liquid starch (Trueliving Heavy Spray Starch) and white kaolin clay provided excellent UV protection of the infective juvenile nematodes. Kaolin clay had been used in orchards in pest protection but had not been used for nematode application; it was found that 5% white kaolin clay had the best protection for sprayable applications. The liquid starch had been traditionally used for shirts after laundry, and this was the first time that its agricultural use had been explored; 50% concentration gave full protection of nematodes from UV radiation. However, in soil applications outdoors, Barricade and hydrogel performed better than liquid starch and white kaolin clay, probably because of the anti-desiccation effects involved. The work was in collaboration with Dr. George Mbata’s lab at Fort Valley State University and was published in the journal Biological Control. Also, in collaboration with Drs. Dana Ment and Guy Mechrez from Volcani Center, ARO in Israel, Wu evaluated the UV protection of novel nanoparticle formulations for both EPN and EPF. For EPN, SiO₂ based nanoparticle oil-in-water pickering emulsions were ineffective, while TiO₂ based nanoparticles protected nematodes from UV radiation, published in the Journal of Invertebrate. Pathology. Meanwhile, for EPF formulations, TiO₂ nanoparticles were effective in protecting M. brunneum conidia, but SiO₂ nanoparticles had no anti-UV effects. In addition, Wu also looked at the persistence of a novel capsule formulation of the EPN Sternernema feltiae ENO2 strain (Nemaplus® Depot) under laboratory and field conditions. In pecan orchards, when nematode products were applied in their recommended methods (traditional aqueous application via soil surface drenching; capsules mixed with soil in 2-inch depth of ditch), capsules persisted longer than aqueous applications. However, when both products were applied in soil ditch, they had similar field persistence, probably because nematodes were better protected by soil or weather conditions (frequent rainfalls) were favorable to nematode activities.

Jimmy Klick asked about the persistence of EPN in foliar spray and was told that the work was going to be conducted in collaboration with Dr. Mbata’s lab. Klick also asked about the cost of hydrogel; Wu shared that costs of various formulations were mentioned in the paper (Wu et al., Biological Control, 2023).

NEW PROJECT REVIEW AND PLANNING

Small Fruits and Vegetables

Julien Levy (Texas A&M University): Research focused on the interactions occurring between Solanaceae crops and Bactericera cockerelli (the tomato / potato psyllid). The potato psyllid is the vector of ‘Candidatus Liberibacter solanacearum’ (CLso), a phloem-limited pathogen associated with multiple economically important diseases of solanaceous crops. In the USA, the main crops affected by this pathogen are potatoes and tomatoes, causing “zebra chip” and “permanente del tomato”, respectively. In this project, we focused on the use of alternative strategies to control insect populations and in particular their effect on the transmission of the pathogen. To reduce pesticide use, bioinsecticides such as entomopathogenic fungi are becoming increasingly attractive to control pests The purpose of this experiment is to determine if exposure to entomopathogenic fungi such as Beauveria bassiana (GHA) and Phialemonium inflatum TAMU490, will affect the ability of psyllids to transmit CLso bacteria to tomato plants.

This research was conducted by undergraduate students’ part of the Aggie Research Program.

Test of two entomopathogenic fungus B. bassiana strain GHA, and P. inflatum TAMU490.

In the first semester the student learned to culture each fungus and prepare the fungal suspension. They learned how to grow and maintain insect colonies, collect insects, and sex tomato psyllids. They also practiced applying the fungus to the plant and to the psyllids. We conducted a pilot experiment by infesting both treated (sprayed with a 10⁷ spore /ml of fungus) and un-treated plants with adult tomato psyllids. One week prior to infestation plants were sprayed twice (and thereafter spraying continued twice a week). Plants were infested by placing 4 insects on one single leaf in a mesh bag, after one-week insects were removed from the mesh bag and mortality was measured. Three weeks post infestation the plant infection by CLso was measured by PCR. In this pilot experiment we did not notice any significant differences between non-treated plants and plants treated by spraying them with a fungus suspension.

We are planning to repeat this experiment with undergraduate students in 2025. A second experiment will test if entomopathogen-treated plants influence the insect host choice.

Jermaine Perier (USDA-ARS): reported on the compatibility of EPN with chemicals, IGR/adjuvant.

Suzanne Wainwright commented that change of chemistry in soil might affect EPN activities, as it was reported that EPN were found to be ineffective to kill fungus gnats, and temperature might have also played a role. Julie Graesch commented on foliar application of EPN. Edwin Lewis added that studies were conducted for smaller infective juveniles (IJs) to be produced in larger quantities. Jimmy Klick asked about breeding of EPN for more persistent and virulent strains. Julie Graesch brought up that between in-vivo and in-vitro production of EPNs, growers mostly used in-vitro produced EPNs, and nematodes carried in a sponge had relatively shorter shelf life. Lewis added that Shapiro-Ilan did some studies on in-vivo and in-vitro production of EPNs.

Jimmy Klick (Driscoll’s): Nematode products (Nemasys® from BASF) were used to control seed corn maggots in strawberry nurseries with 2 billion IJs per acre. Before planting, plants were soaked with nematodes, and nematodes persisted at least one month post planting. Imidacloprid was used in combination with EPN for control of the pest.

Direct application of the fungus Simplicillium sp. had no effect in controlling cyclamen mites. Control of chilli thrips was explored using azadirachtin, B. bassiana and C. javanica. A strain of B. bassiana isolated from grubs was used as a biopesticide in organic strawberry production in Mexico; Brian Lovett asked if the B. bassiana strain was identified. Control of ambrosia beetles in blueberry production with EPF was explored in China.

NEW PROJECT REVIEW AND PLANNING

Urban and natural landscapes, rangelands, and nurseries

Albrecht M. Koppenhöfer (Rutgers University): The annual bluegrass weevil (ABW) is a major insect pest of golf courses in eastern North America with widespread and broad-spectrum insecticide resistance. Previously, we had shown that entomopathogenic nematodes (EPNs) can provide good control of ABW larvae on golf course fairways. Due to the availability of numerous insecticides that are easier to use and mostly cheaper than EPNs, it is unlikely that golf courses will widely adopt EPN use. But golf courses that want to delay insecticide resistance or already have resistant ABW populations may consider using EPNs as an alternative to standard synthetic insecticides.

We tested the effect of insecticide resistance in ABW on the efficacy EPNs in greenhouse and field experiments. In a first greenhouse experiment we found that the efficacies of the EPNs Steinernema carpocapsae, Steinernema feltiae, and Heterorhabditis bacteriophora were only 9-19% lower against moderately resistant ABWs (resistance ratio (RR50) to the pyrethroid bifenthrin: 55x) than against highly susceptible ABWs (RR50: 2x). In a second greenhouse experiment, the efficacy of S. carpocapsae was 30-34% lower against highly (95x) and extremely (343x) resistant ABW than against highly susceptible (1-2x) ABWs. In field experiments conducted twice with ABW populations having resistance levels to bifenthrin of 55x, 95x, and 343x, S. carpocapsae provided similar control against the 55x and 95x populations, but 16-20% control lower against the 343x population.

In the latter field experiment, we also tested a not previously against ABW tested EPN species, Steinernema riobrave, that had provided excellent control against several other weevil species. However, against ABW S. riobrave was significantly less effective than S. carpocapsae, but its efficacy tended to increase with insecticide resistance level, albeit not statistically significantly so. And S. riobrave was as effective as S. carpocapsae against the 343x resistant ABW population.

Our findings suggest that EPNs can be used for ABW control and for insecticide resistance management against insecticide resistant ABW population, albeit their efficacy may be somewhat reduced against extremely resistant populations.

Ann E. Hajek (Cornell University)

Spotted Lanternflies. To evaluate the genetic diversity of Beauveria spp. infecting spotted lanternflies (SLF), in 2018- 2020 we collected SLF and nearby non-target insects killed by Beauveria spp. from 18 field sites in southeastern Pennsylvania. We identified 159 Beauveria isolates from SLF and six isolates from non-targets. Five isolates of B. bassiana and one isolate of B. brongniartii were identified from the non-targets. Based on sequence data from the nuclear B locus (Bloc) intergenic region, all the isolates from SLF were identified as B. bassiana, but there were 20 different strains within this species, grouped into two clades. Three B. bassiana strains (A, B, and L) were found in most field sites and were the most prevalent. Representative isolates for these three strains were used in laboratory bioassays and were compared to a commercialized B. bassiana strain (GHA). Strain B was inferior to A, L, and GHA against nymphs; strains A and L had greater efficacy than B and GHA against adults. We also quantified conidial production on SLF cadavers.

The Hajek lab has been studying entomopathogenic fungi killing spotted lanternflies (SLF; Lycorma delicatula), previously predominantly focusing on Batkoa major and Beauveria bassiana. During studies of the epizootiology of this system two additional species of entomopathogenic fungi killing SLF were collected in 2018 and 2020 and identified. Therefore, when three more probably entomopathogens were isolated from SLF adults in 2021, we decided to sample forested sites containing tree of heaven throughout 2022, especially targeting nymphal stages (June through July and early August). These studies generally included Koch’s postulates to confirm which fungi were pathogens. We have documented 19 species of entomopathogenic fungi killing spotted lanternflies. All are in the Order Hypocreales (Ascomycota) but are from numerous families. During the 2022 season little rain fell during the nymphal stages; most infection occurred among adults when there was more rainfall. However, lack of infection among nymphs is consistent with our previous studies years without a mid-summer drought, and the long-lived adults appear to be more susceptible as they age. Many of the 19 fungal species were not abundant and the most common pathogen was B. bassiana.

Batkoa major caused high levels of infection in SLF in 2017 and questions were addressed about the general biology of this poorly known fungal pathogen, using Galleria mellonella larvae exposed to conidial showers. Death of G. mellonella followed a diurnal cycle with most larvae dying within 4 h before or after the end of photophase. Time for initiation of rhizoid emergence also followed a diurnal rhythm and, on average occurred 3.6 h after host death. While B. major sometimes began producing rhizoids to attach cadavers to substrates while G. mellonella were alive (but moribund), often hosts were dead before rhizoids began emerging. On average, conidial discharge began 18.6 h after host death and was greater 4–8 h before the end of photophase, compared with 4–8 h after scotophase began. At 20 ◦C under high humidity, initiation of conidial discharge was 95% complete within 24 h after host death. To evaluate B. major activity by temperature, we tested percent conidial germination over 24 h from 5 to 35 ◦C. When showered onto water agar, all primary conidia produced secondary conidia. For 20 and 25 ◦C, at 3 h after showering ≥89% of primaries had produced and discharged secondaries and from 10 to 30 ◦C, secondaries were produced by over 75% of primary conidia within 12 h. When cover slips were placed over primary conidia to force production of germ tubes, germination was much slower, with >85% germination from 20 to 30 ◦C only by 24 h. Batkoa major therefore times host death and initiation of conidial discharge for night-time hours and conidial germination occurs within 24 h over a broad temperature range (10–30 ◦C).

With Dr. Stefan Jaronski and Jason Bielski, we asked to what extent contact with B. bassiana on different body parts impacted infection. To investigate contact via walking on spores, trunks of tree of heaven (TOH) were sprayed with BioCeres-WP (containing Beauveria bassiana strain ANT-03). Trees in the vicinity sprayed with water were controls. Spotted lanternflies from nearby trees were then caged over the sprayed areas for 3 days. SLF were then collected, transported to quarantine, and reared on TOH for 21 d. On the 21^st day, any SLF that had not died were frozen, after which these were placed at high humidity to promote fungal outgrowth if they had been infected. Data are presently being analyzed but B. bassiana killed almost all the SLF on sprayed trees and among the survivors, B. bassiana was present within their bodies. Infection among controls was minimal. We then applied B. bassiana GHA to wings versus abdomens of SLF adults; the question was when spraying B. bassiana on adults on a tree trunk, would the wings covering their bodies act as a shield that the fungus would not be able to infect. About 50% of SLF with abdominal inoculations became infected although about 25% of wing inoculations also became infected. Data are presently being analyzed further.

Sirex Woodwasps. Parasitic nematodes and hymenopteran parasitoids have been introduced and used extensively to control invasive Eurasian Sirex noctilio woodwasps in pine plantations in the Southern Hemisphere where no members of this community are native. Sirex noctilio has more recently invaded North America where Sirex-associated communities are native. Sirex noctilio and its parasitic nematode, Deladenus siricidicola, plus six native hymenopteran woodwasp parasitoids in New York and Pennsylvania, were sampled from 204 pines in 2011–2019. Sirex noctilio had become the most common woodwasp in this region and the native parasitoids associated with the native woodwasps had expanded their host ranges to use this invader. We investigated the distributions of these species among occupied trees and the interactions between S. noctilio and natural enemies as well as among the natural enemies. Sirex noctilio were strongly aggregated, with a few of the occupied trees hosting hundreds of woodwasps. Nematode parasitism was positively associated with S. noctilio density, and negatively associated with the density of rhyssine parasitoids. Parasitism by the parasitoid Ibalia leucospoides was positively associated with host (S. noctilio) density, while parasitism by the rhyssine parasitoids was negatively associated with density of S. noctilio. Thus, most S. noctilio come from a few attacked trees in a forest, and S. noctilio from those high-density trees experienced high parasitism by both the invasive nematode and the most abundant native parasitoid, I. l. ensiger. There is little evidence for direct competition between the nematodes and parasitoids. The negative association occurring between rhyssine parasitoids and I. l. ensiger suggests rhyssines may suffer from competition with I. l. ensiger which parasitize the host at an earlier life stage. In addition to direct competition with the native woodwasp S. nigricornis for suitable larval habitat within weakened trees, the large S. noctilio population increases the parasitoid and nematode populations, which may increase parasitism of S. nigricornis.

Dimorphic nematodes in the genus Deladenus have been used or are being considered for use in biological control of the invasive Eurasian woodwasp, Sirex noctilio, which threatens pine (Pinus spp.) trees. Deladenus species that are parasitic on Sirex can kill woodwasp eggs and occupy these same eggs for their own dispersal. These nematodes also have mycophagous phases that feed on the white rot fungal symbionts of Sirex, Amylostereum species. The mycophagous stage of the Hungarian strain of Deladenus siricidicola developed for control of S. noctilio in

Australia feeds exclusively on the species A. areolatum. The mycophagous stage of a North American Deladenus species being evaluated for biological control, D. proximus, feeds on either A. chailletii, or A. areolatum. Amylostereum species and strains associated with Sirex have differential impacts on survival and growth of these nematodes. We investigated whether differences in species and strains of Amylostereum infuence the numbers of Deladenus juveniles adjacent to cultures, as this would impact potential parasitism of Sirex. Fungal species or strain did not influence persistence of juveniles in the fungal vicinity although retention could be influenced by the fungal strain consumed by parents. Investigating D. proximus, we tested whether the most common invasive strain of A. areolatum associated with S. noctilio in North America (IGS D) impacted nematode growth, compared with the common native Amylostereum chailletii. Deladenus proximus increased very slowly when feeding on A. areolatum IGS D, compared with A. chailletii; when provided A. areolatum IGS D, 55 eggs were produced after 4 weeks compared with 8.1 × 10⁴ eggs after 2 weeks when A. chailletii was provided. In summary, behavior of Deladenus juveniles resulted in no or low avoidance of Amylostereum species.

Anamika Sharma (Florida A&M University): In the USA first decapitating phorid was released in 1997 to manage red imported fire ants. Six species (Pseudacteon culltellus, Pseudacteon curvatus, Pseudacteon littoralis, Pseudacteon nocens, Pseudacteon obtusus, and Pseudacteon tricuspis) of phorid flies have been released in North America since then. Along with phorids, microsporidia Kneallhazia (=Thelohania) solenopsae and Vairimorpha invictae and entomopathogenic fungus (EPF) Beauveria bassiana strain 447 were also considered potential biological control agents in the USA. Currently, we are surveying in north Florida (I-10 corridor) to analyze the establishment and efficacy of six species of phorid flies, two microsporidium and one EPF. Four phorid species were found after surveying 10 sites along the I-10 highway in the Northwest part of Florida from Jacksonville to Pensacola in urban and peri-urban locations. Samples with EPFs were obtained from some of the sites and analysis of samples is under process. Detection of the presence of microsporidium and EPF is under process through microscopy and Polymerase chain reaction.

Small undergraduate projects are being established to test the efficacy of IPM traps for urban insects including termites, fire ants, mosquitoes, and bed bugs. We are combining microbial with insecticides in the baiting system, and combination of microbial with pheromone traps is also being tested.

Jimmy Klick commented on commercial fungal baits.

Navneet Kaur (Oregon State University): Exploring endophyte-mediated resistance response against lepidopteran insect pests in cool-season turfgrass systems. The subterranean sod webworm, also known as cranberry girdler (Chrysoteuchia topiaria) is one of the most damaging insect pests in cool-season grass grown for seed crops in Oregon. Chemical control options are limited and require irrigation or rainfall for adequate insecticide incorporation to control C. topiaria larvae. Epichloë endophytes associated with cool-season turfgrass species and their mycotoxin profiles are well-documented in offering plant protection against invertebrates; these fungi may offer sustainable pest management tools. Our objectives were to characterize endophyte-mediated resistance to C. topiaria in 19 commercially available cultivars of tall fescue, perennial ryegrass, and fine fescue grown for seed in Oregon. Endophyte status (presence and viability) of fungal endophytes and their mycotoxin profiles were measured using polymerase chain reaction (PCR); liquid chromatography-tandem mass spectrometry (LC-MS/MS), respectively. No-choice assays were conducted in the laboratory to measure the impact of endophyte status on C. topiaria larvae in two separate no-choice experiments. Our results suggested that increased mortality of C. topiaria larvae (R2= 0.8526, experiment 1; R2= 0.6628, experiment 2) in tall fescue cultivars was most influenced by total peramine and ergot alkaloid, and total ergoline concentrations in experiment 1 and 2, respectively. However, no significant effect on insect mortality was found in the perennial ryegrass and fine fescue cultivars included in this study. Overall, these findings suggest a viable grass-endophyte association can be utilized as a sustainable alternative to foliar insecticides for C. topiaria management in tall fescue seed crops. A manuscript was submitted to the Journal of Applied Turfgrass Science or Crop Forage and Turfgrass Management in 2023, the decision of the journal is still pending.

The winter cutworm Noctua pronuba is another serious lepidopteran pest in Oregon’s grass seed production systems, golf courses, sports fields, and lawns. Currently, there are no cultural or biological control options for this pest; therefore, grass seed growers rely on prophylactic insecticide treatments to prevent crop damage. Noctua pronuba larvae typically emerge in grass seed fields in early winter when field conditions are too wet for equipment operation, and air temperatures are lower than desired for optimal insecticide efficacy. Utilizing symbiotic fungal endophytes in grass seed crops presents a promising non-chemical remedy due to their earlier discovered plant protection benefits against lepidopteran pests. One grad student, Pear Intasin was hired during Fall 2022 to work on these projects for her MS thesis project. Pear just completed her greenhouse experiments and currently is working on data analyses. Findings from this project will be presented at both scientific and regional grower meetings in near future. The information gained from this project will be used to identify the endophyte profiles for cool-season grass cultivars, which could be used for insect management in both turfgrass and grass seed production systems. We intend to continue our research efforts to understand endophyte insect interactions and build partnership with seed companies to incorporate beneficial endophyte populations for insect pest management in cool-season turfgrass species. Funding for this project was received from Oregon Grass Seed Commissions, Oregon State University’s Agricultural Research Foundation, and Western Sustainable Agriculture Research and Extension Grant Program.

Discussions

Stefan Jaronski proposed collaborative review project on biopesticides for crop pest (insect/ arthropod) management.

Theme for next ESA would be discussed via email communication.

Voting was made for the next two-year term:

Anamika Sharma was elected as the Chair

Shaohui Wu as Vice Chair

Pasco Avery and Eric Clifton as Member-at-Large

Lorenzo Rossi as Secretary and Treasurer

It was mentioned that joining the S1070 working group as a hatch/multi-state project could get travel fund for attending the annual meeting.

Minutes Attachment:

S1070 meeting report-Jan 25, 2024.pdf

Accomplishments

Accomplishments/Outcomes 2022-2023   Symposium organized: Tuesday, November 7, 2023, 9:00 AM – 12:00 PM Title Current and Future Impact of Microbial Control Agents; Room: Chesapeake B-C Organizers: Anamika Sharma– Assistant Professor, Entomology, Florida Agricultural & Mechanical University; Julie Ann Graesch– Technical Services Manager, Technical Services, BioWorks Inc; Stefan Jaronski – Retired, Entomology, USDA – ARS

Publications

Microbial related publications (research and outreach) from group members (2022-2023): <ol> <li>Arnoldi, M., Muschweck, L. Duren, E. B., Avery, P. B., and L. Rossi. 2023. Methods of</li> </ol> inoculating citrus rootstock cultivars with Beauveria bassiana as an entomopathogenic fungal endophyte. Proceedings of the Florida State Horticultural Society 135, 78   <ol start="2"> <li>Bardsley, C.A., Chasteen, K.S., Shapiro-Ilan, D., Bock, C.H., Niemira, B.A., and Govindara, D.K. 2023. Transfer of generic Escherichia coli and attenuated Salmonella enterica Typhimurium from the soil to the surface of in-shell pecans during harvest. Heliyon (In Press, Accepted 8-30-2023).</li> </ol>   <ol start="3"> <li>Chavez, A. V., Duren, E. B., Avery, P. B., Pitino, M., Duncan, R. E., Cruz, L.F., Carrillo, D.,</li> </ol> Cano, L. M., and R. D. Cave. 2023. Evaluation of spore acquisition, spore production, and host survival time of tea-shot hole borer. Euwallacea perbrevis, adults after exposure to four commercial products containing Beauveria bassiana. Insects 14, 726; doi.org/10.3390/insects14090726.   <ol start="4"> <li>Clifton, E.H., Castrillo, L.A., Jaronski, S.T., Hajek, A.E. 2023. Cryptic diversity and virulence of Beauveria bassiana recovered from Lycorma delicatula (spotted lanternfly) in eastern Pennsylvania. Frontiers in Insect Science: Focus on Spotted Lanternfly 3: 1127682. <a href="https://doi.org/10.3389/finsc.2023.1127682">https://doi.org/10.3389/finsc.2023.1127682</a></li> </ol>   <ol start="5"> <li>Dara, S. K.  Role of marketing and outreach for the success of entomopathogenic nematodes.  In Entomopathogenic nematodes as biological control agents. Eds. D. I. Shapiro-Ilan and E. E. Lewis, CABI.  Accepted. (Book chapter)</li> </ol>   <ol start="6"> <li>Dara, S. K.  Biopesticides: market, use, research and education needs, and future.  Submitted to West Coast Nut. (Trade journal article)</li> </ol>   <ol start="7"> <li>Dara, S. K.  Entomopathogenic fungi-based biopesticides contribute to more than pest management.  CAPCA Adviser 26(6): 48-51. (Trade journal article)</li> </ol>   <ol start="8"> <li>Geisert, R.W., M.P. Huynh, A.E. Pereira, D.I. Shapiro-Ilan, and B.E. Hibbard. 2023. An improved bioassay for the testing of entomopathogenic nematode virulence to the Western corn rootworm (Diabrotica virgifera virgifera): With focus on neonate insect assessments. J. Econ. Entomol. 116: 726–732. <a href="https://urldefense.com/v3/__https:/doi.org/10.1093/jee/toad052__;!!KGKeukY!0QFvn_JIb3FIu9u1wMvMkd_3hUSqZVAs1-S9QlpRHxLFGI81BSf5dD66eKJIoj9sczrUnD9OsrAQPeMsnPBhyhzNWspn$">https://doi.org/10.1093/jee/toad052</a>.</li> </ol>       <ol start="9"> <li>Gonzalez, J.B., Lambert, C.A., Foley, A.M., Hajek, A.E. 2023. First report of Colletotrichum fioriniae infections in brown marmorated stink bugs, Halyomorpha halys. J. Invertebr. Pathol. 199: 107939. <a href="https://doi.org/10.1016/j.jip.2023.107939">https://doi.org/10.1016/j.jip.2023.107939</a></li> </ol>   <ol start="10"> <li>Hajek, A.E., Brandt, S.N., Gonzalez, J.B., Bergh, J.C. 2023. Entomopathogens infecting brown marmorated stink bugs before, during, and after overwintering. J. Insect Science 23(3): 8; 1-8. DOI: <a href="https://doi.org/10.1093/jisesa/iead033">1093/jisesa/iead033 </a></li> </ol>   <ol start="11"> <li>Hajek, A.E., Everest, T.A., Clifton, E.H. 2023. Accumulation of fungal pathogens infecting the invasive spotted lanternfly, Lycorma delicatula. Insects 14: 912. <a href="https://doi.org/10.3390/insects14120912">https://doi.org/10.3390/insects14120912</a></li> </ol>   <ol start="12"> <li>Hajek, A.E., Harris, C.H. 2023. Diurnal patterns and conidial dynamics of Batkoa major, a generalist entomophthoralean pathogen. Fungal Ecology 65: 101278. <a href="https://doi.org/10.1016/j.funeco.2023.101278">https://doi.org/10.1016/j.funeco.2023.101278</a></li> </ol>   <ol start="13"> <li>Kereselidze, M., Pilarska, D., Guntadze, N., Linde, A., Hajek, A. 2023. Prevalence and distribution of Nosema maddoxi infection in Halyomorpha halys, an invasive pest of hazelnut orchards in Georgia. Acta Horticulturae 1379.64. <a href="https://doi.org/10.17660/ActaHortic.2023.1379.64">17660/ActaHortic.2023.1379.64</a></li> </ol>   <ol start="14"> <li>Koppenhӧfer A.M., Kostromytska O.S., Ebssa L. 2022. Efficacy of entomopathogenic nematodes against black cutworms and split applications and syringing to optimize their performance. Golf Course Management, September 2022, 80-84.</li> </ol>   <ol start="15"> <li>Leite, L.G., Chacon-Orozcoa, J.G., Shapiro-Ilan, D.I., Baldo, F.B., Cardoso, J.M. 2023. Effects of temperature for optimizing production and storage of Steinernema rarum in a novel biphasic process, and efficacy of the nematode against Sphenophorus levis. Biological Control 187, 105381. <a href="https://urldefense.com/v3/__https:/doi.org/10.1016/j.biocontrol.2023.105381__;!!KGKeukY!0QFvn_JIb3FIu9u1wMvMkd_3hUSqZVAs1-S9QlpRHxLFGI81BSf5dD66eKJIoj9sczrUnD9OsrAQPeMsnPBhylynha3K$">https://doi.org/10.1016/j.biocontrol.2023.105381</a></li> </ol>   <ol start="16"> <li>Li, J., Li, Y., Wei, X. Cui, Y., Gu, X., Li, X., Yoshiga, T., Abd-Elgawad, M.M., Shapiro-Ilan, D., Ruan, W., Rasmann, S. 2022. Direct antagonistic effect of entomopathogenic nematodes and their symbiotic bacteria on root-knot nematodes migration toward tomato roots. Plant and Soil. 484(1-2):1-15.  <a href="https://urldefense.com/v3/__https:/doi.org/10.1007/s11104-022-05808-4__;!!KGKeukY!0QFvn_JIb3FIu9u1wMvMkd_3hUSqZVAs1-S9QlpRHxLFGI81BSf5dD66eKJIoj9sczrUnD9OsrAQPeMsnPBhyqI5VV7A$">https://doi.org/10.1007/s11104-022-05808-4</a></li> </ol>   <ol start="17"> <li>Morris, E.E., Harris, D.C., Shen, A., Vermeylen, F., Hajek, A.E. 2023. Impact of Amylostereum (Basidiomycota: <a href="https://en.wikipedia.org/wiki/Russulales">Russulales</a>) diversity on Deladenus (Nematoda: Neotylenchidae) behavior and fitness. Biological Control 179: 105147. Doi: 10.1016/j.biocontrol.2022.105147</li> </ol>   <ol start="18"> <li>Neal, A. S., Avery, P. B., and Cave, R. D. 2023. Survival time, mortality and feeding damage of</li> </ol> adult Myllocerus undecimpustulatus undatus (Coleoptera: Curculionidae) exposed to biopesticides in laboratory bioassays. Applied Microbiology 3(2), 388-399; https://doi.org/10.3390/applmicrobiol3020027.       <ol start="19"> <li>Oliveira-Hofman, C., Steffan, S., Shapiro-Ilan, D. 2023. A sustainable grower-based method for entomopathogenic nematodes production. Journal of Insect Science. 23, 4; 1–5. <a href="https://urldefense.com/v3/__https:/doi.org/10.1093/jisesa/iead025__;!!KGKeukY!0QFvn_JIb3FIu9u1wMvMkd_3hUSqZVAs1-S9QlpRHxLFGI81BSf5dD66eKJIoj9sczrUnD9OsrAQPeMsnPBhyiCqYwRt$">https://doi.org/10.1093/jisesa/iead025</a></li> </ol>   <ol start="20"> <li>Perier, J.D., Kaplan, F., Lewis, E.E., Alborn, H., Schliekelman, P. Toews, and Shapiro-Ilan, D. 2024. Enhancing entomopathogenic nematode efficacy with pheromones: A field study Targeting the pecan weevil. Journal of Invertebrate Pathology. In Press (Accepted 1-23-24).</li> </ol>   <ol start="21"> <li>Rodriguez-Saona, C. and  K. Dara.Entomopathogenic nematodes in berry crops.  In Entomopathogenic nematodes as biological control agents. Eds. D. I. Shapiro-Ilan and E. E. Lewis, CABI.  Accepted. (Book chapter)</li> </ol>   <ol start="22"> <li>Slusher, E.K., Lewis, E., Stevens, G., Shapiro-Ilan, D. 2024. Movers and shakers: Do nematodes that move more invade more? Journal of Invertebrate Pathology. (IN PRESS, Accepted 1-16-2024).</li> </ol>   <ol start="23"> <li>Stevens, G., Erdogan, H., Pimentel, E., Dotson, J., Stevens, A., Shapiro-Ilan, D., Kaplan, F., Schliekelman, P., Lewis, E. 2023. Group joining behaviours in the entomopathogenic nematode Steinernema glaseri. Biological Control.  In Press (Accepted 3/20/2023).</li> </ol>   <ol start="24"> <li>Toledo, P.F.S. Phillips, K., Schmidt, J.M., Bock, C.H., Wong, C., Hudson, W.G., Shapiro-Ilan, D., Wells, L., Acebes-Doria, A.L. 2023. Canopy hedge pruning in pecan production differentially affects groups of arthropod pests and associated natural enemies. Crop Protection (In Press accepted 11/7/23).</li> </ol>   <ol start="25"> <li>van Nouhuys, S., Harris, D.C., Hajek, A.E. 2023. Population level interactions between an invasive woodwasp, an invasive nematode and a community of native parasitoids. Neobiota 82: 67-88. doi: 10.3897/neobiota.82.96599</li> </ol>   <ol start="26"> <li>Wong, C., Oliveira-Hofman, C., Blaauw, B., Chavez, D.J., Jagdale, G., Mizell, R.F., and Shapiro-Ilan, D. 2022. Control of peachtree borer (Synanthedon exitiosa) using the nematode Steinernema carpocapsae: optimization of application rates and secondary benefits in control of root-feeding weevils. Agronomy 12, 2689. <a href="https://urldefense.com/v3/__https:/doi.org/10.3390/agronomy12112689__;!!KGKeukY!0QFvn_JIb3FIu9u1wMvMkd_3hUSqZVAs1-S9QlpRHxLFGI81BSf5dD66eKJIoj9sczrUnD9OsrAQPeMsnPBhylLOinQQ$">https://doi.org/10.3390/agronomy12112689</a></li> </ol>   <ol start="27"> <li>Wakil, W., Gulzar, S., Prager, M., Shapiro-Ilan, D. 2023. Efficacy of entomopathogenic fungi, nematodes and spinetoram combinations for integrated management of Thrips tabaci: a two-year onion field study. Pest Management Science 79: 3227–3238.</li> </ol>   <ol start="28"> <li>Wu, S., Li, Y., Toews, M.D., Mbata, G., Shapiro-Ilan, D.I. 2023. Novel formulations improve the environmental tolerance of entomopathogenic nematodes. Biological Control. 186, 105329. <a href="https://doi.org/10.1016/j.biocontrol.2023.105329">https://doi.org/10.1016/j.biocontrol.2023.105329</a></li> </ol>     <ol start="29"> <li>Wu, S., Toews, M.D., Behle, R.W., Barman, A., Shapiro-Ilan, D.I. 2023. Post-application persistence and field efficacy of a new strain of Cordyceps javanica against the sweetpotato whiteflies, Bemisia tabaci. Journal of Fungi. 9(8), 827. <a href="https://doi.org/10.3390/jof9080827">https://doi.org/10.3390/jof9080827</a></li> </ol>   <ol start="30"> <li>Wu, S., Mechrez, G., Ment, D., Toews, M.D., Mani, K.A., Feldbaum, R.A. & Shapiro-Ilan,I. 2023. Tolerance of Steinernema carpocapsae infective juveniles in novel nanoparticle formulations to ultraviolet radiation. Journal of Invertebrate Pathology. 196, 107851. <a href="https://doi.org/10.1016/j.jip.2022.107851">https://doi.org/10.1016/j.jip.2022.107851</a>.</li> </ol>