Proceedings of the First Regional Workshop on Plant Quarantine

June 17-21, 1996 The University of Guam Mangilao, Guam

1999

Editors: Sponsors: R. Muniappan Taiwan Food and Fertilizer Technology L. Austin Center (TFFTC) O.H. Diambra South Pacific Commission Northern Marianas College College of Agriculture and Life Sciences, University of Guam Proceedings of the First Regional Workshop on Plant Quarantine

June 17-21, 1996 The University of Guam Mangilao, Guam

Edited by: R. Muniappan L. Austin O.H. Diambra

1999

Taiwan Food and Fertilizer Technology Center (TFFTC) South Pacific Commission Northern Marianas College College of Agriculture and Life Sciences, University of Guam

Published with financial support of the University of Guam Endowment Foundation. Preface

It is known that more than 90 percent of the major pests in the Pacific are from exotic introductions. The development of transpacific air and sea traffic since World War II has increased movement of pest insects and diseases. In recent years, the introduction of new pests averages about 3.5 per year in Hawaii and 2.00 per year in Guam. As Micronesia strives to improve its economy by increasing its agricultural productiv­ ity, it is imperative that quarantine programs be strengthened. Under the initiatives of Dr. Kiritani and Dr. Sato, the Food and Fertilizer Technology Center (FFTC) of the Republic of Taiwan funded this First Regional Quarantine Training Workshop in Micronesia. Repre­ sentatives from all Micronesian islands and the Philippines and resource personnel from Taiwan, Japan, the South Pacific Commission, the Northern Marianas College and the College of Agriculture and Life Sciences of the University of Guam actively participated in this workshop. This volume presents all country reports and provides technical information pre­ sented at the First Regional Workshop on Plant Quarantine held at the University of Guam, Mangilao, Guam. One major recommendation is to promote harmonization of quarantine regulation throughout the region. We thank Dr. Jeff Barcinas, Dean/Director of the College of Agriculture and Life Sciences, and the University of Guam for allowing this workshop to take place. Special thanks to Dr. Saito for obtaining microscopes from the Japanese Government for distribu­ tion to country representatives, and also to Continental Air Micronesia for graciously transporting the equipment from Japan to Guam. We also thank the University of Guam Endowment Foundation for providing support for publication.

R. Muniappan L.Austin O.H. Diambra

1 Contents

Plant quarantine regulations of the Commonwealth of the 4 Northern Mariana Islands by Anthony Mufia and Jesse R. Manalo

Plant quarantine regulations in Chuuk 6 by Akissy Edward and Peter Haritos

Plant quarantine regulations in Kosrae 7 by Palikkun Tolenna and Valton Palsis

Plant quarantine regulations in Pohnpei 9 by Solomon Brens and Alex Kim

Plant quarantine regulations in Palau 10 by Jeff Tewid

Summary of the quarantine regulations in the 11 Federated States of Micronesia by Denis Kelly

Plant quarantine regulations in the Marshall Islands 18 by Banner Bwijtak and Stanley Hosia

Plant quarantine regulations in the Philippines 19 by Luben Marasigan

Plant quarantine regulations in Japan 20 by Toshiyuki Kato

Plant quarantine regulations in Guam 26 by Russell Campbell, Marvin P. Chargualaf and Jason T. Paulino

Weeds 28 by. C.T. Lee

Insect and mite damage 31 by Don Nafus

2 Invertebrates of quarantine importance 35 by lise Schreiner

Indexing of plant virus diseases: Serological diagnosis 40 by Norihiko Saito

Quarantine surveillance for fruit flies 44 by Allan Allwood, presented by Russell Campbell

Virus-free nursery system, indexing and micrografting · 48 byHongJiSu

An introduction to plant diseases 57 by George Wall, Robert Schlub and Diane Greenough

Recognition, quarantine surveillance and 63 emergency response planning by Allan Allwood, presented by Russell Campbell

Review and recommendations 67

Participants 69

3 Plant quarantine regulations of the Commonwealth of the Northern Mariana Islands by Anthony Muna and Jesse R. Manalo

Introduction Plant Protection in the CNMI The Commonwealth of the Northern The plant protection function in the Mariana Islands (CNMI) came into exist­ Commonwealth is under the Department of ence on January 9,1978, when the first Lands and Natural Resources headed by elected governor was inaugurated and the Secretary Benigno M. Sablan. Plant Protec­ members of the First Commonwealth tion section comprises the Agriculture Legislature were sworn in. Quarantine, Division of Agriculture and the The population of the Commonwealth assistance of Dr. Moore, associate director froin 1980 to 1995, according to the census of Land Grant. conducted by the CNMI Department of The Agriculture Quarantine office is Commerce, is over 50,000. located in the Saipan International Airport The Commonwealth is comprised of 14 with a branch office at the Saipan Commer­ islands, only four (4) of which are continu­ cial Port. Quarantine officers conduct ously populated. The total land mass is inspections of all incoming and outgoing approximately 184 square miles, with the animals under the Supervision of Asst. northern most island of Saipan (47 sq. mi.), Director, Mr. Mark I. Palacios. The assis­ Tinian (39 sq. mi.), and Rota (32 sq. mi.) tance of Dr. Moore is to identify plant pests. covering over 64 percent of the total land area and have over 99 percent of the Agriculture Quarantine population. The population center and the Quarantine activities slightly increase seat of the Commonwealth government is during typhoon season or when bad on Saipan where 90 percent of the Com­ weather occurs. The number of aircraft and monwealth population resides. ships and volume of cargoes coming into The Commonwealth is located in the Saipan increases. Major development western Pacific Ocean, Mariana Islands ongoing at the airport and seaport is Archipelago, lying between 14' and 21' presently being undertaken by the Com­ North Latitude and 1400 and 1460 East monwealth Ports Authority. This will Longitude. The 14 islands form a chain in a increase the capacity of the airport and north-south axis, stretching over 380 miles seaport operation and, therefore, will from the northern most island, Farallon de increase the amount of work for the Quar­ Pajaros, to the southern most island, Rota. antine Section. Agricultural materials Guam, a separate U.S. territory, is geo­ continue to come through the U.S. Post graphically part of the Mariana Islands and Offices in Chalan Kanoa and Capitol Hill is located about 120 miles south of Saipan. and, therefore, they continue to assign an The CNMI is located in relatively close inspector to each of the facilities. We are proximity to nearly all Asian countries anticipating an increase in passenger ships including Japan, Taiwan, Korea, Philip­ from Guam and Japan making port calls to pines, China and Hong Kong, Indonesia, Saipan and, therefore, continue to send Burma, Thailand, and Laos. inspectors to these areas to board and While considered a remote location provide pre-clearance for these ships. The geographically, with daily direct flights USDA/PHIS/PPQ and U.S. Fish and from the Orient, as well as daily sea flights Wildlife conduct quarantine manual arriving from many Asian ports, the ·training program to update and upgrade Northern Mariana Islands are strategically our inspectors in the use of USDA and U.S. located at a crossroads providing easy Fish and Wildlife quarantine manuals. access to the Orient and the U.S. Mainland.

4 1995 Statistic Report for Airport and Seaport Oan. • Dec. 1995)

Vessels about 461 Inspected incoming ocean freight cargo about 270,161,346lbs. (135,081 tons) Confiscated contraband material about 3522 lbs. ·

Tourism The Commonwealth continues to experience rapid changes in its socioeco­ nomic status with the economic growth rate in tourism. The impetus for this rapid rate of development seems to be the CNMI' s emergence as a mecca for Japanese and Korean tourists, as well as that of a regional leader in the manufacture of textile prod­ ucts. The Plan Protection Quarantine Section ·is very important to the Commonwealth of the Northern Mariana Island due to the tourism industry that continues to be the leading economic activity in the CNMI.

5 Plant quarantine regulations in Chuuk by Akissy Edward and Peter Haritos

Chuuk is one of the states of the inspectors carry out quarantine function on Federated States of Micronesia. The islands a daily basis on an average of 64 flights in a of Chuuk State are in the central Caroline month and 10-15 vessels at the seaport. Islands of the West Pacific, about 3,700 Introduction of any pest into the state kilometers southeast of Tokyo, Japan, and could be easily accomplished without 5,450 kilometer southwest of Honolulu, proper inspection. Chuuk State has no Hawaii. The islands are surrounded by a proper facilities for disposal of contraband barrier reef about 63 kilometers across. nor a fumigation chamber for exported There are 19 high volcanic islands within commodities. The staff requires proper the Chuuk Lagoon, and a total of 24 training in the field of quarantine and inhabited outer islands. knowledge of pest distribution. With this Chuuk is the most populated state training we hope that we could learn a lot among the states of FSM, comprising about and benefit from it. 53,000 people. Agriculture is one of the main primary sectors in the economy. Problems About 80 percent of the state's total popu­ While carrying out law enforcement lation is involved in subsistence agricul­ duties several problems were encountered. ture. Taro and breadfruit are considered the Transportation to airport and seaport is most important staple crops. Sweet potato considered a major problem. The depart­ and cassava are considered important ment has no vehicle assigned to the quaran­ starch foods. Vegetables production has tine section. Cooperation among clearance increased in some of the lagoon islands as parties is lacking, as well as a lack of farmers have started to realize the eco­ knowledge of the risk of introduction of nomic importance of agriculture. pests and under estimation of the impor­ tance of quarantine. The lack of proper Present Situation quarantine facilities is also a major problem Quarantine laws and regulations were in the state. established by the National Government and are enforced by each state in FSM. The Recommendations Department of Agriculture's quarantine To conclude our country paper we staff is responsible in carrying out the recommend the followings: duties. Chuuk State is linked to the world 1. Frequent training on agriculture by our main seaport and airport, situated quarantine. on Weno, the main center in Chuuk. 2. Build adequate quarantine facilities. Quarantine staff in Chuuk consists of five 3. Install a quarantine employees under the supervision of the fumigation chamber. Director of Agriculture. The quarantine 4. Provide a means of transportation. 5. Establish a record keeping system.

6 Plant quarantine regulations in Kosrae by Palikkun Tolenna and Valton Palsis

Introduction dominant attitude which does not allow The Kosrae State Department of other plants chance to grow within a given Agriculture and Land is comprised of four space. It is now widely spread all along the divisions of which Crop Production & roads of Kosrae. In the recent past ship­ Research (CP&R) Division is one. The ments of the parasite, Parachaetes quarantine subsection is under the CP&R pseudoinsulata, were received from the Division. In total there are five subsections COM and UOG Land Grant with the aim to under the CP&R Division. There are eradicate the weed. The use of the parasite currently two staff members managing the to control the weed outside the Kosrae quarantine subsection and ifs activities. Terminal/ Airport was a success while in In the past few years, Kosrae had other areas where the parasite was re­ experienced a number of pests and diseases leased, the monitoring visits later on that led to the introduction of biological­ showed no presence of the parasite activity. control agents from overseas to help It is assumed that the released agents in the alleviate such problems. The College of interior were eaten by small lizards and Micronesia (COM), the University of Guam geckos. There is no presence of lizards at (UOG) Land Grant and South Pacific the Kosrae Airport at this time of control or Commission (SPC) were very generous in when we released this parasite. paying attention and providing technical assistance to solve some of these problems. Citrus Fruitfly Some of these pests and disease prob­ The fruitfly monitoring project, coordi­ lems included the following: nated by entomologist Luc Leblanc, is ongoing. More than 1,000 fruits of lime, Citrus Spiney Whitefly tangerine, orange and other trees were The sooty mold occurring mostly on collected and sampled for fruitfly host the underside of leaves of most plant verification and population. Kosrae now species (including citrus trees, breadfruit, has a small quarantine laboratory where plumeria and other plants) stops the some of these specimens are kept for photosynthetic process, thereby affecting monitoring purpose. All lab results from the reproductive system. With the assis­ these collected and sampled fruits were tance of the UOG Land Grant, a parasite, sent to Luc Leblanc in Pohnpei for final Encarsia smithi, was shipped in from Guam result and confirmation. to help eradicate or at least to lower the population to a controllable level. The sooty Coconut Red Scale mold problem was first observed in the late This is not really a serious problem nor 70s. With the introduction of this parasitic has it spread throughout the island at this wasp, the pest (spiney whitefly) was almost time. The coconut red scale was sighted on totally eradicated. some severely infested coconut trees in Kosrae. For this purpose a shipment of the Devil Weed predator, Adeleyncyrtus oceanicus, was Another problem now facing Kosrae is introduced by Nelson Esquerra, COM, in the unwanted weed called Chromolaena January 1996. This predator was released to odorata or Devil Weed. This particular weed one site in Lelu, Kosrae. Unfortunately was accidentally introduced less than 20 after a lapse of three months, a follow-up years ago (apparently by the Kong Yong check was made and no control establish­ Construction Co., Korea). This weed has a ment or presence of this predator was noted.

7 Kosrae Fumigation travelers into Micronesia should be care­ continued A fumigation chamber which had been fully inspected upon their arrivall' as they lying idle for some years at the agriculture have a habit of bringing in items without station has been fixed and put to operation. bothering to obtain permit or certificate Trials and trainings in the use of this prior to departure from their place. Numer­ chamber were conducted to the quarantine ous ornamental plants and vegetable seeds officers in Kosrae by the Plant Protection are now present on Kosrae and we believe Trainer (PPT) in Micronesia, Dennis Kelly. were illegally imported into Kosrae. This chamber is using methyl bromide gas Local crop productio~ on Kosrae is which, unfortunately, was banned by the increasing tremendously this year and the U.S. and other countries. Therefore it will aim is to increase it by 25 percent annually be used only for treatment of incoming over the next five years. It is hopeful that agricultural materials and/ or those on the the enforcement on the FSM Quarantine island, but not to those for export. Regulations will be improved likewise.

Taro Planthopper Duties and Responsibilities of · Taro planthopper is a major insect pest Quarantine Officers of colocasia taro in Kosrae. And it is widely The duties and responsibilities of the spread on all taro farms throughout the staff of the Plant and Animal Quarantine island. It was not until early part of '90s Section are as follows: when farmers started to concentrate more • Inspect all incoming aircraft and on colocasia taro that they realized the affect vessels in Kosrae. or damage cause by this planthopper on • Inspect all incoming passengers and their farms. Taro planthoppers were under their possessions such as hand carry-on biological control in at least three sites, baggage, luggage and cargoes. If vessel, most commonly by the predator check ship stores, galley, garbage and Cyrtorrhinus fulvus, which was introduced its cargo. to Kosrae in 1993 by Nelson Esquerra, • Collaborate with Customs and Immi­ COM. This predator was very effective in gration officials at airport and/ or controlling this pest. However, due to some seaport in the effective performance of problems, we lost all the predators. duties. • Prepare and submit to division admin­ Airport Quarantine Activities istrator reports on any inconvenience A number of contraband materials during inspection of the day on a daily continue to be seized upon inspection at or monthly basis. the airport in the past. Most often items • Assist agricultural produce exporters in like fruits, seeds, ornamental cuttings and providing clearance for such exports. similar material received from other places • Be responsible for the carrying out of had to be confiscated due to lack of sup­ the biocontrol projects and programs on porting quarantine documents. Some the island. • Perform other duties as may be as­ signed by the Division Administrator or the Department Director.

8 Plant quarantine regulations in Pohnpei by Solomon Brens and Alex Kim

First I would like to thank FFTC and have to work hard to protect our islands the staff of UOG for organizing and spon­ from the introduction of pests and diseases. soring this important workshop so the Export inspections of fresh produce is also participants will have the opportunity to a major part of our service to ensure learn and improve their capabilities in produce exported from Pohnpei complies enforcing the revised regulations that are with other countries' and FSM states' now in existence in the FSM. quarantine requirements. Pohnpei is the capital of the FSM We quarantine officers enforce the (Federated States of Micronesia), located 7 quarantine regulations of our nation in degrees north and 158 degrees east, ap­ order to protect agricultural production proximately 1,000 miles southeast of Guam. and to limit the introduction of exotic pests The main means of transportation in and diseases into Pohnpei as well as to the FSM, both inter-island and internation­ limit the spread of pests and diseases to our ally, are by ships and planes. The prevailing sister states in the FSM. schedule of monthly inbound flights and Recently Pohnpei has constructed a vessels for Pohnpei average 80 flights and Quarantine Inspection Center with methyl 200 vessels, mostly foreign fishing vessels. bromide fumigation chamber. This was Guam serves as the main transit port part of the SPC Plant Protection in for most of the flights and vessels serving Micronesia Project for which we are the FSM, including Pohnpei, to deliver grateful for the help of the SPC and the passengers and cargo from abroad. sponsor, Australia. · Pohnpei Quarantine Service is com­ Also, the FSM has revised the Plant posed of two permanent inspectors with and Animal Quarantine Regulations which some assistance from the chief of Agricul­ became effective June 1, 1996. The FSM ture, the Crops Supervisor and the Senior regulations have been distributed to all of Livestock Officer, all of whom are trained in you within the workshop manual. quarantine procedures. Since our work force is small for quarantine inspections we

9 Plant quarantine regulations in Palau by Jeff Tewid

The islands of Palau, part of the Palau Quarantine Section Caroline Islands, are situated in the north­ The Palau Quarantine section operates em part of the Pacific Ocean. Palau lies under the Division of Agriculture and about 250 kilometers north of the equator Mineral Resources of the Ministry of and 340 km east of the Philippine Islands. Resources and Development. There are six The land area is about 65 square kilometer, quarantine officers who serve as inspectors or 43,830 hectares. The climate of Palau is at the four major entry points or ports into relatively hot and humid with an average Palau. The main task at these entry points is rainfall of about 370 centimeters per year. to monitor the presence of harmful and Major crops in Palau include cassava, taro, transmittable pests or diseases and to sweet potato, banana and coconut. The prevent them from coming into the country. beautiful marine life surrounding the Palau The four major entry points or ports are the Islands is also a main source of food. Palau airport terminal, the sea port, the flight is well known around the world for its cargo warehouse and the postal office. flourishing Rock Islands, sometimes called Palau quarantine officers are trained to "the Emerald Islands," and its magnificent identify and confiscate any fresh produce, marine life. It is this marine life that has plants, livestock products, seeds, live provided growth to Palau's economy, animals and any other items that could based on tourism. Tourists are attracted to pose a potential threat to Palau's fragile Palau's famous underwater diving spots environment. Last year alone, there were such as the Blue Comer and the German over 1,048 recorded flights from commercial Channel. Tourists are also interested in the airlines, cargo planes, military planes and variety of plants that thrive on the Rock private planes into Palau's airport terminal. Islands including unique plants species There were over 275 recorded entries of that are indigenous to Palau. cargo vessels, fishing boats, yachts, tour boats and passenger boats at the seaport. Role of Quarantine Officer Among these, over 1,871 cargo ship con­ The marine ecosystem and the environ­ tainers were reported and released follow­ ment of Palau must be protected, based on ing a careful and thorough inspection of the their importance in the economy. Responsi­ containers for the presence of contaminated bility for protecting Palau's fragile environ­ soils and possible stowaways of brown tree ment falls heavily on the shoulders of snakes and vector rodents. Among the total quarantine officers. As quarantine officers, numbers of recorded airport terminal and we must realize that the most important seaport entries, over 6,422 pounds of policy is to protect the agricultural and various fruits, seafood, seeds, plants and general well being of the people of the animal products were confiscated by Trust Territory. Quarantines are promul­ quarantine inspectors. All contraband items gated as a means of preventing the intro­ were either sent back to their points of duction of and further spread of injurious origin or destroyed (by fire). Live plants or insects, pests and diseases into and within animals are neither confiscated nor de­ the Trust Territory. stroyed if the carriers or passengers pro­ duce authorized documentation. Palau has been fortunate to avoid . serious outbreak of disease and pests and it is the duty of the quarantine officers to keep it that way.

10 Summary of the quarantine regulations in the Federated States of Micronesia by Denis Kelfy

1.0 Prelude to prevent the introduction of the Colorado Quarantine is an instrument used as a beetle. strategy for the purpose of "preventing the The USA set up its first Quarantine introduction and/ or spread of dangerous, Inspection Center at the port of San Pedro or possibly dangerous quarantine pests and in 1891. The State of California introduced diseases." further quarantine laws in 1903 to enforce Agricultural quarantine protects regulations introduced to protect San agriculture production and its associated Pedro's agriculture. The first USA federal crop, forestry, and livestocK industries by law relating to plant quarantine was performing the functions of the Plant and enacted in 1912. Animal Quarantine Inspection Services. To Australia commenced plant quarantine be able to legally operate as a regulatory activities in 1908 when the Commonwealth protection service it is essential that policy Quarantine Service was established. The must first be decided upon and then laws Australian Quarantine Inspection Service must be made and enforced to regulate and (AQIS) is now responsible for all compo­ to control the importation of risk material nents of quarantine, plant, animal and to ensure that the risk material is safe to health. import. Japan established the Export-Import To enable quarantine inspectors to Plant Control Law in 1914 and a plant perform their duties efficiently it is essen­ quarantine system was started. Japan tial that all inspectors have a thorough revised the original law and in 1950 knowledge of their country's plant quaran­ established the "Plant Protection Law." tine laws and regulations. Also, the inspec­ European countries also commenced tors should have a good knowledge of agricultural quarantine activities in the overseas quarantine laws, especially those early part of the 20th century. of their neighboring countries and trade Of course, public health quarantine has partners. been around since the 14th century when Further, quarantine inspectors should controls were introduced to control the understand why the laws are there, not just spread of the bubonic plague, known as the that the laws are there. Also, remember that Black Death. no one is exempt from quarantine. It is the duty of the inspectors to be able to explain 1.2 The International Plant Protection to people why something is contraband Convention (IPPC) and why the contraband must be, for The first international convention of example, destroyed. the Regional Plant Protection Organization was signed in 1881 with the purpose of 1.1 Early Plant Quarantine Laws preventing the spread of "severe" pests. Quarantine dates back a long way. One This convention was called the Phylloxera of the earliest plant quarantine laws was Convention. (Note: Severe pests are now passed in 1873 when Germany prohibited more often referred to as quarantine pests.) the importation of plants and plant prod­ (Note: The Phylloxera aphid destroyed ucts from the USA to prevent the introduc­ over 1 million acres of grapevines in tion of the Colorado beetle, Leptinotarsa France.) decemlineata. In 1951, the International Plant Protec­ In the United Kingdom, the 1877 tion Convention (IPPC) was established Destructive Insects Act was introduced also and came into force in 1952. More than 80

11 FSMsummary countries are now parties to this conven­ requirements and old existing laws should continued tion by which signatory countries agree to be amended to ensure that WTO guidelines work through legislative, organizational are met. and technical means to prevent or at least to minimize the introduction and/ or 2.0 ·understanding and Making Quaran­ spread of dangerous pests and diseases. tine Laws The IPPC provides for immediate and In order for the quarantine rules to be rapid exchange of information on the enforced, it is essential that the enforcing or incidence and outbreak of plant pests and regulatory officers are aw

12 • The making of regulations, quaran­ achieved acts usually provide for the FSM SUII]mary tines, orders, etc., under the act. making of regulations or other legislation. continued • Describing certain things that need to These regulations and other legislation be done. may be described as the machinery by • Authorizing emergency measures. which the act operates. The act states what • Stating actions required to be done may or may not be done and the regula­ under the act. tions prescribe how these things may be •. Stating actions which are offenses done. under the act. Words may be defined in a regulation • Stating penalties for offenses. as well as in an act providing the meaning given does not go beyond the meaning Acts are divided into parts, sections defined in the act. In this case, the defined and subsections. Each section and its meaning only applies to the particular subsections deal with one particular regulation as stated. If the act does not subject.. The sections Within each "part" are define a word then the regulation may numbered separately. When quoting fiom define the word and the meaning given an act the part, section and subsection must may limit or extend the meaning beyond b~ stated. the normal everyday meaning. A word An important section of any act is that defined in a regulation keeps that defini­ dealing with interpretation. When there are tion wherever it appears in the regulation words used in an act are intended to have a unless otherwise stated in the definition. meaning which may differ from the every Another example of a word definition day meaning they are defined. The defini­ is "goods" which means "any movable tion given may limit or extend the meaning property and includes plants and plant beyond the normal everyday meaning. material, animals and animal products." When a word is defined in the act the given meaning must be applied to that word 2.4 Quarantines wherever it is used in th~ act'or,any regula­ Each section of a law should refer to tions, quarantines, orciers, etc. made under only one subject. Quarantines have been the act. However, mostly the interpretation used to list a number of various matters or definitions of words are contained relating to the same subject under the one within regulations. regulation. For example, a regulation may An example of a word definition is cover the impo:t:tapon of plants generally, "person." A person could be defined as then various qua;arttirtes may describe meaning ''both singular and plural and many individual species of plants that may shall include individuals, corporations, have various conditions for import. companies, associations, societies, partner­ ships and natural persons." 2.5 Orders Orders are short-term laws that may be 2.3 Regulations used in emergency situations~ For example, Regulations are adopted following a in relation to plant quarantine, if a serious usually lengthy administration process new pest was introduced, it would be a provided for under a particular administra­ severe disadvantage if the Plant Protection tion act that prescribes the legal processes Service had to wait for a regulation to be for making regulations. Regulations do not made to initiate quarantine actions. To have to tabled before Congress but every overcome this, an order may be issued. person must be informed of the intention to Orders become effective immediately and introduce a regulation. This is not done on remain so for a period of 30 days. After this · an individual basis but by posting the time the order must be reissued and proposed regulations in prominent official administrative processes applied to enact a areas, official registers, radio broadcasts, law such as a regulation through the etc. Once promulgated the regulations normal administrative procedures. remain in force until repealed. To enable the purpose of an act to be

13 FSMsummary 3.0 Interpretation of Quarantine Laws T.T., Title 25. Currently, the Marshalls are continued Laws are made by lawyers and are reviewing the recently introduced- FSM intended for legal use by lawyers. There­ regulations and may elect to adopt similar fore the interpretation of the words of the regulations as used in the FSM. laws are not always easy for non-lawyers to clearly comprehend. This is acceptable, 5.0 Plant Quarantine Laws in the FSM as laws must try to block all loop-holes and The Federated States of Micronesia cover all contingencies. To achieve this (FSM) adopted the revised Plant and using everyday language would require Animal Quarantine Regulations on June 1, the use of many words. Hence the legal 1996. The regulations were issued by the language system survives. Secretary of the Department of Resources The common method of overcoming and Development with the approval of the the interpretation problem is by producing President of the FSM and in accordance operation manuals for use by quarantine with the FSM Administrative Procedure. inspectors. These manuals generally The regulations were made under the Code SUI):llllarize all the requirements of the act of the FSM, Title 22 Agriculture and Live­ and associated regulations into everyday stock, Chapter 4, Quarantines, Section 402. language. However, it is essential that quarantine staff are familiar with the 5.1 Code of the FSM Title 22 quarantine legislation in both the legal and The Code of the FSM, Title 22, Agricul­ everyday languages. ture and Livestock, Chapter 4, Quarantines An example of an interpretation is is the "act" under which the quarantine "soil." Soil is referred to in most plant regulations and quarantines are made. Title quarantine regulations as both soil as an 22 contains sections that provide for: importation and soil as a contaminant. Therefore, when a consignment of "soil" is S401 Purpose (reason for, and objectives of intercepted the inspector would act within quarantine regulations) the regulation referring to soil, for example, S402 Plant & Animal Quarantines: heat treatment. If a vehicle has to be National regulations (authority to imported contaminated with soil, then the make regulations and other controls) inspector would act within the regulation S403 Emergency State orders (Governors referring to the importation of contami­ can make emergency orders.) nated goods, for example cleaning and S404 Plant & Animal Quarantines: destruction of the soil. Translations (vernacular copies) 5405 Administration & enforcement 4.0 Plant Quarantine Legislation in Palau (Secretary of R&D to administer and the Marshall Islands quarantine) The Republic of Palau has recently S406 Agricultural quarantine inspectors - adopted the revised Animal and Plant General appointments (appointed by Quarantine Control Regulations. Prior to Sec. R&D to enforce the regulations this the Trust Territory, Title 25, Animals and controls) and Plants Quarantine Controls were S407 Agricultural quarantine inspectors - enforced. The revised regulations retain a Special appointments (special ap­ lot of the old Trust Territory regulations. In pointments, e.g.. Customs and brief, the regulations rely on importers Immigration officers may inspect for applying for permits which describe quarantine) certain conditions under which goods may S408 Agricultural quarantine inspectors - be imported and for certain imports to be Governors' appointees (Governors accompanied with phytosanitary certifica­ may appoint State inspectors.) tion. The revised legislation retains restric­ S409 Emergency measures authorized tions based on countries rather than on (Sec. of R&D may order immediate scientific justification aligned on pest risk action.) analysis. S410 Inspection (basically everything is The Marshall Islands have retained the subject to inspection)

14 (1) All animals and plants or parts 3. Effective Date (commencement date) FSM sum.mary thereof, entering or transported within 4. Purpose (reasons and objectives) continued the FSM are subject to inspection and 5. Definitions (interpretation of words) may be refused entry if suspected of being, infected or infested with disease or pests. Part II - General Requirement Regula­ (2) All aircraft and vessels or their tions (Plant and Animal) cargoes, including baggage, ship's 6. Points of entry (official, designated first stores, and ballast entering or moving points of entry) within the FSM are subject to inspec­ 7. Requirements of all persons entering tion, etc. the FSM (Every person must comply (3) It shall be a petty misdemeanor for with the regulations. There are no anyone to interfere with or refuse to exemptions.) submit to the inspections, etc. 8. Availability of documents, manifests S411 Manifests and movement informa­ and conveyance movement informa­ tion (made available on request) tion (cargo manifests, aircraft and S412 In-transit material to be kept on vessel arrivals, etc. made available on board (animals, plants, other quaran­ request.) tinable material to be kept on board) 9. Inspection of plants or parts S413 Contraband material (anything thereof, animals, animal products, imported in contravention of the or other goods (all "goods" subject regulations shall be seized and to inspection, treatment, destruc­ destroyed by fire or other means or tion, reshipment, etc.) expelled from the FSM at the 10. Inspection and treatment of any shipper's expense). conveyance arriving from overseas (all S414 Disinsectisation (vessels and aircraft aircraft and vessels are subject to traveling into or within the FSM may inspection, treatment, etc. including all be sprayed with insecticides) cargoes, etc.) S415 Penalties (violations against the 11. Disposal of garbage (controls ship and chapter is a misdemeanor) aircraft garbage) 12. Placement into quarantine (goods may As may be seen by noting the titles of be held in quarantine) each section, quarantine requirements are 13. Entry and exit to and from a quaran­ well covered in a broad sense. However, the tine area, security of quarantine area content within each section do not explain (authorized people only permitted how, in most cases, the requirements are to entry and exit) be enforced, but the contents describe who 14. Removal from quarantine (written will administer the laws and that regula­ permission required) tions and other controls may be issued to 15. Samples, testing, etc. (may take enforce the administration of the chapter. samples, perform tests, treat, etc.) 16. Release of plants, plant material, 5.2 FSM Plant Quarantine Regulations animals, animal products or goods Regulations have previously been (Quarantine Release certificate re­ described as the machinery to operate quired) (enforce) the act. The FSM regulations do 17. Plant and Animal Quarantine Entry just that - the regulations are specific in Permits (requirements for permits) describing what has to be done to enable 18. Any plants, plant material, animals, the safe importation of risk material. animal products, goods or regulated material may be ordered back into quarantine (imported goods always Part 1 - Preliminary Regulations remain subject to quarantine) 1. Short Title (name of the regulations) 19. Seizure and destruction (any contra­ 2. Authority (legal authority by which band may be retained and destroyed) the regulations are made) 20. Disposal of regulated material carrying

15 FSMsummary or suspected to be carrying a pest (any 38. Inspector may employ assistants continued infested, infected goods may be (temporary assistants may be employed) destroyed) 21. Quarantine forms and documents (forms may be introduced at any time) Part IV- Plant Quarantine: Regulations 22. Official imports by Government (inc. and Quarantines (General) biological control agents) (government 39. Quarantines Plant- General may be exempted under certain QP 1 - For entry of fresh fruit and veg­ conditions) etables (general requirements for all 23. Approved places for quarantine to be fruit and vegetables must also refer to performed (any place maybe approved) specific requirements) 24. Quarantine seal (may be applied to QP 2 - Entry of live plants or parts thereof secure anything) (other than seeds or tissue cultures) for 25. Official stamp(s) (used to authenticate propagation (general requirements for documents) most propagation material 26 ..Post-entry I intermediate quarantine (in QP 3 - Importation of seed (general require­ the absence of post-entry facilities ments for most seeds) quarantine may be performed overseas QP 4 - Plant tissue cultures prior to importation) QP 5-Cut flowers and foliage (specifies 27. Treatment (treatments may be ordered prohibited flowers) and fees applies) QP 6 - Timber (treatments, inspection, etc.) 28. Deteriorated or abnormal condition of QP 7 - Grass, bamboo, cane, palm fronds goods deemed to be diseased (broken (specifies prohibited species, etc.) down, abnormal conditions of goods QP 8-Soil, sand and gravel, etc. (specifies may be termed diseased without actual controls) evidence of disease) QP 9-Stored dried food products (subject 29. Transport, storage, unpacking, treat­ to inspection and treatment ment - importer's expense (importer to QP 10 - Packing material, dunnage and meet the cost) bags (requirements, prohibited, permit­ 30. In-transit movement of regulated ted types) material and goods (controls may be QP 11 - Handicrafts made from plant applied to any regulated material in material (subject to inspection, etc.) transit through the FSM) QP 12- Non-plant articles (covers goods, 31. Emergency measures for the eradica­ vehicles, machinery, containers, etc.) tion/ control of a quarantine pest (Sec. QP 13 - Bacteria, viruses, vaccines, cultures R&D or quarantine inspectors have and organisms (import requirements) authority to declare emergency controls) QP 14 - Pre-departure inspections of 32. Fees and charges (costs may be recov­ conveyances and cargo (certification ered for all quarantine services) inspection) 33. Exclusion of liability (no liability for loss or damage) Part V - Specific Plant Prohibitions and Restrictions Part III- Powers of Inspector's Regulations 40. Quarantines Plant- Specific Plant 34. Search and examine (may enter pre­ ·Prohibitions and Restrictions mises, vessels, etc. and search) QP 15 - Banana, Abaca, other Musaceae 35. Access to documents (may examine and Heliconiaceae any document) QP 16-Beans 36. Power to open shipment (may open QP 17-Cacao anything to examine) QP 18 - Cassava 37. Power to examine and certify export QP 19-Citrus consignments (export consignments QP 20- Coconut (Palmae sp. and other may be inspected and refused certifica­ palms inc. Betel nut) tion) QP 21 - Coffee 16 QP 22 - Crucifers 6.0 Summary FSM sum[JJary QP 23 - Ginger The overall objective of agricultural continued QP 24- Maize (inc. popcorn) quarantine is to prevent the introduction or QP 25- Orchids (inc. Vanilla) spread of pests and disease. To facilitate QP 26 - Peanuts (ground nuts) this there must be laws to allow quarantine QP 27- Pepper to function legally. It is important that QP 28 - Pineapple quarantine inspectors are familiar with the QP 29- Potato (Irish) quarantine laws that are based on accept­ QP30-Rice able standards and scientifically and QP 31 -Rubber morally justified. Laws are not easy to QP 32-Sorghum make and they must follow demanding QP 33 - Sugarcane legal procedures. QP 34 - Sweet potato Regulations are made under acts and QP 35 - Taro and edible aroids are the mechanism by which the objectives QP 36- Tomato of the act are met. Other rules may be QP37- Yams quarantines or orders that supplement regulations by giving a wider, more specific description of the regulation. Part VI - Animal Quarantine - Regula­ The interpretation of laws is not always tions and Quarantines (General) easy for non-lawyers to understand. Note: The Animal Quarantine laws Operational or procedural manuals are have been deleted from this document. sometimes published to assist in the interpretation of laws. Everything the quarantine inspector needs to know is Part VIII - Export Requirement contained within the regulations except for Regulations treatments that may be made as supple­ 43. Examination of exports - Plant and ments to the regulations. plant material (issuance of phytosanitary certificates, inspections, responsibilities)

44. Examination of exports -Animals and animal products

Part IX - Penalty Regulations 45. Penalties (provides for penalties for offenses under the regulations as a misde­ meanor)

17 Plant quarantine regulations in the Marshall Islands by Banner Bwijtak and Stanley Hosia

The Republic of the Marshall Islands Islands is imported from the USA, Japan, has two areas of quarantine activities: Australia, and New Zealand. The quaran­ Majuro (airport and seaport), Ebeye and tine section is under the Division of Agri­ Kwajalein (airport and seaport). culture and falls under the Ministry of Kwajalein is under U.S. control and the Resources and Development. There are four quarantine is done by the Marshall Islands quarantine inspectors in the Republic of the Quarantine Inspectors. Certificates and Marshall Islands, assigned in Majuro (two) permits are issued by Marshall Islands and in Ebeye and Kwajalein. Quarantine. Marshall Islands still use the The main plant pests in the Marshall Tiust Territory Title 25 Quarantine Regula­ Islands are the coconut scale, spiralling tions but are currently reviewing the new white fly and Egyptian fluted scale. Biologi­ regulations just introduced in the FSM cal control measures have been successful (Federated States of Micronesia), and on the coconut scale and Egyptian fluted therefore may adopt new regulation soon. scale. The coccinellid beetles have signifi­ The main quarantine confiscation cantly reduced the effect of coconut scale in concerns fruits, vegetables and plants. Majuro and other atolls of the Marshall Most food consumed in the Marshall Islands. Future biological control programs need to be implemented.

18 Plant quarantine regulations in the Philippines by Luben Marasigan

Status of Plant Quarantine for an upgraded quarantine service consid­ Plant quarantine in the Philippines is ering the ROP' s vision toward global enforced by the virtue of Presidential competitiveness which includes the expor­ Decree No. 1433, otherwise known as the tation and importation of agricultural Plant Quarantine Law of 1978 which was products. revised by the Administrative Code of 1987. It has the primary function of preven­ Improvement Plan ting the entry of foreign pests in the A bill re-enacting the Plant Quarantine country and preventing further spread of Law of 1978 is on the second reading in those pests already introduced to places Congress. It seeks to restore under Central where they are not known to exist. Management of Plant Quarantine Service With the provision of Food and Agri­ all plant quarantine laboratories which culture Organization- International Plant have included under the devolved func­ Protection Convention of the United tions as mandated by the Local Govern­ Nations, the Philippines, as a signatory, is ment Code of RA 7610. · committed to comply or follow with the phytosanitary requirements of plants and Strategies/Recommendations plant products for export to ensure accept­ Strategies ability upon entry into the country of 1. Develop inspection sampling rate of destination. different commodities (import/ export) to attain 99.9 percent security. Pest Invasion 2. Establish data base of pests affecting The Philippines islands lie in the heart crops in the Philippines. of Southeast Asia. From the north to the 3. Establish data base of pests occurring south, this tropical archipelago consists of globally. 7,107 islands and islets and stretches for 4. Develop techniques of pest diagnosis more than 1,700 kilometers toward the (laboratory, greenhouse and field). equator. This is one reason why there is a 5. Campaign for plant quarantine aware­ possible risk of pest invasion and dissemi­ ness. nation. One example is the citrus decline in 6. Train manpower to update the knowl­ Batangas in the 1960s solely caused by edge of current trends in the Philippines. virus. The severe decline of abaca was 7. Develop treatment schedule for potential caused by virus disease in the Bicol region. export crops. Commercial Cavendish banana production 8. Upgrade facilities and equipment. in Davao was badly damaged by two (2) major nematode species, sagging the Recommendations banana industry. 1. Additional manpower to manage the Other examples of injurious pets necessary ports I subports of entries. present in the Philippines are as follows: 2. Procurement of modem equipment and • Papaya ring spot virus - Cavite utility vehicles. • Potato decline - Mt. Province 3. Upgrading of facilities. • Socorro wilt of coconut - Mindoro 4. Additional increase budget in Plant • Rice black bug - Palawan, Quarantine. Zamboanga, Catabato 5. Re-enactment of Plant Quarantine Law in • Mango pulp weevil - Palawan conformity with the International Plant Protection Convention' (IPPC). Problems of the Agency The Plant Quarantine Service at present lacks the resources to modernize its equipment and facilities. There is a need

19 Plant quarantine regulations in Japan by Toshiyuki Kato

Plant quarantine in Japan was imported only through the seaports and launched in 1913. At the time, it was only airports which have been designated as for the export of plants on which the ports of entry by the Ministerial Ordinance. United States required a certificate. A Import through other ports is illegal and subsequent plant quarantine system was prohibited. Under the law, the plants to be established in 1914 by the enactment of the imported are grouped into three categories: "Export-Import Plant Control Law." The 1) items prohibited entry, Japanese plant quarantine system has since 2) items subject to import inspection, been gradually intensified as well as 3) items exempted from quarantine. expanded. Now based on the "Plant Quarantine Law," enacted in 1950, Japan is Flow chart of Japanese import plant undertaking every aspect of the plant quarantine system is shown in Fig. 1. quarantine service. Present plant quaran­ tine services in Japan are divided into following categories: Fig. 1 :·Flow Chart of Japanese Import Plant Plant Quarantine Quarantine System · · International Quarantine - Import quarantine • Plants or plant products exported for - Export quarantine Japan Domestic Quarantine • Items prohibited entry - Quarantine of domestic seedling • Items subject to import inspection - Emergency control • Items exempted from quarantine -Monitoring survey for newly • Free from pests invading pests • Export phytosanitary certificate to be . - Designated pests eradication obtained and forwarded to importer programs • Import admissible only at designated port of entry These quarantine services are carried • Application for import inspection out by the Plant Protection Station. The • Import inspection organization consists of five regional center stations located at Yokohama, Nagoya, [PQ official judgment] Kobe, Moji and Naha. Each station hasits To be done at the responsibility of branches and sub-branches at major sea importer under the PQ supervision and air ports where plant quarantine [Importer] officials are stationed. The total number of [Exporter] stations, branches and sub-branches is 89 [Plant quarantine official] with the total of 876 personnel including • Pests discovered 765 plant quarantine officials in 1996.

I. INTERNATIONAL QUARANTINE A. Import Quarantine 1. Import Prohibition Import inspection is made on all plants The import of the following items into and plant products imported to Japan by Japan is prohibited. mean of cargo freight, luggage, and parcel • Pests and pathogens of plants. post. Except for those sent by mail, all • Soil and plants with soil attached. plants and plant products should be • Plants and plant products specifically 20 designated by the Ministerial Ordi­ immersed in sulfuric acid, alcohol, vinegar, Japan nance of the Ministry of Agriculture sugar, or salt, etc. are exempted from plant continued Forestry and Fisheries. quarantine. • Packing materials and/ or containers of the above items. 3. Import Inspection When importing plants and/ or plant Plants and plant products designated products, the importer must submit a in the provisions of the Ministerial Ordi­ written application to the Plant Protection nance are the host plants of more than 15 Station. Such application must be made, kinds of pests. (In this paper, "pest" means without delay, upon arrival of the shipment insect pest and disease such as fungi, concerned. bacteria, and virus) including the Mediter­ In applying for the inspection, the ranean fruit fly (Ceratitis capitata), codling applicant is requested to present the export moth (Cydia pomonella), potato wart phytosanitary certificate. (Synchytrium endobioticum) and tobacco Inspection is carried out at the place blue mold (Peronospora tabacina) which specified by the plant quarantine official in have never been present in Japan and those the seaport or airport where the shipment shipped from or exported through places arrives. The consignment and its content where these pests occur. These pests are are confirmed to be the same with those regarded as topmost quarantine signifi­ declared in the documents. Then, appropri­ cance because they would cause serious ate amount of samples are drawn according damage to the Japanese agricultural to the standards which depend on the industry should they infiltrate Japanese territory. In spite of Table 1: Inspection Record of Import Plants such prohibition, items to be used for scientific research Plants for cultivation 67,677,000 pc's 25,222,000 pc' s can be imported Bulbs for cultivation 193,176,000 pc's 396,836,000 pc's under the special permit given and the Seeds for cultivation 305,840 T 28,140 T conditions set by Cut flowers 362,172,000 pc's 812,263,000 pc's Minister of Agricul­ Fresh fruits 1,488,000 T 1,757,000 T ture, Forestry and Vegetables · 470,000 T 976,000 T Fisheries. Cereals 27,678,000 T 30,748,000 T When the pests Beans 4,966,000 T 5,193,000 T for prohibition have been eradicated in a Miscellaneous articles 7,579,000 T 8,431,000 T country, the prohibi­ Logs 29,350,000 m3 22,650,000 m3 tion can be lifted through legal procedures. nature, quantity, and producing country, etc. Inspection, in general, is done visually, 2. Items Subject to Import Inspection with the aid of magnifying glass or micro­ All plants and plant products such as scopes as needed, by cutting, peeling or seeds, seedlings, ornamental plants, bulbs, sieving the samples. cut flowers, fruits, vegetables, grains, Inspection records of import plants and beans, feeds, spices, herbs, and wood are plant products in shown in Table 1. subject to import inspection. For plants and plant products found Because of negligible pest risks, free of pests upon the inspection, the however, manufactured plant products "Official Certificate" is issued or the such as lumber, preserved log, rattan, cork, "Official Certificate Seal" is stamped. fibrous products, processed tea, fermented vanilla beans, dried fruit, and plants 21 Japan (2) Post-entry Quaran­ Table 2: List of conditionally ban-lifted items continued tine In addition to inspections at ports of entry, the following Australia Sweet orange, lemon, mango propagating materials Canada Cherry are subjected to post­ Chile Grape, Kiwi fruit entry quarantine in China Melon (Xinjiang Uighur), Litchi order to check for the Hawaii Papaya pests such as viruses Israel Sweet orange, Sweetie, Grapefruit that are not detectable Philippines Mango, Papaya by the ordinary Netherland Bell pepper, Tomato import inspection. New Zealand Cherry, Nectarine, Apple These plants are Taiwan Ponkan, Mango, Litchi required to be grown l;'hailand Mango in the national post­ South Africa (Republic) Sweet orange, Lemon, Grapefruit entry quarantine Swaziland Sweet orange, Grapefruit farms for the mini­ Spain Lemon mum of one growing USA (Mainland) Fresh fruit of cherry, nectarine, apple season. Intensive (states of Washington & Oregon), examination for virus Straw of wheat and Agropyron diseases is made species mingled in hay. Walnut in during this period by shell. using various tech- niques as inoculation test, plants, serological diagnosis, electron (1) Preshipment Quarantine at the Origin microscopy, etc. When a country intend~ to export a • Flower bulbs (lily, tulip, hyacinth, etc.). certain prohibited item to Japan and has • Potato tubers (tuber stems) developed for that purpose an applicable • Sweet potato (tuber roots) method which enables complete steriliza­ • Nursery stocks and scions of fruit trees tion of the pests in question, the prohibi­ (citrus, apple, pear, grapevine, etc.) tion may be lifted upon item-by-item basis • Seedlings and scions of strawberry, under the special quarantine conditions sugarcane and pineapple. which have to be agreed upon between the two countries. As of April, 1996,34 plant items from 15 countries are lifted from the ban. Conditionally ban-lifted items are Fig. 2. Flow Chart of Post-entry shown in Table 2. Quarantine System During export seasons, Japanese plant quarantine officials are sent abroad to • Propagating materials subject to post- witness sterilization treatment and inspec­ entry quarantine tion by the authorities in the exporting • Transfer to post-entry quarantine field country. • Post-entry quarantine Another example of preshipment • Free from diseases and pests quarantine is that which is applied to [Plant quarantine official] Dutch cut flowers exported in large • Diseases and I or pests detected quantities to Japan. Japanese plant quaran­ • Delivery to importer tine officials are dispatched to the Nether­ • Destruction lands and carry out preclearance inspec­ tions to expedite the import plant quaran­ Flow chart of post-entry quarantine tine procedures. system is shown in Fig. 2. Flower bulbs can be exempted from the post-entry quarantine in some cases. This

22 system is implemented on the basis of Phosphin fumigation/5-7 days. Japan . bilateral agreement between the plant Ergots, sclerotia and I or soil particles: continued protection agencies of Japan and the Sorting as far as technically feasible. exporting country together with Japanese plant quarantine officials. Another case of (3) Fruits and vegetables exemption is approved for the direct retail Scale insects, thrips, etc.: HCN fumiga­ sale of flower bulbs if sealed one by one. At tion/30 min. present, this system is applied to several Other insect species: Methylbromide kinds of flower bulbs from the Netherlands fumigation/3 hrs. and the Republic of South Africa. (4) Grains, beans, hay bales, etc. (3) Quarantine on Hand Baggage Insect pests: Methylbromide fumiga­ Inspection procedures on incoming tion/24-72 hrs. travellers are follows: quarantine, immigra­ Phosphin fumigation/ 5-9 days or tion, plant quarantine, animal quarantine, Carbon dioxide/10-21 days and customs inspection. Prior to the Diseases: Heat processing at desig­ customs inspection, the plant quarantine is nated factories. conducted on hand baggage in customs at the entry port. (5) Wood logs Wood borers and other insect pests: (4) Quarantine on Postal Matters Methylbromide fumigation/24-72 hrs. Plant imports by mail are only allowed on land or immersion in water I 30 days with parcel post, small packages, and trade in water pools. samples. The inspection is done at the customs clearance postal office. Items B. Export Quarantine passing the inspection are delivered to the Foreign countries have various restric­ addressees. tions on plant imports in order to prevent the introduction of pests. Plants for export 4. Disinfection and Destruction from Japan are inspected to meet the When some pests are discovered in the requirements of the importing countries. import inspection, disinfection is usually Flow chart of Japanese export quarantine ordered. If there is no means of disinfection system is shown in Fig. 3. which can completely kill the detected pests or diseases or when the shipment is found to be import-prohibited, the im­ Fig. 3 Flow Chart of Japanese porter is allowed to choose either to Export Plant Quarantine System destroy or reship it. The method of disinfection involves • Application for export plant inspection fumigation, chemical spraying, hot water • Export inspection and sterilization treatment and other heat therapies, sorting, subject to the requirements of import etc. The treatment schedules are prescribed country according to the types of plants and the • Agreement to the requirements of species of pests detected. Examples are as import country follows. • Non-agreement to the requirements of (1) Seedling & nursery stocks, scions, cut import country flowers • Issue of plant quarantine certificate Scale insects, thrips etc.: HCN fumiga­ tion/30 min. Other insects: Methylbromide fumiga­ Recently many countries require a field tion/2 hrs. inspection on nursery plants and seeds prior to shipment from Japan. The main (2) Seeds for planting items are seeds, bonsai trees and bulbs Weevils, Pyralid moths, etc.: destined for North America and Europe, Methylbromide fumigation/3 hrs. or and fresh fruits and vegetables for Asian 23 Japan countries and North continued America. Export of Table 3: Inspection Record of Export Plants Unsyu orange, bound for the United States, and "Nijisseiki" Plants for cultivation 17,060,000 pc's 15,130,000 pc's (Japanese pears), 7,840,000 pc's 5,550,000 pc's bound for the United Bulbs for cultivation States and Australia, Seeds for cultivation 1,737T 2,826 T are under particular Cut flowers 110,000 pc's 200,000 pc's quarantine items and Fresh fruits 4,969 T 3,516 T conditions. These Vegetables 2,931 T 1850T include the designation Cereals 55,063 T 1,244 T of the production area, 255T joint site inspection at Beans 73T the growing site, Miscellaneous articles 22,083 T 16,950 T packing house, etc. The Logs 31,222m3 12,117m3 Unsyu orange requires further measures such as sterilization treatment and a bacterioph­ during their growth. The seed potato is age test for the citrus canker. Bonsai, such only designated plant up to the present. as the Japanese white pine, etc. destined for Unless the seedlings have pass the inspec­ EC countries, are quarantined for two tion, their movement to the end-users is years at the growing site for pine helminth prohibited. In Hokkaido and 10 other rust prior to export inspection. prefectures, seed potatoes are inspected for Inspection record of export plants and freedom from nine kinds of pests including plant products is shown in Table 3. ring rot and virus diseases. Mother stocks of fruit nursery plants, from which scions are taken for propaga­ II. DOMESTIC QUARANTINE tion, are also subject to plant quarantine Domestic quarantine undertakes inspection for virus diseases. The plant inspections on seed potato and major fruit items include citrus, apple, grape, pear, nursery plants to certify freedom from the peach, cherry and plum. VItus and other harmful diseases in order to provide healthy propagation materials 2. Emergency Control to the farmers. Harmful pests such as the To prevent the spread of harmful pests sweet potato weevil and the giant African in Japan, the government undertakes snail occur in the Southwestern Islands emergency control. The Plant Protection (Okinawa and Amami) and Ogasawara, Station joins in the operation and confirms but are not inhibited in the other region of the eradication for any further decisions to Japan, Movement of these pests and their be taken on the program. The emergency host plants out of the habitat regions are control programs in the past are as follows: legally prohibited. Also, in case when a pest has invaded Japan and seriously • Potato tuber moth (1954-65) endangers domestic agricultural produc­ • Sweet potato weevil (Eradicated in tion, the movement of host plants is Kagoshima in 1965) restricted and the emergency control is • Citrus burrowing nematode (Eradi­ initiated by the government. cated in Hachijou Is., Tokyo in 1967) • Sweet potato weevil (Eradication 1. Quarantine of Domestic Seedling underway in Nishino-omote city In order to secure healthy propagating (Taneagashima Is.), Kagoshima since materials for farmers, plant quarantine 1991) officials inspect a certain designated plants • Bacterial shoot blight, Hokkaido eradication in progress smce 1995.

24 3. Monitoring Survey for Newly Invad­ pests and diseases. However, the increasing Japan ing Pests diversification in the Japanese diet in recent continued It is vitally important to detect newly years and the development in modern introduced pests in early stages of outbreak transportation technology has brought in order to take immediate control mea­ about an influx of wide varieties and a sures. In an attempt toward such purposes, tremendous amount of plants and plant the Plant Protection Station installs lure-bait products. As a consequence, the risk of traps at the major ports of entry and invasion of noxious pests accompanying international post offices. In the inland theses plants and plant products has areas, prefectural pest control centers also become increasingly more serious. There­ join in the monitoring survey programs for fore, plant quarantine is required to be the newly invading pests, enforced more effectively. Moreover, internationally Agreement 4. Eradication Programs on Designated on the Application of Sanitary and Pests Phytosanitary Measures (SPS) has been Against the pests which are specifically evoked to introduce the concept of plant designated and regulated by domestic quarantine pest and to develop interna­ quarantine, prefectures undertake eradica­ tional standard on phytosanitary measures tion programs with subsidies from the under the framework of International Plant government. The Plant Protection Station Protection Convention. Japan is required to cooperates in the programs, monitors the harmonize with the international move­ results of the eradication control, and gives ment. the final decree on the success of the In accordance with such national and programs. The oriental fruit fly, which had international movement, the Japanese inhabited the Southwestern Islands government also has been examining a (Okinav1Ta and Amami) and Ogasawara partial revision of the Plant Protection Law. Island, was eradicated in 1986 with the total The major revised articles include: expenditure of 5 billion yen over 18 years. • Introduction of the concept of quaran­ The melon fly in the Southwestern Islands tine pests, was exterminated in 1993 at the total cost of • Request of exporting countries to 20.4 billion yen over 22 years. inspect certain designated plants At present, a pilot test program aimed during growing season, at the eradication of the sweet potato • Exemption of the phytosanitary weevil is underway in the Southwestern certificate attachment for certain Islands. designated plants, • Others. III. REVISION OF PLANT PROTEC­ TION LAW As mentioned above, Plant Protection Law, enacted in 1950, has accomplished a great role in preventing economic losses to useful plant resources from the invasion of

25 Plant quarantine regulations in Guam by Russell Campbell, Marvin P. Chargualaf and Jason T. Paulino

Introduction • Pesticide certification (University of Plant Protection and Quarantine (PPQ) Guam), is charged with enforcing Guam laws and • US Fish and Wildlife Service (Aquatic & regulations concerning the importation of Wildlife Division). all agricultural commodities. The inclusion of sand and quarrie~ ptoduct was recently In addition to the new recruits and passed into la\y. PPQ officers perform upon identification of funds, four passive inspections at all ports of entry, collect and beagle brigade dogs will join the force. convey to th~ entomologist any pest found, However, the Guam Department of Agri­ maintain 'luatantine integrity on any culture must provide the required kennels commodity found infested, and follow up in order for USDA to tum over custody of in all quarantine commodities. the dogs and provide the training. The Department of Agriculture PPQ Section consists of 11 PPQ Officers and one Plant Protection Function professional entomologist. Recently, The plant protection function of PPQ interviews were conducted to hire 24 consists of: additional PPQ Officers. • Research • Control and eradication efforts. Training The new recruits will undertake an All these efforts require the active intensive two to three months of training, participation of a doctorate-level entomolo­ which will includes both Guam Agriculture gist with support from technical personnel. laws and regulations as well as USDA­ As noted above, the Department of Agricul­ APHIS-PPQ manuals and other pertinent ture currently employs an entomologist but training provided by the USDA. The is lacking biologically trained support staff recruits will be closely supervised by a to devote to these activities. A primary senior officer while undergoing training. concern of the entomologist is to provide This training will consist of: rapid taxonomic determinations of insects • Propagative manuals, so that their pest status is immediately • Non-propagative manuals, available. There is a need to increase the • Airport I maritime operation manuals, resources devoted to surveying for and • Animal product manuals, cataloging of pests that currently exist. This • Selective baggage training, work should be closely coordinated with • Fumigation and other types of scientists at the University of Guam. treatment, • Profiling of passengers and their PPQ Headquarters baggage. Currently PPQ headquarters issues Phytosanitary Certificates of Origin. With In addition to the PPQ training, other the recent hiring of a clerk and cashier, PPQ agencies will provide training to PPQ Headquarters will be handling all process­ officers since we closely interact with the ing of Import Permits, however we are in Guam Customs and Quarantine Agency. the transition period of undertaking this Such training will consist of: responsibility. • Drug identification (Guam Police Guam recently exported large quanti­ Department/ Guam Customs and . ties of fruits and vegetables, live plants to Quarantine), various areas including the continental • Health awareness (Public Health and USA, CNMI, FSM and Palau. In order to Social Services), protect other countries from infestation of

26 pests that currently exist on Guam, the inspected and found suitable for the PPQ Section conducts a through inspection Inspection Station. However, we are still of all agricultural commodities being working with the airport officials on the exported out of Guam. All fruits and lease contract agreement. vegetables being exported are placed in a chilled box to insure no infestation occurs Melon Fly Project prior to departure and certified with a The melon fruit fly control program Phytosanitary Certificate. started in March 1996. A total of 45 surveil­ lance traps were set around the island. Two Fines and Penalties different traps were placed at various farms Currently, PPQ officers issue civil and backyard producers and consist of: penalties to passengers, garbage violations, • A yellow cup coated with a non-toxic, etc., but only through federal authority. non-drying glue, or The Guam Department of Agriculture is • An attractant mixture of malathion, currently working on the possibility of honey, yeast and water for attracting implementing its own fines. female flies.

Inspection Station This is an ongoing project the Depart­ The Department of Agriculture is ment of Agriculture has undertaken and working with the Guam Airport Authority will likely continue as long as the public· to lease out a building for the Plant Inspec­ interest remains high. tion Station. Recently a building was

27 Weeds by. C.T. Lee

What is A Weed? water, nutrients, light, carbon dioxide, and A weed is a pest. It is an enemy of space. Weeds reduce the yield and quality agriculture and many related branches of of crop and forage species. They restrict human activity. Man has been at war movement of water, reduce the efficiency of . against weeds since the beginning of irrigation systems, cause navigation agriculture. problems, reduce fish availability, and Although we have a general concept as contribute to poor water quality for culi­ to what a weed is, there is no single nary and industrial uses. definition, because the concept of a weed is Eradicating weeds from crop fields relative and the situation whereas a plant is increases costs of production. Weeds considered a weed depends mainly upon reduce the quality of crop products. Also, the economic importance attributed to it by weeds harbor insects and diseases by people acting as alternate hosts for insects and There are many useful plants that have organisms causing diseases. Some weeds turned into pestiferous weeds simply spread pollens that cause allergies or hay because of the relative value that man has fever, resulting in misery and loss of work attributed to them. A weed, however, may time and efficiency. Weeds may also cause be described as follows: skin irritation. They obstruct the vision of • A plant growing out of place. motorists and become fire hazards around • A plant growing where something else buildings. should grow. • A plant growing where nothing should grow. Weed Control Methods • A plant that is noxious, useless, or Once a weed is properly identified, poisonous. possible methods of control can be sug­ • A plant whose economic importance is gested. yet to be discovered. I. Preventive Weed Control Thus many plants that affect the The old adage "an ounce of prevention beneficial utilization of the environment by is worth a pound of cure" is especially man fall under the category of a weed until applicable to weed control. Initially, many such time he discovers some use or need important weeds were introduced uninten­ for that _plant. Some crop plants can tionally by man and his activities. As the become weeds when they appear where farmer moved from one area to another, he they are not wanted. On the other hand, a trailed weeds as contaminants in his crop number of plants usually thought of as seeds and animal feeds. If this accidental weeds may actually be useful under some introduction could have been prevented, conditions or in some areas. For example, -· many weed control programs would not be Tangantangan (Leucaena leucocephala Lam.) needed. Therefore, a basic rule to any weed may help to control soil erosion or may control program is to prevent the introduc­ serve as firewood. tion of new species of weeds into the country, state, farm, or garden and to prevent the reproduction and proliferation Why Are Weeds Troublesome? of existing weeds. Generally, weeds interfere with man's . Introduction by animals and birds can use of land and compete with his crops for be prevented by controlling their move-

28 ment into a given area. If new animals are introduced it was essentially freed of its Weeds brought to the farm, they should be held in natural predators. To initiate control, the continued an enclosure for at least 48 hours to allow area of origin must be visited and natural passage of seeds in the intestinal tract. _The . enemies or predators must be found. These manure should be collected and allowed to - -:W.tural enemies must be specific in their ferment before use. Also, animals should be., eatin$ habits and not cause harm to crops 7 cleaned to remove all seeds adhering to the of economical importance growing in the skin, fur, or hair. Feeds and feedstuffs same.. area. These biological control agents should be checked for foreign weeds. must be introduced free of their own Tools, machinery, and supplies all are natural predators in the area in which they potential carriers of weeds. All hand tools, will be introduced. planting equipment, cultivating equipment or harvesting equipment should be thor- oughly cleaned and inspected as they are V. Chemical Weed Control moved from field to field. Chemicals used to control weeds are called herbicides. Herbicides are classified according to either chemistry or method of II. Mechanical Weed Control application. Herbicides are labeled for Mechanical weed control is the physical specific use in specific crops or for specific removal of weeds from the soil or the weed control situations. Labels should be manipulation or stirring of soil to destroy read, understood, and followed explicitly. weed growth. A number of weed-control methods falling into this category are hand pulling, hand hoeing or hand cultivation, CLASSIFICATION OF WEEDS deep plowing, mechanical cultivation (such as harrows, cultivators, rotary hoes, rod - I. Based on Morphology weeder, etc.), dipping or mowing, burning Grass Weeds and mulches. Grass weeds are monocots and have the following distinct properties: two-ranked leaves that are usually long, narrow and III. Cultural Weed Control upright with parallel venations, protected Various cultural practices common to apical meristem (growing point), and farm management can be used effectively hollow stem. for controlling weeds. Practices such as use of weed-free seed and clean equipment fall BroadleafWeeds under the category of preventive weed Broadleaf weeds are mainly dicots and control, as discussed earlier. Other practices have the following distinct properties: such as the use of cover crops, smother leaves usually broad with netted venation, crops, and crop rotation can be worked into exposed apical meristem. farm management programs in such a way as to reduce many weed problems. II. Based on Life Cycle Annual Weeds These plants complete their life cycle in IV. Biological Control one year or less. They grow from seed, The use of natural enemies to control mature, and produce seed for the next certain weeds is known as biological weed generation in one year or less. control. In most cases, considerable re­ search must be done· before an effective Biennial Weeds control can be found, because usually the Plants with a two-year life cycle. They weed to be controlled is not a native species grow from seed and develop a heavy root but has been introduced. When it was and compact cluster of leaves the first year.

29 Weeds In the second year they mature, produce a. Creeping perennials continued seed, and die. Plants produce seeds but also produce rhizomes or stolons. Perennial weeds Plants living for more than two years b. Simple perennial and may live indefinitely are perennials. Plants normally reproduced by seeds. During the winter, many lose their foliage But a root piece may produce new plants and the stems of others may die back to the following mechanical injury during cultiva­ ground. Others produce tubers, bulbs, tion. rhizomes (below ground root-like stems), or stolons (above ground stems that Bulbous perennials produce roots). Plants may reproduce by seed, bulblets, or bulb.

Each weed has one scientific or Latin name. Each may also have a number of common names depending on where it was collected and the person who identified it.

30 Insect and mite damage by Don Nafus

Movement of injurious insects by man which are small and hidden. Quantities are is a matter of considerable concern. The generally too much to search all items, so a continental United States receives about selection is usually chosen. Plants with nine new species of foreign insects each obvious damage should always be year, Hawaii about two dozen, Guam two checked, but be aware that the insect to three, and the FSM, Marshalls, and Palau causing the damage may have shifted to about one to three (Schreiner and Nafus). another plant, a corner of a box or crate, or Many of these become serious pests, some may no longer be present. damaging crops or stored food, while There are steps that should be taken in others damage structures such as homes or inspection. First, when opening a box or other buildings. Others may feed on man or container, watch for movement. Insects other animals causing annoyance or may fly out or move towards the light. transmitting diseases. Next, examine plants for insects or for These exotic pests enter on aircraft or insect damage. This should include a visual ships, sometimes as hitchhikers in cabins or examination of the outside, and movement holds, or sometimes as additional passen­ of leaves or other plant parts that may be gers on items moved by man. The primary covering insects. Then shake or brush some defense against these unwanted immi­ plants over a white sheet. Often insects that grants is quarantine procedures. Quaran­ are hard to see may fall off or be dislodged. tine cannot prevent all new introductions, Also examine corners, edges, or other but it can slow down the rate of introduc­ hidden places to see if insects have taken tion substantially, depending on effective­ refuge in these spots. ness. The more effective quarantine is, the Some of the types of damage you may fewer the problems of new pests will be. see are: One of the tools used in quarantine is • holes or cuts in leaves, inspection of imported food and plants. • skeletonized spots, Detection of pests, along with proper • folded over leaves, treatment or disposal, can go a long way • twisted or distorted leaves, toward helping slow the flood of new • dead tips, organisms entering our islands. • discolored spots, (sometimes white, Detection of pests is not always easy yellow, bronze, red, or brown. Can and may require substantial training. Many include large patches, or a few spots), of the insect pests that have entered • tracks in leaves, Micronesia in the last 50 years are scales, • blisters, mealybugs, leafminers or other insects that • bumps or odd protrusions, are cryptic and easily overlooked. The • holes in stems, fruits, buds, roots, purpose of this section of the course is to corms familiarize you with some of the kinds of • swollen buds or stems, insects that historically have evaded • discolored area on fruits, bulbs, roots, quarantine. We will also look at various • nodules or lumps on roots, types of insect damage, so that you can • tunneling in enlarged roots, reducing develop an idea of the potential causal crop, agents you may need to search for. • distortion or blemish of fruit, and • chewed plant rem.ains, insect feces, or cast insect skins. Plant Inspection Imported fresh vegetables may have a Look at the damage carefully to wide variety of insects on them, most of determine if it is fresh or not. Fresh damage

31 Insects and should not have dried or brownish looking damaged leaf, but would check other mites continued edges. Also check to see if traces of the hiding spots on the plant. insect such as feces are present. If you find Grasshoppers, other beetles and these, you need to check for freshness also. Lepidoptera can create very similar types of Caterpillar feces should be moist looking damage, but the type of holes, placement, and intact if fresh. If the damage is old, and other features usually vary. Grasshop­ most likely the insect has left the leaf, or pers tend to create larger holes, and often perhaps was killed. Many insects will feed from the edge of the leaf rather than move away from the feeding site to pupate. the interior. Lepidopter(l damage can vary Also, sometime damage is done by insects greatly, ranging from edge feeding to such as grasshoppers, which eat and then scraping off tissue, to rolled leaves, to move on to another site. mines. Sometimes even Hymenoptera can If the damage is fresh, it is possible that create damage to leaves. A leaf-cutter bee some kind of insect is present. Knowing cuts semi-circular chunks of leaf out and something about the kinds of insect uses them to line its nest. In this case, the 9-amage may help you make decisions damage would not be of significance to about what to look or not look for. It is an quarantine, because the causal agent would aid to quarantine efforts. never be found on the plant. Chewing insects also tunnel or bore in twigs, stems, roots, fruits or scrape away TYPES OF INSECTS tissue on the outside. Many beetles and The different types of mouth parts Lepidoptera tunnel in stems or twigs. The found in insects offer clues to the type of adult longhorn beetle girdles the twig damage that they create. There are actually consuming the bark, and the larvae feed several types of distinct mouth parts; the along the cambium underneath the bark. most important of which for our purposes Often the tunnels are packed with frass, or are chewing, sucking, and rasping-sucking. fecal matter. In other cases the frass is expelled, as in the case of powder-post Chewing Insects beetles. On the outside of dry wood, in this Insects with chewing mouth parts case bamboo, you see tiny shot holes. include the beetles, butterfly and moth Below, there are piles of what looks like fine larvae, and grasshoppers, crickets, and grained sawdust. The internal galleries, as other related orders. Damage from these shown in the cut-away, are usually empty. groups is variable. On leaves they tend to Lepidoptera may also expel their frass, create holes of one kind or another, or particularly those that feed on succulent tracks or mines. tissue. On the outside you see a hole, and The type of damage caused by an sometimes a pile of excreted material, and insect is often quite specific, and sometimes on the inside a large cavity. We see similar you can even tell which species created it. cavities excavated in roots or tubers at­ For example, damage by rose beetles tacked by Lepidoptera. In these cases there consists of distinctive ragged holes in the are visible holes, but in other cases the only inside of the leaf. The holes typically have symptoms may be swollen areas, lumps, irregular edges, are inside the leaf margins, discolored spots, or at times no symptoms and tend to be somewhat the same size. are visible. This is also true with root or The size and shape of the holes is charac­ tuber feeding beetles. In the case of the teristic, and you can usually identify rose sweet potato weevils, you may or may not beetle feeding without seeing the beetle. find a hole outside the tuber. If you do find This is useful because you normally do a hole, it is usually an exit hole. If you cut not find the beetle on the plant. It feeds at the tuber open, you will find numerous night and hides elsewhere during the day, tunneling tracks. Scalloping can be done by often in leaves folded over by a spider or a · either mole crickets or termites. leaf-folding insect, or in leaves pressed Beetles and Lepidoptera also tunnel in together for some other reason. In this case fruits. Sometimes you may see exit holes, or you would not look for the insect on the frass removal holes, but at other times, 32 symptoms are less conspicuous. In the case and are not noticed by the general public. /nseds and of soybean tunneling, there may be an exit Some adult Lepidoptera will pierce mites colrtinued hole, but more often there is only a discol­ fruit, and others, which cannot themselves ored spot, or the area where the seed is create holes, will feed on fruit juices if the developing may be marked by slight fruit is pierced by another insect. This type brown discoloration. In mangoes, seed of damage can easily be confused with weevils often leave no trace while they are damage caused by oviposition of fruit flies. developing. Fruit-piercing moths pierce the skin, and suck juices out of the fruit, leaving an Sucking Insects empty cavity and a hole that allows access The most important group of sucking for rot organisms. Scouring beetles and insects is the Heteroptera (aphids, scales, other insects may also use these holes for whiteflies, leafhoppers, plant hoppers, and access to the fruit. other true bugs). Damage caused by these insects may be very difficult to spot. It Diptera ranges from feeding spots,.which are tiny Leafminers, stalk miners and other holes in the leaves and may be virtually kinds of flies may bore in leaves or stems of invisible, to small circular yellow spots, plants. Serpentine leaf miners tunnel inside burned yellowish areas, distorted curled the leaf leaving a whitish track with black areas, or dead vines or tips, Sometimes the feces. On Guam, Yap and Palau, the round most visible symptom is sticky or shiny circular holes found in mango leaves are leafs covered with sugary excretions created by a fly larvae that is present only a voided by the insects. Commonly a black few day. Flies feed with a mouth hook mold, called sooty mold, grows on the which rakes in the tissue. Fruit flies are a sugar making the leaf appear black. serious problem in fruits. These maggots Some species of insects may create very tunnel in fruit creating a rotten spot, or distinctive growths on the leaves called perhaps causing the entire fruit to rot. galls. The insects live inside the lumps. Fruits with maggots are easy to overlook, These come in a wide variety of sizes and and if only punctures from oviposition and shapes, and each species creates its own eggs are present, they may be nearly distinctive shape. Other animals may also impossible to spot. For this reason it is cause galls including wasps, flies, mites, better not to allow entry to any fruit from and nematodes. areas that have fruit flies you do not want Yellow spots, burned looking areas, in your island. dead tips and similar damage is often caused by the saliva being toxic to the plant. This causes cell death in areas Insect Eggs beyond the initial feeding site, magnifying Insect eggs are a serious problem for the feeding damage and making the attack quarantine inspections. Many eggs are very more visible. small, even when the insect is large, and Homoptera are probably the group many are cryptically colored or located in that has moved around Micronesia the spots where they are hard to find. Eggs most. This is principally because they are may be laid between scales in a bud, in hard to recognize. Most species do not punctures in a leaf, in cracks or crevices in cause enough damage to be visible when in bark, or in other cryptic places. This makes low numbers, and the insects themselves them nearly impossible to find, and is one are tiny and often covered with wax scales of the reasons all plants being introduced that make them easy to overlook. Some should go though a period of quarantine whiteflies are transparent and flat in the after entry. juvenile stage, and are almost impossible to Flowers are a signifjcant source of see. We have had several species of white­ insect introductions. Thrips and other flies moving around the Micronesian insects often visit flowers to feed on nectar, region in recent years. Many scales look and they go inside the flower where they more like parts of the plant than insects, are not visible. Thrips feed by rasping open 33 Insects and cells and sucking up the contents. They One of the best ways to learn is to go out mites continued cause scraped looking areas with brown or into the garden, forest, farm, or other areas white scars, and sometimes can cause and look for damage on plants. Once you distorted or twisted leaves. Mites can also find it, you can then try to find the cause. cause this kind of damage. Observing the natural world can be a It takes a lot of time to learn how to pleasant and diverting way to enhance your spot some kinds of insect damage, and to job skills. recognize which ones may be the cause.

34 Invertebrates of quarantine importance by lise Schreiner

Overview of the life cycle and distin­ herbivores feeding on leaves and other guishing characteristics of orders of vegetative matter. Mole crickets feed on insects roots. Eggs are laid in soil, or in the case of katydids, in slits under tree bark. In Guam, Collembola: springtails in past 40 years at least two katydids and 1. No metamorphosis. Life cycle: egg, two grasshoppers have been introduced. multiple instars, continues to molt after One grasshopper has become an agricul­ adulthood. tural pest feeding on vegetable crops. The 2. Since they have no metamorphosis, two katydids occasionally damage trees adult are wingless an<;i look just like laying their eggs. immatures. 3. Chewing mouthparts. Orthoptera: mantodea 4. Springtails get their name because Mantids are predators. Their most most species have a forked structure outstanding feature is their striking legs. (furculum) on their abdomen that There are no native species in Micronesia, enables them to jump. At rest the but several species have become established structure is folded under abdomen and are spreading. These are probably held in place by a clasp-like structure. moving as eggs. Mantids lay their eggs in 5. Mostly or no quarantine significance as foam cases which are attached to a variety they feed on organic material or fungi of things including shipping containers. in the soil. A few species damage They may have deleterious effects on native gardens, greenhouses and mushroom insects, and should not be introduced by cultures. These species tend to have carelessness. round rather than elongated bodies. Orthoptera: blattodea Orthopteroid orders Cockroaches are general feeders. A group of related insects which used Several species live in houses with man. to be grouped into one order, the Ortho­ Human-associated cockroaches are the ones ptera, but has now been divided into most likely to reach new islands. Eggs are several groups as described below. laid in little cases which are left in dark 1. Incomplete metamorphosis. Life cycle: comers. egg, several instars as a nymph, adult. 2. Most of these insects are fairly large, lsoptera: termites even smallest crickets tend to be at 1. Termites are similar to the other ortho­ least 3-4 millimeters. pteroid orders, but are social, living in 3. Often have elongated front wings huge colonies. which are thickened and leathery. 2. Termite life cycle. An individual has 4. All species in this group of order have incomplete metamorphosis. All ter­ chewing mouthparts. mites are social and can only live in 5. Phasmida stick and leaf insects are groups, so there is also a colony life disguised as twigs or leaves. Eggs cycle. Colonies are founded by a mated are hard and seed-like, and dropped at winged adult male and female. These random. No recent quarantine prob­ then rear non-winged workers and lems in Micronesia. soldiers which do f!.ll the jobs in the colony. Eventually a large colony will Orthoptera: crickets, grasshoppers , mole produce winged males and females crickets and katydids which leave to form new colonies. These insect for the most part are

35 Invertebrates 3. Termites eat wood or sometimes dried 3. Very diverse shapes. Many do not look continued grass. like insects (scales, mealybugs,.5pittle- 4. Termites look like ants, which also live bugs), but resemble bits of a plant. This in colonies. The differences are: can be stem-shaped, leaf-shaped, or a) Termites have four wings of about simply small and_inconspicuous equal size in adults, while ants have without a recognizable animal shape. four wings but the first pair is much 4. Piercing sucking mouthparts. larger than the second pair. 5. All Homoptera are plant feeders. They b) Termites are relatively soft bodied, plug into the sap or into. cells, causing '1 often white or brownish white. They leaf curling, discoloration and yield are not hard or metallic like ants. Ants loss. I are active in sunlight and are darkly 6. Excrete excess sugar in sap in their pigmented as a result. feces. It evaporates creating a sticky c) Termites have their waist rather mess on the leaf. Fungus grows on it, broadly joined to the thorax. In ants, turning the leaf black (sooty mold). the waist is ve,ry slender and narrow. 7. Transmit many plant diseases, espe- 5. No recent quarantine problems in dally viruses and mycoplasma. Micronesia, although the worst pest, 8. Many species reproduce asexually. the "Formosan" termite, was intro- Only one female is necessary to popu- ducedto Guam around the end of late an island. World War II and has since moved to 9. Eggs often laid under scale of body of some or all of the other islands of female, if the species do not move Micronesia. much, otherwise inserted into stems of plants. Adult female aphids give birth Hemiptera: true bugs including stink bugs to first instar nymphs. 1. Incomplete metamorphosis. Life cycle: 10. In scales and mealybugs, the mobile egg, several instars as a nymph, adult. stage is the first instar nymphs. Adults 2. Variable size. Some may be several do not move much or at all. centimeters long as adults, while 11. Serious quarantine threat. About one others are less than 1 millimeter. third of the new pests introduced to 3. Wings are half leathery, half membra- Guam in the last few decades have nous. been Homoptera. As they cannot live 4. Mouthparts are modified for sucking. long without their host plants, it is 5. Variety of feeding habits, including likely most are introduced on omamen- plants and other insects. A few species tal plants being brought in. are blood suckers. Some are facultative predators meaning they will eat other Thysanoptera: thrips insects when available, but will eat (Thrips always ends in "s". If you have plant tissue if no insects are to be only one, it's still called a thrips). found. 1. Thrips are very small insects 6. A fairly recent introduction throughout (1/2 to 5 mm). Micronesia is a small shiny black stink 2. Feed by scraping plant cells and bug (which is often confused with sucking up the cell contents. This leaves beetles). Feeds primarily on various a rough surface behind and can cause types of beans and related species. brownish patches on leaves and fruits. 3. Most thrips eat p1ants, but a few are Homoptera: cicadas, hoppers, psyllids, predators. whiteflies, aphids, scale insects, mealybugs 4. Life cycle intermediate between incom- 1. Incomplete metamorphosis. plete and complete metamorphosis. 2. Most have two pair of membranous Immatures look like the adults, but they wings. The wings may be clear, col- also have pupal stage (a resting stage ored, or covered with wax. Many where they change from the immature species are wingless, particularly the to the adult form). females. 5. The melon thrips recently introduced 36 into Micronesia has become a serious 10. Some butterflies, especially the Invertebrates problem in the Marianas and some birdwing butterflies of Australia and continued other islands. It seriously damages New Guinea, are listed on the CRMS vegetable crops, but also is found in list. Importing these species may be orchid flower, the likely route by which illegal. it reached Guam. Western flower thrips, a serious pest in Hawaii, is often found Coleoptera: beetles in vegetables and flowers coming into 1. Complete metamorphosis: egg, larva, Guam, but has so far been successfully pupa, adult. intercepted by quarantine officers. 2. Beetle adults are characterized by having the first pair of wings hard, Lepidoptera: moths and butterflies leathery or brittle. The hind wings are 1. Complete metamorphosis. Life cycle: membranous. These are used for flight, egg, larvae (caterpillar), pupa (chrysa­ and are folded under the protective lis), adult. first pair when not in use. 2. Larvae have chewing mouthparts. 3. Beetle larvae are quite variably shaped, Adults have a long sucking tube which but are characterized by having a is coiled up when not in use. distinct head region and three sets of 3. Caterpillars are wormlike, but have a true legs (no false ones). distinct head, three true legs Gointed 4. This is the largest group of insects. legs) and variable numbers of false legs Over 250,000 are known; many more (fleshy legs). remain to be found and described. 4. Adults have four wings, which are 5. Variable size, ranging from less than 1 covered with scales which give them mm to 125 mm in size. The goliath their color. beetle outweighs a mouse. 5. Many moths are attracted to lights 6. All have chewing mouthparts in larval (such as the lights in the hold of an and adult stage. airplane on the runway being loaded 7. There are species that eat specialized to after dark). almost everything plant related (leaves 6. Caterpillars have chewing mouthparts. or flowers or stem borers or root Almost all are plant feeders. They eat feeder). Seed eaters may become leaves, bore in wood, or eat grain or stored-product pests. Stem feeders cereal products. A few are predators or include wood pests. Scavengers eat scavengers feeding on material such as carrion, dung, mold, fungi, organic fur (wool) or bee's wax. matter of various types. Many are 7. Adults in general feed on flower nectar. predators, eating other insects or snails. An exception are the fruit-piercing 8. Several species have become estab­ moths. lished in Guam in last couple decades. 8. Variable size. Biggest moths have Two are notable because one is con­ wingspans up to 25 em. Smallest fined (so far) to Andersen Air Force leafminers may only be 2 mm. Base and one to the Navy base (indicat­ 9. Several species of moths have become ing military vessels also have a role in established in Guam in the last few importing pests). So far, neither has decades. Many of these have caterpil­ turned out to be much of pest, though lars which feed on trees (such as the both have the potential if they become flame tree looper). Because ofthis it is very abundant. unlikely they are introduced while feeding on ornamentals, and more Diptera: flies, mosquitoes, midges likely that they came in as an adult in 1. Complete metamorphosis. Life cycle: the holds of airplanes. However, a egg, larva (usually .called maggot), butterfly which recently became pupa, adult. established on Saipan probably came 2. Adults have only one pair of membra­ as an egg or caterpillar stage on an nous wings. The second pair is reduced ornamental cycad (federico palm). to halteres which spin around in flight

37 Invertebrates and act as gyroscopes. social systems; that is, they live in large continued 3. Fly mouthparts are built for sucking or groups with a queen who lays eggs and sponging. In many cases, the mouth many workers to do the work in the parts telescope into the head of the fly nest or hive. when not in use. 8. Not of great quarantine significance 4. The larvae are legless maggots. The because they are not plant feeders, but mouthparts consist of pair of mouth social insects can be very disruptive to hooks. island ecosystems. Care must be 5. Most flies feed on rotting organic exercised by quarantine officers as, for material as larvae. However, some feed example, Africanized bee nests have on plant tissues such as fruit or as been discovered in shipping containers. leafminers inside leaves. Some feed on 9. Recent accidental introductions which other insects, and some on flesh. have become established on Guam 6. Adult flies feed on nectar or other include several parasitic species which . ·juices, or on blood. may have come in with new plant pests 7. Mosquitoes and a few other kinds of (which have been beneficial more than flies are most damaging as carriers anything) but also big headed ants (vectors) of disease. which are a great nuisance in houses 8. Recent accidental introductions of and have destroyed to forest ecology of Diptera to Guam include mosquitoes many islands of Eastern Polynesia. (which probably came in as adults in passenger holds), and several leafminer pests of vegetables which II. NON-INSECT ARTHROPODS probably came in as eggs or larvae in an ornamental plant. Quarantine Acari: mites and ticks officers have successfully prevented 1. Body usually oval and compact, with fruit flies from entering Guam despite little differentiation between the two constant attempt by the public to body segments. Newly hatched ones smuggle fruit. have three pairs of legs and acquire fourth pair after first molt. Hymenoptera: ants, bees, and wasps 2. Ticks are fairly large, and feed on the 1. Complete metamorphosis. blood of vertebrates 2. Adults have four membranous wings. 3. Ticks carry a few important diseases, The hind wings are much smaller than such as Rocky Mountain Spotted Fever the front and often hooked together so and Lyme Disease. they can appear as one. Except for one 3. Mites are small to minute. They are group (sawflies), adults have "wasp" scavengers and predators, or feed on waists. plants. Many are parasitic on verte­ 3. The larvae are legless except for one brates or invertebrates (mange, ear group which looks like caterpillars mites). (sawflies). 4. Dust mites feeding on dust in houses 4. Adult mouthparts vary from chewing are one of main causes of asthma. to lapping-sucking. 5. Some mites induce their host plants to 5. Many are beneficial species. Either they make galls and they live inside the feed on pollen and nectar (bees) and shelter of the gall. are useful pollinators or they feed on 6. Not well studied in Micronesia, so not other insects (wasps). Sawflies (which much is known about recent introduc­ .are extremely uncommon in tropics) tions. feed on plants as larvae. Ants feed on a great variety of things. Spiders 6. Females, but not males, can sting. The · 1. Eight legs, two body regions: cephal sting can be painful, and for some othorax and an unsegmented abdomen. people with allergic reactions, deadly. 2. All spiders have spinnerets on abdo­ 7. Many Hymenoptera have advanced men to spin silk. 38 3. Use silk to make webs, entangle prey. III. MOLLUSKS Invertebrates Young spiders float on the wind at the continued end of a long strand of silk (can cross Gastropoda: slugs and snails the ocean that way). 1. In snails, the most obvious parts of the 4. All are predators. anatomy are the mantle and the shell. 5. Some have very poisonous bites. 2. In slugs, the shell has been reduced to 6. No recent quarantine problems in a small plate covered by the mantle. Micronesia, but black widow spiders 3. Snails and slugs move on a trail of were accidentally introduced to some mucus produced by glands near the remote islands the Air Force used in front of the body. the Atlantic. 4. In most snails the mouth opening is surrounded by a set of small teeth-like Scorpions structures called a radula, and most 1. First set of appendages behind cheli- snails feed by scraping. cerae are long and clawlike, posterior 5. Snails can be plant feeders, predators, end of creature terminates in a sting . scavengers or parasites. 2. Largely nocturnal in habit. Feed largely 6. Snails live on land and in the water. on insects and spiders which they 7. Freshwater snails are the intermediate capture with their claws and kill with hosts of some serious human diseases their sting. such as schistosomiasis and blood, 3. The young are born alive, and are liver and lung flukes. carried by their mother for some time. 8. Many of the commonly encountered Some require several years to mature. snails are hermaphrodites (both male 4. Effect of the sting on man depends on and female). the species, mostly just painful and 9. Snails lay eggs. In land snails and slugs, accompanied by local discoloration these are generally placed inside a and swelling. One species known in capsule. In fresh water snails, they are the U.S. can be fatal. often laid on plants. 5. One species on Guam is native; the 10. Some snails are eaten. Freshwater other introduced through human species eaten in Asia have been ille- commerce some time ago. gaily imported and released into the streams of Guam. 11. The most serious recent quarantine problem on Guam has been a slug probably introduced in potting mate- rial with ornamental plants. This slug has become a serious pest of gardens.

39 Indexing of plant virus diseases: Serological diagnosis by Norihiko Saito

Important plant diseases pathogens, • Serological diagnosis. viruses are composed of nucleic acid (DNA • Gene diagnosis. · or RNA) and coat proteins. They are too small to detect with light microscope and Each of these methods mentioned incapable to culture artificially. Some above has its merits and demerits. The methods are used singly or in combination symptom observations are easy to do, but for detection and identification of plant are difficult to identify infected virus. The viruses. Serological diagnosis is one of electron microscopy made it possible to them and is very effective method. Espe­ observe virus particles, but equipment is cially, enzyme-linked immunosorbent expensive. The transmission test needs . assay (ELISA) is widely applicable for many kind of herbaceous or woody indica­ many plant viruses because of its high tor plants, but are able to identify infected utility. Many samples can be indexed at virus. Serological diagnosis needs specific once with high sensitivity. And time anti-serum for each virus, but it becomes requited for indexing is very short. So possible to index many samples at one test ELISA can be said one of the most suitable with high sensitivity by a development of virus indexing method for plant quarantine ELISA. Gene diagnosis, including CDNA inspections. probe method and PCR, is the newest method and in development, but it is becoming more common. In this paper, the Introduction serological diagnosis is described. Pathogens of infectious plant diseases are divided into two groups. One is a microbe group including bacteria and Serological Diagnosis fungi. Another one is a virus group includ­ The outer coat of plant viruses consists ing viruses and viroids. of protein subunits characteristic to each Viruses are composed of nucleic acids virus. The virus particle, therefore, because (DNA or RNA) and coat proteins. Viroids of its specific three-dimensional shape and are composed of RNA only. They are very size, forms an ideal antigen following .. small pathogens which are not visible by injection into the blood system of a suitable any optical microscope. Virus and viroid,s animal, such as rabbit or mouse. The particles are able to be observed by electron injection of antigen (virus particle) causes microscope only. So we are limited to the production of antibodies. Such antibod­ observations of symptoms produced by ies, present in the serum fraction of the viruses or viroids on various host plants blood, react only with the injected virus or with the naked eye. They also are incapable with very closely related viruses. The to reproduce by culture artificially. There­ specificity of this antigen (virus) - antibody fore, the following methods are used singly (antiserum) reaction can be used in a or in combination for detection and identi­ variety of in-vitro tests to index viruses. fication of these pathogens. In case of viroids, these pathogens are • Observations of symptoms appeared naked RNAs without coat protein. For lack on host plants. of coat protein, serological diagnosis is • Electron microscopy to detect virus ·inapplicable to indexing of viroids. Re­ particles. cently, RNA sequences of most viroids have • Transmission tests using indicator been established and gene diagnosis such plants. as PCR are used for viroids indexing.

40 Many plant viruses can be purified ELISA with comparative ease and the production Many disadvantages encountered by of specific antisera of a high quality has the user of the serological tests involving become routine procedure. precipitation or agglutination reactions are To obtain antiserum, at first, virus overcome by labelled antibody techniques purification is required. Virus purification such as the enzyme-linked immunosorbent follows a number of ba6ic steps. The first assay (ELISA) .. A frequent requirement is step is virus inoculation to host plants in the routine detection of plant virus pres­ which virus can multiply by systemic ence in large number of samples. The invasion. Having the time at which maxi­ ELISA test has the following advantages. mum virus content occurs, the material is • Extreme sensitivity. harvested. The harvested virus-infected • Applicability to large numbers of material is homogenized using suitable samples. buffer. The crude sap may be clarified by • Economy in use of high cost antisera. low speed centrifugation, filtration, the • Semi-automatic. addition of active carbon,.etc. The next step, • Quantitative. deferential centrifugation, is used for concentration of virus; many other methods But it needs purification of IgG and conju­ are also available. Finally, rate-zonal gation of IgG with enzyme. density-gradient centrifugation should The enzyme-labeled antibody tech­ remove all containing materials, leaving a niques were first developed in the medical pure virus preparation. field. ELISA tests adapted to detection of The purified virus is injected into plant viruses by using polystyrene rabbits or other suitable animals intrave­ microplate was described by Voller et al. nously or intramuscularly or in combina­ (1976). Clark and Adams (1977) improved tion of both at intervals of several days. this method. Recently, ELISA become the After 1-3 weeks of immunization, the most common serological diagnosis. And antibody level become sufficient. Blood many ELISA kits of important viruses, such samples are allowed to coagulate and the as potato viruses, have come onto the serum may be decanted and clarified by market. In the case of Japanese plant low speed centrifugation. quarantine, ELISA is used for post-entry The specific component of antisera is quarantine of fruit tree nurseries, potato made up of globulins, and the type and tubers, flower bulbs, and for official amount of these determine the quality of inspection of domestic fruit tree mother the antiserum. The essential immunoglobu­ stocks. lin is IgG. IgG exists as a Y-shaped mol­ ecule, of which the arms of the Y are the combining sites which are specific for each 4. ELISA Protocol antiserum. They combine with the protein There are basically two different forms of the target virus. of ELISA tests. Many in-vitro tests have been devel­ Inthe first, microplate wells are coated oped which use the specific relationship with the specific IgG and incubated with between an antigen and its (homologous) the plant sap followed by adding "conju­ antiserum. These tests can be conveniently gate," a specific IgG that is directly conju­ divided into four different types: precipita­ gated with an enzyme. In most instances, tion, agglutination, immunodifusion, and the enzyme used to label the specific IgG labelled antibody techniques. The first three has been alkaline phosphatase. Then, types of tests involve the visualization of enzyme substrate is added and resolved by antigen/ antibody precipitates. Disadvan­ the enzyme. The resolved products appear tages of these tests are that a large amount yellow in color. Colore~ products are in of antisera is required, their sensitivities are proportion to concentration of virus. This is comparatively low, not quantitative, etc. called "direct sandwich" or "double But these tests are able to use antiserum sandwich (DAS)" ·ELISA. itself without any processing. In the second, "indirect ELISA," plant

41 Serological sap is added to wells and incubated, Reagents continued followed by a primary IgG specific for the Phosphate buffered saline pH 7 .A (PBS) targeted virus and secondary IgG-enzyme 8.0 gNaCl

conjugate specific for the primary IgG are 0.2 gKH2P04 added. Then, enzyme substrate is added. Make up to 1litre with distilled DAS ELISA is more common than indirect water

ELISA at routine inspections in Japanese 2.0 g Na2HP04 .12Hp to give normal plant quarantine. Abbreviated method of concentration for ELISA use. DAS ELISA is described below. 0.2gKCI Make up to 1litre with distilled water to 1. Viral antibody (IgG) adsorbed to plate. give normal concentration for ELISA use. 2. Virus added. 3. Enzyme-labelled antibody added PBS may be conveniently handled and (conjugate). stored as 10-times concentrate (adapt above 4. Enzyme substrate added. dilution accordingly) which can be diluted as appropriate to 2 x , 1 x or 1 I 2 x strength, as required. Such concentrated stock Abbreviated Method of DAS ELISA solutions need no preservative or antibacte­ 1. Add 200 ml purified IgG diluted in rial additive such as sodium azide. Diluted coating buffer, to each well of the PBS, however, will accumulate bacteria microplate. rapidly if it kept without preservative. It is 2. Incubate for 2-4 hours at 37 o C . best to make a fresh supply daily from the 3. Wash wells with PB5-Tween. Leave at 3 10-times concentrate. min. Repeat wash three times. Empty plate. PBS-Tween 4. Add 200 ml aliquots of test sample to Add 0.5 ml of Tween 20 to 11itre PBS. two wells lined side by side. 5. Wash plate three times as in (3) above. Coating buffer pH 9.6 6. Add 200 ml aliquots of enzyme­ 1.59 gN~C03 labelled IgG to each well. Incubate at 2.93 g NaHC03 37 ° C for 3-6 hours. Dissolve in 1litre distilled water.

7. Wash plate three times as in. (3) above. 0.2 gNaN3 8. Add 200 ml aliquots of freshly pre­ NaN3 (sodium azide) is toxic and pared substrate to each well. Incubate solutions containing it should not be at room temperature for one hour, or as mouth pipeted. long as necessary to observe reaction. 9. Stop reaction by adding 50 ml3M Dissolve in 1litre distilled water. NaOH to each well. 10. Assess results by: Substrate buffer (a) visual observation; 97 ml of diethanolamine (b) measurement of absorbance at 800 ml distilled water

405 nm of light wave length using 0.2gNaN3 microplate reader. Mix together. Then add HCI drop-wise to bring pH 9.8. This should bring volume almost to 1litre. Top up to 1litre with distilled water.

Equipment 96 wells microplate 200 ml micro pipette microplate reader mortars and pestles

42 Linskens, H.F. and J.F. Jackson. (1986). Serologi~a/ M.F. and A.N. Adams. (1977). Immunology in Plant Sciences. Springer­ continued of the microplate method of Verlag. Berlin, Germany ,.,-,lll'-'""'"" immunosorbent assay for letedi<>n of plant viruses. Journal of Hillt S.A. (1984). Methods in Plant Virology, Virology 34,475-483. Blackwell Scientific Publications. London, Great Britain. A. et al. (1976). The detection of by enzyme-linked immunosorbent (ELISA). Journal of General Virology 67 l;{egenn1ort:eJ M.H.V. (1982). Serology LITUnrurrut:neJliLI·~m of Plant Viruses. Press. New York, U.S.A.

43 Quarantine surveillance for fruit flies by Allan Allwood*, presented by Russell Campbell

Introduction No amount of quarantine, with the _ Fruit flies (finnily Tephritidae) are exception of a complete prohibition of classified as one of the most important movement of produce and people which is insect pest groups of horticultural produc­ not acceptable to any country, will prevent tion and export throughout the world. the introduction of exotic fruit flies or other Over 4,500 species occur worldwide. About · pests and diseases. Therefore, effective 50 species are regarded as major pest early warning systems and emergency species; at least another 30 species may be response plans must be in place so that of minor economic importance. Twenty­ quarantine and plant protection services two· of the major species occur within the are able to respond quickly to any incur­ South Pacific region. sion of an exotic fruit fly in a sound techni­ Fruit flies are very mobile and, as a cal way. result, pose significant threats to quaran­ This paper aims to introduce partici­ tine security and inevitably to the national pants to the issues that may be considered export trade in fresh fruits and fleshy . in setting up quarantine surveillance for vegetables in all South Pacific countries. exotic fruit flies. It should be remembered Relatively recent introductions of economi­ that quarantine surveillance is only one ccilly important species, such as papaya · component of overall quarantine prepared­ fruit fly (Bactrocera papayae) into Papua ness to cope with outbreaks of exotic fruit New Guinea, Torres Strait Islands and the flies that will eventually happen. This Cairns area of Queensland, melon fly (B. paper is not meant to present a comprehen~ cucurbitae) into the Perth area of Western sive quarantine surveillance system Australia and the Honiara area of Solomon because it is recognized that each country Islands, and Mediterranean fruit fly in the South Pacific has its own require- ·(Ceratitis capitata) into the Auckland area of . ments and conditions and should design New Zealand, demonstrate their mobility. systems that are appropriate to its specific Also, recent records of exotic fruit flies in needs. fruits collected from amnesty bins at airports a,nd confiscated from passengers in New Zealand clearly show that serious Purpose of Quarantine Surveillance fruit fly pests are capable of moving Quarantine surveillance encompasses around the Pacific. activities that assist in the early detection The greatest quarantine threat is via of unwanted exotic fruit flies and other fresh fruits and vegetables carried by pests and diseases. It is not restricted to pa,~_:;engers on aircraft or as food on board trapping systems alone. It includes bprder boats. In the South Pacific where bearing · inspections, passenger profiling, profiling gifts for friends and relatives is common of airline and boating routes with respect and culturally expected, the risk of intro­ to level of risk, assessments of the contents ducing dangerous fruit fly species is very of amnesty bins, trapping, host surveys, real. Commercial consignments, on the · and public and government awareness. It other hand, are not likely to carry damag­ is designed as an early warning system so ing fruit fly species, providing quality that authorities are able to mobilize assurance schemes during production are resources quickly to deal with any incur­ adhered to and off-shore quarantine qion or establishment that may occur procedures are followed. before the introduced species becomes too

*Allan Allwood is chief technical advisor, Regional Management of Fruit Flies in the Pacific Project, Secretariat of the Pacific Community (South Pacific Commission), Suva, Fiji. Paper used with his permission. 44 widely distributed. ensure penalties imposed by magis- _ .. Having an effective quarantine surveil­ trates are appropriate to the serious­ lance in place gives importing countries ness of the breaches of quarantine greater confidence in data on the presence regulations. (cir absence) and economic importance of fryit fly species that already exist in each country. Quarantine surveillance is a Establishment of Amnesty Bins/Quaran­ prerequisite for overseas trade in fresh tine Bins and Assessment of their contents fruits and vegetables. The voluntary surr~nder and disposal of fresh fruits and vegetables arid other animal and plant products into amnesty Some Components of Quarantine bins or quarantine bins is a painless way of Surveillance Systems gaining the cooperation of the travelling public. On the way to achieving vollll1fary Public and Government Awareness disposal of fresh produce, there is an ideal ..... Tourism has become an important opportunity to educate the travelling money earner within the Pacific Region. public on the importance of plant aJ1d Pacific destinations have become very animal quarantine and particularly the popular amongst tourists from Asia and ramifications of illegally moving ltntreated Europe. European and northern Asian fruits and vegetables between countries. tourists are often fascinated by tropical Amnesty bins need to be easily acces­ fruits, such as guavas, starfruits and sible and clearly identifiable to the traver:.· mangoes, but are not familiar with the strict lers. ·Clear signs and posters need to quarantine restrictions of island countries. emphasize to the traveller the dangers of Consequently, to reduce the risk of spread bringing in fresh produce and the penalties of exotic fruit flies, these groups of travel­ ·involved. The bins should be located in . r lers need to be better informed than they several areas of the arrival concourse or are at present. wharf. Coupled with the practice of bearing The contents of the amnesty bin may gifts of fruit when Pacific Island people be sampled regularly by quarantine and travel between countries, improved trans­ plant protection services to detenp.ine the port throughout the region means that the origin of the fresh produce, the amount of risk of fruit flies moving between countries each type of produce, the incidence of fruit is higher now than it was, say, 10 years ago. flies or other pests, and the species of frttit National and regional programmes need to flies. This information then may be used to focus on this group of travellers as well. profile airline or shipping routes. This In developing a quarantine surveillance assessment also defines the level of risk · system, consideration may be given to the posed by passengers from particular ports use: of origin and consequently may assist in • of in-flight videos depicting the determining the percentage of passengers importance of fruit flies as quarantine whose luggage may be searched from these and production pests and what travel­ routes. lers should do; • posters at airport and sea terminals; • clearly marked amnesty bins I quaran­ Airline and Boat Route Profiling tine bins at airports and sea terminals; By using amnesty bin assessments as • pamphlets to be included with tickets mentioned above, it may be possible to by travel agents-and--airline companies; identify the high risk routes~ In the case of • awareness projects in schools; and trying to combat papaya fruit fly spread • fostering of specific awareness educa­ throughout the South Pacific, it may be tion within government, the horticul­ feasible to focus attention of passenger tural industry and the legal fraternity luggage from flights originating or passing to facilitate the funding of quarantine through ports in northern Queensland, preparedness programmes and to particularly Cairns, and through PNG.

45 Fruit flies Similarly, because of the serious fruit flies species together with lists of other pests continued that occur in Nauru, flights to Solomon and diseases. An assessment of the eco­ Islands and Fiji from Nauru may be nomic importance of each species may also targeted. be required and this will require data on the commercial and wild host ranges of each species. Host surveys should complement Passenger Profiling the trapping system. The trapping system It is well known that some classes of should be designed to detect, as early as passengers from certain areas in the world possible, the incursion of an exotic fruit fly are more likely to carry quarantinable species, thus facilitating early eradication products than others. As mentioned above, attempts or other actions. some Asian and European tourists are very Ideally, and based on the system that interested in tropical fruits such as man­ operates in New Zealand with some minor goes, guavas and starfruit and may carry modifications, the system may comprise the these between countries. Unfortunately, following components: these commodities are highly susceptible to • an array of traps (modified Steiner fruit flies and could result in significant traps) that are serviced every two numbers of eggs or larvae being brought weeks in urban areas, particularly into a country unintentionally. By combin­ those associated with tourism and ing the airline route profile with passenger trade activities, and in horticultural profile, it may be possible to focus quaran­ production areas; tine inspections so that the chance of • for trimedlure and Cue-lure baited intercepting the high risk commodities and traps, approximately eight traps per carriers may be improved. square kilometer, placed about 400- SOOmapart; • for methyl eugenol baited traps, Border Inspections approximately one trap per square From a fruit fly quarantine perspective, kilometer, placed ca. 900 to 1100 m the border inspections of greatest impor­ apart; tance are passengers and their luggage, • traps to be placed in host plants with parcels sent by post or air freight, and good foliage cover; commercial consignments. The sections • traps to be hung about 2m above the above have addressed many of the issues ground; associated with passengers and their • flies to be identified within about two luggage. days after the clearance of traps; For post and air freight and commer­ • nomination of a reference entomologist cial consignments, it is important to be and procedures for confirmation of aware of the country of origin and inspec­ identification of suspect flies; and tions should be geared to the level of risk. • reporting system to the chief plants For example, a parcel coming from the officer or chief quarantine officer. Cairns area should be assessed as higher risk than one coming from Melbourne in the middle of winter. In this respect, there Host Surveys is a need to educate Quarantine and Valid information on the host ranges of Customs officers so that the level of fruit flies in each country is a prerequisite inspection is appropriate for the degree of for negotiating quarantine agreements with risk. prospective importing countries. Lack of this information makes negotiation difficult and often results in conservative decisions Trapping being made out of scientific ignorance. A trapping system is essential if Biological data on the host ranges of fruit countries wish to export fresh fruits and fly species, on the stages of maturity at fleshy vegetables because importing which hosts become susceptible to fruit fly countries will require a list of fruit fly infestation, on the location of larval feeding 46 and rates of development within the host, Conversely, if these fruits and vegetables Fruit flies and on differences in susceptibility of were regularly sampled in Fiji and papaya continued different varieties to fruit flies are prerequi­ fruit fly was not recov~ed from the sites to undertaking research into quaran­ samples, it would indicate strongly that tine treatments to establish trade. Host this species does not occur in Fiji. Similarly, surveys also provide valuable information if cucurbit crops are regularly sampled that may substantiate the taxonomic together with Cue-lure trapping, there description of new species in a complex. would be a very high chance that melon fly Host surveys complement the trapping would be recorded soon after it arrived in a systems designed for quarantine surveil­ country. lance, especially for species not attracted to To ensure adequate coverage of the the normally available range of attractants. high-risk fruits and vegetables throughout Solanum fruit fly (Bactrocera rations), for the year, it is advisable to draw up a example, is not attracted to any known lure calendar of fruiting times for these com­ so host surveying is the only method modities. This can then be matched with a available to determine area freedom from list of fruit flies of high risk to ensure that this species. appropriate high risk hosts are being By regularly surveying the most regularly sampled. Negative results from susceptible fruit and vegetable species as these surveys may then be added to they come into season, it is possible to trapping data to provide a highly reliable provide additional information to prospec­ assessment of the area of freedom status for tive importing countries to support trap­ fruit flies. ping data, showing area of freedom status During the training course, one of the for various species of fruit flies. By regu­ group exercises is to develop these fruiting larly sampling guava, mango, banana, calendars for several countries and match Pacific almond (Terminalia catappa), chili them up to the exotic fruit fly species that and peach mango in the Cairns area, countries are monitoring. papaya fruit fly would have been recorded.

47 Vir~s-free nursery system, indexing and micrografting byHongJi Su

Abstract virus-free foundation stock of citrus and Virus and virus-like diseases have banana cultivars in Taiwan (Su and Chu, become among the most serious constraints 1984; Wu and Su, 1991). The precise and for the production of tropical fruits in the rapid in~exing techniques are indispens­ Asian and Pacific regions. These diseases able for management of pathogen-free ar~ tr~smitted by vegetative propagation nursery system. Development and applica­ such as sucker, cutting and grafting as well tion of molecular diagnostic probes includ­ as insect transmission. This means that ing monoclonal antibodies and DNA probes these diseases might readily prevail if the were made for formulating indexing mother tree stock or seedling would be techniques (Wu and Su, 1990; Tsai and Su, infected latently with these pathogens. 1991; Yeh, Su and Chao, 1994; Su, Hong and Establishment of virus-free nursery system Wu, 1995). The virus-free nursery system ' seems the only way to prevent prevalence for controlling virus and virus-like diseases of these diseases. Micrografting of citrus of citrus and banana has been performed combined with heat treatment will be also through micrografting, tissue culture and presenteci as the most promising method to indexing in Taiwan. The concerned tech­ make disease plants virus-free. The precise niques will be discussed during the work­ and rapid indexing techniques are indis­ shop. pensa"~:?le for management of pathogen-free ~ nursery system. 1. National scheme of pathogen-free citrus foundation stock and healthy Introduction ~ seedling propagation. Such tropicaJ. fruits as citrus and In order to control the systemic diseases banana are the most important fruits for caused by aphid-borne virus (citrus tristeza local consumption and export in Taiwan virus = CTV), non-vector-borne virus (citrus and some other Asian countries. Some tatter leaf virus= CTLV and citrus exocortis systemic virus and virus-like diseases have viroid = CEV) and psyllid-borne fastidjqus been causing considerable damage to fruit bacteria (greening), the present national yield and quality and are becoming a scheme of citrus bud-wood certificate limiting factor in fruit production. These program was initiated and granted by the diseases are transmitted by vegetative Council of Agriculture. It is conducted and propagation of seedlings such as sucker, jointly subsidized by Taiwan Provincial cutting and bud-scion for grafting as well Department of Agriculture and Forestry as insect transmission. These diseases are under the Guidance Committee of Taiwan generally controlled by integrated control Citrus Industry according to the following measures including cultivation of patho­ systems since 1983. gen-free seedlings, elimination of inoculum sourcoes,·andprevention of secondary 1,1 Pathogen-free Citrus Foundation. infection by vector insects. Stock Establishment of pathogen-free nurs­ 1.1.1 The sto~ks are produced by means of ery system is primarily important for improved method of shoot-tip grafting and preventing prevalence of these diseases (Su · heat-therapy in Plant Virus Laboratory of and Chen, 1991). Shoot-tip micrografting of National Taiwan University (PVL/NTU) citrus and tissue culture combined with (Project leader Prof. H.J. Su). heat therapy have been used for obtainiri.g 1.1.2 The stocks indexed to be free of the

48 pathogens by PVL/NTU are transferred to The solid and liquid media were Virus-free the Provincial Citrus Foundation Stock prepared according to Murashige et. al. continued Collection (PCFSC) in screenhouses of the (1972). The practical techniques for micro­ Chlayl Agricultural Experimental Station of grafting and double grafting will be Taiwan Agricultural Research Institute demonstrated in detail during workshop. (CAES/TART) (Charged by Miss Lin, horticulturist).

< 3. Nature and detection of greening 1.2 Foundation Parent Trees pathogen by DNA probe and PCRtech­ Foundation scions propagated from niques foundation stocks in PCFSC green houses The heat-tolerant form of greening are provided for growing potted parent known locally as Likubin in Taiwan has trees for propagating pathogen-free scion seriously affected the citrus industry by woods in green houses of TARI and CAES. shortening tree life-span. Most citrus cultivars except pummelo were susceptible 1.3 Pathogen-free Citrus Seedlings to the Asian form of greening organism Pathogen-free seedlings are produced (GO) before the 1970s. However, pummelo by using pathogen-free scions derived from has became infected by GO in Taiwan since the parent trees in the Healthy Nursery 1970, and pummelo trees grown in the Green houses operated by each of five Philippines, East Malaysia, Southern branch offices of Taiwan Fruit Marketing China, and Parao were found infected. Cooperative. ·Multiplication of GO was detected in graft­ inoculated seedling of Chinese box orange 1.4 Citrus Growers (Sevefinia buxifolia), Thailand woody apple The healthy citrus seedlings trans­ (Lemonia acidissima) and curly leaf (Murraya planted to the orchards protected from euchrestifolia), suitable hosts of vector psylla reinfestation by biological and chemical (Diapbofinia citii). DNA probe and poly­ control of psyllid vector and cross-protec­ merase chain reaction (PCR) with adequate tion with CTV mild-strain. primer pairs were developed through DNA cloning and sequencing of greening fastidious bacteria pathogen for detection 2. Micrografting for obtaining pathogen• and identification of GO. The schedule of free citrus foundation stocks dot hybridization with DNA probe and An improved method of shoot-tip PCR test with primer pairs are as follows. micrografting (STG) was developed by using triangle-hole cut (Su and Chu, 1984) instead of inverted-T cut described by 3.1 Diagnosis and detection of fastidious Murashige et. al. (1972) The outline sched­ bacteria (G-FB) causing citrus greening ule of STG is as follows: disease by DNA probe • Etiolated stock seedling (Troyer citrange two-week-old in vitro) I. Extraction of G-FB for probing: • Micrograft shout-tip into a rectangular triangular hole or V-cleavage of 1-1. Collect plant leaves and twigs. Strip the decapitated seedling midribs from leaves and peel the bark from • Culture on liquid medium, incubation in twigs. Collect the midribs and bark for air­ dark for two days. drying. • Grow in growth chamber or green house (28-30 °C) under cheese cloth 1-2. Freeze the chipped dried tissue (1-5 g) shading two weeks. in liquid nitrogen and grind to fine powder • Growing under uncovered condition for in coffee blender or a mortar and pestle. two-three weeks. • Double grafting to potted stock seedlings 1-3. Transfer frozen powder to a beaker and (rough lemon, citrange) and cultivate in immediately add 2-4 ml of FB-extraction green house. buffer* per gram tissue. Stir the tissue- 49 Virus-free powder suspension. gram tissue). Store at -20 °C. continued I-4. Filter the tissue suspension through * FB-extraction buffer (pH 7.4): gauze to remove grass debris that may be 300 mM mannitol subjected to the second extraction. 50 mM Tris-HCl

10mMKH2P04

I-5. Centrifuge the filtrate at 4,000 rpm for 5MMMgC12 10 min. and the supernatant is spun at 5mMEDTA 1.7-fOOO rpm for 30 min. at 4 ° C. Step 5 may 4 mM mercaptethanol be repeated. *DNA-extraction buffer (pH 8.0): I-6. Re-suspend pellet in DNA-extraction 100 mM Tris-HCl, buffer* (1 ml per 1-2 grams tissue) plus 1 100mMEDTA percent Sarkosyl (wt/vol), incubate at 55 ° 250mMNaC1 C/30-60min. and spin at 6,000 rpm/10 100 J.Ll/ ml proteinase K, may be omitted. min. at4° C.

I-7. Save supernatant, add 0.6 volume II. Dot hybridization on nylon (or NC) isopropanol, and gently mix. A nucleic acid paper precipitate will be visible; if not, place at- 20 ° C for 30-60 min. Il-l. Transfer 5 Jll purified G-DNA into eppendorf and add 5 Jll 6X SSC*, and I-8. Centrifuge at 12,000 rpm/ 30 min at 4 ° C. denature G-DNA in boiling water bath for 10 min. Chill quickly on ice. I-9. Re-suspend pellet in TE buffer (1 ml per 3 grams tissue. Add 25 J.Ll 20 percent II-2. Blot 3 Jll DNA samples on nylon or (wt/vol) SDS and 20 Jll proteinase K (5 NC membrane. (Make dilution series of mg/ml) perl ml of NA-TE buffer. Keep at DNA samples with 6X SSC if necessary.) 37 ° C for 30-60 min. II-3. Treat the membrane in UV Linker I-10. Add 175 Jll of 5M NaCl per 1 ml TE . (120,000 Jl joules) or vacuum oven (80- and mix thoroughly. Add 140 Jll of CTAB/ 85 ° C/2 hrs.) for cross-linkage. NaCl per 1 ml/TE and mix thoroughly, and incubate at 65 ° C I 10 min. II-4. Place membrane in hybridization bottle and add 2X SSC for rinsing. Add pre­ I-11. Remove aqueous supernatant to hybridization solution* (10 ml/bottle) and eppendorfs and add equal volume of incubate at 68 °C/2 hrs. in rotary hybridiza­ chloroform/isoamyl alcohol (24:1), mix tion oven. thoroughly and spin at 6,000 rpm. I 5 min. II-5. Pour out pre-hybridization solution I-12. Repeat step 11. and add hybridization solution (10 ml), incubate at 68 ° C/ 12-16 hrs. in hybridiza­ I-13. Add equal volume of phenol/ tion oven. chloroform/isoamyl alcohol (25:24:1). Mix thoroughly and spin at 8,000 rpm/ 5 min. II-6. Pour out hybridization solution* (save for further use). I-14. Add 0.6 volume of isopropanol to precipitate the nucleic acid. Gently shake * Pre-hybridization solution and place tubes at -20 ° C for 60 min. or 5XSSC overnight. 0.1 percent (w /v) N-lauryl sarcosine, Na-salt (Sigma) I-15. Wash pellet with 70 percent ethanol 0.02 percent (w /v) SDS to remove residual CTAB. Briefly dry pellet Add to the freshly prepared solution and re-suspend in TE buffer (20 Jll per 1 percent (w /v) blocking reagent (BM) 50 * Hybridization solution: 3.2 Diagnosis and Detection of Fastidious Virus-free Add 1 J.!l of boiled dig or biotin­ Bacteria (G-FB) Causing Citrus Greening continued labeled DNA probe to 10 ml pre­ Disease by PCR (Polymerase Chain hybridization solution. Reaction) with primer pairs.

final cone. III. Detection and coloring 10 X Taq buffer 5J.!l 1x

25mMMgC12 4J.!l 2mM ill-1. Wash filters 2 X 5 min at room 10mMDATP 1J.!l 0.2mM temperature (RT) with at least 50 ml of 2X IOmMDTTP 1 J.!l 0.2 mM SSC, 0.1 percent (w /v) SDS, per 100 cm2 10mMDCTP 1 J.!l 0.2mM filter and 2 X 15 min at 68 ° C with at least 10mMDGTP 1J.!l 0.2mM 100 m1 of 0.1 X SSC, 0.1 percent (w /v) SDS. Taq polymerase 0.3J.!l 1.5 units Forward primer 2J.!l 100ng ill-2. Wash filters briefly (1 min.) in buffer (50 ng/J.!l) #1. Buffer #1: 100 mM Tris-HCl, 150 mM Reverse primer 2J.!l 100ng NaCl; pH 7.5. (50ng/J.!l) Template DNA 100ng 100ng ill-3. Incubate for 30 min. with about 100 ml buffer #2. Buffer #2: 0.5 percent (w I v) Add sterile H 20 to 50 J.!l and 30 J.!l mineral blocking reagent (BM) in buffer #1. oil. ill-4. Wash again briefly with buffer #1. PCR cycles: Step 1: 94 °C 4 min; 47 °C 1 min; 72 °C ill-5. Dilute anti-dig-AP solution (BM) for 2 min. 1 cycle. dig-labeled DNA probe or Streptavidin-AP Step 2: 94 °C 1 min; 47 °C 1 min; 72 °C solution (BRL) for biotin-labeled DNA 2 min. 30 cycles probe to 150m J.!l/ml (1:5000) in buffer #1. Step 3: 72 °C 10 min. Incubate filter for 30 min. with about 20 ml Step 4: 4 ° C soaking of diluted AP solution. ill-6. Remove unbound antibody-conjugate 4. Nature and detection of CTV by by washing 2 X 15 min. with 100 ml of ELISA with monoclonal antibodies buffer #1. Citrus tristeza virus (CTV) is the most common and destructive virus in the ill-7. Equilibrate membrane for 2 min. with Western hemisphere and has a worldwide 20 ml of buffer #3. Buffer #3: 100 mM Tris­ distribution. Various strains of CTV ; HCl, 150 mM NaCl, 50mM MgCl2 pH 9.5. referred to as seedling yellows, tristza, stem pitting, dwarf, and mild type, have been ill-8. Incubate filter in the dark at room wide spread in Asia. Several stable hybri­ temperature with about 10 ml freshly doma cell lines secreting CTV-specific prepared color solution. Color solution: monoclonal antibodies were obtained (Tsai 50J.!l NBT (50mg/ml), 25J.!l BCIP (50mg/ and Su, 1991). The same strains of CTV are ml), in 7 ml buffer #3. detected and identified by the following DAS-ELISA with the monoclonal antibod­ ill-9. When the desired spots are detected, ies. stop the reaction by washing the mem­ brane for 5 min. with 50 ml of stopping 4.1 A Protocol of CTV detection by direct solution to terminate reaction. Stopping OAS-ELISA with AP-conjugation of solution: 10 mM Tris-HCl, 1 mM EDTA, pH McAb (3EIO) 8.0. 4.1.1. Microplates coated with purified IgG monoclonal antibody (McAb) against CTV: Add 100 J.!l of McAb-3EIO (1 ppm) to each

51 Virus-free well [dilute the McAb-3EIO (orange vial) KH2P04 0.24 g continued 1000 times by coating buffer], and incubate Tween 20 0.05 ml • plates within a moistened box at 37 o C for Distilled water 1 L two hours. Adjust pH to 7.4, then store at room tem­ perature. 4.1.2. Washing the plates with PBS-T, three times. 4.2.2. Coating Buffer (50 mM bicarbonate buffer)

4.1.3. Add 100 J.Ll/well of sample extracts* Na2C03 1.51 g

to each well, then keep the microplates NaHC02 2.93 g within a moistened box at 37 ° C for two Distilled water 1 L hours. Adjust pH to 9.6, then store at 4 ° C.

4.1.4. Washing the plates with PBS-T, three 4.2.3. Substrate Buffer times. Diethanolamine 97 ml - MgC12 12Hp 100 mg 4.1.5. Add 100 J.Ll/well of alkaline phos­ Distilled water 1 L phatase (AP)-conjugated McAb to each Adjust pH to 9.8, then store at 4 ° C. well [dilute the McAb-3ElO (blue vial) 5000 times by PBS-T containing 0.2 percent 4.2.4. CTV-Extraction Bugger (Tris-DIECA BSA]. Incubate plates within a moistened buffer, pH 7.5) box at 37 ° C for two hours. Tris base 1.18g Tris-HCl 6.35 g 4.1.6. Washing the plates with PBS-T, three Na-DIECA* 0.1 percent times. Distilled water 1L Store at 4 ° C. 4.1.7. Add 100 Jll/ well of p-nitrophenyl­ * sodium, N, N-Diethiocarbamate phosphate (1 mg I ml) in substrate buffer, Trihydrate and incubate in a moistened box at 37°C for 30-60 minutes. 5. Regeneration of healthy banana 4.1.8. Read ELISA values of absorption at plantlets from virus-infected tissues 405 nm or observe with naked eyes. culture at high temperature. Exposure of banana bunchy top virus *Sample extract: [It is better to process the (BBTV)-infected plantlets of Cavendish samples under lower temperature (4 ° C).] banana to 40 ° C for 16 hours daily for (1) Weight 0.5 g of citrus tissue sample periods of up to five weeks did not free (bark or midrib of leaf), put in a them from BBTV. However, BBTV was less tube and add 5ml of sample buffer. readily transmitted by aphids from treated (2) Chop into pieces in the sample plants than from untreated controls. The buffer with scissors. same heat treatment applied for up to 12 (3) Homogenize the sample by weeks also failed to eliminate the virus homogenizer with 17,000 rpm for from BBTV-infected tissue. When similar 0.5 min. BBTV-infected tissue was cultured for three (4) Centrifuge the sample with 3,000 months at 35 ° C some of the buds started to rpm for 5 min. and take the produce healthy looking plantlets. Five out supernatant as test sample. 11 cultures produced healthy looking plantlets in six months. Crude extracts from 4.2.1. PBS-T (phosphate-buffered saline the leaves of these plantlets did not react with Tween 20, pH 7.4) . with a monoclonal antibody prepared against BBTV. NaCl 8.00g CMV causing banana mosaic was also KCl 0.20g eliminated by tissue culture at 35 ° C for

Na2HP04 1.44g three· month. The tissue culture has been 52 operated in commercial scale for propagat­ 6.1.6. Washing the plates with PBS-T, 3-5 Virus-frf!e ing healthy banana plantlets from the virus­ times. continued free foundation banana stocks. 6.1.7. Add 100 J.Ll/well of p~nitrophenyl phosphate (1 mg/ml) in substrate buffer, 6. Nature and detection of banana and incubate plates in a moisture box at bunchy-top virus (BBTV) by ELISA with 37 °C for 30-60 minutes. monoclonal antibody and strain identifi­ cation by PCR technique primer pairs. 6.1.8. Read ELISA values of absorption at The bunchy-top disease caused by 405nm. BBTV has been the common and destruc­ tive virus disease of banana since the *Sample extract beginning of this century in Taiwan and (1) Collect 0.5 g blade pieces from some banana-growing areas in the Pacific basal parts of three younger leaves Regions. Using direct ELISA with mono­ per plant; chop into pieces in clonal antibodies, and polymerase chain extraction buffer with scissors. reaction (PCR) test with primer pairs, (2) Homogenize the sample by ho­ different BBTV strains including severe to mogenizer 20,000 rpm for 20 sec. mild bunchy-top symptoms, dwarfing and (3) Centrifuge the sample 5,000 rpm symptomless, were identified. Garland for 5 min. Take the supernatant as flower (Hedychium coronarium) and canna, test sample. hosts of banana aphid, were found to be the alternative hosts of the virus. The indexing It is better to process the samples under techniques of DAS-ELISA and PCR test are lower temperature (4 °C). as follows: 6.2.1. Coating buffer (50 mM bicarbonate 6.1 Protocol of BBTV detection in banana buffer) by direct DAS-ELISA with AP-conjugate Na2C03 1.59 g of McAb (2H6) NaHC03 2.93 g Distilled water 1 L 6.1.1. Microplates coated with purified IgG Adjust pH to 9.6, then store at 4 °C monoclonal antibody against BBTV: Add 100 J.Ll of McAb (5 ppm) in coating buffer to 6.2.2. PBS-T (phosphate-buffered saline each well, and incubated plates in a mois­ with Tween 20) ture box at 37 °C for 2 hours. NaO 8.00g KCl 0.20 g

6.1.2. Washing the plates with PBS-T, 3-5 Na2HP04 1.44 g times. KH2P04 0.24 g Tween 20 0.05 ml 6.1.3. Add 100 J.Ll/well ofBBTV extracts Distilled water 1 L from banana samples* in Tris-DIECA­ Adjust pH to 7.4, then store at 4 °C Sucrose buffer + 0.5 percent skim milk at 1 I 5-1/20 (W /V) dilution, then keep the 6.2.3. BBTV-extraction buffer (Tris-DIECA­ microplates in a moisture box at 37 °C for Sucrose buffer) two hours or at 4 °C overnight. Tris base 1.18 g Tris-HCI 6.35 g 6.1.4. Washing the plates with PBS-T, 3-5 Na-DIECA 0.1 percent times. (Sodium N,N-Diethiocarbamate Trihydrate) 6.1.5. Add 100 J.Ll/well of alkaline phos­ Sucrose · 5 percent phatase (AP)- conjugate in PBS-Tat 200 X Distilled water 1 L (0.5 ppm) dilution. Incubate plates in a Adjust pH to 7.5, then store at 4 °C + 0.5 moisture box at 37 °C for two hours. percent skim milk ·

53 Virus-free 6.2.4. Substrate buffer II. Buffer solutions continued Diethanol amine 97 ml MgC12 6Hp 100 mg * BBTV-extraction buffer (pH 7.4): Distilled water 1 L (Tris-DIECA-Sucrose buffer) Adjust pH to 9.8, then store at 4° C Tris base 1.18 g Tris-HCI 6.35 g NA-DIECA 0.1 percent 6.3 Diagnosis and detection of BBTV (Sodium N-Diethiocarbamate strains by PCR technique Trihydrate) Sucrose 5 percent I. Extraction of BBTV DNA. Distilled water 1 L ·Adjust pH to 7.5, then store at 40 ° C + 0.5 I-1. Freeze the chipped tissue (0.5 g) in percent skim milk liquid nitrogen and grind to fine powder in coffee blender or a mortar and pestle. * DNA-extraction buffer: 0.02 M Tris (pH 9.0) I-2. Transfer frozen powder to a beaker and 1mMEDTA immediately add 2 ml of BBTV-extraction 4 percent SDS buffer*. Stir the tissue-powder suspension and centrifuge at 6,000 rpm/ 10 min. for obtaining supernatant (SN). 7. Nature and detection of cucumber mosaic virus (CMV) causing banana I-3. Add equal volume of N.A. extraction mosaic by DAS-ELISA with monoclonal buffer* to SN, and mix thoroughly. antibody The banana mosaic caused by CMV I-4. Add 1751!1 of 5M NaCl per 1 ml and was commonly found in banana areas, and mix thoroughly. Add 140 Ill of CTAB/NaCl particularly has become epidemic in the per 1 ml and mix thoroughly, and incubate orchards growing tissue-culture plantlets. at 65 ° C/10 min. Three strains of CMV were identified and characterized by inducing different symp­ I-5. Remove aqueous supernatant to toms including severe mosaic with leaf eppendorfs and add equal volume of distortion, necrotic mosaic with heart rot, phenol, mix thoroughly and spin at 10,000 and mild chlorotic streak. The protocol of rpm/10 min. CMV detection is as follows:

I-6. Add equal volume of chloroform/ 7.1 Protocol of CMV detection in banana isoamyl alcohol (25:24:1) to SN. Mix by direct DAS-ELISA with AP-conjugate thoroughly and spin at 10,000 rpm/ 10 min. of McAb (7 -1)

I-7. Add 0.6 volume of isopropanol to SN , 7.1.1. Microplates coated with purified IgG for precipitating the nucleic acid. Gently monoclonal antibody (7-1) against shake and place tubes at -20 ° C for 30 min. CMV: Add 100 Ill of McAb (2 ppm) in or overnight. coating buffer to each well, and incubated plates in a moisture box at 37 ° C for two I-8. Wash pellet with 70 percent ethanol to hours. remove residual CTAB. Briefly dry pellet and re-suspend in 100 Ill of distilled H20. 7.1.2. Washing the plates with PBS-T, 3-5 times. I-9. Spin at 6,000 rpm for 5 min. and collect supernatant (SN). 7.1.3. Add 100 Ill/well of CMV extracts from banana samples* in 0.2M potassium phosphate buffer (pH= 7.4) + 0.5 percent

Na2S03 at 1/5-1/20 (WN) dilution, then keep the microplates in a moistened box at 54 37 ° C for two hours or at 4 ° C overnight. ~HP04 27.86 g Virus-free KH P0 5.44 g 2 4 continued 7.1.4. Washing the plates with PBS-T, 3-5 Distilled water 1 L times. Adjust pH to 7.4, then store at 40 ° C +

0.5 percent Na2S03 7.1.5. Add 100 J.Ll/well of alkaline phos­ phatase (AP) - conjugate in PBS-T at 500 X 7.2.4. Substrate buffer (0.5 ppm) dilution. Incubate plates in a Diethanolamine 97 ml moisture box at 37 ° C for two hours. MgC12 6H20 100 mg Distilled water 1 L 7.1.6. Washing the plates with PBS-T, 3-5 Adjust pH to 9.8, then store at 4 ° C times.

7.1.7. Add 100 J.Ll/well of p-nitrophenyl 8. Nature and detection of banana streak phosphate (1 mg/ml) in substrate buffer, badnavirus by PCR with primer pairs and incubated plates in a moisture box at Banana streak badnavirus (BSV) has 37 ° C for 30-60 minutes. spread to banana areas in the Pacific and Africa regions. The BSV infected Mysore 7.1.8. Read ELISA values of absorption at (AAB) banana originally and was transmit­ 405nm. ted to Cavendish by citrus mealybug (Planococus citri) on which severe fine * Sample extract: chlorotic streak with necrosis was devel­ (1) Collect 0.5 g blade pieces from oped. The virus was detected by PCR basal parts of three younger leaves followed electrophoresis analysis. per plant and chop into piece in extraction buffer by scissors. (2) Homogenize the sample by homog­ 8.1 Detection of banana streak enizer with 20,000 rpm for 20 sec. badnavirus by polymerase chain reaction (3) Centrifuge the sample with 5,000 (PCR) with primer pairs (J. Thomas). rpm for 5 min. Take the superna­ tant as test sample. It is better to Primer F1: CAA process the samples under lower CTC temperature (4 ° C). AAG AGC 7.2.1. Coating buffer (50mM bicarbonate CTA buffer) GTA

Na2C03 1.59 g TGC

NaHC03 2.93g Distilled water 1L Primer R2: TAC CTC Adjust pH to 9.6, then store at 4 ° C CGA CCG 7.2.2. PBS-T (phosphate-buffered saline TAT with Tween 20) TTC NaCl 8.00 g CAG KCl 0.20 g

Na2HP04 1.44 g PCRcycles: KH2PO4 0.24 g Step 1: 94 ° C 4 min; Tween 20 0.05 ml 44°C 1 min; Distilled water 1 L 72 °C 2 min; Adjust pH to 7.4, then store at 4 ° C cycle.

7.2.3. CMV-extraction buffer (0.2M potas­ sium phosphate buffer) 55 Virus-free Step 2: 94°C 1 min; Su, H. J. and M. C. Tsai. 1991. The etiologi­ continued 44°C 1 min; cal nature and transmission of fruit virus 72 °C 2 min; and virus-like diseases in Taiwan. Inte­ 30 cycles grated Control of Plant Virus diseases. 1991 FFTC Supplement No. I pp 83-87. Step 3: 72 °C 10 min. Su, H. J., T. H. Hong, and M. L. Wu. 1995. Step 4: 4°C soaking Preparation and application of diagnostic DNA probe on the fastidious bacteria causing citrus greening. Proc. Symposium References Research and Development of Citrus in Mirashige, T., W. P. Bitters, E. M. Naver, C. Taiwan. p 167-176. N. Roistacher, and P. B. Holiday. 1972. A technique of shoot-tip grafting and its Tsai, M. C. and H. J. Su. 1991. Development utilization towards recovering virus-free and characterization of monoclonal anti­ citrus clones. Hort. Science 7: 118-119. bodies to citrus tristeza virus (CTV) strains in Taiwan. Proceeding of the Eleventh Su, H.J. and C. N. Chen. 1991. Implementa­ IOCV Conference 1991. p46-50. tion of IPM of citrus virus and greening (likubin) diseases. Integrated Control of Wu, R. Y. and H. J. Su. 1990. Production of Plant Virus Diseases. 1991 FFTC Supple­ monoclonal antibodies against banana ment No.1 pp 3-11. bunchy top virus and their use in enzyme­ linked immunosorbent assay. J. of Phytopa­ Su, H. J. andJ. Y. Chu. 1984. Modified thology 128: 203-208. technique of citrus shoot-tip grafting and rapid propagation method to obtain citrus Wu, R. Y. and H. J. Su. 1991. Regeneration budwoods free of citrus viruses and of healthy banana plantlets from banana likubin organism. Proc. Int. Soc. bunchy top virus-infected tissues cultured Ciriculture, 1984. Vol.2: 332-334. at high temperature. Plant Pathology 40: 4- 7.

Yeh, S. H., H. J. Su, andY. C. Chao. 1994. Genome characterization and identification of viral-associated DNA component of banana bunchy top virus. Virology 198: 645- 652. An introduction to plant diseases by George Wall, Robert Schlub and Diane Greenough

A plant disease is broadly defined as most common cause of plant disease. These any condition in which a plant is in some diseases are not contagious and their way different from a normal plant in either control depends solely on correcting the structure or function. In other words, a conditions causing the diseases. diseased plant is one that is growing or developing abnormally. The plant may be II. Parasitic diseases are caused by living shorter, have more branches, or fewer organisms which derive their food from leaves than a normal one. In such a case, it growing as parasites upon plants. The most differs in structure. Or it may wilt and die common causes of parasitic diseases are prematurely, or not produc~ flowers or infections by fungi, bacteria, viruses, and fruit, in which case it differs in function. nematodes. Other less common plant Plant diseases have had an important pathogens include mycoplasmae, rickett­ role in history. The holy fire of the Bible is siae, viroids, protozoa, and some seed­ believed to have been caused by the ergot producing plants, such as dodders and fungus growing in the heads of grain, mistletoes, which are called phanerogams. which made bread poisonous. The great famine in Ireland in the mid-1800s was Fungi caused by the destruction of the potato crop A. Fungi are classified as plants that lack by late blight. The American chestnut has the green pigment (chlorophyll) found in been virtually eliminated as a commercial seed-producing plants, and therefore tree in the United States by chestnut blight, cannot manufacture their own food. There and the American elm is now similarly are more than 100,000 different species of threatened by the Dutch elm disease. The fungi of many types and sizes, some 8,000 emigration of thousands of Cubans in 1980 of which are plant parasites. Most are (the Mariel incident) to the U.S. was microscopic in size, but some produce large triggered by several new diseases in Cuba reproductive structures, such as the mush­ (sugarcane rust, sugarcane smut, and blue rooms. Most fungi reproduce by spores. mold of tobacco) which affected the island's Millions of spores may beproduced by a top export crops. single diseased plant. These spores are In the U.S. alone, estimates of yearly either airborne or they are dispersed by crop losses were $9.1 billion from diseases, splashing rain and wind. Some fungi can $7.7 billion from insects, and $6.2 billion also become seedborne and travel with the from weeds, towards the end of last decade seed of their host plant. Thus, some fungal (the '80s). Such figures show why plant plant pathogens can reach far away. Rust diseases, and other plant pests, are impor­ spores can become airborne; they can be tant. carried thousands of feet up in the air, and may travel thousands of miles in the air currents. Other fungi survive in the soil What causes Plant Diseases? and may be carried by surface water and Plant diseases are generally divided farm implements. into two groups based on their cause. Some characteristics of fungi include the following: I. Non-parasitic diseases are induced by 1. They are not green. some genetic or environmental factor such 2. They may exist as individual micro­ as nutrient deficiencies, extreme cold or scopic cells (spores).or exist as heat, toxic chemicals (air pollutants, weed thread-like cells called hyphae that killers, or too much fertilizer), mechanical weave together to form mycelium. injury, or lack of water, which is the world's 3. They are generally hidden from sight 57 Plant diseases with only their fruit bodies (structures Blotch -Areas of discoloration on leaves or continued that contain spores) visible. fruits that are irregular in shape and size. 4. They disintegrate organic matter and most pathogenic forms require an Canker - an area on a stem resulting from elaborate food source. shrinking and dying of stem tissue bor­ 5. They usually infect living tissue only dered on one or both sides by health tissue. after predisposition. 6. Plants have acquired resistance to Curl- a puff-like distortion of a leaf. native fungal pathogens through coevolution. Damping-off- Rapid death and collapse of very young seedling. Quarantines and inspections are the major means by which governments can Die back - Slow progressive death of reduce unexpected plant disease epidem­ shoots, branches, and roots generally ics. Plant pathogens introduced into an starting at the tip. area where they did not exist before are more likely to cause catastrophic epidemics Downy mildew - mycelium and spores of than are existing pathogens because the the downy mildew fungus appear as white native plants would not have evolved fluffy growth on the underside of leaves defenses necessary to survive attack from and other tissues. that particular foreign pathogen. Some of the worst plant fungal disease epidemics Ergot- the grain (seed) of a grass plant that have occurred were the result of which has been replaced with a hard black accidental introductions, such as downy mass of mycelium. mildew of grapes, chestnut blight and Dutch elm disease. Generally the most Fruiting body - a small speck which serious of these pathogens can only grow contain spores and is usually black. and reproduce on a particular living host. This is why many quarantine regulations Galls - a localized area of swelling due to a restrict all importation of certain host tissue abnormality. because by restricting the host you are also restricting the pathogens that attack only Gummosis - sap that has broken through to that host. These host-specific pathogens are the surface of a stem or fruit. among the downy mildews, powdery mildews, smuts and rusts. Honeydew - a shinny appearing sticky Preliminary identification can be made substance excreted during feeding by by observing the host (food source for the certain insects. fungus such as fruits, stem, roots, flowers or leaves of plants) for signs of abnormali­ Mosaic - leaves with a broken pattern of ties (disease). Observations of the diseased dark and light green patches. with a hand lens often reveal the presence of fruiting bodies or spores which can lead Mummy - a dried, shriveled fruit. to positive identification of the fungus and its classification. Diseases regardless of the Powdery Mildew - yellowish areas on cause may be loosely classified according leaves, stems and fruits covered by a to how the diseased host looks such as: grayish to white layer of mycelium and spores. Anthracnose- a leaf- or fruit-spot type disease caused by fungi that produce their Rot - a state of decomposition caused by asexual spores on a saucer-shaped struc­ microorganisms. ture called an acervulus. Rust - messes of spores from one of the rust Blight- rapid dying of young tissue such fungi that appears as "rusty" spots on as leaves, shoots and flowers. leaves and stems. 58 Scab- tissue that has developed a crust-like Allum spp. is of particular concern in our Plant diseases appearance. region. continued Imperfect fungi are like the asco­ Spots - any small portion or dot of discol­ mycetes but do not have sexual spores; oration on a leat stem or fruit resulting therefore, they do not have asci. This group from dead or dying tissue. of fungi usually produce conidia in abun­ dance and are responsible for most of the Smut - masses of dark, powdery spores fungal diseases. If an ascomycete has a caused by a smut fungus. distinct asexual spore cycle like that of imperfect fungi then the pathogen may be Stipple - few to numerous tiny shallow referred to by two names: one based on its bleached spots. sexual spore stage, and the other on its asexual spore stage (imperfect stage). Wilt -lack of water in the leaves and stems Sigatoka disease of banana has the sexual causing the plant to droop. stage name of Mycosphaerella musicola and imperfect stage name of Cercospora musae. Fungi that are responsible for white This group constitutes many of the spots, rust, down mildews and damping-off are blights, and rots that inspectors encounter. classified as lower fungi. They are primitive Banana imports into Guam are mainly and often exist in the soil. Some do not limited to fruits because of the many form hyphae and are referred to as slime diseases that banana harbor one of which is molds. The hyphae that are produced are Panama wilt disease cause by Oxysporum f primitive and have no cross-walls dividing cubense. Those imperfect fungi that do not the hyphae into smaller cells. Those that normally produce any type of spores are produce hyphae may lack well developed placed in the group Mycelia sterilia. Rhizoc­ mycelium and produce their life cycle tonia is an example of such a fungus. The within a host cell. Those that do produce importation of sweet potato roots and vines well developed mycelium produce large for planting are often restricted to prohibit sexual spores call oospores. Some of the Pellicularia rolfsii which has the imperfect lower fungi produce swimming spores stage of Sclerotium rolfsii. (zoospores) and are commonly referred to The basidiomycetes are considered the as water molds. Only a few species of these most highly evolved group of fungi. The fungi are encountered by inspectors. A distinguishing feature of this group is the water mold that is of concern to many taro presence of a basidium (a club-shaped or growers is Phytophthora colocasiae, the cause tubular cell bearing four basidiospores ). of taro leaf blight. This group is commonly encountered by The ascomycetes are the class of fungi inspectors and constitutes the rust and responsible for some of the powdery smuts which are some of the most damag­ mildews, ergots, scabs, blights, spots, ing plant pathogens known. anthracnoses and wilts. The main feature of this group is the formation of the sac-like Bacteria ascus which usually contains eight sexual B. Bacteria are very small, one-celled spores. Its mycehum has crossways and plants that reproduce by simple binary most species produce asexual spores called fission. They divide into two equal halves, conidia. Some produce chlamydospores each of which becomes a fully developed (resting spores, modified from existing bacterium. In this way, they are capable of cells); stromata (cushions of hyphae form­ duplicating their population every 15-20 ing a mat, in or on which fruiting bodies of minutes, so they can spread at an alarming the fungus are born); and sclerotia (masses rate. Some 1,600 bacterial species are of mycelium with a dense dark cortex known, and of these, 80 are plant patho­ which functions as resting bodies). Quaran­ genic. Bacterial plant pathogens do not tine inspectors frequently encounter these produce spores of any kind; they spread fungi as the cause of leaf and stem spots. mostly by splashing rain and wind, a few Leaf tip and die-back disease of onions and of them are soilborne, and some may also

59 Plant diseases become seedbome. 3. Some viruses are also capable of being continued transmitted in the seed of the infected 1. Bacteria are classified according to the plant. Blackeye cowpea mosaic virus is very types of nutrients they can utilize for common on yard-long bean. Farmers who growth, and also by their shape and size, use their own seed to plant next season's the color of their colonies, whether they are crop can end up with very high disease capable of infecting. incidence in their fields because this virus is seed-transmitted, besides being also carried 2. Plant parasitic bacteria can cause infec- by aphids and being spread mechanically. tions on leaves, stems, fruits and roots of plants. Some of them cause leaf and fruit Nematodes spots on their host plants, like the bacterial D. Nematodes are small eel-shaped worms leaf spot of pepper and tomato, caused by that reproduce by eggs. Several thousand Xanthomonas campestris pathovar vesicatoria. species are known, with a few hundred of Others may induce wilting, like the infa- them being plant parasitic. The number of mous Burkholderia solanacearum, which eggs produced by one female nematode causes Moko disease of bananas and also and the number of generations in a season bacterial wilt of pepper and tomato. depends largely on soil temperature. Therefore, nematodes are usually more of a Viruses problem in warmer areas of the world. c. Viruses are so small that they cannot be Most nematodes feed on the roots and seen with the ordinary microscope. They lower stem of the plants, but a few attack consist of nucleic acid coated with protein, the leaves and flowers. and cannot reproduce except inside living cells. Some 2,000 viruses are know today, 1. Plants infested with root-knot nema- but new ones are discovered quite often. todes, for instance, have a reduced root Around 500 viruses are plant parasitic. system with few root hairs and many Many of the viruses that cause plant tumors on their primary and s~condary diseases are transmitted from one plant to roots. Immature nematodes swim about another by insects, usually aphids or freely in the soil water, and when females leafhoppers. Virus infections are almost find a host root, they settle down to feed; always systemic, which means that they their feeding causes the plant root cells to infect the plant tissues throughout. There- grow and multiply abnormally, forming the fore, viruses are very serious problems in characteristic tumors. This abnormal plants that are propagated by bulbs, roots, growth of the roots interferes with the and cuttings because the virus is easily translocation of water and nutrients in the I; carried along with the propagating mate- plant, causing substantial yield losses. As . ! rial. the female nematodes grow, they enlarge and produce millions of eggs, which may 1. In addition to being almost invisible, survive in the soil for next season. viruses are difficult to handle because they can also be transmitted by contact, or 2. Nematodes find their way from one field mechanically. Tobacco mosaic virus, for to another by the movement of soil in farm example, which is found in cigarette implements, in water, even in shoes. One tobacco, can be transmitted to tomatoes must be careful when purchasing seedlings and peppers by smoking workers. from a nursery because if their soil contains nematodes, these may find their way 2. Unlike fungi, bacteria and nematodes, wherever the seedlings are transplanted. viruses do not respond to pesticide appli- cations. We have no pesticides capable of 3. Nematodes are classified according to controlling virus-induced plant diseases. their body morphology and sexual struc- Management of viral diseases is mostly tures. The type and shape of their based on the use of genetic resistance on aesophagus is important. Also, the presence ·..;,,. ~·~ the part of the host plant. or absence of a stylet must be determined. 60 The stylet is a needle-like part of nema­ same for all. Plant diseases todes' mouthparts, which they use to pierce continued plant cells and suck their juices out. Plant Important note: parasitic nematodes possess a stylet. Guam, the Northern Marianas, Palau, Yap, Truk, Ponape, Kosrae and the 4. Reproduction in nematodes takes place Marshalls all are in the tropical zone. That by eggs. Eggs can survive quite adverse means that the influence of temperature conditions, such as dessication in the soil. (often quite hot) and moisture (humidity or These give rise to a live larva, which is the amount of wetness that's almost always capable of moving about at will. There are in the air) are quite pronounced; they affect sedentary and migratory types of nema­ very much the amount of disease that's todes, and also endoparasitic and ectopara­ present in our crops and ornamental plants. sitic, depending on whether they remain When it's hot and/ or wet, chances are inside or outside plant tissues. good that there is going to be some pres­ ence of disease.

Disease Development In order for disease to occur, there has How do We Recognize Plant Diseases? to be a susceptible host, a virulent patho­ The fact that diseased plants are in gen, and conducive environmental condi­ some way different from normal plants tions. These three factors form what is indicate that there are ways of recognizing known as the disease triangle. plant diseases. Symptoms are outward A plant may be predisposed for infec­ expressions of plant diseases and include tion by improper nutrition and/ or environ­ three general types: mental conditions. But in order to have • Over-development of tissue, such as disease, there must still be a virulent tumors or galls; pathogen and a favorable environment. • Under-development of tissue, such as Otherwise, the mere presence of a pathogen yellowing or dwarfing; and does not guarantee dis~ase development. • Death of tissue, such as blights and For example, a spore of the black leaf streak cankers. fungus has landed on the leaf of a banana plant during an overcast and humid day, Signs are outward growths of the but it is too old to germinate, and therefore pathogen, and are an important key to their does not result in an infection site. Or a identification. Often it is necessary to look viable and virulent spore lands on another at these signs under a microscope, to be leaf, but the hot sun and dry wind dessicate able to determine size and shape of spores, it before it can germinate and infect. The for instance. point is made. All three components are necessary for infection and disease devel­ opment to occur. How are Plant Diseases Identified? After infection takes place, the invad­ The extremely large number of plant ing pathogen colonizes the host tissue. diseases (tens of thousands) makes it Then it can move ahead and begin produc­ impossible for any one person to be ing propagative structures, whether eggs or familiar with very many of them. However, spores. These propagules must now be there are certain facts that help in the disseminated, and they may have to endure identification of plant diseases. The most hardships in order to survive until next important fact is to know the name of the season, when new host tissue will again be plant that is affected. This can be used to go available for infection, then colonization, through lists of known plant diseases to see propagation, dissemination, and endur­ if a similar condition has ever been re- ance, ad infinitum. Thus, this is what ported on the plant. · constitutes the disease cycle. Details of the When previous descriptions cannot be disease cycle vary from one pathogen to found or do not exist, it is necessary to go another, but the general scenario is the through a series of steps known as Koch's

61 Plant diseases postulates to determine the causal agent of disease cycle, thus breaking it. This obstacle continued a disease. Briefly, an organism should be: may be the use of disease-resistant plants on which the disease organism finds it 1. Associated with the malady all the difficult to grow. It may be to create envi­ time; ronmental conditions that are unfavorable 2. It should then be isolated and cultured for disease development, or it may involve (if it is not an obligate parasite); placing a protective chemical over the 3. Healthy plants should then be inocu­ surface of the plant. lated to see if the organism will infect The three things needed for plant and reproduce the symptoms; disease (a virulent pathogen, a susceptible 4. Finally, the organism needs to be host, and a favorable environment) are also reisolated from the inoculated plants . the key for disease control. Take away any and compared to the original. Comple­ of these, and plant disease does not occur. tion of these postulates-constitutes This, of course, is easier said than done, but proof of pathogenicity. nevertheless, in this disease triangle is the key to control. We can group the various control How the Disease Developed is also techniques into the following categories: Important. 1. exclusion of the pathogen, H it occurred in a very short time 2. eradication of the pathogen, or (overnight) and over a large area it is reduction of the inoculum, probably not a parasitic disease, but is 3. protection of the host, more likely due to some unfavorable 4. adjustment of the environment, environmental condition or chemical. How 5. use of plant resistance. is the condition distributed in the field? Is it general over the area or only in one or The first category is the first choice for two spots? Parasitic diseases usually do not keeping a disease out of an area, if it does affect a large percentage of the plants in the not exist there yet. This may involve legal early stages, but start in one area and measures. Eradication may be a very costly gradually spread to the other plants. And endeavo~ and is usually attempted only parasitic diseases usually do not affect when the pathogen has invaded a small several different kinds of plants in one area area. Reduction of the inoculum, that is, the at one time, even though some disease spores or other propagules that can start organisms can attack many different new infections, is quite valuable in the case plants. Knowing the past history of the of endemic diseases (those already estab­ field and what chemicals have been lished in an area). Adjustment of the applied may help identify a: problem. environment may involve a number of A knowledge of the growing require­ activities, such as cultural practices. And ments and habits of the affected plant also finally, to the grower, disease resistance is helps to understand certain conditions. the cheapest way to fight plant diseases, There is often a tendency for gardeners to although the development of a resistant immediately suspect a disease when a leaf variety may require thousands of dollars turns yellow or brown. But it may just be a and several years to achieve. normal response. Even evergreens lose Other factors to consider in the choice their leaves eventually. Remember that of practices to control a plant disease are plant disease is the abnormal growth or the secondary consequences or side-effects. development of plants. Fortunately in most cases there are things to do for controlling diseases that do not pose a threat to the environment. However, each How are Plant Diseases Controlled? .particular situation must be carefully The key to controlling parasitic dis­ evaluated on an individual basis to choose eases is to introduce an obstacle into the the best method of disease control.

62 Recognition, quarantine surveillance and emergency response planning by Allan Allwood*, presented by Russell Campbell

Introduction flies for South Pacific island countries. In Quarantine authorities are charged preparing this summary, application has with the responsibility of main quarantine been made from the plans that are cur­ security and, consequently, keeping un­ rently being used by the New Zealand wanted pests and diseases out of countries. Government in eradicating Mediterranean Unless there is a complete prohibition on fruit fly (Ceratitis capitata) from the the movement of people and goods, this is Auckland area. Based on the effectiveness an impossible task. As a re~ult, quarantine of recording incursions of fruit flies in New authorities operate under a risk manage­ Zealand and the resultant actions, the ment regime, whereby activities and response plans are obviously workable and regulations focus on reducing the risk of may be used as a model by other countries unwanted pests and diseases gaining entry in the South Pacific. and becoming established. As part of the armoury to protect horticultural production against invaders, quarantine authorities Definition of Emergency Response for and plant protection services need to fruit flies operate efficient and early warning systems An Emergency Response for fruit flies to record, as early as possible, incursions or may be defined as the actions taken as a establishment of exotic pests and diseases. result of the detection of fruit flies to: This system should allow authorities to • demonstrate that the unwanted exotic recognize quickly an introduction and to pest is not established; or to take remedial action to eradicate or sup­ • contain and eradicate it and then press the unwanted pest or disease before it demonstrate that the eradication was spreads. successful. The emergency response, in the event of an incursion or outbreak, should begin within hours of the confirmation of the Emergency Response components exotic pest or disease. For this to happen with minimum delay, confusion and 1. Definition of Response Zones disruption to commerce, it is essential to Zone A - 200 metres radius of the find have in place a well thought out, techni­ Zone B - 1. 5 kilometres radius of the cally sound emergency response plan. The find operation of the plan should be tested Zone C - as defined by the Chief Plants under simulated conditions so that faults Officer (CPO) during the response may be ironed out prior to an actual incursion or outbreak. Clear lines of 2. Definition of Response Level responsibility and co-operation between Three response levels have been government, the agriculture sector, the defined by New Zealand, these being media and the scientists need to be enunci­ categorized by specific activities, viz., ated. (table on next page). This document identifies some of the As a guide to timeframes, Appendix 1 issues that may be considered in develop­ contains the activities associated with each ing an emergency response plan for fruit response level and the suggested

*Allan Allwood is chief technical advisor, Regional Management of Fruit Flies in the Pacific Project, Secretariat of the Pacific Community (South Pacific Commission), Suva, Fiji. Paper used with his permission. 63 Emergency Table 2: Definition of Response Level response Technical Req\).irements planning 1. Maps continued • Topographical with Mapping + + + scale of 1:250,000 or Trapping + + + 1:500.000 Fruit Monitoring + + + • Town Planning with Fruit Collection/Disposal + + scale of J:2,000 or 1:5,000 Protein Bait Spray + + Ground Spraying (Larvae) + + 2. Information to be put Cover Spraying + + on Maps Fruit Movement Control + + + The following informa­ Alternative Methods tion is needed urgently so Male Annihilation should probably be SIRM + completed within one working day of the timeframes for each activity. Table 2 and response commencing: Appendix 1 are taken almost directly from • Response Zones the MAF Regulatory Authority Standard • Properties in the Response Zones 153.61: Specification for Fruit Fly Response. • Exact location of infested properties • Distribution of hosts, if known 3. Establishment of Control Centres • Reference location names, e.g., street • National Response Centre (NRC) numbers, streets, suburbs, towns, rivers - Policy I strategy decision-making information release, resource and 3. Trapping logistic support. 3.1 General • Fruit fly Headquarters • Determine current distribution and - Responsible for investigations and changes to distribution eradication within zones • Trap types - modified Steiner traps • Regional Response Centres • Bait trap type for species not attracted - Support on investigations and contain­ to lures- McPhail or Nakagawa traps ment and eradication outside the zones • Methods of setting up traps • Suppliers of traps and lures 4. Investigation, containment and eradica­ tion 3.2 Placement of Traps The following is a summary of the trap . 5. Validation of successful eradication or placements used by New Zealand: containment

A 1 per property As for 10-15 per sq.km. As for with hosts Trimedlure as distributed Trimedlure

B 40-50 I sq.km. 20-30 I sq.km. 10-15 I sq.km. 30-40 I sq.km. . . c 400m apart 400m apart 1200mapart 400m apart 8persq.km. 8persq.km. 1 persq.km. 8persq.km.

64 3.3 Trap Placement Schedule compliance Emergency • Public awareness response Traps in Zone A- within 3 working days planning Traps in Zone B - within 3 workin,g days Traps in Zone C - within 7 working days Other Issues for Consideration continued 1. Nomination of Emergency Response Team 3.4 Selection of trapping sites Nomination of the composition of the Emergency Response Team needs to be 3.5 Servicing of Traps done very early after the confirmation of the exotic incursion or outbreak. It needs to 3.6 Specimen Identification and Data comprise a manager, a headquarters Recording controller, a reference entomologist and a technical adviser. 3.7 Reporting Systems 2. Adequacy ofLegislation and Regulations 4. Fruit Monitoring The adequacy of legislation and • Sampling programme - species, regulations to allow restrictions to be frequency, quantity applied to movement of fruits, entry to • Fruit transportation security properties, treatment of host fruit trees and • Incubation of fruit produce, destruction of fruits, prosecution • Disposal of fruit samples of offenders and others needs to be as­ • Laboratory colonies and security sessed now, not when an incursion or • Reporting outbreak occurs.

5. Fruit Collection and Disposal 3. Information on Technical Aspects (Response 2 and 3) Technical information on the following • Fallen fruit collected within 5 days of topics need to be collated and be readily commencement of Response 2 or 3. available: • Repeated weekly. • Exotic species to be targeted in the • Fruit transport security emergency response plans; • Fruit disposal • Life cycles and rates of development in • Reporting system fruits; • Host ranges; 6. Insecticide Use (Response 2 and 3) • Geographical distributions of target • Notification of property owners species; • Protein bait composition • Damage symptoms • Protein bait application rate and • Results of pest risk analysis, if avail­ application able; • Protein bait treatment schedule • Eradication methods, such as male • Ground spraying for larvae annihilation, sterile insect release (diazinon) under trees method (SIRM), protein bait spraying • Cover spraying for eggs and larvae and combinations of these. (dimethoate or fenthion) • Exemptions of properties from 4. Suppliers of Trapping and Eradication insecticide usage Equipment and Materials • Reporting system 5. Public Awareness and Relations 7. Movement Control of Fruits Co-operation between government • Fruits from individual properties departments (Agriculture, Trade, Planning, • Control of export fruits Finance), the private sector (farmers, • Commercial operators and industry exporters, industry groups and associa­ involvement tions), and environmental groups is • Sale of host material to the public essential to the success of an emergency • Vehicle inspections and monitoring of response for fruit flies. It is important to 65 Emergency keep all of these groups informed and to be transparent in the information disseminated. response The information must, above all, be technically correct. planning continued APPENDIX 1: Timeframes for activities for Response 1, 2 and 3

RESPONSE 1, 2 AND 3 Area of zones mapped

Trapping intensified ·For trimedlure/ capilure and Cue-lure 1 day I I responsive species All A zone traps placed 3day All B zone traps placed 3 days Traps placed within 10 km from the periphery of the B zone 7 days

For methyl eugenol responsive species Traps placed in A and B zone properties 3 days

For species that do not respond to lures All A zone traps placed 3 days All B zone traps placed 3 days

Fruit monitoring commenced in the A zone

RESPONSE 2 AND 3

Bait applied To all properties with suitable vegetation 2 days . within the A zone

To all properties with suitable vegetation 4days around the outer boundaries of the B zone (covering 30% of the B zone)

To all properties with suitable vegetation in 4days the remaining area of the B zone

Collection of windfall fruit commenced from all 3 days A zone properties with host trees

66 Review and recommendations

The Food & Fertilizer Technology Following discussions with many of Centre of Taiwan (FFTC) funded the the participants, both during and on University of Guam (UOG) to host a conclusion of the workshop, all indicated workshop on Plant Quarantine for the that they now have a better understanding Micronesian region. of quarantine generally and a much better knowledge of the various countries within the region and their quarantine regulations Workshop Agenda and requirements which will enable more Dayl harmonization and insurance of compli­ • Country reports by participants ance of quarantine controls. Further, the • Plant quarantine regulations participants have acquired the essential • Safety of agricultural products in Japan, knowledge of scientific aspects of plant technology and regulation protection, e.g. entomology, plant pathol­ • Phytosanitary certification ogy and virus indexing. An association now exists between the front-line inspec­ Day2 tors and the laboratory scientists. • Insects - identification and control The workshop was an ideal forum for • Recognition of pests in consignments of representatives of the region's plant fruit, vegetables, cut flowers, & plants protection services to correlate and gener­ • Recognition & control of pests in stored ally get to know one another. The idea of products & wood having professionals (plant pathologists, • Fumigation virologists, entomologists), extension • Control of introduced pests agents and quarantine officers all together enabled the framework of plant protection Day3 and each body's roles to be better under­ • Plant pathology stood by all. • Plant disease identification The workshop was considered a great success by all. A future workshop is plan Day4 for September, also sponsored by FFTC and • Plant indexing for virus diseases hosted by UOG. • Virus-free nursery system and A report in the form of a manual is to micrografting be produced by the UOG which will enable the participants and their partners to utilize DayS as a reference for future needs. • Identification of weeds • Weeds of quarantine importance • Weed control Accommodations (not prioritized) • Endangered Species Act (CITES) 1. That the plant protection workshop • Review & recommendations planned for September include as many of • Closing ceremony the same participants as possible.

2. That all countries submit an Exotic Fruit Review Fly Response Plan, and that fruit fly As may be seen from the workshop surveillance is enhanced or in the very least agenda, the content was very intensive. maintained. Excellent presentations were given by the visiting resource personnel on all subject 3. That all countries communicate regularly matters. in regard to quarantine regulations and

67 that all countries review and revise the be produced and distributed as suggested regulations to promote harmonization of by UOG and that the manual be a~ techni­ quarantine throughout the region. cal as possible. FFTC could be approached to fund the production. 4. If possible, a follow-up workshop be conducted to concentrate on inspection 7. That the FFTC be approached to consider procedures, i.e. field work, airport and supplying much needed inspection equip­ seaport inspections of passengers and ment, e.g. light microscopes. cargo, post office inspections and quaran­ tine treatments. Congratulations and thanks are ex­ tended to FFTC, principally Dr. Sato, and to 5. That FFTC communicate with the SPC to the UOG, principally Dr. Muniappan and determine if the Plant Protection Networks his staff for the manner in which the being developed by the SPC and others workshop was presented and also for the may be utilised by FFTC and resources way the UOG hosted and assisted the combined. participants. Congratulations and thanks, of course, are also extended and due to all the 6. That a manual on the workshop subjects resource personnel and the participants.

68 Participants

South Pacific Commission Republic of the Philippines Mr. Denis Kelly Mr. Luben Q. Marasigan Plant Protection Trainer (Micronesia) Officer-in-charge SPC - Division of Agriculture Plant Quarantine Service P.O. Box PS 12 Bureau of Plant Industry Palikir, Pohnpei 96941 Ninoy Aquino International Airport Tel: (691) 320-2646 Manila, Philippines Fax: (691) 320-5854 Tel: (02) 832-29-82

Food and Fertilizer Technology Center for Republic of the Marshall Islands the Asian and Pacific Region Mr. Banner Bwijtak Dr. Tohru Sato Quarantine Inspector Deputy Director, FFTC Ministry of Resources and Development 5th Floor, 14 Wenchow Street P.O. Box 1727 Taipei, Taiwan Majuro, Marshall Islands, MH 96960 Tel: 886-2-3626239 Tel: (692) 6265-3206/3352 Fax: 886-2-3620478 Fax: (692) 625-3218

Mr. Stanley Hosia Japan Quarantine Inspector Dr. Norihiko Saito Ministry of Resources and Development Director, Entomology and Nematology P.O. Box 1727 Section Majuro, Marshall Islands, MH 96960 Senior Plant Pathologist Tel: (692) 6265-3206/3352 Plant Pathology Section Fax: (692) 625-3218 Research Division Yokohama Plant Protection Station., MAFF 1-6-10 Shinyamashita, Naka-ku, Yokohama Pohnpei State, Federated States of 231 Japan Micronesia Tel: +81-45-662-8892 Mr. Brens Solomon Fax:+8145-621-7560 Quarantine Officer Division of Agriculture Dr. Tohiyuki Kato P.O. Box 1028 Director, Entomology and Nematology Pohnpei, FSM 96941 Section Tel: (691) 320-2400/2401 Research Division Fax: (691) 320-5887 Yokohama Plant Protection Station, MAFF 1-6-10 Shinyamashita, Naka-ku, Yokohama Mr. Alex Kim 231 Japan Quarantine Inspector I Tel: 81-45-662-8892 Division of Agriculture Fax:81-45-621-7560 P.O. Box 1028 Pohnpei, FSM 96941 Tel: (691) 320-2400/2401 Republic of China (Taiwan) Fax: (691) 320-5887 Dr. Hong Ji Su Department of Plant Pathology National Taiwan University Taipei, Taiwan, R.O.C. Tel & Fax: 886-2-362-5717

69 Republic of Palau Yap State, FSM Mr. JeffTewid Mr. Patrick Sogaw Agricultural Quarantine Officer I Quarantine Supervisor Division of Agriculture Division of Agriculture and Forestry Bureau of Resources and Development P.O. Box463 P.O. Box460 Colonia, Yap, FM 96943 Koror, Republic of Palau 96940 Tel: (691) 350-2183/3833 Tel: (680) 488-2504 Fax: (680-9) 488-1725 Mr. Ignathio Tithinfall Quarantine Supervisor Mr. Haru-Ichi Telmetang Division of Agriculture and Forestry Agricultural Quarantine Officer I P.O. Box463 Division of Agriculture Colonia, Yap, FM 96943 Bureau of Resources and Development Tel: (691) 350-2183/3833 P.O. Box460 Koror, Republic of Palau 96940 Tel: (680) 488-2504 Commonwealth of Northern Mariana Fax: (680-9) 488-1725 Islands Mr. Leo Reyes Agricultural Quarantine Inspector I Chuuk State, FSM Division of Agriculture Mr. Akissy Edward P.O. Box 7696 Plant Quarantine Inspector SVRB, Saipan, MP 96950 Department of Agriculture Tel: (670) 288-1045/1046 Chuuk, Eastern Caroline Islands 96942 Fax: (670) 235-9001 Tel: (691) 330-2755 Fax: (691) 330-2233 Mr. Anthony Muiia Agricultural Quarantine Inspector I Mr. Peter Haritos Division of Agriculture Plant Quarantine Inspector P.O. Box 10007 Department of Agriculture Saipan, MP 96950 Chuuk, Eastern Caroline Islands 96942 Tel: (670) 288-1045/1046 Tel: (691) 330-2755 Fax: (670) 235-9001 Fax: (691) 330-2233 Dr. Aubrey Moore Associate Director and Entomologist Kosrae State, FSM Northern Marianas College Mr. Palikkun Tolenna Saipan, MP 96950 Quarantine Officer Tel: (670) 234-9023 Department of Agriculture and Land Fax: (670) 234-0054 P.O. Box82 Kosrae, FSM 96944 Dr. Chao-Hon Chiu Tel: (691) 370-3017 Entomologist Fax: (691) 370-3952 Northern Marianas College P.O. Box 1250 Mr. Val ton Palsis Saipan, MP 96950 Quarantine Officer Tel: (670) 234-9023 Department of Agriculture and Land Fax: (670) 234-0054 P.O. Box82 Kosrae, FSM 96944 Dr. Diana Greenough Tel: (691) 370-3017 Plant Pathologist Fax: (691) 370-3952 Northern Marianas College P:o. Box 1250 Saipan, NT 96950 Tel: (670) 234-9023 Fax: (670) 234-0054

70 Mr. Anthony Tudela Guam Research Assistant Dr. Jeff D.T. Barcinas Northern Marianas College Dean/Director P.O. Box 1250 College of Agriculture & Life Sciences (CALS) Saipan, UT 96950 University of Guam Tel: (670) 234-9023 UOGStation Fax: (670) 234-0054 Mangilao, Guam 96923 Tel: (671) 735-2000 Mr. David Evangelista Fax: (671) 734-6842 Research Assistant Northern Marianas College Dr. John Brown P.O. Box 1250 Associate Director Tinian, MP 96952 Agricultural Experiment Station Tel: (670) 433-0444 CALS, University of Guam Fax: (670) 234-0054 UOGStation Mangilao, Guam 96923 Mr. Isidoro T. Cabrera Tel: (671) 735-2000 Agricultural Extension Agent Fax: (671) 734-6842 Northern Marianas College P.O. Box 1250 Dr. R. Muniappan Saipan, MP 96950 Professor Emeritus Tel: (670) 234-9025 Agricultural Experiment Station Fax: (670) 234-0054 CALS, University of Guam UOG Station Mr. Bill Cabrera Mangilao, Guam 96923 Agricultural Extension Agent Tel: (671) 735-2067 Northern Marianas College Fax: (671) 734-6842 P.O. Box 1250 Saipan, MP 96950 Dr. C.T. Lee Tel: (670) 234-9023 Professor Emeritus Fax: (670) 234-0054 Agricultural Experiment Station CALS, University of Guam Mr. Marcello Romolor UOGStation Agricultural Extension Agent Mangilao, Guam 96923 Northern Marianas College Tel: (671) 735-2000 P.O. Box 1250 Fax: (671) 734-6842 Saipan, MP 96950 Tel: (670) 234-9023 Dr. Donald Nafus Fax: (670) 234-0054 Professor of Entomology Agricultural Experiment Station Dr. Lewis MacCarter CALS, University of Guam Horticulturist UOGStation Northern Marianas College Mangilao, Guam 96923 P.O. Box 1250 Tel: (671) 735-2068 Saipan, MP 96950 Fax: (671) 734-6842 Tel: (670) 234-9023 Fax: (670) 234-0054 Dr. lise Schreiner Professor of Entomology Mr. Lucas C. Mendiola Agricultural Experiment Station Department of Lands & Natural Resources CALS, University of Guam P.O. Box839 UOGStation Songsong Village Mangilao, Guam 96923 . Rota, MP 96951 Tel: (671) 735-2068 Tel: (670) 532-3415 Fax: (671) 734-6842 Fax: (670) 532-5320

71 Dr. George Wall Dr. Russell Campbell Associate Professor, Plant Pathology Entomologist Agricultural Experiment Station Department of Agriculture CALS, University of Guam 192 Dairy Road UOG Station Mangilao, Guam 96910 Mangilao, Guam 96923 Tel: (671) 734-3942 Tel: (671) 735-2070/2000 Fax: (671) 734-6569 Fax: (671) 734-6842 Mr. Marvin Chargualaf Dr. Robert "Bob" L. Schlub Department of Agriculture Extension Agent III, Plant Pathology 192 Dairy Road Cooperative Extension Service Mangilao, Guam 96910 CALS, University of Guam UOGStation Mr. Jason T. Paulino Mangilao, Guam 96923 Department of Agriculture Tel: (671) 735-2000 192 Dairy Road Fax: (671) 734-6842 Mangilao, Guam 96910

Dr. Lee Yudin Mr. Joey Lopez Associate Professor of Entomology No Ka Oi Termite & Pest Control Cooperative Extension Service P.O. Box 24426 CALS, University of Guam GMF, Guam 96921 UOG Station Tel: (671) 734-1773 Mangilao, Guam 96923 Fax: (671) 734-1777 Tel: (671) 735-2000 Fax: (671) 734-6842

72 COLLEGE OF AGRICULTURe AND LIFE SCIENCES -•• -IIIII!'

Workshop participants, University of Guam campus, July 1996.

73 Dean/Director: Dr. Jeff D.T. Barcinas

The Guam Agricultural Experiment Station is an equal opportunity employer. All information gained through its research program is available to anyone without regard to race, color, religion, sex, age or national origin.

Trai!le names of products are used to simplify the information. No endorsement of named products is intended.

Funding for this publication has been provided by the Uniyersity of Guam Endowment Foundation.

"Any opinion, findings, conclusions or recommendations expressed in this publication are those of the author(s) and do not necessarily reflect the views of the University of Guam.

Publication design and production: Linda Austin, February 1999. 74