PROTECTING NATIVE AND ENDANGERED CACTI IN DRY FORESTS FROM A NEW INVASIVE MEALYBUG, THE HARRISIA CACTUS MEALYBUG HYPOGEOCOCCUS PUNGENS

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: COMPLETE
• Funding Source: MCINTIRE-STENNIS
• Reporting Frequency: Annual
• Accession No.: 0213788
• Project Start Date: Apr 1, 2008
• Project End Date: Aug 31, 2012
• Program Code: [(N/A)]- (N/A)

Recipient Organization
UNIVERSITY OF PUERTO RICO AT MAYAGUEZ
P. O. BOX 9000
MAYAGUEZ,PR 00681

Performing Department
Crops and Agroenvironmental Sciences 2

Non Technical Summary
Biological information obtained by funding this project is likely to result in effective mitigation of the environmental and economic impacts caused by the recent introduction of the Harrisia cactus mealybug, Hypogeococcus pungens into Puerto Rico. If not undertaken, most dry forest columnar cacti populations will continue to be negatively impacted, and rare autochthonous species may be lost. HCM has so far demonstrated its aggressive nature in our island environment. Serious ecological impact has probably occurred. At this moment, there is no quantification of the economic impacts of this pest on natural ecosystems, or evaluations for potential damage to the production of agriculturally important host plants. Thus, our project will pave the way for a rapid, safe, and effective establishment of a sound classical biological control program. We believe that activities under this project represent an essential foundation to a classical biological control program for the HCM. Objectives are designed to facilitate environmental and biological evaluations needed before responsibly adding exotic natural enemies into fragile dry forest ecosystems. Our team of collaborators has proven their expertise by having worked on international mealybug biological control programs for almost a decade. Others are experts in the ecology and biodiversity of dry forest ecosystems. We believe this is a remarkable mixture of expertise, well suited to tackle the current menace posed by HCM to dry forest ecosystems. Finally, we believe that funding this project will result in the design, and the eventual implementation of a state-of the-art classical biological control against HCM in Puerto Rico. We are confident that such program will serve as a model and as a starting point to other biological control programs worldwide against HCM. We know that there is great concern about the eventual HCM invasion into important centers of cactus biodiversity in Mexico and Central America. A pioneering project of this nature could serve as exemplary to other eventual projects in these countries, and Puerto Rico may again serve as a trusted reservoir for biological control agents against HCM, as it has been against other tropical invasive pests of global significance over many decades.

Animal Health Component

(N/A)

Research Effort Categories

Basic

(N/A)

Applied

(N/A)

Developmental

(N/A)

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
123	0640	1070	50%
211	3110	1130	50%

Knowledge Area
211 - Insects, Mites, and Other Arthropods Affecting Plants; 123 - Management and Sustainability of Forest Resources;

Subject Of Investigation
3110 - Insects; 0640 - Tropical forests;

Field Of Science
1130 - Entomology and acarology; 1070 - Ecology;

Keywords

tropical dry forest

biocontrol

harrisia cactus mealybug

hypogeococcus pungens

puerto rico

pilos ocereus royenii

invasive species

extinction

endangered species

Goals / Objectives
With this proposal we intend to address key pre-introduction research needs leading up to the establishment of a classical biological control program for the Harrisia cactus mealybug (HCM), Hypogeococcus pungens. The classical approach has proven effective against many invasive mealybugs. Nevertheless, important foundation studies are necessary before undertaking a classical biocontrol program to assure success and minimal impacts to biodiversity in fragile ecosystems. Thus, our objectives are to: 1. Establish life history, ecology, and effective population monitoring methods for HCM. 2. Identify indigenous natural enemies attacking cactus mealybugs in dry forests. 3. Establish mass rearing protocols for HCM, and for other endemic mealybug species. 4. Determine host suitability of economically important crops and endangered cacti. 5. Design a classical biocontrol program to introduce natural enemies from abroad.

Project Methods
Objective 1- Establish life history, ecology and efficient population monitoring methods for HCM. A- Developmental biology and life history studies. Most life history and biological studies will conducted. During initial research stages, gravid females raised on Royen's cactus will be kept under greenhouse conditions. Instar descriptions and duration, and sex ratio will be obtained from these data, and comparisons among regimes will be done using analysis of variance (ANOVA). B- Seasonal phenology and population monitoring. We will select three HCM-infested sites. Two trap locations will be used: three traps at breast height, and three traps at 10 cm to stem apex. C- Spatial and geographic distribution. Patches of Royen's cactus will be selected at different dry forest locations in Southern Puerto Rico. Ten to fifteen locations will be scouted and sampled within one/two weeks. These locations will be visited 2-3 times per year to note changes in attack severity. Comparisons between locations and sample dates will be done with analysis of variance (ANOVA). Objective 2. Identify indigenous natural enemies attacking HCM in dry forests. We will select three locations with high HCM populations to sample for indigenous natural enemies. Each individual HCM collected will be placed in separate gelatin capsules. Percent parasitism will be calculated from these collections. Identification will be done by comparing with reference collection specimens, and by confirmation by outside sources. Objective 3. Establish mass rearing protocols for cactus mealybugs and for other endemic species. Using data acquired in Objective 1, we will develop a mass rearing protocol for HCM. Initially, several rearing methods will be tried. The following mass rearing protocols will be developed: (1) mealybug development, (2) reproductive efficiency testing, (3) colony sanitation and disease prevention, and (4) host plant management. We should be ready to begin mass rearing trials by the end of the second year. Objective 4. Determine suitability of economically important crops, endemic alternate hosts, and endangered cacti. Alternate host plant suitability testing will be done at the Lajas Experiment Station. Using development we will attempt to rear HCM on alternate hosts. Host plant species are chosen because (1) they are environmentally sensitive species; (2) they are crops or ornamentals; or (3) they are common non-cactaceae alternate hosts. Objective 5. Design a classical biocontrol program to introduce natural enemies from abroad. This objective is intended to prepare and design a program for the importation of biological control agents, likely from South America, and that includes the following elements: Foreign exploration, clearing and needs assessments for importation permits, identification of environmental requirements, quarantine screening, parasitoid biology studies, field release, parasitoid/enemy performance evaluation, and molecular studies. A foreign exploration trip to visit HCM endemic areas is planned for the 3rd year.

Progress 04/01/08 to 08/31/12

Outputs
OUTPUTS: HCM has been found attacking most columnar cactus species in Puerto Rico. A census has demonstrated that the invasive pest is present in most important dry forests on the main island. HCM male and female larval instars have been characterized as to their morphological characteristics and development at different temperatures. Among the best characters for rapid determination of development stage are the length of metathoraxic tibiae, the antenna, and the presence of circuli. Development was fastest at 32 C than at 27 C or 24 C, with reductions of 3 to 4 days. The best monitoring method to determine field population fluctuations in male flight patterns and abundance of crawlers was the use of yellow sticky traps. Traps were also useful to monitor parasitoid and predator populations. A year of weekly trapping results in older infestations -Guanica Biosphere Reserve - yielded the lowest captures, whereas newly infested sites - FWS Cabo Rojo Wildlife Reserve yielded higher captures. Natural enemies have been identified as follows: the predacious fly Leucopis bella; the ladybeetles Cryptolaemus montrouzieri, Decadiomus sp.n. seini, and Symnobius bilucernarius; the beetle Cybocephalus kathrynae; and the parasitoids Leptomastidea nr. antillicola and Anagyrus sp. Mass rearing of HCM was attempted by using potted live cactus, live cactus chunks, and Althernanthera or Portulaca plants. Best results were obtained by using potted Alternanthera. Other hosts evaluated include sprouted potato and squash, but these were not useful. Development and laboratory rearing of Leptomastidea nr. antillicola was successful, with development from egg to adult in about 25 days. Leptomastidea sp. host range with nine mealybug species resulted in no attacks on species other than on H. pungens. PARTICIPANTS: Participants from the UPR were Aristides Armstrong, Alex Segarra (PI), and graduate students Emmanuel Velez and Adriana Lopez. Partner organizations were USDA-ARS (Dr. Dave Jenkins); USDA-APHIS (Dr. Matt Ciomperlik, Ms. Leyinska Wiscovich); USDA-FS (Dr. Ariel Lugo/Christian Torres); USFWS (Mr. Oscar Diaz, Mr. Omar Monsegur, Dr. Susan Selander), Puerto Rico Department of Agriculture (Ms. Aixa Ramirez); and Puerto Rico Department of Natural Resources (Mr. Jose Sustache). TARGET AUDIENCES: Target audiences were students, land managers, regulatory agencies, and environmental organizations. PROJECT MODIFICATIONS: Not relevant to this project.

Impacts
Activities in this project have accomplished breakthroughs in the areas of HCM population monitoring, natural controls, breeding, life cycle descriptions, and alternate host breadth. Comparison of likely monitoring techniques demonstrates that the use of yellow sticky traps is a good method for determining natural HCM population fluctuations, measuring both dispersing crawlers and adult males. Identification of a variety of natural control agents provides informational and biological baselines for classical biological control options for future infestations elsewhere, or for possible exotic introductions in Puerto Rico. Activities within the project have also demonstrated the feasibility of using Alternanthera plants as host for mass rearing operations. Work has also been directed toward determining preferred HCM development stages for two natural enemies occurring in Puerto Rico. Descriptions of instars and stadia are now complete giving insight into generational times and likely outbreak conditions. Finally, host range of promising parasitoid Leptomastidea nr. antillicola is apparently very narrow, and should be further considered as a good classical biological control candidate outside Puerto Rico. More research is needed to determine host-parasitoid interactions and temporal synchrony.

Publications

• No publications reported this period

Progress 01/01/11 to 12/31/11

Outputs
OUTPUTS: HCM has been found attacking most columnar cactus species in Puerto Rico. A census has demonstrated that the invasive pest is present in most important dry forests on the main island. HCM male and female larval instars have been characterized as to their morphological characteristics and development at different temperatures. Among the best characters for rapid determination of development stage are the length of metathoraxic tibiae, the antenna, and the presence of circuli. Development was fastest at 32 C than at 27 C or 24 C, with reductions of 3 to 4 days. The best monitoring method to determine field population fluctuations in male flight patterns and abundance of crawlers was the use of yellow sticky traps. Traps were also useful to monitor parasitoid and predator pupulations. A year of weekly trapping results in older infestations -Guanica Biosphere Reserve - yielded the lowest captures, whereas newly infested sites - FWS Cabo Rojo Wildlife Reserve yielded higher captures. Natural enemies have been identified as follows: the predacious fly Leucopis bella; the ladybeetles Cryptolaemus montrouzieri, Decadiomus sp.n. seini, and Symnobius bilucernarius; the beetle Cybocephalus kathrynae; and the parasitoids Leptomastidea nr. antillicola and Anagyrus sp. Mass rearing of HCM was attempted by using potted live cactus, live cactus chunks, and Althernanthera or Portulacca plants. Best results were obtained by using potted Alternanthera. Other hosts evaluated include sprouted potato and squash, but these were not useful. Development and laboratory rearing of Leptomastidea nr. antillicola was successful, with development from egg to adult in about 25 days. Leptomastidea sp. host range with nine mealybug species resulted in no attacks on species other than on H. pungens. PARTICIPANTS: Participants from the UPR were Aristides Armstrong, Alex Segarra (PI), and graduate students Emmanuel Velez and Adriana Lopez. Partner organizations were USDA-ARS (Dr. Dave Jenkins); USDA-APHIS (Dr. Matt Ciomperlik, Ms. Leyinska Wiscovich); USDA-FS (Dr. Ariel Lugo/Christian Torres); USFWS (Mr. Oscar Diaz, Mr. Omar Monsegur, Dr. Susan Selander), Puerto Rico Department of Agriculture (Ms. Aixa Ramirez); and Puerto Rico Department of Natural Resources (Mr. Jose Sustache). TARGET AUDIENCES: Target audiences were students, land managers, regulatory agencies, and environmental organizations. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
Activities in this project have accomplished breakthroughs in the areas of HCM population monitoring, natural controls, breeding, life cycle descriptions, and alternate host breadth. Comparison of likely monitoring techniques demonstrates that the use of yellow sticky traps is a good method for determining natural HCM population fluctuations, measuring both dispersing crawlers and adult males. Identification of a variety of natural control agents provides informational and biological baselines for classical biological control options for future infestations elsewhere, or for possible exotic introductions in Puerto Rico. Activities within the project have also demonstrated the feasibility of using Alternanthera plants as host for mass rearing operations. Work has also been directed toward determining preferred HCM development stages for two natural enemies occurring in Puerto Rico. Descriptions of instars and stadia are now complete giving insight into generational times and likely outbreak conditions. Finally, host range of promising parasitoid Leptomastidea nr. antillicola is apparently very narrow, and should be further considered as a good classical biological control candidate outside Puerto Rico.

Publications

• Segarra-Carmona, A.E., A. Ramirez-Lluch, I. Cabrera, A.N. Jimenez. 2010. First record of a new invasive mealybug, the harrisia cactus mealybug, Hypogeococcus pungens (Hemiptera: Pseudococcidae). J. Agric. U.P.R. 94: 183-189.
• Curbelo-Rodriguez, J.C., E. Melendez-Ackerman, J. Rojas-Sandoval, A. Segarra-Carmona. 2012. New record of Cybocephalus kathrynae (Coleoptera: Cybocephalidae) on Mona Island, Puerto Rico. Scielo (accepted).
• Hypogeococcus pungens, a new cactus pest. 2011. Brochure (multi-agency sponsorship)APHIS, ARS, USDA/FS, USFWS, PRDA, Dept. Natural Resources.

Progress 01/01/10 to 12/31/10

Outputs
OUTPUTS: Studies to determine life cycle duration and immature instar identification of Harrisia Cactus Mealybug (HCM) under laboratory conditions have been completed. Four immature instars and adults were described and illustrated. Salient among morphological findings was the presence of trilocular pores in 1st instars. Development time from 1st instar to adult was 24 d (27 C) to 17.5 d (35 C). Population monitoring with yellow adhesive traps proved to be a good method for population monitoring. Population monitoring was established weekly at two sites between Nov 2009 and Oct 2010. Male flights correlate well with crawler dispersal, and are more frequent following the August rainy season, through December. Several natural enemies have been identified attacking HCM. The most abundant seem to be an encyrtid parasitoid, Leptomastidea sp., and a ladybeetle Decadiomus sp. These were also monitored using yellow traps. Mass rearing protocols were developed to rear HCM, using a method adapted from mass rearing in Barbados. This method rears HCM in Alternanthera plants. Classical biocontrol agreement was established between APHIS-CPHST and USDA-ARS to conduct foreign exploration for HCM enemies. PARTICIPANTS: Alejandro E. Segarra-UPR-AES Aristides Armstrong-UPR-AES Victor Gonzalez-UPR TARGET AUDIENCES: IPM Specialists Forest Managers PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
1. Morphological studies now allow identification of HCM stage attacked by natural enemies. 2. Population dynamics of HCM indicate up to a 5 day difference in development time between winter and summer generations. 3. Yellow sticky traps may be used to monitor yearly HCM reproductive population peaks. 4. Rearing HCM in Alternanthera allows the establishment of easily maintained laboratory rearing colonies. 5. Two important natural enemies have been identified and thus become candidates for introduction in other areas invaded by this pest.

Publications

• No publications reported this period

Progress 01/01/09 to 12/31/09

Outputs
OUTPUTS: HCM has been found attacking most columnar cactus species in Puerto Rico. Census demonstrated that the invasive pest is present in most important dry forests on the main island. HCM Male and female larval instars have been characterized as to their morphology. Twenty variables are being measured per individual and best correlation with stage will be used for rapid determination of development stage. Best monitoring method was determined to be the use of yellow sticky traps. Twelve weekly-censused traps are now in place at four sites in the Guanica Biosphere Reserve and in the FWS Cabo Rojo wildlife reserve to determine population fluctuations in male flight patterns and abundance of crawlers. Natural enemies have also been identified as follows: the predacious fly Leucopis bella; the ladybeetles Cryptolaemus montrouzieri, Decadiomus nr. hughesi, and Symnobius bilucernarius; the beetle Cybocephalus sp.; and the parasitoids Leptomastidea sp. and Anagyrus sp. Mass rearing of HCM was attempted with potted live cactus, live cactus chunks, and Alternanthera or Portulaca plants. Best results are obtained using potted Alternanthera. Other hosts being evaluated include sprouted potato and squash. Attempts to rear Leptomastidea sp and Decadiomus nr. hughesi are under way. Also testing Leptomastidea sp. host range by using other mealybugs is being tried. PARTICIPANTS: Two graduate assistantships are granted from this funding. Participants are Aristides Armstrong, Alex Segarra (PI), and Graduate Students Emmanuel Velez and Adriana Lopez. TARGET AUDIENCES: Target audiences were students, land managers, and environmental organizations. PROJECT MODIFICATIONS: Not relevant to this project.

Impacts
Activities in this project have accomplished breakthroughs in the areas of HCM population monitoring, natural controls, breeding, life cycle descriptions, and alternate host breadth. Comparison of likely monitoring techniques demonstrates that the use of yellow sticky traps is a good method for determining natural HCM population fluctuations, measuring both dispersing crawlers and adult males. Identification of a variety of natural control agents provides informational and biological baselines for classical biological control options for future infestations elsewhere, or for possible exotic introductions in Puerto Rico. Activities within the project have also demonstrated the feasibility of using Alternanthera plants as hosts for mass rearing operations. Work has also been directed toward determining preferred HCM development stages for two natural enemies occurring in Puerto Rico. Descriptions of instars and stadia are now complete, giving insight into generational times and likely outbreak conditions.

Publications

• No publications reported this period