Overall sampling protocol

1. Overall sampling protocol The aims are to carry out the 9 sampling programs (along with individual programs) at (as far as possible) each of the 9 study sites (3C, 3R, 2B, 1I). The sampling procedure should be identical or similar at each study site, to allow statistical comparison of the results between the sites. The main sampling programs are (c = targets canopy, s = targets soil): Vegetation Light trapping (c) Sticky traps (c) Soil/Canopy microarthropods (s, c) Flight interception traps (c) Winkler - litter (s) Beating (c) Pitfall traps (s) Fogging (c) Individual programs (s, c)

2. Sampling program: Vegetation Managers: S.J. Wright, F. Hallé, S. Ribeiro, M. Samaniego & Andres Hernandez Aims: compare the vegetation (species richness, frequency) in the understorey and upper canopy of 12 sites (9 + 3 surrogate crane sites) Technique: a 20m x 20m plot is grided on the ground. The vegetation up to 2m is measured and mapped using standard techniques. Similarly a 400m2 plot is surveyed in the upper canopy, and the vegetation recorded within the last meter. Methods: 100 random points (or within reach) in where encounters between leaves and a PVC stick are recorded. Measurement of canopy height by range finder. Notes: 1. Bubble. Upper canopy = transect line; understorey = both 20 x 20m plot plus transect line 2. Ikos. Upper canopy = simplified survey; understorey = 20 x 20m plot 3. Vertical point-transects may also be organised by S. Ribeiro at all sites 9 sites (3C, 3R, 2B, 1I) to survey in the understorey and canopy 3 sites (3C’) to survey in the understorey only (no canopy access) Thus 9+9+3 = 21 ‘plots’ (canopy+understorey) Schedule for one site: 1 field day for establishing ground plot and marking large trees/shrubs 1 field day for canopy plot 1 field day for further work at the ground plot

3. 25m Sampling program: Sticky traps Managers: Y. Basset, G. Curletti & R. Harrison Aims: compare the flying activity of small insects in the understorey and upper canopy Technique: yellow sticky traps, set up either in the understorey (n = 25, at 1.3m, DBH) and upper canopy (n = 25) at each site. In addition, if possible, 3 vertical transects of 8 traps (0m, 1.3m (DBH), 7m, 14m, 21m, 28m, 35m and 42m [top]) will be set up at each site. Correlation with crown openness and incident light may be possible. The traps will be running for 5 days at each site. Position of traps on the raft; in crane subplots, traps are similarly set up in the periphery. Total 74 samples (traps) per site, 25 upper canopy + 25 understorey + 3 x 8 traps in transects Ikos: only one transect of 8 traps 9 sites (3C, 3R, 2B, 1I): Total (8 x 74) + 8 = 600 samples Position of sticky traps at 1.3m (DBH) at crane and raft sites (20mx20m): mostly inside the plot Schedule for one site: 1 field day for set up of traps 3 days laboratory BCI/University Panama 1 field day survey traps + measurement variables 9 sites x 5 = 45 days (out of 40) Overlap for Ikos site and one crane site, so that 7 x 5 = 35 days Position of traps in one bubble transect; 1 sample each meter; understorey samples are vertical to canopy samples Material needed: 600 traps, benzina, luxmeters, 3-6 weighted line transects, etc.

4. Sticky traps: transects Aims: compare the flying activity of small insects at selected transects (3 per site) and height: 0m, 1.3m, 7m, 14m, 21m, 28m, 35m and 42m (top). Climbing rope: not established at random (security) Transect line lowered with sand weight, close to the climbing rope, and secured up and down. This line includes pre-measured distance and attachment points for the traps. Traps are set up when one person climbs and activated when abseiling down. Simultaneous recording of incident light at all transect levels (2 luxmeters) Survey: Before surveying each trap, incident light is measured and two fisheye pictures of the canopy above the trap are shot. Simultaneously the incident light at the brightest point in the upper canopy is recorded. Similar measurements are performed at the 25 traps in the upper canopy and 25 traps in the understorey. These measurements will be used for the sticky trap program, but also to characterize overall the site Crown openness determined at all levels by hemispherical pictures

5. Sampling program: Flight interception traps Managers: R. Didham & L. Fagan Aims: compare the flying activity of insects in the understorey and upper canopy Technique: FITs set up in transects of 8 traps each. 24 traps at each sites, working for two weeks. Crane sites first, bubble, icos and (possibly) raft sites later on. Care is needed about emplacement of FITs (use sticky trap transects whenever possible) and possible interaction with fogging. 3 transects per site Total 8 x 3 = 24 samples (traps) per site 9 sites (3C, 3R, 2B, 1I): Total 9 x 24 = 216 samples Position of transect below the raft or crane subplots Schedule for one site: 1 field day for set up of traps 2 days laboratory BCI/University Panama 1 field day survey traps 9 sites x 4 = 36 days (out of 40) Position of transects in one bubble site As far as possible, FITs will be set up on the same transects than sticky trap transects, but after sticky traps are surveyed

6. 40m Sampling program: Beating Managers: H. Barrios & F. Ødegaard Aims: compare the foliicolous fauna in the understorey and upper canopy Technique: quantitative beating (1 sample = 0.85m2 beating sheet full of foliage = 1 bag) During day-time (night work is to be considered). Identical beating sheet should be employed. Position of beating samples on the raft Total 40 samples (beating) per site, 20 upper canopy + 20 understorey Ikos: impossible to use this site 8 sites (3C, 3R, 2B): Total (8 x 40) = 320 samples Position of beating samples in the understorey plots and crane subplots (20m x 20m): at a peripherical zone extending beyond the plot. Beating is done preferably early in sampling sequence Schedule for one site: 1 field day for canopy+understorey Samples 2 days laboratory BCI/University Panama 1 field day miscellaneous (or night work?) 9 sites x 4 = 36 days (out of 40) Position of beating samples in one bubble transect; 1 sample every 2 meters; understorey samples are vertical to canopy samples Material needed: 2 beating sheets, 120 ziplock plastic bags (reusable 3 times)

7. Sampling program: Fogging Managers: A. Floren & J. Schmidl Aims: compare the arboreal fauna at 9 sites Technique: fogging (xx trays) at each site after all other sampling programs are finished at this site 9 sites (3C, 3R, 2B, 1I) Total 9 fogging including xx trays (samples) at each site Schedule for one site: 2 field days for preparation and fogging 2 days laboratory BCI/University Panama 9 sites x 4 = 36 days (out of 40) Observations: 1. The 3 crane sites will be surrogate sites outside the crane perimeter and plot 2. For raft sites, fogging is performed after the raft has been removed; access lines to the canopy must remain after this operation. Fogging will be performed partly at the raft site 3. Fogging should be performed away of FITs... Material needed: 2 foggers, trays, containers, white oil, pyrethrum, etc.

8. d d d 20m +20m +20m Sampling program: Light traps Manager: R. Kitching Aims: compare the moth fauna flying in the understorey and upper canopy Technique: 6 UV light traps, automatic collecting (bucket), no manual collecting 6 traps running in the understorey (3) and in the upper canopy (3), simultaneously 3-4 hours of trapping (21h00 - 01h00?) Total 6 samples (light traps) per site, 3 upper canopy + 3 understorey Ikos: only 1 trap upper canopy + 1 trap understorey 9 sites (3C, 3R, 2B, 1I): Total (8 x 6) + 1 + 1 = 50 samples Position of light traps on the raft d = 24m (flat) 20m Position of light traps in the understorey plots and crane subplots (20m x 20m); or directly vertical to upper canopy traps Schedule for one site: 1 field night for canopy+understorey samples 2 days laboratory BCI 1 field day miscellaneous (or second field night?) 9 sites x 4 = 36 days (out of 40) Position of light traps in one bubble transect; understorey traps are vertical to them Batteries re-charging: sent to STRI in the city Material needed: 6 light traps with rain shield, 10 batteries, etc.

9. Sampling program: Soil/Canopy microarthropods Managers: N. Winchester (c), K. Jordan (c), A. Dejean (s), B. Corbara (s), M. Leponce (s) & Y. Roisin (s) [c = canopy cores, s = soil cores] Aims: compare the soil microfauna of the canopy and forest ground Technique: coring (3 x 5 cm) and extraction with 48 Berlese extractors (48 hours) Total 16 samples per tree, 8 canopy + 8 soil 3 trees per site: Total 24 canopy + 24 soil = 48 samples 9 sites (3C, 3R, 2B, 1I): Total 9 x 48 = 432 samples One tree: 2 paired cores (samples) away/close from the trunk 4 samples in the upper canopy 4 in the lower canopy Soil: 4 directions N - S - E - W 1 sample close/away (10m) in each direction Schedule for one site: 1 field day for canopy+soil samples 2 days laboratory BCI 1 field day miscellaneous 9 sites x 4 = 36 days (out of 40) Material needed: 48 Berlese, ziplock plastic bags (reusable 3 times n= 150, 6 corers, etc.

10. Sampling program: Winkler - litter Managers: H.-P. Aberlenc, B. Corbara & J. Orivel Aims: compare the active and passive macrofauna of the litter at 9 sites Technique: sifting litter and extraction for 48 hours with mini-Winkler bags 60 samples (1m2 of liter = 2 liters or less) per site (max 120 liters or 60kg) 9 sites (3C’, 3R, 2B, 1I): Total 9 x 60 = 540 samples Note: the crane sites are surrogates, outside the crane plot, as the method is too destructive. At each site, 3 transects of 20m each Take 20 samples (1m2 = ca. 2 liters) with a one-square-meter frame Pitfall traps should be activated and surveyed before the Winkler survey Schedule for one site: 1 field day for performing 3 transects 2 days laboratory BCI for extraction 1 field day miscellaneous activities 9 sites x 4 = 36 days (out of 40) Position of sticky traps at 1.3m (DBH) 20m Winkler transect (n = 3) litter totally removed here Position of pitfall traps at one site. Distance between traps = 1.3m Material needed: 3 sifters, 60 Winkler bags, plastic bags (reusable 3 times, n = 200), large shopping plastic bags to carry samples, 3 one-square-meter frames

11. Sampling program: Pitfall traps Managers: A. Tishechkin, L. Cizek & E. Medianero Aims: compare the active macrofauna of the litter at 9 sites Technique: pitfall traps running for 3 days (Y. Basset to do a pilot study) Rain protection: 2 BBQ sticks through a plastic soda cover 15 samples (pitfall traps) per site 9 sites (3C, 3R, 2B, 1I): Total 9 x 15 = 135 samples Plastic cup: diameter ca. 6cm depth ca. 15cm (0.5 liter) Solution: salt (100g), water and detergent, possibly ethanol (total 0.175ml) Pitfall traps should be activated and surveyed before the Winkler survey Schedule for one site: 1 field day for emplacing traps + miscellaneous 2 days laboratory BCI 1 field day for surveying traps + miscellaneous 9 sites x 4 = 36 days (out of 40) Position of sticky traps at 1.3m (DBH) 20m Winkler transect (n = 3) litter totally removed here Position of pitfall traps at one site. Distance between traps = 1.3m Material needed: 50 (reusable) plastic cups, 50 (reusable) plastic soda cover, 100 (reusable) BBQ sticks, 2 plant corers, 2 wash bottles, 10-20 containers with handy box.

12. Sampling programs: Individual programs Social insects: A. Dejean, B. Corbara, J. Orivel, A. Floren, M. Leponce & Y. Roisin Pollinators: D. Roubik & D. Frame CO2 and soil respiration: C. Potvin & students Ground FITs: A. Tishechkin & G. Curletti Wood borers: F. Ødegaard & G. Curletti Gall-makers: S. Ribeiro Note: at crane sites, individual programs need to be performed outside the crane perimeter AND outside the botanical plot. That is near the sites C1’, C2’ and C3’.

13. Summary: ground plots Most samples will concentrate on the 20x20m ground plot, but not all, as this would bias samples. The inner area of 20m x 20m (see diagram next page) will be restricted to a few sampling programs. More destructive programs, including individuals programs would need to be performed at some distance of the plot. This is indicative only, as some programs need to overlap between sites (sticky traps). Sequence of sampling: Day 1: Griding and vegetation survey; beating Day 2: Set up of sticky traps, pitfall traps Day 3: Light trapping; microarthropods Day 4: Individual programs Day 5: Survey of pitfall traps; Winkler transects Day 6: Individual programs Day 7: Survey of sticky traps Day 8: Fogging (not close to FITs, may be scheduled later); set up of FITs on sticky trap transects Later: survey of FITs (two weeks later); refine vegetation survey if needed And at all time: work on individual programs outside the plot permitted

14. Microarthropods (n = 3 trees) situation variable, maybe outside the plot Ground plots: summary Transects Sticky and FIT traps ONLY vegetation, sticky traps, light traps & microarthropods 1 20m Beating: extends 5-10m beyond plot 2 All individual programs: extends 50-100m beyond the plot 3 20m Pitfall traps (n=15), each 1.3m Winkler transects Light traps (n = 3) Walking and access area (for raft sites) Sticky traps at 1.3m (n=25), may extend beyond the plot PVC marks for vegetation study Fogging: preferably in area 3 Not to scale

15. Summary: canopy plots Most samples will concentrate on the 20x20m canopy plot, but not all, as this would bias samples (see diagram next page). This is indicative only, as some programs need to overlap between sites (sticky traps). Sequence of sampling: Day 1: Vegetation survey; beating Day 2: Set up of sticky traps Day 3: Light trapping; microarthropods Day 4: Vegetation/individual programmes Day 5: Vegetation/individual programmes Day 6: Vegetation/individual programmes Day 7: Survey of sticky traps Day 8: Fogging (not close to FITs, may be scheduled later); set up of FITs Later: survey of FITs (two weeks later) And at all time: work on individual programs below the sites or adjacent to them The Icos site will be more limited

16. Canopy plots: summary (Example of one raft site) Light traps Sticky traps: 25 traps on the periphery and 3 vertical transects Beating samples: 20 samples on the periphery Microarthropods: 3 trees below/adjacent to the raft Sticky and FIT traps transects Individual programmes: on the raft or adjacent Fogging: preferably adjacent to vertical projection of the raft