Lab Report 1


Comparison of Biotic Factors: Biodiversity, Productivity, Benthic Cores, And Abiotic Factors: Temperature, pH

Among Two Locations in the Palo Verde Lagoon and One Location in the Quebrada Mula Wetland in Palo Verde National Park

Submitted by: Hal Groeneboer, Mary Moreira, Carol Paine, Stacy Weaver
  1. Introduction
Palo Verde National Park is primarily a dry forest area, which was formerly a cattle ranch. It was set up by the Costa Rican government in 1977 as the Palo Verde National Wildlife Refuge. There are numerous wetland areas throughout the park, but little is known about the natural ecosystem prior to the introduction of cattle ranching over 300 years ago. We chose two wetland areas to survey and compare: the Laguna Palo Verde which is a large seasonally flooded herbaceous marsh along the eastern bank of the Tempisque River and the Quebrada Mula wetlands which is near the eastern boundary of the park adjacent to cattle pastures. Both of these wetlands have some incidence of cattle grazing during part of the year, but this is limited compared to full time cattle grazing in the pasture areas.

The Palo Verde lagoon is on the floodplain of the Tempisque River. It is bounded by the river on the south and by limestone hills on the other sides. "The lowland areas between the hills and the wetland are composed mainly of montmorillonite clay with 2 x 2 lattice structure of the clay micelles which permits considerable incorporation of water molecules during the wet season and a substantial dry season loss of water." (Jansen, p. 131) During cattle ranching times, an opening was made between the lagoon and the river which allowed continual exchange of water between the river and the wetland, especially during the wet season. The river drains numerous rice fields along its route and as a result carries much sediment and agricultural chemicals. When the park was set up, the opening between the river and the lagoon was closed. At present, the main sources of water entering the lagoon are rainfall and runoff from the higher areas, primarily forested areas on the limestone cliffs. However, there is still flooding on a regular basis (usually at high tides) about 12-18 times per year. All areas of the lagoon seemed to have consistent water levels of about 0.5 to 0.7 meters within 10 meters of the edge.

Two areas in the lagoon were surveyed. One is being invaded by Typha dominguensis (cattails) which is becoming the dominant plant in that area. The other area, while not open water, is covered by primarily low growing species, also invasive – primarily Eeichhornia crassipes (water hyacinth), Nymphaea ampla (a water lily with small, rounded leaves) and another water lily having larger leaves with toothed margins. There are mangrove areas along the river bank and an area of Parkinsoniia aculeata (palo verde tree) in a parallel band between the mangroves and the lagoon.

The Quebrada Mula site, in addition to rainfall, run off from surrounding areas, and much less frequent river flooding, could also be fed by surface water (quebrada means small stream) although this is not documented. It is bisected by a concrete bridge and a filled in road area, with a narrow, 4.5 meter opening between them. Under and on both sides of the bridge, as well as one other small area, there was open water, probably indicating deeper water (>2 meters). The water level in the marsh varied between very shallow (15-20 cm) to water probably over our heads. This wetland had numerous areas with significantly different characteristic vegetation. Some areas were covered in primarily low growing floating vegetation while others had tall grasses and arrowroot.

The park management has concerns that the invasive species found in the wetland areas are decreasing the biodiversity which could eventually limit the ability of the wetland to maintain a healthy ecosystem.
 
 

II. Methodology
 
  1. Diversity – In Laguna Palo Verde, there seemed to be two distinct areas with characteristic vegetation. (There was also a shoreline area with distinct vegetation which we did not sample.) We marked off a plot 2 by 2 meters (4 square meters) about 60 meters from shore in approximately 80-90 cm. of water, where there was no discernible effect from grazing cattle. We took one sample far enough in so that Typha was the dominant plant and another where water hyacinth and water lilies were dominant. Each species of plant in the marked area was counted.

  2. In Quebrada Mula, because there was a large difference in depth accompanied by a wide difference in type of dominant vegetation, we chose to use 4 separate 1 square meter plots. The procedure here was basically identical to that followed at the other side.

    Plant identification was done at the OTS station from various books and articles which listed vegetation found in these wetland areas.
     

  3. Biomass – We utilized a clip plot in all areas. In the lagoon we marked a one meter square plot, while in Quebrada Mula we used four (4) - 0.25 meter plots to reflect the widely different vegetation noted above. All vegetation was cut off immediately above the substrate and collected in plastic bags. Holes were punched in the bottom of the bags to drain all the water, and allowed to rest for about 24 hours before being massed.
  1. Benthic Cores – We took 4 samples at the lagoon sites and 5 samples at the QM site (of which we dropped one). Each sample was collected by gently placing a 60 cm. PVC pipe (7.5 cm. diameter) on the substrate and then quickly pushing it in about 8-10 cm. or until it was too hard to push further. The pipe was then loosened, and a hand placed under it while it was lifted to ensure that no material fell out. Water which was collected in the tube above the substrate was poured out and quickly examined for evidence of larger organisms. The benthic core was then loosed from the PVC pipe and placed in a plastic sealable bag.
  1. Temperature and pH – A small vial was used to collect water at the surface, approximately half way to the bottom and about 1 cm above the bottom at each of the sites. This water was brought back to the laboratory and measured with pH paper.

  2. Temperature was measured by placing the thermometer about 1 cm below the surface of the water and about 1 cm above the substrate. The thermometer remained at each position for about 2 minutes.
     

  3. Live Organisms – We took each water sample which had been collected less than 2 hours previously, poured it into a petri dish, and examined it under a microscope. We used the DAFOR (dominant, abundant, frequent, occasional, rare) characterization scale.
  1. Presentation of Data
    1. Plant Diversity (4 square meter plots)
Note: Thalia geniculata (Arrowroot) and Mimosa pigra was found in numerous places in the lagoon, but not in either of the areas surveyed. There is also a tall (1m) round reed along the shore in 15-20 cm. water, with darker green segments about 2.5 cm apart. Palo Verde – typha area Name Quantity Common name or description
Typha dominguensis 18 Cattails

Sagittaria guyanensis 17 Water plaintain

Aeschynomone sensitiva 9 Pinnately compound leaf, small yellow flowers, floating white rope like, tuberous main root surrounded by mats of hair roots.

Nymphaea ampla 8 Small leafed, smooth edged white flowered water lily Grass A 50-60 Segmented grass with alternating leaves, reddish stem, 30 cm. above water, long underwater stems

Pistia stratiotes about 200 Small duckweed

Floating plant A 2 Water lettuce

Eleocharis mutata or 5 Sedge (triangular stalk), no seeds or Cyperus articulatus flowers. (found mostly along shore in C. digitatus 15-20 cm water)

Palo Verde – Low vegetation

Eichhornia crassipes 14 Water Hyacinth

Nymphaea ampla 24-30 Small leafed, smooth edged water lily stems and bottom of leaf reddish colored Water lily A 12-25 Larger, toothed edged lily with typical white Paspalum sp. < 5% Grass (Phillipe identified)

Potomogeton sp. <5% Fern like aquatic plant (seaweed)

Aeschynomone sensitiva 9 Pinnately compound leaf, small yellow flowers, floating white rope like, tuberous main root surrounded by mats of hair roots.

Minute yellow flower on 5-6 cm stalk with no leaves, rooted in floating roots of other plants
 

Quebrada Mula (4 separate 1 meter square lots)

We found Canna lutea (canna lily) in the wetland, but not in the area surveyed.

Eichhornia crassipes 100 Water Hyacinth

Grass A 50 Segmented grass with alternating leaves, Reddish stem, 30 cm. above water, long underwater stems
? 570 Fuzzy leafed (similar to Venus Flytrap), clumped with root ball, floating plants

Pistia stratiotes <1% Small duckweed

? 60 Floating seaweek (milfoil) type plant

Grass C 16 Sharp leafed, light green segmented grass

Grass D 23 Sharp leafed, dark green, segmented grass

Grass E 37 Hairy leafed and stem, medium green segmented grass
Mimosa pigra? 2 Herbaceous, thorned mimosa type legume

? 6 Water lettuce
? 10 Fuzzy deeply lobed palmate leafed, tall (1 m.) single stemmed, herbaceous plant
Vine with trifoliate leaf

Plants supposedly found in the lagoon (Crow), but unidentified by us are: Ludwigia inclinata, Najas guadalupensis, Nymphaea prolifera, Heteranthera limosa, Bacopa repens, , Neptunia plena, Paspalidium germinatum
 

  • Plant Biomass (1 meter square clip plot) – biomass was massed following day after water was drained but was not dried. (+/- 0.1 kg.)

  • Palo Verde Lagoon – Typha area - 7.6 kg.

    Low vegetation 2.7 kg

    Quebrada Mula 6.0 kg.
     
     

  • Benthic Core Samples

  • Palo Verde Typha

    Core Sample #1

    Mollusk #1 4 Small Snail Shells Planaspiral- light brown

    Mollusk #2 6 Small fragments - white

    Core Sample #2

    Mollusk #1 4 Small Snail Shells Planaspiral -light brown

    Mollusk #2 8 fragments -white

    Core Sample #3

    Mollusk #1 1 Large Snail Shell Planaspiral, White

    Mollusk #2 1 Large Snail Shell Planaspiral Brown

    Mollusk #3 6 Small Snail Shells 1 brown, 5 white

    Mollusk #4 9 Small White Fragments

    Mollusk #5 5 Small Brown Fragments

    Core Sample #4

    Mollusk #1 1 Large Snail , Planaspiral, Light Brown

    Mollusk #2 3 Tiny White Shells Planaspiral

    Mollusk #3 12 Shell Fragments -White helispiral

    Mollusk #4 3 Shell Fragments Brown helispiral

    Mollusk #5 Small white Bivalve
     
     

    Palo Verde Low Vegetation- Core Sample #1

    Mollusk #1 Large Half Snail, planaspiral light brown

    Mollusk #2 3 Small Snail 1 helispiral white

    2 planasprial light brown

    Mollusk #3 Fragments 8 light brown

    13 white

    brown bivalve

    Core Sample #2

    Mollusk #1 2 Large Snail Shells both hosting aquatic plant growth

    white helispiral

    brown striped helispiral

    Mollusk #2 2 Small Snail Shells planaspiral 19.0 mm. brown

    8.3 mm. White

    Mollusk #3 Fragments 3 helispiral brown

    27 planaspiral white

    Core Sample #3

    Mollusk #1 1 Large Snail

    Mollusk #2 1 Tiny Snail

    Core Sample #4

    Mollusk #1 1 Large Snail

    Mollusk #2 3 Small Snail

    Quebrada Mula

    Core Sample #1

    No living or dead mollusks were found

    Core Sample #2

    Mollusk #1 Large large snail Helispiral grey and white - Live

    Mollusk #2 Large Snail Shell helispiral brown

    Mollusk #3 2 Small Snail Shells helispiral brown

    Core Sample #3

    No living or dead organisms

    Core Sample #4

    Mollusk #1 2 Large Snails Striped dark brown and white helispiral - Live

  • Microbial Infestation Top Middle Bottom

  • Palo Verge Lagoon – Typha Very Rare Rare Occasional

    Low vegetation None Very Rare Rare

    Quebrada Mula Occasional Frequent Frequent

    (Insect, protist)

  • PH (Used Hydrion Paper) Top Middle Bottom(1 cm above substrate)

  • Palo Verge Lagoon – Typha 7.5 7.5 8.0

    Low vegetation 7/0 7.5 8.5

    Quebrada Mula 5.5 5.5 5.5

  • Temperature (Ambiente air –30.6 C (87 F)
  • Top of water Bottom (1 cm above substrate) Palo Verge Lagoon – Typha 30.6 C (87 F) 27.2 C (81 F)

    Low vegetation 31.1 C (88 F) 26.6 C (80 F)

    Quebrada Mula 31.1 C(88 F) 26.6 C (80 F) (27.8 C on shallow side)

    1. Analysis of Data
      1. Plant species biodiversity - We used the Shannon-Weiner Biodiversity Index to compare the 3 sites. This index provides a measurement of both the number of different species as well as the proportion of each type of species as compared to the total number. The calculations are shown below:

      2.  

         

      Palo Verde Typha
      Spp f P plogp flog^2f flogf
      1
      18
      0.0573248
      -0.0711778
      28.362763
      22.594905
      2
      17
      0.0541401
      -0.0685674
      25.738077
      20.917632
      3
      9
      0.0286624
      -0.0442171
      8.1952089
      8.5881826
      4
      8
      0.0254777
      -0.0406074
      6.5245722
      7.2247199
      5
      55
      0.1751592
      -0.1325197
      166.58743
      95.719948
      6
      200
      0.6369427
      -0.1247768
      1058.9478
      460.206
      7
      2
      0.0063694
      -0.0139866
      0.1812381
      0.60206
      8
      5
      0.0159236
      -0.0286299
      2.4427953
      3.49485
      SUM
      314
      H'-->
      -0.5244828
      1296.9799
      619.3483
      s^2
      0.0007642
      Ho: H'1 = H'2
      Ha: H'1 NE H'2
      Contrast s t df p
      PVTvsPVO
      0.0386541
      6.083321
      282.07458
      <0.001 reject Ho
      PVTvsQM
      0.0338445
      1.1788094
      648.19504
      >0.1 accept Ho
      Palo Verde - low-vegetation
      f flog^2f flogf
      14
      0.1590909
      -0.127011
      18.390533
      16.045792
      28
      0.3181818
      -0.1582397
      58.639458
      40.520425
      19
      0.2159091
      -0.143737
      31.069005
      24.296318
      4
      0.0454545
      -0.0610192
      1.4499049
      2.40824
      4
      0.0454545
      -0.0610192
      1.4499049
      2.40824
      9
      0.1022727
      -0.1012746
      8.1952089
      8.5881826
      10
      0.1136364
      -0.1073276
      10
      10
      88
      H'
      -0.7596281
      129.19401
      104.2672
      s^2
      0.0007299
      Quebrada Mula
      flog^2f flogf
      100
      0.1129944
      -0.1069992
      400
      200
      50
      0.0564972
      -0.070507
      144.32495
      84.9485
      570
      0.6440678
      -0.123061
      4329.0623
      1570.8487
      9
      0.0101695
      -0.0202648
      8.1952089
      8.5881826
      60
      0.0677966
      -0.0792401
      189.70931
      106.68908
      16
      0.0180791
      -0.0315087
      23.198479
      19.26592
      23
      0.0259887
      -0.0411977
      42.648962
      31.31974
      37
      0.0418079
      -0.0576423
      90.992496
      58.023464
      2
      0.0022599
      -0.0059795
      0.1812381
      0.60206
      10
      0.0112994
      -0.0219994
      10
      10
      2
      0.0022599
      -0.0059795
      0.1812381
      0.60206
      879
      H'
      -0.5643791
      5238.4942
      2090.8877
      s^2
      0.0003812

      The Shannon-Wiener diversity index can be used to compare the distribution of observations among numerous categories. It takes into account both the diversity (types of vegetation in this study) as well as the number of examples of each category (species richness). The diversity value – H’ – is highest with many categories (species) and an even number of examples in each category. In our samples, the Typha area had a diversity index H’ =0.52448 (the absolute value is used), the low vegetation area –H’ = 0.75963, and the Quebrada Mula site was H’= 0.57908. From this index you can see that the low vegetation type has a much larger diversity index, while the other 2 sites (Typha and QM) have almost identical ones.

      With the diversity index, H’, we can only make a general statement. In order to test the null hypothesis that there is no difference in the diversity at any of the sites, we will use a "t" test to measure the variance between the indices.

      First we compared the PV Typha site and the PV low vegetation site. The calculations are shown on the first chart above. PV-Typha versus PV low vegetation – t = 6.08 which indicates significance at p<0.001. PV Typha versus Quebrada Mula – t = 1.63 which does not indicate significance at p>0.1. Therefore, the low vegetation site in the lagoon is significantly more diverse than the Typha site in the lagoon- (Reject null hypothesis that sites are the same). However, the QM site is not significantly more diverse than the PV Typha site- (Accept null hypothesis that sites are the same).
       

    1. Plant biomass – The PV Lagoon low vegetation area had only 36% percent biomass of the Typha area of the lagoon. The QM wetland had 79% of the biomass of the Typha area. This shows that the invasive specie has greatly increased the amount of biomass produced in the PV lagoon compared to the low vegetation area. It also produced more than the QM site, but only a small amount more.
    1. Benthic Core samples - Four core samples were taken from each site. The benthic core samples were very limited and only general observations were made from them. A count of shells and fragments, which were basically the only thing found in the samples showed that the Palo Verde lagoon sites had very little indication of any fauna. Some whole snail shells and numerous shell fragments were found at both sites, with no apparent difference between the 2 sites. The Quebrada Mula site had a few live snails, but otherwise the cores were quite similar to the lagoon sites. One might conclude the conditions near the bottom of all these wetlands are basically anaerobic and not supportive of a rich ecosystem. However, more extensive research would have to be done to support this conclusion.
    1. Microorganism survey - After seeing the results of the benthic core samples, we took water samples to see what protozoans might be in the water. Samples from three levels – top, middle, and bottom - at each site showed some difference, but no statistical studies were done due to the limited samples. We used the DAFOR scale to characterize the number of live organisms in each sample, with D (dominant) > 75% of total volume of sample occupied by live species, A (abundant) < 75% and > 50%, F (frequent) < 50% and >25%, O (occasional) < 25% and >5%, and R (rare) < 5%.

    2. As you can see from the data, all of the lagoon samples showed little to no microorganisms with the bottom of the Typha area showing only occasional protists. The Quebrada Mula site showed more microorganisms but the limited sample site does not allow us to draw any conclusions.
       

    3. Ph and Temperature- Since we did not have equipment to measure dissolved oxygen which would have given us an indication if the reason for the paucity of organisms shows by both the benthic cores and the water samples, we decided to use pH measure. This could theoretically be more basic (pH > 7) if dissolved oxygen were present. Further, significantly different temperatures among the three sites could affect microorganisms.
    Temperatures were virtually identical at all sites, with the surface temperature almost the same as the ambient temperature, and the bottom temperature about 2-3 degrees lower than the surface.

    The pH testing showed that the QM site was slightly acidic (pH = 5.5at all levels) and that the lagoon areas were neutral to slightly basic (pH = 7.0 to 8.5). While this could indicate the presence of dissolved oxygen in the lagoon, it is much more likely that it reflects the water runoff from the limestone cliffs, which border the lagoon.
     

        V. Final discussion The question of biodiversity among the three sites and the effect this has on the health of the ecosystem cannot really be answered without further study. The continual effect of flooding from the Tempisque River, which brings in significant sediment and chemical fertilizers, as well as possibly pesticide and herbicide residue, may be the major factor in the health of the ecosystem. The dominance of Typha in part of the lagoon may be affected by the nutrient rich flooding which allows it to out compete other indigenous flora. The finding that the Typha area is not significantly more diverse than the less impacted Quebrada Mula site should be viewed with skepticism due to the limited and different samples taken in the two areas. Observation of the low vegetation area in the lagoon, as well as similar parts of the Quebrada Mula site, showed that one common bird species, the jacana, prefers the lily pad/water hyacinth environment to the taller type of vegetation.

    (Note: in a further observation of the Typha dominated area, plant diversity was noted along a 150 meter straight line perpendicular from shore. This observation seemed to show that the Typha dominated area continues to have significant biodiversity. The Typha seems to grow in patches interspersed with significant low vegetation. Within the Typha patches most of the species represented in our 2 x 2 meter plot can be observed in similar proportions. In determining the future growth of Typha and its effect on biodiversity, it would be valuable to know how long the Typha has been present in the lagoon. Is the present state an equilibrium which has been reached among the plants presently found in the lagoon, or will Typha become more widely spread and choke out the other species?

    This issue is of importance to virtually any ecosystem impacted by human activities and by the introduction of exotic or non-indigenous vegetation. The development of a generalized model which would take into account all the diverse factors which affect the health of an ecosystem, particularly which are crucial or indicative factors, would be valuable.

    Bibliography – "Aquatic Plants of Palo Verde Park". Crow, Garrett and Rivera, Dora. Uniciencia, p. 71-78, 1986

    Flowering Plants of the World, Oxford Press, 1993

    Woody Plants of Northwest South America, Gentry, Alwyn, Conservation International, 1993

    .
    The Woodrow Wilson National Fellowship Foundation
    CN 5281, Princeton NJ 08543-5281 - Tel:(609)452-7007 - Fax:(609)452-0066
    Technical contact: lpt@woodrow.org