Department of Agriculture, Fisheries and Forestry

The tsunami in December 2004 that devastated Aceh and other provinces in Sumatra resulted in a delay in project implementation as planned because the resources of both collaborating agencies in Indonesia were directed towards relief and reconstruction in the tsunami affected areas for the next few months. Nevertheless, two senior Indonesian project staff from the Department of Horticulture protection (DHP) and the Agency for Agricultural Quarantine (AAQ) were brought to Brisbane from 31 January to 7 February 7 2006 to plan the fruit fly survey work in Indonesia and also to undergo training with Griffith University staff on trapping and surveillance procedures. On their return to Indonesia, the project staff then conducted a training workshop from 26 - 30 June 2005 in Yogyakarta, on fruit fly surveillance techniques, identification to species, field pest management and techniques for training farmers. The training course was attended by 47 participants representing 18 provinces in Sumatera, Java and Kalimantan. Following the training workshop, field staff from DHP and AAQ established a trapping network in the provinces of Banten, West Java, DKI Jakarta, Central Java, D.I. Yogyakarta, East Java, West Kalimantan, Central Kalimantan, East Kalimantan and South Kalimantan. Major fruits and vegetables of economic importance such as mango, citrus, guava, mangosteen, chilli, tomato, gourds, and many others have also been collected to determine the infesting fruit fly species. Trapped and host reared specimens have been sent to Griffith University where they are being identified, pinned and labelled. A full set of accurately identified specimens are to be returned to Indonesia. The first year of implementation of the project has greatly strengthened the capacity of Indonesian quarantine to meet WTO-SPS requirements in relation to fruit flies.

All the samples of fruit flies collected in Java and Kalimantan during Year 1 of the fruit fly survey and that were sent to ICMPFF at Griffith University have been identified and a list of fruit flies occurring in Java and Kalimantan has been compiled. This list together with two sets of authoritatively identified fruit fly specimens have been returned to DHP and AAQ in Indonesia respectively for their reference collections.

A training workshop on "Fruit Flies of Indonesia: Their Identification and Pest Status" was held in Darwin at the AQIS Training Facility from 27 - 31 March 2006. The aim of the workshop was to provide training on the identification and pest status of flies collected during Year 1 of the survey in Kalimantan and Java. The workshop was attended by 17 Indonesian field staff who had participated in the Year 1 fruit fly survey in Kalimantan and Java. The training workshop was also of benefit to another 5 Australians, 4 of whom were from the Northern Australian Quarantine Strategy (NAQS), AQIS and 1 from the Northern Territory Department of Primary Industries, Fisheries and Mines.

Project staff in Indonesia organised the second training workshop on fruit fly surveillance techniques for project field staff from who were to be involved in the Year 2 fruit fly survey in the islands of Bali, Sulawesi and Sumatera. The training workshop was conducted in Bali from 9 - 14 July 2006 and was attended by 32 project field staff from Bali, Sulawesi and Sumatera. Following this training workshop, the field staff have established a fruit fly trapping network using methyl eugenol and cue-lure traps in 17 provinces in Bali, Sulawesi and Sumatera.

Crop damage assessments have been initiated in West, Central and East Java as well as in East Kalimantan. A controlled environment fruit fly rearing facility has been established at the Pest Forecasting Centre in Jatisari, West Java with flies reared from mango and citrus.

All samples of fruit flies collected in 16 provinces from the islands of Bali, Sulawesi and Sumatera during Year 2 of the fruit fly survey, and that were sent to ICMPFF at Griffith University have been identified and a list of fruit flies occurring in these regions has been compiled.

Project staff in Indonesia organised the third and final training workshop on fruit fly surveillance techniques for staff involved in Year 3 of the fruit fly survey. The workshop was conducted from 11 - 16 May, 2007 in Mataram, West Nusa Tenggara and was attended by 13 staff from the Food Crops and Horticulture Protection Centre. Following this training workshop, the field staff have established a fruit fly trapping network using methyl eugenol and cue-lure traps in 6 provinces, i.e. West Nusa Tenggara, East Nusa Tenggara, North Maulku, Maluku, Papua and West Irian Jaya. A separate training workshop on Indentification of Fruit Flies in Indonesia, funded by the Indonesian Ministry of Agriculture, was also conducted at Jatisari, Java, for 10 technical staff from the Food Crops and Horticulture Protection Centre.

In conjunction with the ACIAR project, a workshop on Fruit Fly Pest Free Areas was conducted in Jakarta from 18-22 June 2007. Invited speakers from Griffith University, Biosecurity Australia, NSW and Victoria Departments of Primary Industries and AQIS, provided in-depth presentations on fruit fly area freedom as practised in Australia. The workshop was attended by 44 senior scientists and quarantine staff from various agencies in Indonesia.

Crop damage assessments have also been initiated in West Nusa Tenggara, East Nusa Tenggara, North Maulku, Maluku, Papua and West Irian Jaya.. The controlled environment fruit fly rearing facility established earlier at the Pest Forecasting Centre in Jatisari is now able to supply laboratory reared flies for attractancy testing of protein baits.

Fabrication and installation of the protein bait plant at PT MultiBintang Brewery in Tangerang, Java began in July 2007 and is expected to be completed by early 2008.

All samples of fruit flies collected during Year 3 of the fruit fly trapping survey conducted in the provinces of Nusa Tenggara, Maluku, Papua and West Irian Jaya and sent to Griffith University have been identified, and a list of species occurring in these provinces has been compiled. Fruit fly damage assessments on fruits of economic importance is still in progress in these provinces.

The second training workshop on "Fruit Flies of Indonesia: Their Identification, Pest Status and Pest Management" was held in Darwin from,10 - 14 March 2008. The workshop provided training to 21 staff members from the Department of Horticulture Protection and the Agency for Agricultural Quarantine in Indonesia, particularly on the identification of fruit flies collected in the nationwide survey of fruit flies. With the aid of an illustrated key that was prepared and provided, trained staff now has the independent capacity to carry out accurate identifications of fruit flies in Indonesia. In addition to the Indonesian staff, the training workshop was also of great benefit to 3 participants from the Northern Australia Quarantine Strategy, Darwin, Broome and Mareeba, and 3 participants from the Northern Territory Department of Primary Industry and Mines, all of whom were able to obtain the most up to date knowledge on fruit flies of quarantine importance to Australia.

In conjunction with the ACIAR project, a workshop on Fruit Fly Pest Free Areas was conducted in Jakarta from 18-22 June 2007. Invited speakers from Griffith University, Biosecurity Australia, NSW and Victoria Departments of Primary Industries and AQIS, provided in-depth presentations on fruit fly area freedom as practised in Australia. The workshop was attended by 44 senior scientists and quarantine staff from various agencies in Indonesia.

Construction of the protein bait plant at PT MultiBintang Brewery in Tangerang, Java was completed in February 2008 and the plant was commissioned in March 2008. Once registration has been obtained, production of the protein bait will begin in Indonesia.

A list of fruit fly species occurring in West Nusa Tenggara, East Nusa Tenggara, North Maluku, Maluku, Papua and West Papua has been completed after extensive sorting and study of thousands of fruit fly specimens sent to ICMPFF at Griffith University during the third year of the project. This survey exercise now provides a complete picture of the fruit flies of the Indonesian archipelago. The 7 species of economic importance are Bactrocera albistrigata, B. carambolae, B. cucurbitae, B. latifrons, B. occipitalis (only in Kalimantan), B. papayae, and B. umbrosa.

The third and final training workshop on "Fruit Flies of Indonesia: Their Identification, Pest Status and Pest Management" was held in Brisbane from,16 - 20 February 2009. The workshop provided training to another 14 staff members from the Directorate of Horticulture Protection and the Agency for Agricultural Quarantine in Indonesia, particularly on the identification of fruit fly collected in the nationwide survey of fruit flies. With the aid of an illustrated key that was prepared and provided, Indonesian project staffs now have the independent capacity to carry out accurate identifications of fruit flies in Indonesia.

The protein bait plant at PT Multi Bintang Brewery in Tangerang, West Java which was commissioned in March 2008 is now producing fruit fly bait for the field control trials in Indonesia. The bait has been registered under the trade name Indo Prima and is selling at a price of 40,000 Rupiah per litre. Imported protein baits cost more than 3 times this amount, making Indo Prima very affordable to Indonesian farmers.

Since Project start-up on 1st June 2007 and the conduct of Project Inception meeting on 30 July - 3 August 2007 at the UP-MSI Bolinao Marine Laboratory, Philippines, the following has been accomplished and they are elaborated in the Annex sections of this report:

Completed upgrading of the hatchery and initiated production of sandfish juveniles at the UP-MSI Bolinao Marine Laboratory as well as sea ranching activities in two villages (i.e., Victory and Silaki) in Bolinao, Philippines;

Initiated production of sandfish juveniles for Davao node, preparation of High ponds for larval/juvenile rearing, and initial consultations with communities in Barangay Bato, Davao, Philippines);

Initiated construction of hatchery at NIFTDC in Dagupan, Philippines and aided NFRDI to produce sandfish juveniles;

Site assessments for potential restocking experiment were conducted in Northern Luzon, Philippines;

NT node coordinator continued contact with project key personnel in Australia (i.e., Warruwi Community, DAFF and Tasmanian Seafoods) while awaiting project funding from the Aboriginal Business Account;

Organization and conduct of the "Hatchery Operations Training" from 24 March to 15 April 2008 at NIFTDC and UP-MSI Bolinao Marine Laboratory. The training was participated by key project staff from the Philippines, i.e., NFRDI, UP-MSI and UP-Min including BFAR Regional staff;

Organization and conduct of the "Release Strategy Workshop" on 14-18 April 2008 at NIFTDC in Dagupan and UP-MSI Bolinao Marine Laboratory. The workshop was participated by key research partners in the Philippines and Australia including scientist from Indonesia, Malaysia and Vietnam; and

Participation of key project staff in various national meetings/reviews and planning sessions in support of the National Extension and Advisory Group.

Annex 7 indicates the Projects' Financial semi-annual report covering the period 1 December to 31 May 2008.

Hatchery production of sandfish
Significant progress has been made with the production of juvenile sandfish for sea ranching trials. In year 2 of the project, this has involved initiating new production nodes, as well as improving methods and removing 'bottlenecks' in existing systems. The Bolinao node produced ca. 26,300 juveniles (>3g) between May 2007 to September 2008. Typhoon damage to the newly constructed hatchery at Dagupan (National Fisheries Research and Development Institute) delayed commissioning until temporary repairs could be affected in May 2008. About 6,300 juveniles were produced from July - December 2008, of which about 4,000 were used for sea ranching trials in the northern Luzon area.
Due to difficulties with transporting broodstock and juveniles from Bolinao to Mindanao, a production facility was established in Mindanao. Spawning trials also conducted with Alsons Corporation (a private hatchery). From an approved objective of producing 10,000 juveniles per year, this node has produced 2,000 sandfish to-date, however it is clear here also that significant progress in shifting production bottlenecks has been made. All production nodes reported significant dividends in hatchery survival following a visit to the RIA 3 hatchery facility in Nha Trang, Vietnam. The largest constraints (bottlenecks) for production are now post-hatchery, and relate to appropriate technology and facilities for juvenile on-growing.
Establishing sea ranching trials
Good progress was made in engaging communities and local government units in the project. Five sea ranching sites have been established in collaboration with People's Organisations (POs) or other local actors, and are in varying stages of development. Partners in four sites have permits from the local government to manage and exclusively harvest sea cucumbers in the 5 hectare sea ranching area. Socio-economic and biophysical assessments, and monitoring of growth and survival of released juveniles were conducted at the four more advanced sites.
The first site (Victory, Bolinao Pangasinan) was formally established in September 2007. It is managed by a local fisher organization. Following on from the first year releases of 5,011 hatchery-reared juveniles (> 3g), a further 11,106 juveniles were released in year 2 (target 10,000). Monitoring of growth and survival is progressing well, although the staining techniques adopted to identify released individuals are not proving to be satisfactory. Alternatives will be investigated. Releases of between 5,500 and 2000 juveniles have occurred at 3 other sites. The least advanced site (Barangay Libuak, Mindinao) is currently being surveyed, and the process of obtaining permits has commenced.
The initial plan to conduct sea ranching trials with Aboriginal communities in Australia has not come to fruition at this stage. This component was initially contingent on substantial additional funding from sources within Australia, and this funding has not eventuated. Alternative approaches to working with these communities are currently being investigated.
Restocking trials
Following significant effort on site assessment and selection, it was clear that conducting valid, controlled trials to assess the success of stock enhancement would not be feasible. It was therefore proposed that the restocking experiment not be pursued as part of this project. This decision was endorsed recently by the mid-project review panel.
Project Review
On 4 - 5 June 2009, a mid-project review was conducted in Manila to assess progress against milestones, and provide an adaptive implementation process to re-focus the research on issues identified to be limiting progress. A mid-term review panel comprising representatives from the Scientific Advisory Committee (SAC) and Philippine national institutions involved in resource management and development recommended a number of revisions to the planned program and for ACIAR to continue to invest in the project for a further 2 years. Among the most significant changes was the inclusion of a new partner (SEAFDEC Aquaculture Department) in the Philippines, implementing new research on pond production of sandfish, and including a Vietnam node in the project with a focus on pond-based production.

Hatchery production and nursery systems:
Capacity to regularly and repeatedly produce sandfish juveniles continues to improve at varying rates across the 6 production nodes (4 in Philippines, 1 in Vietnam, 1 in Australia). The new partnership with RIA 3 (Vietnam) has contributed significantly to the capacity of the group in this area, with several exchanges taking place between Vietnam and Philippines. The recent completion of a purpose-built hatchery facility at SEAFDEC Aquaculture Department, Iloilo, Philippines, will add significantly to the capacity of the project for hatchery production, training of partners and scaling out to industry. At the Vietnam node, pond-based nursery production of both 5g and 30-50g juveniles for provision to industry participants has been very successful, with farmers keen on larger juveniles due to shorter grow-out periods. A continued and important focus is possible alternative systems for nursery production where ponds are not available.
Sea-ranching trials:
Some 65,000 hatchery produced sandfish have now been released in cooperation with local community groups across 4 sites in Philippines (3 in northern Luzon, 1 in Mindanao). Monitoring has continued showing variable growth and survival rates, and shedding new light on the viability of different models of sea ranching. Negative growth was seen following storm action from 2 typhoons at the Victory site in Northern Luzon, and it appears that the loss of organic components in sediments following the storm may be the driver of this decrease. Significantly, sampling of chemically tagged individuals at one site has shown substantial movement beyond the gazetted ranching site, raising some concerns about the size of ranching areas, and the 'uncaged' model of sea ranching. Small numbers of sandfish have been harvested from sea ranching areas although low wet:dry weight ratios (about 3%) have been observed. Security issues at 3 sites (loss due to poaching) have largely been overcome though combinations of improved guarding and community dialogue. Following complications that rendered a chosen site inappropriate, a final site for the Mindanao node (Barangay Tambo) has recently been selected, and discussions with the People's Organisation have been positive and productive.
Spawning of hatchery-produced stock:
Successful spawning of hatchery-produced sandfish (male and female) was observed at ranching sites in northern Luzon, with synchronised spawning at 2 spatially isolated sites. This confirms that hatchery-produced sandfish can generate viable spawning stocks, and lends support to the possibility of effective stock enhancement.
Australian trial - Goulburn Island:
A redesigned Australian pilot trial has commenced, with successful community consultations and site selection visits with aboriginal groups on Goulburn Island and Groote Eylandt. Sampling protocols have been designed, and pilot trials will soon commence. Permits are currently being sort to allow local communities to harvest and process sea cucumbers. The industry partner (Tasmanian Seafoods Pty Ltd) has agreed to supply processing equipment and train community members in processing methods. Delays with hatchery production have seen the schedule for large-scale releases moved back to early 2011, with smaller pilot releases to be conducted from 3rd quarter 2010.
Pond and co-culture trials:
Preliminary tank-based co-culture trials with five species of finfish (at SEAFDEC AQD) and vannamei shrimp (at RIA3, Vietnam) have been completed. Four species of finfish proved to be viable for early stages of co-culture, while some issues were encountered with shrimp trials; shrimp at semi-intensive farming densities were deleterious to sandfish growth and survival, and further trials are being undertaken. Rotational culture is being trialled by several cooperating farmers in Vietnam, with good results to date. A co-ordinated harvest of 5 t of cultured sandfish from a number of ponds resulted in keen interest from buyers, and a good price for product.
Value-chain analysis and bio-economic modelling:
A value chain analysis in Philippines is mostly complete. This includes a detailed analysis of the 'value added' at each node of the chain, and leads to a comprehensive characterisation of the industry at all levels. Options for value-chain upgrading are highlighted. This analysis will be repeated in Vietnam later this year, with distant market surveys (Singapore, Hong Kong and China) completing the analysis.
Preliminary bio-economic models as developed in FIS-2007-117 were presented to a group of researchers, government agencies and industry participants for input and comment. Suggestions have been incorporated, and a second draft of these models is now available to be circulated to project nodes for testing. Incorporation of empirical growth, survival and cost data will be incorporated in the next iteration of models.

Objective 1: To develop improved recommendations for integrated pest management and judicious pesticide use.
Objective 2: To improve control and detection of seed and pulp weevils.

Objective 3: To identify and trial improvements to current practices and conditions for managing mango supply chains.
Integrated Pest Management
In Australia, nine orchard sites producing the main commercial varieties 'Kensington Pride' and 'R2E2', near Bowen and near Mareeba have been surveyed fortnightly, finding some high fruit spotting bug populations actively feeding on emerging flower panicles at two locations.
Fruit spotting bug is causing reduction in yield because of damage to flowers and by causing premature fruit drop and quality reduction by causing damage to fruit. It is believed that feeding damage caused by fruit spotting bug establishes entry points for the development of anthracnose and the combination of these two factors is a major contributor to the problem.
A DPI&F facility to detect and analyse pheremones is currently being established within other projects. This will be used for the major mango pests when operational.
In the Philippines the project team has set up experimental sites on commercial farms. Two farms in each of the provinces of Guimaras, Davao del Norte and Davao del Sur have been chosen as areas for the IPM and supply chain improvement project. Mangoes grown in Guimaras are exported to the US while fruit from Mindanao are sent to Hongkong and Japan as well as to Manila. In Mindanao the trial sites take in the two distinct climatic condition encountered in the Davao region.
A survey to obtain initial baseline data on farmer production management practices ha been commenced in Guimaras, a, involving both well-managed orchards and backyard-type mango production. The survey is collecting information on farmers' practice and proposed management intervention at pre-bearing stage. Data is also being obtained on yield and production cost.
The project team also conducted a workshop to familiarize the field staff in Guimaras and Davao with project design and strategies, proposed intervention, commitment/roles and responsibilities, use of pest monitoring sheets, and coordination mechanisms established for the project. Although pest monitoring has now commenced, it has been found that those doing the monitoring do not have adequate resources to perform the task effectively. The development of a pest monitoring kit to assist anyone involved in monitoring to improve effectiveness, is under discussion.
Mango pulp weevil and seed weevil
Planning has been completed and field trials commenced on Palawan on pulp weevil biology and control strategies using chemical and particle films.
A complementary project on mango seed weevil and pulp weevil under the AusAID Asia Public Sector Linkages Program (PSLP) is currently running.
Mango seed weevil, which has been considered a minor pest of Australian mangoes, has over the last year become a significant pest, as it's presence is now having a substantial detrimental effect on mango export opportunities to China, UAE, Malaysia and South Korea. This project will contribute to a suite of new projects to investigate the problem, which will commence in the coming season.
Supply Chain Improvement.
In the Philippines, fruit damage caused by poor packaging and handling practices have been identified as major causes of quality loss. Changing practices will require some parts of the industry to adopt significant changes. Strategies to facilitate this are being developed.

In Australia, the mango industry is undertaking a major review of its export strategy. When this is complete in mid-2006, the export supply chain work within this project will commence.

To develop improved recommendations for integrated pest management and judicious pesticide use. (This objective has been expanded to include the additional outputs of providing pest monitoring kits and training workshops for Philippines project personnel and industry stakeholders).
To improve control and detection of seed and pulp weevils.
To identify and trial improvements to current practices and conditions for managing mango supply chains.

Integrated Pest Management. In the Philippines field infestation by insects cause losses and damage levels ranging from 10 to 40 percent, or limit market access, particularly for small-scale farmers. Systems approaches that integrate field control with postharvest treatments will reduce costs and the risk of quarantine failures and other regulatory penalties.
Using sites in Davao del Norte, Davao del Sur and Guimaras as pilot demonstration areas, collection of baseline data on farmer pest management as well as reviews of existing cultural management practices were conducted. Current yield and problems encountered by farmers were identified, to develop specific interventions. In each demonstration site, cooperators were identified and backyard and orchard mango trees were chosen. The pre-bearing activities include pruning, sanitation, fertilization and flush management. Interventions for bearing trees were flower induction, foliar fertilization, insecticide/fungicidal sprays, flower brisking, tagging and proper harvesting and postharvest handling. Familiarization of stakeholders with the major insect pests per site, damage symptoms and pest monitoring has also been done.
In Australia, for the second season, populations of fruit spotting bugs have been found to increase from mid- flowering through to early fruit-set in both the Atherton Tablelands and Burdekin regions. This season also experienced a high population of green vegetable bugs (Nezara viridula) during the flowering and fruit set period.

Mean orchard infestation levels of fruit spotting bugs began rising in August in the Burdekin and peaked at 13% in September for R2E2's and 10% for KP's. On the Tablelands fruit spotting bug populations began rising in mid September and peaked at 17% for R2E2's and 13% for KP's. Population levels were lower on all farms then the previous season.

The level of fruit drop and damaged fruit caused by fruit spotting bugs were lower on the Tablelands then the previous season being 17% and 15% for Oct and Nov for R2E2's and 21% and 10% for KP's of all collected fruit drop samples. The fruit damage results may have also been skewed higher then normal because of the similar feeding style of the green vegetable bugs that were active during the same sampling periods.
Pulp weevil and seed weevil. An IPM work plan for pulp weevil has been developed for Palawan to reduce usage of insecticides by 40% and to improve control of leafhoppers, pulp weevil and fruit fly. Field studies on pulp weevil have just been conducted and the data is being summarized. The method of fruit volatiles collection will be modified as initial analysis did not yield good results. Crude extracts from fruits, flowers, and weevils were collected and will be used in bioassays to determine attractancy and synergistic effects of blends.
Insect semiochemistry. Following earlier work in collaboration with the USDA Beltsville Laboratory, a trial pheromone blend for Amblypelta nitida was deployed in sticky plastic traps. However this proved unsuccessful. Further work is required to refine the formulation with respect to constituents and the ratios required. For Amblypelta lutescens, a particular isomer of one of the identified substances is required and this will be supplied by the Belysville laboratory when available.

SPME sampling of volatile emissions of major tropical fruits is continuing to investigate attractancy. Preliminary analysis has been commenced and identification of key components will be completed when resources become available.

Compounds released by both mango seed weevil (Australia) and mango pulp weevil (Philippines) have been collected by solid-phase microextraction (SPME) field collectors to analyse for possible pheromones. They are being analysed by DPI&F.
Supply chain. In the Philippines, deficiencies in the supply chain severely affect fruit quality and market opportunities. Accurate analysis will provide a 'road map' from which technical or business priorities can be identified and strategies implemented to bring about improvements. The detailed methodology for the Philippine mango supply chain has been developed, as well as a preliminary outline describing in general term the country's mango industry and information relevant to the development of specific supply chain maps were also initiated. The supply chain reviews marketing information more specific to focus areas in Davao del Norte, Davao del Sur, and Guiamaras.. The initial supply chain map is being validated during the series of field visits to the production areas. This component is providing an overview of the major technical postharvest problems
With the support of the Australian mango industry, a Queensland Government initiative and commercial exporters, several major supply chains were monitored from farm to retail in Singapore and Hong Kong. A range of problems have been identified including unsatisfactory product handling before export, unsuitable facilities for product handling in target markets as well as unfamiliarity with Australian fruit. Preliminary training activities have been commenced in both markets to address some of the issues. As a result of the monitoring activities in Australia, deficiencies in handling and preparing fruit for export were identified. Procedures to improve practices have been discussed with supply chain members and training has been provided to their staff.

Development of well validated and standardised serological assays for the detection of Leptospira, Trichinella and A. cantonensis infection in livestock (Objective 1)

ELISA tests are under development for the detection of infection with Leptospira, Trichinella and Angiostrongylus cantonensis in livestock in the Pacific islands. Sources of antigen and reference serum have been identified and each assay will be optimised and validated in the next twelve months.

In addition, PCR-based tests for the detection of Leptospira and A. cantonensis infection have been developed that will be used to test samples collected on surveys conducted in Pacific island countries in 2003.

Transfer of skills to partner institutions necessary to perform Trichinella detection techniques and post mortem examination of rodents to collect parasites and serum (Objective 1)

A workshop to train technicians from NAQIA and SPC in the skills necessary to detect Leptospira, Angiostrongylus and Trichinella infection was postponed from November 2002 until mid-2003 because a survey on the Bula Plain earlier in the year failed to provide a new isolate of T. papuae was that was to be used for experimental studies at the NVL. The workshop will be conducted once a sample of T. papuae is made available by the International Trichinella reference Centre in Rome.

Information on the prevalence and distribution of Trichinella, Leptospira and A. cantonensis in the Pacific Islands (Objective 2)

Two villages in Fiji have been comprehensively studied to date. Serum was collected from every animal in each village and anti-Leptospira antibodies detected using the microscopic agglutination technique (MAT) at the WHO reference centre for leptospirosis in Brisbane. Results show that prevalence of Leptospira infection was higher in cattle in Naduri compared to Burebusaga. In Naduri, the prevalence of infection in cattle and dogs was higher than in pigs. Although not significant the same trend appears to exist in Burebusaga. At this stage it is not possible to interpret these data fully. This will be done once information on infecting serovars and data from testing the human population are available.

Collection and identification of new isolates of Trichinella from pigs and rodents in an area in PNG where human infection is suspected (Objective 4)

One survey was conducted in collaboration with the Northern Australia Quarantine Strategy (NAQS) to recover a new isolate of T. papuae from pigs on the Bula Plain, Western Province. One pig sampled was found to be infected but only with approximately 1 larvae per gram of muscle. This larval burden was too low to enable passage to a rat for further experimental studies. Future experimental work at the NVL will be done using a sample of T. papuae supplied by the International Trichinella Reference Centre in Rome, which holds the original isolate recovered in 1997.

Information on the impact of A. cantonensis infection in ruminants (Objective 4)

The life cycle of A. cantonensis has been established at Murdoch University including maintenance of infected snails (intermediate host) and infection of rats (definitive hosts). Two experiments have been conducted to determine the pathogenesis of A. cantonensis infection in sheep. Results to date are not conclusive. No evidence of eosinophilic meningitis has been observed in experimentally infected sheep This could be due to a number of factors including; the isolate used is not infective for sheep, the dose given is not sufficiently high to elicit an immune response, the method of preparation of larvae reduced their infectivity, sheep are refractory to A. cantonensis infection. We do not have sufficient data to fully interpret our data as yet. Experiments are planned for 2003 to attempt to address some of these issues.

Research conducted by at Murdoch University showed that a large number of snail and slug species commonly found in Perth are able to act as intermediate hosts for A. cantonensis. Snail species included small aquatic snails that are likely to be accidentally ingested by ruminants grazing near sources of standing water.

Year 2 (01/01/2003-31/12/2003)
An ELISA test was successfully developed for Leptospira pomona. This ELISA has now been standardised for use in cattle and pigs. The results from testing serum collected cattle on Numundo Farm in PNG show that the test has good agreement with the standard serological test and that it had high specificity. Further work is required to test sufficient samples from PNG to determine the diagnostic sensitivity and specificity compared to the standard test. The test is now in a format suitable for transfer to partner laboratories.
A two-day workshop on diagnostic techniques for Trichinella infection was held at the National Veterinary Laboratory (NVL) in PNG.
Results of a retrospective serological survey of 479 people living in the Balimo area showed that the prevalence of infection with Trichinella of 29.5%. Antibody prevalence did not differ according to sex but there was a significant difference between age groups, and also appeared to differ significantly according to geographic region and age. These results (and those from other surveys) show that T. papuae is probably widely distributed in wild pigs (the most likely source of infection for humans) in the south coastal area of PNG at least as far east as Gulf Province.
At the NVL, examination of muscle digests from 31 pigs and 15 rats from the Goroka area found no evidence of T. papuae.
Following the discovery of Trichinella zimbabwensis in crocodiles in Zimbabwe AQIS has required PNG to confirm freedom from Trichinella infection in its crocodile meat exported to Australia from Mainland Holdings Crocodile Farm in Lae. A total of 112 samples of crocodile muscle have been tested at the NVL from captive bred and wild-caught animals. The majority of wild-caught crocodiles originate from the Kikori area (n=52) in Gulf Province. Sixteen of the 52 (30.8%) samples of meat taken from crocodiles from Kikori contained T. papuae larvae.
Surveys to determine the prevalence and identity of infecting Leptospira serovars began in 2003. Results of a cross-sectional survey of breeding cattle on Numundo Beef (West New Britain Palm Oil) show that there is a high prevalence of Leptospira infection in cattle older than two years of age and that the predominant infecting serovars were hardjo, pomona, swajizack and medanensis. These data confirm that active Leptospira infection exists in cattle herds in PNG and may be a factor in the infertility observed in cattle in PNG.

Development of the capacity for effective surveillance of Trichinella, Leptospira and A. cantonensis infection in the South Pacific region through the development and transfer of diagnostic technologies

Training was provided to a technician from the NVL in the use of an antibody detection ELISA to detect infection with Leptospira spp and Trichinella spp during a workshop at Murdoch University.

Peter Wai'in from the NVL is completing a PhD on techniques to detect infection with Leptospira (PCR, culture and ELISA) and the epidemiological principles required to design, implement and analyse data from surveys to determine the incidence of infection and its association with infertility. Dr Nime Kapo (NAQIA) has also received hands-on training in the sampling strategies for disease surveys in PNG.

To determine the prevalence of Trichinella, Leptospira and A. cantonensis infection in livestock in Fiji and Kiribati and risk factors associated with their transmission

Results of surveys conducted in PNG show that there is a high prevalence of infection with Leptospira spp in cattle in PNG and it is a significant cause of infertility in affected herds. These results show that the prevalence of infection is higher in large commercial herds and that there is evidence that attempts to control infection using vaccination have failed. However, the reasons for this apparent failure may relate to management issues and not the failure of vaccination per se. Further work is required to determine the optimal vaccination strategy.

A comparison of data from testing humans and animals in two villages in Fiji show that in the village that experienced human cases of leptospirosis the pattern of serovars affecting the human population most closely resembles the profile found in domestic dogs. This opens up the possibility of interventions by vaccination of village dogs or by modifying the contact between the species.

Results from testing a total of 108, 48 and 73 serum samples collected from pigs from Fiji, Tonga and Kiribati respectively showed that the prevalence of antibodies to Trichinella were low. Only three pigs and one mongoose from Fiji were shown to have high levels of antibodies, which suggest that infection, if present is sporadic and may be present in Mongoose. Further work is required to determine the presence or absence of Trichinella in Mongoose because they are abundant and relatively easy to trap.

A field visit to Kikori in April 2004 to investigate the potential source of Trichinella isolated from farmed crocodiles in Lae revealed that crocodiles held in the villages are fed them on wild pig meat and fish prior to transport to the crocodile farm in Lae. It is likely that pigs are the source of Trichinella because larvae were recovered from the meat of a wild pig sold at Kikori market.

To determine which species of Trichinella are present in Fiji and Kiribati

No further progress was made towards fulfilling this objective due to staff shortages in the SPC

To investigate the pathogenesis of T. papuae in pigs and Angiostrongylus cantonensis in ruminant

Experimental studies in PNG showed that the larvae of T. papuae remain infective in pig meat for up to 26 days at 5C and for 9 days in meat maintained outdoors at ambient temperature (32C). This is important because it shows that surveys to recover Trichinella from animals in remote locations are possible if tissue samples can be refrigerated.

There was no evidence of clinical or subclinical disease induced by experimental infection of two merino cross-bred ewes with at least 1,000 L3 larvae of A. cantonensis. This suggests that A. cantonensis is not a significant pathogen of sheep.

The majority of project activities are due to begin in 2006. There has been limited progress towards Objective 1 mainly because of delays in the finalisation of project documents for this extension. Preliminary discussions with cattle producers in the Markham Valley have been conducted and their cooperation achieved. The first survey is planned for May 2006.

Research activity to perform a cross-sectional survey of domestic pigs in PNG to prove freedom from Trichinella infection will commence in 2006. In 2005 a total of 327 crocodiles were screened for the presence of infection with Trichinella using the digest method. A total of 44 crocodiles were shown to be infected with Trichinella larvae, of which 27 came from Kikori, 12 from other locations and 5 were born on the farm. Larvae from the crocodiles from Ambunti (Sepik) were analysed at the International Trichinella Reference Centre, Rome and found to be genetically identical to larvae isolated crocodiles from Kikori. These data suggest that Kikori is the only source of infected crocodiles at Mainland Holdings. In addition, meat from 3 wild pigs from Ambunti was also negative for Trichinella infection.

Progress towards project milestones has been hampered by technical issues in PNG related to the mergence of avian influenza as a high priority disease, which reduced the time project staff had to devote to project activities.

Longitudinal and cross-sectional surveys to determine the prevalence of Leptospira infection in animals in smallholder villages and to determine the dynamics of transmission on commercial cattle properties have progressed well. This work has so far demonstrated that there is a very low prevalence of Leptospira infection in animals in smallholder villages associated with commercial properties. This means that the animals do not pose a risk to humans and that leptospirosis is unlikely to be a major constraint to livestock production. Samples collected from commercial properties are in the process of being tested.