machote




Protocols: The following are the main protocols selected by the Diptera TWIG for the present project. Protocol for Setting Up Malaise Traps and Handling Samples Protocols for Culicidae (Darlene Judd) Protocols for Ceratopogonidae (Art Borkent) Protocols for Dolichopodidae (Daniel Bickel) Protocols for Asilidae (Eric Fisher) Protocols for Sarcophagidae (Thomas Pape) Protocol for Acalyptratae (Stephen Marshall and Matthias Buck) Protocol for Setting Up Malaise Traps and Handling Samples The Diptera TWIG agreed to use several Malaise traps in each of the five conservation areas. For each conservation area at least one trap is located in each of the following elevations or habitats: Coast (estuaries and mangrove swamps) Lowland forests (0-700 m) Midland forests (700-1800 m) Highland forests (1800-3000 m) Paramo. Sorting out focal taxa from alcohol samples In each of these elevations, traps are located near the following habitats: - On the edge of undisturbed forest, with the length of the trap perpendicular to that edge. - On the banks of streams, rivers, or standing water with the length of the trap perpendicular to the bank. If available, different types of aquatic habitats should be sampled. - On hilltops or at the highest elevation available. - Placed in the middle of primary forest. Samples are collected monthly or more frequently if it is necessary. The samples of diptera in alcohol, into bottles filled up to its capacity, are stored in a dark and cool site as much as it is possible, before being transported from the conservation area to INBio. Specimens are separated according to the following instructions: All Ceratopogonidae into one vial All Culicidae into one vial All the rest of the diptera, series of all present morphospecies are selected, avoiding large series of very common specimens in the sample. These specimens are separated into 3 different vials, one for Nematocera, another for small diptera such as Phoridae and Acalyptartae, and another for large diptera such as Tachinidae, Asilidae, Syrphidae and others. The remainder of the sample is stored in refrigeration or preferably frozen during the period of the project. The separated specimens and the remainder of the sample are monthly taken to INBio where a collecting number or lot numer is assigned to each sample and subsamples. Vials containing Ceratopogonidae and Culicidae are directly giving to parataxonomists or technicians working with these groups. The remainder of the sample is kept in refrigeration for its subsequent processing. Periodically, small diptera are processed by using the Critic Point Dryer (CPD), while large diptera are processed by using Etylene-Glycol. Then, these specimens are pin-mounted and labeled, sorted and identified. Psychodidae dryed by the CPD are placed between layers of soft paper towelling and then sent to Dr. Quate who will prepare them for their study. Some traps will be run dry and these will be emptied twice a day (but may be run only once every 2-4 weeks). Others will be filled about 3/4 full with 75% ethanol and the sample removed every 2-4 weeks. To prevent sloshing, which damages some specimens, the sample bottle will be filled to capacity before being transported back to the laboratory. Each sample will be fully labeled to indicate location, date, collector, elevation and malaise trap code and this information will be printed with archival quality ink or with a dark pencil on a large label (2 X 6 cm). As an example: COSTA RICA: Puntarenas: Cerro Rincón, 8 deg, 30 min N, 85 deg 10 min W (not accurate, just an example) 20-30.viii.1999, C. Godoy, 700 m Malaise trap OSA-1 Each malaise trap receives a number reflecting the conservation area and specific malaise trap location (for example, OSA-1, referring to location 1 in the Osa Conservation Area). The exact location and description of the surrounding habitat are recorded for each malaise trap and each one is photographed. This information (including the photograph) will be available at INBio as a web page, so that researchers may easily retrieve pertinent ecological information for the taxa they are studying. If future needs indicate that a trap should be moved, it will receive a new number (hence, ultimately a given conservation area may have records for 10 or more localities, indicating that a number of the traps have been moved during the course of our study). Samples from dry malaise traps are pinned in the field, including all the families which are of interest to the group. Dry Psychodidae are placed between layers of soft paper toweling, with a label indicating full locality data. Mounting with ethylene glycol:* 1. Transfer specimens from 75% alcohol to ethyl acetate for 1 hour. 2. Transfer specimens to ethyl acetate to which 1% or less of ethylene glycol has been added; 1 drop of glycol to 5-6 cc of acetate seems to be satisfactory. Leave for 1 hour or longer. 3. Remove, dry on blotting paper for a few seconds and mount. Small specimens may then be glued to pins. Larger specimens are treated in the same way except that they are pinned upon removal from alcohol. They are then returned to the ethylene glycol and taken through the rest of the process. Diptera TWIG / Sampling site selection / Taxa selection / Products / Protocols