1. Invertebrate and Algae
2017 PISCO UCSC
Invertebrate and Algae
Sampling Methods

2. Overview of today’s training
PISCO Annual Survey Design
Swath survey method
UPC method
Benthic transect protocol
Organism ID slides

3. Annual Survey Design When: June – September 1999 – present Where:
Different oceanographic regimes
Lower upwelling – southern Monterey Bay
Higher upwelling – Point Lobos, Carmel Bay
Marine Protected Areas and paired comparison sites
Lover’s Point SMR, Ed Ricketts SMCA, Pacific Grove Marine Gardens SMCA
Point Lobos SMR, Pescadero Point, Soberanes

4. Example of permanent, stratified random sampling design
Pacific Grove Marine Gardens SMCA
Benthic transects at 5, 12.5 and 20m
Fish transects at 5, 10, 15 and 20m
Asilomar SMR
Existing PISCO long term monitoring unit
New randomly selected monitoring unit
’89 ’99 ’02 Composite Kelp
2005 Kelp
100m buffer from reserve boundary

5. Depth-stratified sampling design: fishes
Transects: 30 m (n=3 per depth)
Depths: 5, 10, 15 & 20 m
Total = 12 transects per Area

6. Benthic transects: 30 m (n=2 per depth)
Depth-stratified sampling design: inverts & algae
Benthic transects: 30 m (n=2 per depth)
Transect depths: 5, 12.5 & 20 m

7. Annual Benthic Surveys – Nested Hierarchy
1999 ……
20??
Time
Sites 1 2 3 …… ?
Cells 1 1 2
Zones
(5 , 12.5 , & 20 m)
A cell (occasionally in past years called an Area) is a sample within a site, and a site can have one or two cells in it. S is shallows, M is mids, and D is deeps. S M D
Transects:
(n=2)

8. Annual Benthic Surveys: Depth Stratification
Depth zones:
Shallow (S) = 5 m, Mid (M) = 12.5 m, Deep (D) = 20 m
designed to assure that all depth zones are representatively sampled for description of invert and algae assemblages.
2. Allows representative sampling from outer to inner edges of the reef.
If reef depths are constrained (e.g., 5-10 m), then sampling is distributed from outer to inner edges of reef including comparable depths (when possible) to those above (e.g., 5 and 10 m), plus additional depth strata to assure coverage from inner to outer edges of the reef.
Note that divers descend at known GPS locations, not where kelp is present or not. In some years, the kelp is very thick, and in others it can be absent. We have GPS points to tell us where to go for each depth zone and we return to them year after year.

9. Annual Benthic Surveys: Depth Stratification
Algae/invertebrate transect distribution
Coastline
North
Upcoast cell
Zone
Downcoast cell
5 m (shallows)
12.5 m (mids)
20 m (deeps)

11. Annual Benthic Surveys: Methods
PISCO-UCSC divers use three methods:
(1) Uniform point contact (UPC) to estimate percent cover of benthic organisms and to characterize the substrate
(2) Swath transects to estimate the density of kelps and targeted macroinvertebrates
(3) Size frequencies of abalone and urchins (AbU)

12. Annual Benthic Surveys: reeling tapes out
Priorities: reef > depth > heading
- If your pre-designated compass heading will take you off reef, make a systematic deviation and maintain the new heading for the remainder of the transect.
- Maintain constant depth (continuously watching depth gauge) by contouring around large rocks and avoid stringing the transect tape over very large crevices.
- Use kelp, rocks, etc. to wrap/anchor the transect tape along the bottom.
**If you ever complete a transect and feel that something was “off”, speak up. We can always re-do transects.**

13. Annual Benthic Surveys: tape tension
Do not lay tape with high tension
Lay tape so it generally contours bottom

14. Annual Benthic Surveys: tapes and crevices
Side view of reef with red tape: too taut
Less tension on red tape
Upper diagram shows a taut tape.
Lower tape shows some droop.
All depths into reef are 1 m, but angles differ leading to various amounts of droop, and switching from a crack/crevice to a contour/valley.
1 m

15. Swaths should extend one meter on either side
Swath Surveys: 2 m wide
Swaths should extend one meter on either side
of the transect line.
Top view of transect
1 meter
Transect line
Maintain reasonable speed (~20 minutes per transect) and only count invertebrates 2.5 cm or larger in diameter
(except urchins and certain sea stars)

16. Swath Surveys: 1 m body measurement
1 m is measured by diver extending arm and moving body over line to the point on chest where 1 m ends.
Each diver must know where on their body 1 meter from the tip of their fingers stops (endpoint). This is often near armpit, but it varies.

17. Swath Surveys: swath width
Diver uses “arm to endpoint” to measure 1 m on either side of the transect tape.
End-on view of transect —
tape (red dot) is into and out of slide
1 m
1 m
1 m
1 m
Transect tape

18. Swath Surveys: mandatory flashlights
Use a flashlight at all times when surveying for swath invertebrates to make sure organisms in cracks and
crevices are counted

19. Swath Surveys: search area
Reef 1m
Flat
Transect tape
(into/out screen) 1m 1m
Crevice
Flat and vertical

20. Green line indicates area searched1m 1m
Transect tape
(into/out screen)
Reef
Ridge
Wall 1m
Green line indicates area searched 1m 1m 1m
Overhang

21. Sub-sampling protocol: proper counting
Cover symmetric sides when subsampling
You always collect data from quadrangles!!

22. Distance covered, not meter mark
For a sub-sample, record the distance sampled within the 10 m segment to the nearest 0.1 m.
Example: this diver started at 30 m and counted 33 Pterygophora by the 26.2 m mark, thereby sampling a rectangle 3.8 m long.
Record this as: 3.8 m.

23. Swath Surveys: invert data sheet
Siren
330
Lonhart
Malone
36’-34’
Four genera are counted and three genera are measured:
Count = Patiria, Dermasterias, Henricia, & Mediaster
Measure = Pisaster, Pycnopodia and Orthasterias.

24. Sea star measurement
Measure length (nearest cm) for TARGET sea stars encountered in the 2 m wide swath.
Measure along the aboral surface, from the center of the medial disc to the tip of the single longest ray.
Remove the star if easy to do, otherwise measure in place.
Note disease status (* = mild, ** = dissolving)

25. When measuring, follow the curve of the longest ray

26. Measure to the nearest cm (without bias)
Measure to the nearest cm (without bias)

27. Sea star disease and recording
Note health status for sea stars: * = mild disease ** = severe
*(II, 3, E) translates to 16 bat stars that display signs at mild stage of disease
Swath diver measures sea star ray length for certain species.
III, 7, 5, **(2,I)
II, 2, E, *(II, 2)
Some stars counted, others measured.
7, 12, E, 8*, 10, 14** 8(III, 3) 8*
3, 7, E, 5*
Ortkoe 5, 9,

28. Swath Surveys: invert data sheet
These species must be
≥2.5 cm diameter/length

29. Swath Surveys: invert data sheet
These species must be
≥2.5 cm diameter/length
Do not forget the write-ins:
Anthopleura xanthogrammica = giant green anemone, no lines on oral disc
Metridium spp. = white anemones, giant plumose anemone (M. farcimen)
Lytechinus pictus = white urchin, has brown patches
Aplysia californica = red sea hare
Aplysia vaccaria = black sea hare
Megastrea undosa = wavy turban snail (never been seen)
Loxorhynchus grandis = sheep crab, greenish, spines, few epibionts
Neobernaya spadicea = chestnut cowry

30. Swath Surveys: recording observations
III
IIIIIII
III
If you ever want to write the number eleven, do not write II, instead use E. There is no eleven other than E. Anything else is “2”.
III, 7, 22 I I
I, 4, IIIIII, E

31. Swath Surveys: recording observations
III
IIIIIII I 32 22 3 7 12 II
IIIIIII
III
Prior to next dive, translate and sum your data in each row and circle that number. Overall total on extreme right.
III, 7, 22
I, 4, IIIIII, E

32. Swath Surveys: kelp counts
Count all individuals >30 cm, except Macrocystis and Nereocystis (>1 m) and Cystoseira (>6 cm diameter).
>6 cm
We’ll address this further during the algae identification portion of the class.
1 m
Efird

33. Swath Surveys: recording data
Fuller’s
330
Lonhart
Carr
38’-30’
1, 3, 37, 15, 8, 5, 2, 45
III, 4, 2, III III I 16
III, 12, E 4.3 m
6.3 m
2, 4, 6, 2, 14, 5, 6.3 m
III, E, 2, III 19
Record # of stipes for each individual Macrocystis, separate with commas
Record number of Nereocystis
Count stipes of Pterygophora, Eisenia, L. setchellii, & Pleurophycus
Sum all non-Macrocystis counts upon surfacing

35. UPC Surveys: tape meter marks

36. UPC Surveys: tape meter marks
Data collected every 1 m on a 30 m transect tape
1 meter increments
10-m segment
Transect tapes are marked at 1 m increments. Every 10 m segment has a single color of tape at each 1-m mark.
Be sure to count 10 points per segment. For example, if you start at 30 m mark you stop at 21 m mark, and 20 is your next segment’s starting point, thus you finally end at 1 m mark and not 0 m (30 to 1). If you start at 29 m, you end at 20 m, and at end of tape, end at 0 m mark (29 to 0).
0-9 is ten digits, just like 1-10 is ten digits. Think about it….

37. UPC Surveys: tape to bottom
Remember not to bias your data!
Look once and pick the point, do not “look around.”
If the tape is suspended above the bottom, then be sure to hover over the point and look straight down. While deploying the tape, avoid too much tension that causes suspended tapes and covers more than 30 m of actual bottom.

38. UPC Surveys: 30+ counts per 10 m
Substrate
Relief
Superlayer
(special category)
Laminaria superlayer
Write – ins
Point
(cover)

39. UPC Surveys: substrate
Substrate type
You should know 10 cm based on your hand. For example, on my hand, 10 cm is the width from pinkie to trigger finger, or across the knuckles of a fist.
Barlotti

40. UPC Surveys: sand vs. bare sand
Sand can be substrate (Sand) and a point (Bare sand category)
What happens when bedrock has a pocket of sand?
If the point lands on a pocket of sand that cannot be waved away with the brush of a hand, but you can poke your finger into the sand and touch the rock under it, then
Substrate = Bedrock and Point = Bare sand
If the pocket of sand is deep enough that you cannot touch the rock under it, it is potential habitat, so then
Substrate = Sand and Point = Bare sand
If you are on bedrock and come across a sand-filled crevice full of Diopatra, and your point hits the worm, then
Substrate = Sand and Point = Diopatra

41. UPC Surveys: ignore epibionts
If your point falls on a mobile invertebrate or epibiont, ignore it and make sure you are recording the organism underneath it that is directly attached to the substrate!
(Note, it need not be attached under the point, elsewhere is ok)
Colonial tunicate
on a solitary tunicate
Lonhart/MBNMS

42. UPC Surveys: ignore epibiontsA C B
Sponge
Green
algae
Red
branching
Scenario A: Alga is an epibiont so ignore. Orange sponge is attached to substrate, as far as you can tell.
Scenario B: Alga could be counted, follow it back to determine if it is attached to substrate. Epibiont, so ignore.
Scenario C: Alga could be counted, follow it back to determine if it is attached to substrate. It is, so use it.

43. UPC Surveys: superlayer
Drift algae and juvenile laminariales (e.g., <30 cm tall Laminaria, Eisenia, Macrocystis, Pterygophora) can be the first thing you encounter at your point. Record the superlayer, move it aside and then record the primary substrate holder.
Figurski
Figurski

44. UPC Surveys: epibionts & superlayer
Red branching and superlayer if it is a juvenile laminariales
Sponge
Red
branching
*Stephanocystis (nee Cystoseira) is Order Fucales.
Dictyoneurum is Laminariales. Dictyotales is not Laminariales. Desmarestia is Desmarestiales.
If it is not a juvenile laminariales then it is either Cystoseira*, Dictyota, Desmarestia, Sargassum spp. or Other Brown

45. UPC Surveys: superlayer species or not?
Laminariales genera
in central California:
Laminaria
Costaria
Pleurophycus
Egregia
Eisenia
Alaria
Pterygophora
Macrocystis
Nereocystis
Dictyoneuropsis /
Dictyoneurum
Non-laminariales genera
that are brown and
in central California:
Cystoseira
Dictyota
Desmarestia
Sargassum
Other Brown
*Stephanocystis (nee Cystoseira) is Order Fucales.
Dictyoneurum is Laminariales. Dictyotales is not Laminariales. Desmarestia is Desmarestiales.

46. UPC Surveys: ID superlayer spp.
Key is to determine
what this is!
Juvenile Nereocystis above, and likely juvenile Macrocystis below it
Nereocystis and likely Macrocystis below it.

47. UPC Surveys: ID superlayer spp.
Key is to determine
what this is!
Juvenile Eisenia arborea
Eisenia juvenile.

48. UPC Surveys: ID superlayer spp.
Key is to determine
what this is!
Juvenile Macrocystis pyrifera
Macrocystis pyrifera juvenile.

49. UPC Surveys: ID superlayer spp.
Key is to determine
what this is!
Adult Dictyoneurum / Dictyoneuropsis, and therefore it is NOT a superlayer.
Dictyoneurum/Dictyoneuropsis. Very short stipe, flattened, and two emerging from one holdfast. Rectangular reticulation.

50. UPC Surveys: determining relief
Relief is the absolute difference in elevation between the highest and lowest point within a 1 m by 0.5 m rectangle
Top view of transect
0 m
1 meter
0.5 meter

51. UPC Surveys: determining relief
Relief is determined within non-overlapping rectangles (1 m by 0.5 m) as depicted below
Top view of transect
1 meter 1 2
0.5 m
The rectangle is centered on the UPC point. Each rectangle extends 0.5 m on both sides of the tape, and 0.25 m in front of and behind the point along the tape.

52. UPC Surveys: determining relief
Imagine the rectangle is suspended above the bottom, as if it had been lowered from the surface and was perfectly level.
Top view of transect
1 meter 1 2
0.5 m
Within each rectangle find the highest and lowest points, then determine the vertical distance between the two.

53. UPC Surveys: high and low points
Find the high and low points in each rectangle. Remember it is 0.5 m to either side of the meter mark, and 0.25 m in front and behind it.
0.5 meter
End-on view of transects

54. UPC Surveys: relief categories
Relief is binned into four categories. Basically they are: flat, knees to head, standing, and wall.
0-0.1 m m 1-2 m >2 m

55. UPC Surveys: review Substrate sand, cobble, boulder, bedrock Relief
At the end of the 30 m you should have 30 points in 3 categories (i.e. substrate, relief, and point—90 total) and possibly some superlayer data.
Substrate
sand, cobble, boulder, bedrock
Relief
0-0.1, , 1-2, >2 m
Superlayer
(special category)
Drift or juvenile laminariales
Laminaria superlayer
Write – ins
What was under the point, live or dead
Point

56. UPC Surveys: recording data
Hopkins
330
Lonhart
Saarman
16’-14’
IIII II
II II II
IIIIIIIIII
II II
II II
I I I II I I I
I I I I I I I
I I II II I III 2 I I
Did you find the error?

57. UPC Surveys: summing rows
Hopkins
330
Lonhart
Saarman
16’-14’
IIII II
II II II
IIIIIIIIII
II II
II II 22 4 4 E 19
I I I II I I I
I I I I I I I
I I II II I III 2 I 8 3 7 10
Between dives, sum all rows and circle total at right.

59. Abalone & Urchin surveys
Count abalone and purple & red urchins on the swath transect (2 m x 30 m) in 10 m segments.
Use flashlight to look in crevices.
Count and size (cm) what you can see.
Record to nearest cm.
Sub-sample urchin species if 30 counted, pooling across all size classes in that segment.

60. Abalone & Urchin sizing
Measure greatest shell length (nearest cm) for each abalone encountered in the 2 m wide swath.
Urchin diameter is for the test (i.e. shell) and does not include the spines.

61. Length
Greatest shell length for abalone

62. Diameter

63. AbU Surveys: recording data
McAbee
9-8-17
270
Lonhart
Vylet
46’-50’
I, 4, II, 4, IIII, 2, 2, III)
III
IIIII, 5, 3, IIII
5 (II, 2, 4) @ 7.3 m 33

65. 2017 Benthic Transect Protocol
Three diver system: UPC diver, swath diver, and AbU diver
There will be three divers. In addition to UPC (black) and swath (green) divers, there is also an abalone-urchin (purple) diver, aka AbU.
The AbU diver collects size-frequency data for both red and purple urchins as well as three abalone species.

66. Phase 1: Reeling out transect #1
Swath diver reels out tape, follows heading but maintains target depth;
The UPC diver collects data;
The AbU diver collects data while following UPC.

67. Phase 2: Heading to 0 m on transect 1
Swath diver secures tape, then starts counting swath inverts;
UPC diver reaches end of tape, heads back to 0 m counting swath algae;
The AbU diver reaches end of tape and reels it in, making sure both swath and UPC are ahead;
Always check in with buddies as you pass.

68. Phase 3: Reeling out transect #2
Swath diver reels out tape, follows heading but maintains target depth;
The UPC diver collects data, transitioning from tape #1 to #2;
The AbU diver collects data while following UPC.

69. Phase 4: Swath algae data on transect 2
If the swath diver is done collecting invert data on transect #2, then that diver is now at the 0 m end. The swath diver can collect swath algae data from 0 m out to the end of transect #2.
The UPC diver may be working back from the 30 m end of transect #2, also counting swatch algae. When they meet, they ensure that the area between them is counted.
If the swath diver is still on inverts and passed by the UPC diver, then the UPC diver finished algae on transect #2.
The AbU diver reels in the tape since they are typically slowest.

70. Ideal scenario for swath algae on transect 2
Swath is working back to 0 m doing inverts. UPC is going to 30 m doing UPC.
UPC is heading to 0 m doing algae, swath is heading to 30 m doing algae.
UPC and swath divers meet, ensuring all algae counted between them. At the surface they put all data onto one data sheet in between dives.

72. Sub-sampling reminder!
Within each 10 m segment, if you reach 30 or more individuals of a single species, note how many were counted followed by the distance covered (e.g., 32 at 2.7 m). M. pyrifera excluded.
You start over in the next 10 m segment.
Be sure to finish the entire ‘rectangle’ so we capture an accurate density estimate.

73. Sub-sampling protocol: 10 m segment
Within each 10 m segment, once ≥30 individuals of a species are counted, indicate the distance you sampled within that 10 m segment (NOT THE METER MARK!) and stop counting that particular species until you reach the next 10 m segment.
For example, if you count 34 Styela from the 20 m mark to the 17.5 m mark, then you record that as 2.5 m” on the data sheet.
Be sure to count all individuals in the rectangle you sample, even if it is slightly over 30, since density estimates are based on a rectangular area.
Count all Macrocystis.

74. Safety Considerations
Dive profiles - approved by DSO - well within no-decompression limits - maximum depth 20 m min safety stops mandatory on dives >10 m
Air consumption - safety over data collection (500 surface)
Buddy system - members of a dive team are always in contact via the transect line
Sea conditions - diving will only be conducted in good conditions

75. Dive Safety Brief 1. Dive log - sign out and in
2. Dive procedures ascent rate 1 ft per 2 sec deep-shallow maintain dive team
3. On board communications: cell phone, VHF Ch 16 (Paragon), & SPOT system
4. First Aid & Oxygen - Shore and vessel
5. Evacuation plan: this is developed for each site and must be covered prior to the dive

76. Locations of CHOMP and PG chamber

77. Locations of LML, the SC harbor and Dominican Hospital

80. Transect 2
Transect 1
Invert 30 30
3. Reel out
1. Reel out
4. Collect data
2. Collect data
5. Collect algae data
UPC
3. Collect data
1. Collect data
4. Collect algae data
2. Collect algae data
AbU
1. Collect data
3. Collect data
4. Reel in
2. Reel in