Protocol for Quarantine and Environmental Risk Assessment

1

ANNEX I

PROTOCOL FOR QUARANTINE AND ENVIRONMENTAL RISK ASSESSMENT

1. Background conditions.

1. A permit would be always required from competent local bodies for the corresponding import, in line with the legal standards in force in the country once an interest has been notified by a legal or natural person. In every case, the protocol for the request should include information on the origin of the material, the preliminary treatment applied to the said import material for the reduction of ecological risks, environmental control measures which would be considered, and the general features of the site where the material would be released.
2. Introduction within the frame of government actions to develop and broaden fisheries in the country
3. When the introduction is not an isolated action for the benefit of a fishery involved community, authorized bodies would carry out an early inventory of the places with a potential for the development of the related field of activities, according to natural and socio-economic conditions.
4. Once an inventory of the regions or areas with a potential for the development of mariculture has been made, only those where there are no usages linked with tourism or conservation that contradict extensive mariculture would be selected. This depends on the results of an analysis of fitness and real or potential conflicts.

1. On the introduction of biomass

1. If possible, the extension of the biomass for farming ends to other sites of the ecoregion or biogeographical unit would be made from material previously introduced there and adapted to its ecological conditions.
2. If a foreign source is needed, the best choice would be to introduce juvenile apical portions (2-5 gm) which would stay at the lab under supervised conditions until farming. If such logistics is not available or rapid extension is required, only small amounts of biomass would be introduced (5-20 Kg) for in situ reproduction goals after the appropriate quarantine term. The use of limited amounts provides for a previous treatment that would minimize the possibilities for the introduction of an exotic epibiota, diseases or endophyte aliens to the receiving environment.
3. Under no circumstance, the biomass should come from places where there are reports of an independent spread. Selection would be based on criteria such as vitality, intense and homogeneous color, no metabolic stress (ice-ice), and absence of epiphytes such as Ceramium or Polysiphonia and healthy apices with no evidence of damage caused by grazing pressure. If possible, the biomass selected should match the sterile lineages being farmed.
4. Before transportation from the place of origin, the material should be weeded out and cleaned with sea water or drinking water for no more than 4 minutes to prevent osmotic imbalances that might cause apex necrosis. Transportation should be in the open air using a container where relative humidity should be close to 100% and temperatures up to 30-310C. If these conditions are met, the biomass may be preserved for a term from 6 to 8 hours with no loss of turgidity.
5. Upon arrival at the destination, rehydration would take place at least for one hour in sea water filtered in one micron sized filters (1mm/1000) and appropriate containers. If possible, aeration should be provided. The water used in rehydration would be discarded after completion of the process. It would be substituted by a solution of copper sulphate under a concentration of 2 mg L-1 (20 mg in 10 L) so as to remove remains of epibiosis. The material should stay in the solution for three minutes, and then, it would be washed again with sea water. Afterwards, it would be taken to quarantine containers.

1. Keeping under quarantine

1. Any move of the biomass out of the biogeographic unit would require a strict application of quarantine procedures.
2. The essential means for a safe quarantine imply:

2.1 Physical isolation from any other activity so as to prevent biological pollution.

2.3 Set of instruments allowing for disinfection.

2.4 Facilities to filter or UV-sterilize the water used.

2.5 Discharge system that avoids direct discharge in the sea.

1. When the bulk of biomass introduced requires being kept in tanks, the quarantine of the material introduced to avoid any spread of exogenous biological pollution would take place for no less than ten days. The following conservation conditions should be observed:
2. The volume of tanks should be the right one so as to keep a balance of one litter of sea water per gram of biomass. Water flow would be secured through aeration. Salinity should be neither below 30 ng L-1 nor above 39 ng L-1. Temperature should be stable, ranging from 25 a 300C, with no sharp changes. Lighting must be intense but with no direct sunlight. Filtered coastal water use is recommended so as to avoid fertilization of tanks and the spread of algae there into.
3. Water in the tanks should be changed every day. Cleaning the tanks should be made at least twice a week, washing with a broom. When necessary, sodium hypochlorite would be used under the concentration used for marketing. Rinsing off is finally done with running water. The material would have to be removed temporarily from the tanks to carry out cleaning. Drying is prevented in this way.
4. The material under quarantine would be manually checked daily so that necrotizing damage may be detected. Whiting, evidence of infection caused by fungi or bacteria, abnormal composition or density in the epibiosis and unusual or unexplained morphological changes may be detected as well in this way. Samples of the material should be removed for analysis with a magnifying glass or a microscope.
5. All of the material showing evidence of infection or significant changes should be removed from the tank and incinerated. If infection or changes are observed in 25% of the biomass under quarantine, a strict selection of the material would be made, and then the complete process of the quarantine starts over again. If more than 25% of the biomass has been affected, it would be discarded and incinerated.
6. The water used during the quarantine and the procedures involved would not be directly discharged into the sea. Instead, it would be discharged in a drain above the highest tide levels after a solution of sodium hypochlorite with a concentration of 125 ml/m3 of water has been added and after no less than three hours so that disinfection has been made by the solution.
7. A solution of sodium hypochlorite commercially available would be used for the disinfection of the equipment employed in the manipulation and control of the material under quarantine.

1. On the implementation of procedures for the evaluation of ecological risks caused by the uncontrolled spread of the material in a natural environment.

1. The potential for expansion and the feasibility of Indo-Pacific carrageenophytes to turn into an ecological risk factor in the competition for the substrate with focal organisms, considering its role within the food web or engineering effect in communities where introduction has taken place, will depend on the interrelation of four factors: hydrodinamic forces, regularity in the biomass removal depending on the commercial use, grazing pressure and the intrinsic growth rate derived from the trophic conditions of the place.
2. Whenever there is an exotic carrageenophyte in situ release, there would be a continued ecologic monitoring for no less than two years if it is a pilot plan with a production on a small scale (up to 10 t p.s./year). Monitoring in this case would cover a radius up to two km from the farming area. It would extend up to five years in no less than five km surrounding the culture if it extends to a commercial scale (˃ 100 t p.s./year)
3. Carrageenophyte commercial farming is the cause of a significant impact on the underlying benthic communities. There are effects such as shadowing, changes in the rate of siltation of materials suspended in the column and possible changes in the granulometry and the concentration of biogenetic elements in bottom sediments. The selection of farming sites should consider the previous facts based on the ecologic consequences and the economic profits in every site.
4. There are two ways to monitor the ability of geographic propagation in the new site: determination of the existence of reproductive bodies in the material artificially propagated or through the quantification of the colonization rate in the substrate available. This is done though two alternative procedures: placing artificial substrates and/or direct observation of propagule attachment or material that has fallen off from rocks, pavements or any other hard substrate.
5. The ratio between the consumption rates by herbivores and the generation of biomass based on daily growth rates allows for the calculation of the remainder biomass in the system. It also contributes to determine the spread potential of the material introduced if it is not removed through commercial use in the site.
6. Farming tests were not made in places close to coral reefs, turtle nesting sites, manatee feeding sites or protected areas, natural parks or places with recognized natural values.
7. Although it is easy to reach these conclusions and most of them are based on the application of well-known estimators such as the coefficient of variation or central trend measures such as the mean or median to contrast growth rates recorded under exposure or protection from predators, they all require calculation based on an appropriate test design. It is only under this premise that appropriate reproductiveness and representativity may be accomplished. Therefore, it is recommended that academic advice should be sought to check any experimental protocol derived from the introduction of exotic carrageenophytes.