Biological Control Laboratory Research Articles

Field Efficacy of Steinernema Carpocapsae Against Lima Bean Pod Borer, Etiella zynckenella Treitschke, Attacking Groundnut

Lima bean pod borer (LBPB), Etiella zynckenella Trietschke, is major insect pest of soybean in Indonesia. Currently it was also reported attacking groundnut in some parts of Sumatera causing serious yield loss. Insecticide treatment is not only ecologically unsafe, but also is not practical due to concealed behavior of the larvae within pods below soil surface. Systemic insecticide might be so but unsafe as it might leave unaccepted residue in harvested pods. It is clear that safer more effective technology such as biological control is badly needed. A Field study was conducted at the University of Bengkulu Research Station in 2012-2013. The objectives was to determine the effectiveness of non local strain of entomopathoegen nematode (EPN), Sterinenema carpocapsae , to control LBPB attacking groundnut. Groundnut seeds were sewed in plots which were later treated with S. carpocasae produce by Biological Control Laboratory University of Jember at the dose of 500.000 JI m -2 . EPN was sprayed upon plants and soil surface twice at plant age of 45 and 65 days after sewing (DAS). There were insecticide treatment and control plots as comparison. Observations of damaged pods and the presence of LBPB larvae were done at plant age of 63, 77, 90 DAS and at the haevesting date. Data were subjected to analysis of variance and the means of damaged pods were separated with least significant different (LSD). The result revealed that in two years study, damaged pods consistently tended to be less in plots treated with S. carpocasae or insecticide than those in controls, indicating that it might be effective against PLPB. The lack of significant different between EPN treated and control plots might be as a result of low percentages of EPN larval survival due shipment problem (i.e. inappropriate packaging and long transportation that toke several days) or it might be that the strain was less adapted yet to local conditions. I speculate that local strain of EPN might be more effective to control LBPB attacking groundnut.

Development of Superior Strain of <I>Trichogramma chilonis</I> Ishii for Temperature Tolerance

Adaptation to climatic conditions (more importantly the temperature) of an area play a decisive role in influencing the parasitoid efficiency in controlling pests. Besides, the optimal conditions for inundative release of parasitoid e.g. Trichogramma , may also depend on strain. A strain of Trichogramma chilonis tolerant to high temperature was developed in the Biological control laboratory of IARI (PUSA), New Delhi and effect of temperature was studied on parasitoid efficiency. The biological traits viz., fecundity, emergence, survivability (male&female) and female populations were studied at 25, 27, 29, and 31oC. The fecundity and emergence were recorded highest at 27 oC. An increase of 107.23% (fecundity), 184.50% (emergence) and 15.65% (female survival) was observed from base to 35th generation, while male survival and female population decreased by 8.2 and 1.2%, respectively. The survival of immature stages of tolerant strain of Trichogramma proved their ability to withstand the temperature.

New Record of Predation on AdultDiaprepes abbreviatus(Coleoptera: Curculionidae) byEuthyrhynchus floridanusandPodisus maculiventris(Heteroptera: Pentatomidae)

The feeding responses of the predatory stink bugs E. floridanus and P. maculiventris to the citrus root weevil D. abbreviatus were studied at the Florida Department of Agriculture and Consumer Services, Division of Plant Industry’s Florida Biological Control Laboratory in Gainesville, FL. This work was done in quarantine at temperatures of 25 ± 3°C with a 16-hour photoperiod (16:8 L/D) and a relative humidity of 50-55%. The E. floridanus and P. maculiventris males, females and 3rd-5th instars were kept individually in a Petri dish with a bean pod and moistened paper for 24 h. They were then exposed to a single D. abbreviatus male or female in Petri-dishes for 24 h. Twenty replications were made. At the end of the experiment percent predation was determined. Euthyrhynchus floridanus was more efficient than P. maculiventris at feeding on D. abbreviatus adults in the lab with no choice tests. This particular stinkbug can be easily mass reared and therefore has potential as augmentative biological control agent for the citrus root weevil. However, more testing is necessary to determine the overall impact of this predator in the field. Las repuestas alimenticias de los chinches E. floridanus y P. maculiventris depredadores del picudo de la raiz de citricos fueron estudiadas en la cuarentena de Gainesville a una temperatura de 25 ± 3°C, 16 horas de fotoperiodo (16:8 Luz/Oscuridad) y una humedad relativa de 50-55%. Machos, hembras y ninfas del 3ro-5to estadio de E. floridanus y P. maculiventris fueron mantenidos individualmente en platos de Petri con papel humedecido y una vaina de frijol comun sin presa durante 24 h. Luego de este periodo, los depredadores fueron expuestos a un macho o una hembra adulta de D. abbreviatus por 24 h. Se utilizaron veinte repeticiones. Al final del experimento, se determino el porcentaje de depredacion. Euthyrhynchus floridanus fue mas eficiente que P. maculiventris alimentandose de adultos del picudo de la raiz de citricos. Euthyrhynchus floridanus podria criarse masivamente para ser utilizado en programas de control biologico del picudo de la raiz de citricos por aumentacion y conservacion en los Estados Unidos de Norte America. Sin embargo, estudios adicionales son necesarios para determinar el posible impacto de este depredador en el campo. View this article in BioOne

Zamorano y sus Contribuciones a la Agricultura Sostenible a Través del Control Biológico de Plagas

El Centro para el Control Biológico en Centro América (CCBCA) fue creado en 1989 por la Escuela Agrícola Panamericana, financiado por la United States Agency for International Development (USAID) en Honduras. El CCBCA se enfocó en el control biológico clásico, el aumento y la conservación de enemigos naturales. En el 2000 cambió el nombre de Laboratorio de Control Biológico y se enfocó en el aprender haciendo de los estudiantes de la Escuela Agrícola Panamericana y la producción y comercialización de enemigos naturales de plagas agrícolas. El único parasitoide conocido que ataca al picudo de las bromelias (Metamasius quadrilineatus), fue encontrado en las investigaciones del CCBCA; resultó una especie nueva nombrada Lixadmontia franki que fue liberada en Florida en el 2007 para el control biológico del picudo mejicano de las bromelias (Metamasius callizona). Se determinaron los organismos parasíticos de las plagas Plutella xylostella, Mocis latipes, Spodoptera frugiperda, Leptophobia aripa y Liriomyza spp. Cuatro especies de avispas parasíticas exóticas, un baculovirus, un hongo y tres picudos fueron introducidos a Honduras para control biológico clásico: Cotesia plutellae (parasitoide asiático de P. xylostella); Diadromus collaris (parasitoide pupal de P. xylostella); Telenomus remus (parasitoide que ataca a los huevos de 30 especies de lepidópteros); una especie de Eretmocerus (originaria de India, para controlar Bemisia tabaci); los picudos Neochetina bruchi y Neochetina eichhorniae y el hongo Cercospora piaropi (para el control de la maleza acuática lirio de agua (Eichhornia crassipes); y el picudo Neohydronomous affinis (controlador biológico de la maleza acuática lechuga de agua (Pistia stratiotes). En el 2000, Zamorano cambió la estrategia del uso de control biológico y empezó a incursionar en la producción comercial de microrganismos para el control de plagas, debido a una demanda no satisfecha. Los controladores biológicos que ha producido son el hongo antagonista Trichoderma harzianum (Trichozam™) para combatir hongos en el suelo, Beauveria bassiana (Bazam™) para controlar lepidópteros y coleópteros, Lecanicillium lecanii (Verzam™) para el control de áfidos y mosca blanca, Metarhizium anisopliae (Metazam™) para el control de salivazo (Aeneolamia spp.) y larvas de coleópteros en caña de azúcar, y Purpureocillium lilacinum (Pazam™) para controlar nematodos. Además, ha reproducido y vendido la chinche depredadora Orius insidiosus para el control de trips, áfidos y mosca blanca, el ácaro depredador Neoseiulus longispinosus para el control de la arañita roja (Tetranychus spp.) y el nematodo entomófago Heterorhabditis bacteriophora para el control de insectos del suelo, especialmente contra Phyllophaga spp., Cosmopolitus sordidus, larvas de lepidóptera, picudo del camote (Cylas formicarius) y termitas en el suelo. Posiblemente, el mayor aporte es en la enseñanza de las técnicas y tecnologías del control biológico de plagas que se han distribuido por América Latina a través de los graduados de Zamorano que estudiaron y fueron entrenados en Zamorano.DOI: http://dx.doi.org/10.5377/ceiba.v52i1.966

Field Demonstration of Trichoderma harzianum as a Plant Growth Promoter in Wheat (Triticum aestivum L)

A three year study (2008-2011) on the validation of the Trichoderma technology for growth promoting ability of Trichoderma harzianum (Th3) was evaluated in the popular wheat (Triticum aestivum L.) variety Raj 3765 at farmer’s field through TIFAC-DST project entitled “On Farm Demonstration and Commercial Production of Trichoderma as Biopesticide and Growth Promoter”. The biological formulation was developed at Biological Control Laboratory, Division of Plant Pathology, IARI, New Delhi, and was successfully demonstrated in two districts of Rajasthan viz., Jaipur and Kota belonging to different agro climatic zones. Rhizospheric Competence Index along with its growth promotion effect on rootlets, tillers, weight of grains and grain yield were evaluated by using it at three stages of crop viz., seed, flowering and preharvesting @ 4g/kg and @ 4ml/L along with soil treatment with a mixture of farm yard manure and formulation @ 50:1 before sowing. Compared to the first year where the farmers were unaware of Trichoderma in 2008-09, a significant increase in yield of wheat from 36.25 to 46.73Q/ha (29% in Jaipur) and from 36.88 to 50.12Q/ha (36% in Kota) has been observed after continuous application for three years (2008-2011) The total income and the benefit cost ratio of farmers increased both at Jaipur (Rs 56242/ha, 1:1.8) and Kota (Rs 60332/ha, 1:1.9).

Performance OfPsyttalia Humilis(Hymenoptera: Braconidae) Reared From Irradiated Host on Olive Fruit Fly (Diptera: Tephritidae) In California

The parasitoid Psyttalia humilis (Silvestri) was reared on Mediterranean fruit fly, Ceratitis capitata (Wiedemann), third instars irradiated at 0-70 Gy at the USDA, APHIS, PPQ, Moscamed biological control laboratory in San Miguel Petapa, Guatemala, and shipped to the USDA, ARS, Parlier, CA. Irradiation dose did not affect the parasitoid's offspring sex ratio (53-62% females), percentage of unemerged adults (12-34%), number of progeny produced per female (1.4-1.8), and parasitism (19-24%). Host irradiation dose had no significant effect on the forewing length of female P. humilis and its parasitism on olive fruit fly, Bactrocera oleae (Rossi) and offspring sex ratio, but dissection of 1-wk-old female parasitoids reared from hosts irradiated with 70 Gy had a significantly lower number of mature eggs than females from nonirradiated hosts. Longevity of P. humilis adults decreased with increased temperature from 15 to 35°C, regardless of food provisions, gender, and host irradiation dose. Females survived 37-49 d at 15°C with water and food, and only 1-2 d at 35°C without food, whereas males lived shorter than females at all temperatures and food combinations tested. Adult P. humilis reared from fertile C. capitata and aspirated for dispensing in cups lived significantly longer after shipment than those specimens chilled and dispensed by weight. At 21 and 32°C, 50% of parasitoids departed release cages after 180 and 30 min, respectively, but none departed at 12°C. Thirteen shipments of P. humilis (2,980-21,922 parasitoids per shipment) were received between September and December 2009, and seven shipments (7,502-22,560 parasitoids per shipment) were received between October and December 2010 from San Miguel Petapa, Guatemala. Daily number of olive fruit fly adult and percentage female trap captures ranged <1-19 and 8-58% in 2009, and <1-11 and 0-42% in 2010, respectively. The number of parasitoids released ranged 848-12,257 in 2009 and 3,675-11,154 in 2010. Percentage parasitism of olive fruit fly third instars at all locations ranged 0-9% in 2009 and 0-36% in 2010.

Field Release in Florida ofDiaphorencyrtus aligarhensis(Hymenoptera: Encyrtidae), an Endoparasitoid ofDiaphorina citri(Homoptera: Psyllidae), from Mainland China

The Asian citrus psyllid, Diaphorina citri Kuy awama (Hemiptera: Psyllidae), is a serious pest of citrus that vectors the phloem-limited non culturable bacterium Candidatus Liberibacter asiaticus Jagoueix, Bove and Gamier (Jagoueix et al. 1994). The resulting disease is commonly referred to as citrus greening or huanglongbing (HLB) (Bove 2006). The psyllid was first reported from Florida in 1998 and citrus greening in 2005 (Halbert 1998, 2005). Two species specific parasitic wasps, Tamarixia radiata (Waterston) (Hymenoptera: Eulophidae) and Diaphorencyrtus aligarhensis (Shafee, Alam and Argarwal) (Hymenoptera: Encyrtidae), were imported into Florida from Taiwan and Vietnam and released beginning in 1999 as biological con trol agents for D. citri (Hoy & Nguyen 1998). Tam arixia radiata has established in Florida (Qureshi et al. 2009), while D. aligarhensis has not. An additional population of D. aligarhensis was collected in Guangdong province, China and sent to Gainesville, Florida in 2006. A colony was initiated in the Maximum Security Room of the Florida Biological Control Laboratory, Florida De partment of Agriculture and Consumer Services, Division of Plant Industry. Parasitoids were held in quarantine through 20 generations before re lease permitting was complete and mass rearing began. Sixty-seven releases were made at 10 sites in 6 Florida counties (Alachua, Collier, DeSoto, Hendry, Orange and Polk) between 8/2007 and 10/2009, deploying a total of approximately 11,100 wasps (Table 1). Sites included commer cial and private citrus groves as well as Murraya paniculata hedges. Wasps were either tapped out of holding tubes directly onto emerging plant shoots infested with immature psyllids or were released into cages consisting of fine mesh bags tied around new shoots infested with nymphs. Shoots were carefully inspected and predators removed prior to tying cages closed. Mesh size al lowed parasitoid wasps to move freely in or out of the cage while preventing most predaceous in sects from entering. Released wasps successfully parasitized nymphs in the field and second generation wasps emerged. Evidence of this includes laboratory emergence of adult wasps from field clippings, D. aligarhensis parasitized mummies found on shoots covered with mesh cages, and several adult wasps sighted foraging on D. citri infested M. paniculata plants (Table 1). All citrus groves used for wasp release, including certified organic groves, used pesticide sprays dur ing the release period. The use of insecticides target ing D. citri likely reduced both psyllid and natural enemy populations (Qureshi & Stansly 2007). Mur raya paniculata release sites were not subjected to any form of insect control, although occasional plant trimming by the property owners removed much of the new soft growth that I). citri uses for feeding and oviposition, consequently leaving few imma ture hosts for D. aligarhensis reproduction. Adult psyllids were present at all locations throughout the release period although their incidence appeared to decline over the >2 year release period, likely from increased insecticide use to reduce the spread of citrus greening dis ease. During many trips to various locations for parasitoid release there was little to no host plant flush or immature psyllids present, particularly during late summer and fall. Tamarixia radiata wasps were present at all release sites utilized in this study. Through the combined effect of host feeding and parasitism, a single female T. radiata can kill nearly twice as many psyllids in her lifetime as can a female D. aligarhensis (500 vs. 280, respectively) (Chien 1995). Additionally, T. radiata has a shorter gen eration time and a higher reproductive rate (Skel ley & Hoy 2004). Furthermore, any hosts parasit ized by T. radiata before or within 5 d following parasitism by D. aligarhensis will result in T. radiata offspring (E. R. unpublished data). Thus, competition with T. radiata could present another impediment to establishment of D. aligarhensis. Coccinellid predators were seen at all release sites. Generalist predators can induce high lev els of psyllid mortality (Qureshi & Stansly 2009). Curinus coeruleus Mulsant, Cycloneda sanguinea (L.) and Harmonia axyridis Pallas readily con sumed D. citri nymphs parasitized by D. aligar hensis (E. R. unpublished data) causing wasp mortality.

Occurrence of Atopozelus opsimus preying on nymphs and adults of Glycaspis brimblecombei

The stink bugs genus Atopozelus (Hemiptera: Reduviidae) is native in the Americas, with five described species of which only three have been recorded in Brazil. In August 2007, Eucalyptus camaldulensis leaves infested by lerp psyllid were collected and taken to the Forest Pest Biological Control Laboratory situated at Sao Paulo State University. A small green stink bug, identified as Atopozelus opsimus Elkins (Hemiptera: Reduviidae) that fed on nymphs and adult psyllids, was found on the leaves. These stink bugs presented phytophagy, omnivory and ability in opening the lerp (shell) that protect the psyllid. A. opsimus has demonstrated unusual particularities, different from many biological control agents found in nature; this predator thus deserves more attention for its potential to be evaluated.

Fire Ants (Solenopsisspp.) and Their Natural Enemies in Southern South America

We review the fire ant research conducted by the ARS-South American Biological Control Laboratory (SABCL) since 1987 to find a complex of natural enemies in southern South America and evaluate their specificity and suitability for field release as self-sustaining biological control agents. We also include those studies conducted by the ARS-Center for Medical, Agriculture, and Veterinary Entomology in the United States with the SABCL collaboration. Ecological and biological information is reported on local fire ants and their microsporidia, nematodes, viruses, phorid flies, eucharitid wasps, strepsiptera, and parasitic ants. Their biology, abundance, distribution, detrimental effect, field persistence, specificity, and phenology are discussed. We conclude that the objectives of the ARS program in South America are being achieved and that the pioneering studies have served to encourage further investigations in the United States and other countries and advanced the implementation of biological control programs to decrease imported fire ant densities and damage. Still, several promising organisms should be further investigated for eventual field release in the near future.

The Biological Stain Commission's Quality Control Laboratory operations and improved traceability of certified stains

The Biological Stain Commission (BSC) is a quality control laboratory that certifies biological dyes for staining cells and tissues. Originally, a single lot of a certified dye was sold to histologists. Today, companies frequently change their lot numbers as part of regulatory efforts. When a certified dye undergoes a lot number change, the BSC must re-certify this dye to verify that it is identical to the one certified earlier. The BSC has improved how these lot changes are monitored using a redesigned BSC certification label. Certification labels always have been issued by the BSC and are attached to every bottle of “BSC certified dye” that is sold. The new BSC certification label has added security features and currently bears both the BSC certification number and the manufacturer batch lot number. The result is improved security and traceability of certified dyes.

Field performance and fitness of an olive fruit fly parasitoid, Psyttalia humilis ( Hymenoptera: Braconidae), mass reared on irradiated Medfly

Adult Psyttalia humilis (Silvestri) were produced from irradiated larvae of Mediterranean fruit fly, Ceratitis capitata (Wiedemann), at the USDA-APHIS-PPQ, Moscamed biological control laboratory, San Miguel Petapa, Guatemala, and shipped to the USDA-ARS, SJVASC, Parlier, from September 2008 to January 2009 for biological control of olive fruit fly, Bactrocera oleae (Rossi), in California. In total, 230,908 individuals were shipped to California and 202,328 were released in the field, a 12.4% mortality rate from shipment to release. Thirteen sample sites were located in nine counties; 11 sites received parasitoids from September 2008 to January 2009 (range: 3114–55,826 per site). Olive fruit fly population density, as measured by adult traps, varied greatly among sites and sample dates – ranging from <1 fly per day (sites in San Joaquin Valley and Sacramento Valley) to 29 individuals per day in (San Jose, CA). Adult emergence of olive fruit fly and P. humilis from >10,000 fruit collected, across all sites and sample dates, showed parasitism ranging from 0–17%. Laboratory studies were conducted to determine the fitness of P. humilis reared on irradiated and fertile C. capitata. Results showed no difference in parental size, progeny produced during a 48 h period, or the number female: male offspring produced; there were significantly fewer eggs in the dissected ovaries of irradiated than fertile female P. humilis. Continuous flight in different air temperatures and air speeds for female and male parasitoids showed no difference in female or male P. humilis reared on irradiated C. capitata or female P. humilis reared on fertile olive fruit fly. Results are discussed with respect to the establishment and effective use of P. humilis for control of olive fruit fly in California’s varying climatic regions where olives are grown.

Biological Control of Tropical Soda Apple (Solanaceae) in Florida: Post-Release Evaluation

Foreign exploration to search for natural enemies of the invasive plant tropical soda apple, Solarium viarum Dunal (Solanaceae) were conducted initially in 1994 by University of Florida researchers in collaboration with Universidade Estadual Paulista, Jaboticabal campus, Brazil. Sixteen species of insects were found associated with tropical soda apple in Brazil and Paraguay (Medal et al. 1996). A biological control program was initiated by the University of Florida in 1997 with funds provided by the United States Department of Agriculture, Animal and Plant Health Inspection Service, and the Florida Department of Agriculture & Consumer Services. Explorations in the place of origin (southern Brazil, northeast Argentina, south Paraguay, and Uruguay) of the target weed were conducted in collaboration with Brazilian university researchers, and the USDAAgriculture Research Service Biological Control Laboratory in Hurlingham, Argentina. During exploratory surveys, the leaf beetle Gratiana boliviana Spaeth (Coleoptera: Chrysomelidae) was found on tropical soda apple plants in southern Brazil. A high level of specificity and significant defoliation of tropical soda apple were demonstrated in host-specificity tests conducted at the Florida Biological Control Laboratory quarantine facilities in Gainesville, the USDA-ARS (Agriculture Research Service) South American Biological Control Laboratory in Argentina, and the USDAARS quarantine facilities in Stoneville, Mississippi, as well as in extensive field surveys and open-field tests conducted in South America. After five years of intensive plant feeding and oviposition tests, the South-American leaf beetle G.boliviana was the first biological control agent approved for field release (7 May 2003) by the USDA-APHIS-PPQ in the southeastern United States to join the battle against tropical soda apple also known by the nickname the 'plant from heir (Coile 1993). This exotic weed has invaded over half a million hectares of grasslands and natural areas in at least 6 states (Florida, Alabama, Georgia, Mississippi, South Carolina, and Texas). Intensive host-specificity tests including 126 plant species in 35 families were conducted with G. boliviana from 1998 to 2001 (Medal et al. 2002, 2003, 2004). Initial releases of G boliviana in Florida began in summer 2003, and currently approximately 200,000 G. boliviana immature and adults have been released in 37 Florida counties, 14,000 in 2 counties in Georgia, 18,000 in 3 counties in Alabama, and 825 in 1 county in Texas. In th s article we report the establishment, post-release monitoring, and initial impacts of G. boliviana on tropical soda apple plants in Sumter County, Florida. A field release of G. boliviana was made in a

A New Species ofChaenusa(Hymenoptera: Braconidae) Reared fromHydrellia pakistanaeandHydrellia Sarahae Laticapsula(Diptera: Ephydridae) InfestingHydrilla verticillata(Alismatales: Hydrocharitaceae) in India and Pakistan

Chaenusa glabra Kula, new species from India and Pakistan is described, and sexual dimorphism is reported and discussed. A diagnosis is provided to differentiate it from all other species of Chaenusa Haliday sensu lato. It was reared from Hydrellia pakistanae Deonier and Hydrellia sarahae laticapsula Deonier infesting Hydrilla verticillata (L. f.) Royale during foreign exploration for natural enemies of H. verticillata, and is in quarantine at the Florida Biological Control Laboratory in Gainesville. It is the first species of Chaenusa described from the Oriental Region and the only species of Chaenusa known to attack species of Hydrellia Robineau-Desvoidy infesting plants from a genus other than Potamogeton L.

PRODUÇÃO DO PARASITÓIDE Cotesia flavipes (HYMENOPTERA: BRACONIDAE) PARA CONTROLE BIOLÓGICO DA BROCA DA CANA-DE-AÇÚCAR (Diatraea saccharalis) (LEPIDOPTERA: CRAMBIDAE)

Since the introduction of Cotesia flavipes (larval parasitoid) in Brazil, in 1971, researches has been developed to obtain the best laboratorial method of massal rearing, aiming the most important plague control in sugarcane, the sugarcane borer Diatraea saccharalis. The actual methods of massal rearing of Cotesia flavipes were verified in this work, as well as the rooms needed in a biological control laboratory, with the propitious environment to rear and maintain host (Diatraea saccharalis) and parasitoid (Cotesia flavipes) during the year. This work also discuss the necessity of a quality control in the beneficial insects massal rearing, as the pathogen contamination is one of the highest obstacle to get a quality product with desirable performance in field.

Neomusotima Fuscolinealis (Lepidoptera: Pryalidae) is an Unsuitable Biological Control Agent of Lygodium japonicum

In its native Japan Neomusotima fuscolinealis Yoshiyasu feeds on and damages the leaves of Lygodium japonicum (Thunberg ex Murray) Swartz, an invasive weed in Florida and the southeastern U.S. Larvae and pupae of the moth were imported into the quarantine facility at the Florida Biological Control Laboratory, Gainesville, Florida, to establish a colony for preliminary host range studies and to define its lifecycle and reproduction parameters. Larvae of the moth did not feed significantly nor develop on 5 tested rare, native Florida ferns. The rare North American native climbing fern, Lygodium palmatum (Bernhardi) Swartz, however, supported complete development of N. fuscolinealis, and 6 continuous generations of the moth were reared on the fern. Because the rare L. palmatum and the invasive L. japonicum co-occur in the US, the release of N. fuscolinealis could result in the harm to L. palmatum, a risk that makes the moth unsuitable as a potential biological control of L. japonicum.

Impact of Insecticides Recommended for Sugarcane on Parasitization and Emergence of High Temperature Tolerant and Ludhiana Strains of Trichogramma chilonis Ishii

The studies to evaluate the effect of insecticides recommended on sugarcane in Punjab, on parasitization and adult emergence of high temperature tolerant (PDBC, Bangalore) and Ludhiana (Punjab) strains of Trichogramma chilonis Ishii conducted during April-July 2004 at Entomological Research Farm and Biological Control Laboratory of Punjab Agricultural University, Ludhiana revealed that Ludhiana strain of T. chilonis was more tolerant to insecticides as compared to the high temperature tolerant strain. Among the insecticides, endosulfan (Thiodan 35EC) @ 395g a. i./ha was the safest followed by imidaeloprid (Conlidor 200 SL) @ 20g a, i./ha, triazophos (Hostathion 40EC) @ 600g a. i./ha, chlorpyriphos (Dursban 20EC) @ 175g a. i,/ha and malathion 50EC @ 1250g a. i./ha. The latter two at the applied doses were highly deleterious to the parasitoid.

First Report of Leaf Spot Caused by Cladosporium herbarum on Centaurea solstitialis in Greece.

Centaurea solstitialis L. (yellow starthistle), family Asteraceae, an invasive weed in California and the western United States, is targeted for biological control. In the summer of 2003, an epidemic of unknown etiology on dying C. solstitialis plants was observed near Kozani, Greece (40°22'07″N, 21°52'35″E, elevation, 634 m). Plants had necrotic light brown leaf spots on the lower leaves and the decurrent leaf bases along the stems. Often, necrotic lesions extended along the stems to the capitula. Virtually all plants in a solid stand of C. solstitialis (approximately 0.5 ha) showed disease symptoms. Diseased plants were collected, air dried, and sent to the quarantine facility of the Foreign Disease-Weed Science Research Unit (FDWSRU), USDA/ARS, Fort Detrick, MD. On the basis of culture growth (45-cm diameter after 2 weeks at 25°C on malt extract agar), fungal morphology (1), and comparison with 21 internal transcribed spacer sequences in GenBank, the putative causal organism was identified as Cladosporium herbarum (Pers.:Fr.) Link. (teleomorph = Davidiella tassiana (De Not.) Crous & U. Braun). Sixteen C. solstitialis plants in the rosette stage and 16 plants in the bolted stage were inoculated with an aqueous suspension of spores (106 conidia ml-1) and placed in an environmentally controlled chamber at 25°C with 8 h of dew and 12 h of light daily. Plants in the rosette stage were resistant, but the fungus was very aggressive on bolted plants. Within 4 to 6 days of inoculation, necrosis developed on leaves and stems and then spread up the stems to the capitula, often resulting in plant death. The fungus also infected developing flowers. Cladosporium herbarum was reisolated from each of the 16 bolted C. solstitialis plants in two separate tests at the FDWSRU and from all bolted inoculated plants at the European Biological Control Laboratory (EBCL) in Greece. In the greenhouse at the EBCL, the pathogen readily spread to (and was isolated from) another 10 noninoculated C. solstitialis plants in close vicinity to the inoculated C. solstitialis plants. Results of host range tests will establish if this isolate of Cladosporium herbarum has the potential as a biological control agent of C. solstitialis in the United States and does not pose a threat to other Centaurea spp. used in horticulture. A voucher specimen has been deposited with the U.S. National Fungus Collections (BPI 863446). Live cultures are being maintained at the FDWSRU and EBCL, Greece. To our knowledge, this is the first report of a disease caused by Cladosporium herbarum on C. solstitialis. Reference: (1) M. H. M. Ho et al. Mycotaxon 72:115, 1999.

The European Biological Control Laboratory: an existing infrastructure for biological control of weeds in Europe

The Agricultural Research Service of the United States Department of Agriculture has maintained a continuous European effort in classical biological control of exotic pests in the USA. The European Parasite Laboratory was established in France in 1919, while the European Weed Laboratory began operations in Rome in 1958. The two laboratories were merged in Montpellier in 1991 as the European Biological Control Laboratory (EBCL), becoming the primary overseas biocontrol laboratory of the USDA. The management of weeds, insect pests, and pathogens is an important feature of agricultural research programmes worldwide. These invasive species can lead to vast financial losses for countries engaged in agriculture. The overall goal of research at EBCL is to develop biological control technologies to be used to suppress invading weeds and insect pests. This is done through expeditions to find natural enemies (insects, mites and pathogens), or phytophagous, parasitoid or predator species. These are characterized in careful experimentation in quarantine facilities and eventually developed as biological control agents. Current weed projects include studies on the Centaurea spp., Arundo donax, Vincetoxicum spp., Isatis tinctoria, Taeniatherum sp. and Dipsacus sp. Insect projects research Lygus bugs, the olive fruit fly, the vine mealybug, and the Asian long‐horned beetle. The EBCL team is international and interdisciplinary. Entomology, plant and insect pathology, molecular biology and ecology are the main approaches of our biocontrol research. The team cooperates with universities and agencies worldwide.

First Report of Anthracnose of Salsola tragus Caused by Colletotrichum gloeosporioides in Greece.

In early October of 2005, dying Salsola tragus L. (Russian thistle, tumbleweed), family Chenopodiaceae, plants were found along the Aegean Sea at Kryopigi Beach, Greece (40°02'29″N, 23°29'02″E, elevation 0 m). All of the 30 to 40 plants in the area were diseased and approximately 80% were dead or dying. All plants were relatively large (approximately 1 m tall × 0.5 m diameter), and living portions of diseased plants were flowering. Dying plants had irregular, necrotic lesions extending the length of the stems. Leaves of these plants were also necrotic. Lesions on stems and leaves were dark brown and usually coalesced. Diseased stem pieces were taken to the European Biological Control Laboratory, USDA, ARS at the American Farm School in Thessaloniki, Greece. There, diseased stem pieces were surface disinfested for 15 min with 0.5% NaOCl and placed on moist filter paper in petri dishes. Numerous, waxy subepidermal acervuli with black setae were observed in all lesions after 2 to 3 days. Conidiophores were simple, short, and erect. Conidia were one-celled, hyaline, ovoid to oblong, falcate to straight, 12.9 to 18.0 × 2.8 to 5.5 μm (mode 16.1 × 4.5 μm). These characters conformed to the description of Colletotrichum gloeosporioides (Penz.) Penz. & Sacc. in Penz. (2). Conidia were placed on modified potato carrot agar and axenic cultures from these isolations were sent to the quarantine facility of the Foreign Disease-Weed Science Research Unit, USDA, ARS, Fort Detrick, MD for testing. On the basis of DNA sequences, two variants within S. tragus have been described in California and named "Type A" and "Type B" (1). Conidia were harvested from 14-day-old cultures grown on 20% V8 juice agar, and healthy stems and leaves of 18 30-day-old plants of S. tragus Type A and 10 Type B plants were spray inoculated with an aqueous conidial suspension (1.0 × 106 conidia/ml plus 0.1% non-ionic surfactant). Three control plants of each type were sprayed with water and surfactant only. Plants were placed in an environmental chamber (18 h of dew in darkness at 25°C). After 1 day, all plants were transferred to a greenhouse (20 to 25°C, 30 to 50% relative humidity, and natural light augmented with 12-h light periods with 500-W sodium vapor lights). Lesions developed on stems of inoculated Type A plants after 5 days. After 14 days, all inoculated Type A plants were dead. Lesions on Type B plants were small and localized; all plants were diseased but no plants died. No symptoms occurred on control plants. C. gloeosporioides was reisolated 14 to 21 days after inoculation from stem pieces of all inoculated plants of both types of S. tragus. This isolate of C. gloeosporioides is a destructive pathogen on S. tragus Type A and is a potential candidate for biological control of this weed in the United States. To our knowledge, this is the first report of anthracnose caused by C. gloeosporioides on S. tragus in Greece. A voucher specimen has been deposited with the U.S. National Fungus Collections, Beltsville, MD (BPI 871126). Nucleotide sequences for the internal transcribed spacers (ITS 1 and 2) were deposited in GenBank (Accession No. DQ344621) and exactly matched sequences of the teleomorph, Glomerella cingulata.

ATTEMPTS TO ARTIFICIALLY PROPAGATE THE FIRE ANT PARASITE SOLENOPSIS DAGUERREI (HYMENOPTERA: FORMICIDAE) IN ARGENTINA

Since the 1960s, the workerless (permanent) parasitic ant Solenopsis daguerrei (Santschi) has been considered part of a complex of potential candidates for introduction into the United States for the biological control of the red and black imported fire ants, Solenopsis invicta Buren and S. richteri Forel respectively (Lofgren et al. 1975, Jouvenaz 1990, Wojcik 1990). Pioneering work on S. daguerrei was conducted in Uruguay and Argentina by Silveira Guido et al. (1973; unpublished reports). More recent studies have been conducted at the USDA-ARS-South American Biological Control Laboratory (SABCL) since 1988 and more intensively since 1995. Studies included records on its occurrence and abundance in southern South America (Briano et al. 1997) and field and laboratory observations on its specificity, biology and behavior (Calcaterra et al. 1999, 2000, 2001). Also, many fire ant colonies from Argentina parasitized with S. daguerrei were shipped into quarantine at the USDA-ARSCenter for Medical, Agricultural, and Veterinary Entomology (CMAVE), Gainesville, FL, where complementary studies were conducted (unpublished information). Although detrimental effects on fire ants were reported in Argentina (Calcaterra et al. 1999), further progress on the project was not achieved because of (1) the difficulty in rearing S. daguerrei under laboratory conditions and (2) the failure to introduce and establish the parasite into new host colonies. A number of attempts to introduce and propagate S. daguerrei in laboratory and field host colonies were conducted in Argentina from 1996 to 1999 following different approaches. Some of this work was reported by Calcaterra et al. (2001). Although some tests showed limited success, S. daguerrei did not establish permanently in any of the target host colonies. Despite this failure, we believe that reporting the different approaches used in our studies will contribute to future research efforts. Parasitized and nonparasitized colonies used in the tests were mostly multiple-queen colonies of S. richteri collected in four areas of Buenos Aires Province: (1) San Eladio (60 km W of Buenos Aires), (2) Saladillo (180 km SW of Buenos Aires), (3) Mercedes (100 km W of Buenos Aires), and (4) Suipacha (125 km W of Buenos Aires). The colonies were excavated in the field, brought to the laboratory in 10-liter buckets dusted with talc, separated from the soil by flotation (Banks et al. 1981) and placed in trays. The basic approaches used in the laboratory tests were: #1) Transference of queens: Target host colonies were fragmented into microcolonies (n = 34) composed of one host queen, 50 workers and approximately 1 gr. of brood. These microcolonies were maintained in artificial nests (Bishop et al. 1980) in plastic rearing trays (40 x 25 x 7 cm) coated with Fluon? and fed adult house flies and/or canned Vienna sausage. A water source was always present. One active S. daguerrei queen separated from a parasitized colony was transferred to each microcolony. For about one month, the colonies were kept at different temperatures as follows: 10?C (five colonies), 10-15?C (eight colonies), 15?C (11 colonies), and 30?C (10 colonies). In five of these colonies kept at 30?C, the S. daguerrei queens were sprayed before transference with an extract of macerated workers from the target (receptor) host colony. The tests were monitored daily for the presence of the parasite in the receptor microcolonies. #2) Transference of sexuals: Similarly, target host colonies were fragmented into microcolonies (n = 22) as above and kept in trays at 15 'C. A total of 92 sexuals (alates) of S. daguerrei (3-9 per microcolony) was transferred and monitored daily. #3) Transference of pupae: Test 1. Target host colonies were fragmented (n = 6) as above and kept at 150C. Six pupae of S. daguerrei were transferred to each microcolony. Test 2. Host colonies were fragmented into microcolonies (n = 4) composed of six host queens and 1 gr. of workers and maintained between 19-30?C. A total of 200 pupae of S. daguerrei was transferred to the microcolonies (six pupae per colony every 3-4 days for three weeks). #4) Entire colonies in contact: Test 1. A small plastic tray (40 x 25 x 7 cm, coated both inside and outside with Fluon?) with a parasitized colony was placed in a larger tray (50 x 40 x 12 cm) with a parasite-free colony. The colonies were monitored daily to observe the short flights of S. daguerrei sexuals into the larger tray and their survival. Test 2. Two trays (50 x 40 x 12 cm), the first with a parasitized colony and the second with a parasite-free colony were interconnected through plastic tubes (1 cm in diameter) to a third