United States
                           Environmental Protection
                           Agency
    Environmental Research
    Laboratory
    Duluth MN 55804
                           Research and Development     EPA/600/M-85/011  May 1985
                           ENVIRONMENTAL
                           RESEARCH   BRIEF
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Experimental A cidification of a Stream Ecosystem

Initial L ong- Term A cidification

The purpose of this research was to investigate the effects
of increased acidity on a stream ecosystem. Qualitative and
quantitative effects of experimental pH  manipulation in
Norris Brook within the Hubbard Brook Experimental Forest,
New  Hampshire, were studied. Field experiments were
designed to test the hypothesis that lowered pH (increased
acidity) has  detrimental effects on microorganisms, peri-
phyton,  macroinvertebrates,  and fish. A  summary of the
experimental research design in Norris Brook from 1977
through 1980 is presented in Table 1.

In 1977 the pH in Norris Brook (see Figure  1) was experi-
mentally decreasedto levelsfound in incident precipitation
(pH 4) by the addition of dilute concentrations of sulfuric
acid. This experimental depression lasted for five months
(see Hall et al., 1980 for further details). Following are the
main  results from this study:

1.  Aluminum, calcium, magnesium, and potassium were
    mobilized into the stream water during acidification.
2.  Emergence of adult mayflies (Ephemeroptera), some
    stoneflies (Plecoptera), and some true flies (Diptera)
    decreased at the lower pH.
3.  More immature aquatic invertebrates in  collector,
    scraper, and predator functional groups drifted out of
    the acidified relative to the control area.
                                                4.  Fewer adult insects assigned to collector functional
                                                   groups were found in  emergence samples from the
                                                   acidified reach.
                                                5.  Chironomids,  tipulids,  certopogonids,  and mayflies
                                                   decreased by 75% in  the benthos of the acidified
                                                   relative to the reference area.
                                                6.  Periphyton biomass increased and fungal (hyphomy-
                                                   cetes) densities decreased at low pH.
                                                7.  A basidiomycete fungus increased in the acidified area
                                                   relative to the reference section.
                                                8.  A loss of nutrients (e.g.,  N) occurred in the invertebrates
                                                   and organic matler but not in the water; this loss was
                                                   significant in terms of the total stream ecosystem.
                                                9.  Stream acidification altered biotic structure, metabo-
                                                   lism,  and  biogeochemistry of  this poorly buffered
                                                   mountain stream ecosystem.

                                                Subsequent Short- Term  Acidic Episodes

                                                In a significant result from the 1977 field work, the largest
                                                amount of invertebrate drift occurred within the first five
                                                days after acid addition (Figure 2), the peak occurring within
                                                two days. Natural pH depressions in streams during spring
                                                snowmelt are often only  a  few  days  in  duration. For
                                                example, episodic input of hydrogen ions  during spring
                                                thaw caused pH of stream water to drop to near 4.0 in first-
                                                to third-order streams and  remained at that level for three
                                                to  seven days in  the Adirondack Mountains, New York
                                                (Schofield,  1977). A  similar phenomenon occurred in
                                                southern Norway (Leivestad and Muniz, 1976). The ramifi-
Table 1.     Summary of Experiments Conducted in Norris Brook Within the Hubbard Brook Experimental Forest from
            1977 Through 1980
Year    Season
                  Stress
         Duration Applied
                                         pH Values:
                                         Reference/
                                         Treatment
                                                          Stream
                                                          Order
                                                                           Chemical
                                                                        Determinations
                                                        Biological
                                                     Determinations
1977
Spring/
A utumn
                5 mo
                                         6-6.4/4.0        3rd       Ca, Mg, K, Na, Al.
                                                                    DOC, POC, etc., SO4~,
                                                                    NH4+, NO3~, Cr, SiOi,
                                                                    Pb, In, Cd, Cu, Mn, Fe
                                                 Invertebrate density in
                                                 drift, emergence, and
                                                 benthos. Standing crop
                                                 of periphyton. Aquatic
                                                 fungi species. Fish
                                                 behavior.
1978
1979
       Spring/
       Autumn
Spring/
Summer
          1 day
          each
                 1 -hr to
                 2-day
                 periods
                          NO STRESS
                          followed
                          recovery
 1980   Spring   1-4 hrs  HCI
        Spring    1-4 hrs  AIC/3
6-6.4/6-6.4
                                         6.4/4.O
                                         6.4/3.0
                                         6.4/5.0
                                         6.4/4.5
                                         6.4/3.5

                                         6.3/4.O
                                         6.3/4.0
                                         6.0/4.0

                                         6.3/5.0-5.25
                                         6.3/5.0-5.25
                                                   3rd
                 3rd
                 3rd
                 3rd
                 3rd
                 3rd

                 3rd
                 2nd
                 1st

                 3rd
                 2nd
                                                                     Same as above
Same as above
plus TP and different
species of AI
                                                                     Same as above
                                                                     Same as above
                      Invertebrate drift and
                      emergence.
Invertebrate and
vertebrate drift.
                                                                                   Invertebrate and
                                                                                   vertebrate drift.
                                                                                   Invertebrate and
                                                                                   vertebrate drift.

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     Norris Brook
          Acid
          Addition
  0  3
           meters
        Legend

   II  Drift nets

  l~~l  Plastic house

  ^  Black plastic house

   4  Direction of flow
        Hubbard Brook
     Experimental Forest
West Thornton, New Hampshire
                                                                                    /  • Weather station
                                                                                   '   A Operating weir
                                                                                       2 Gaged watershed

                                                                                              Mile
                                                                                          0     5    1
Figure 1.     Norris Brook study area located in the Hubbard Brook Experimental Forest, New Hampshire. Reference area (site A) was
             located 5 m above the acidification point. Sampling sites in the treatment area were located at 15 m (site B), 50 m (site CJ, 75
             m (site Dj, and 100 m (site E) below the point of acid addition.

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    Acid addition
  800
  eoo
•Q

I 400]
g 200
Q>
Q
     15
Figure 2.
           20    25
             April
                         30
                                      10
                                     May
                                             15
                                                   20
            Drift of invertebrates in relation to experimental
            acidification of Norris Brook, New Hampshire. Total
            number of organisms collected in drift nets per 24 '-h
            period from 16 April through 17 May 1977. Closed
            circles represent mean for two drift nets; vertical
            bars represent range.
cation  of the results  of the first five days of our 1977
experimental acidification, then,  becomes very clear. The
following questions were formulated after analyses of that
work:

1.   Does episodic input of hydrogen ions during the initial
    phase  of  the snowmelt  period  significantly alter
    community structure and metabolism of poorly buffered
    mountain streams?
2.   What is the effect of short-term pulses of acidity and
    aluminum on  community  structure and metabolism
    and on nutrient cycling and energy flow in streams?
3.   How does the impact of acidity alter nutrient flux during
    high (spring) and low (summer) discharge, as well as
    during storm events?

To answer the above  questions,  additional research was
completed. The results are as follows:

1.   Hall, R. J. and G. E. Likens. 1 980. Ecological effects of
    experimental acidification on a stream ecosystem, pp.
    375-376. In: D. Drablefs and A. Tollan (eds.). Proceed-
    ings of the  International  Conference on  Ecological
    Impact of Acid Precipitation. SNSF Project, Sandefjord,
    Norway, March 11-14, 1980.

    Experimental acidification to pH 4.0 in a small mountain
    stream in the Hubbard Brook Experimental Forest, New
    Hampshire, stressed the biotic and abiotic components
    of  that system. The biotic structure, metabolism, and
    biogeochemistry of this stream ecosystem underlain by
    siliceous bedrock were  quantitatively altered  at  pH
    levels (4.0) commonly found  during snowmelt as well
    as in ambient precipitation.

2.   Hall, R. J. and G. E. Likens. 1 981. Chemical flux in an
    acid-stressed stream. Nature, 292:329-331.

    A third-order section of a small mountain stream in the
    Hubbard Brook Experimental Forest, New Hampshire,
    was experimentally acidified  to pH 4.0 to measure the
    effects of increased acidity on chemical and biological
   export in the stream. The most significant inorganic
   component affected by the experiment was aluminum.
   A significant net flux of organic carbon and nitrogen
   occurred in the biologically bound forms but not in the
   dissolved substances. The net flux of phosphorus was
   significant only in the biologically bound forms. The
   increased loss  of nutrients in the particulate organic
   fraction was also important, particularly if scaled to the
   total stream ecosystem.

3.  Pratt, J. M. and R. J. Hall. 1981. Acute effects of stream
   acidification on the diversity of macroinvertebrate drift.
   pp. 77-95. In: R. Singer (ed.), Effects of Acidic Precipita-
   tion on Benthos. Proceedings of a Symposium on the
   Effects of Acidic  Precipitation on Benthos.  NABS,
   Hamilton, N.Y.

   A third-order reach of Norris Brook, a small stream in
   the Hubbard Brook Experimental Forest, was experi-
   mentally acidified for five months to determine what
   effects acid precipitation may have on the ecology of a
   poorly buffered lotic ecosystem. The initial six days of
   stream acidification simulated a lowpH regimethatcan
   occur in a small mountain stream receiving meltwater
   from a rapidly  thawing snowpack contaminated with
   acidic deposition.

   The first two to three days of acid addition constituted a
   period of acute I-T stress (pH 4) that elicited a tenfold
   increase in the daily drift rate of benthic macroinverte-
   brates. This increased drift leaving the  acidified reach
   was also more diverse overall  in terms of major taxa
   (orders),  trophic functional groups, and  behavioral
   groups but  less diverse at the  generic  level than the
   drift entering. In addition, the macrofauna abandoning
   the acidified  area compared  to that  entering was
   particularly more diverse generically in mayflies and
   midges, collector-gatherers, and clingers and swim-
   mers.

   Ecological implications of the  acute changes in drift
   rate and diversity  extrapolated throughout the low-
   order (1 -3) tributaries of a weakly buffered stream are
   discussed. Recommendations for monitoring the effects
   of acid-sensitive lotic ecosystems are offered.

4.  Hall, R. J., J. M. Pratt, and G. E. Likens. 1 982. Effects of
   experimental acidification on macroinvertebrate drift
   diversity  in a mountain stream. Water, Air, and Soil
   Pollution, 17:1-15(1982).

   A small stream (Norris Brook) within the Hubbard Brook
   Experimental Forest was acidified to determine what
   effect elevated H+ stress may have on the ecology of a
    mountain stream.  The experiment was designed to
   simulate a pH  level (4.0) that can occur during initial
   snowmelt (acute period) and during long-term (chronic
    period) acidification. Daily  macroinvertebrate drift
   samples were  collected from treatment and reference
    areas of Norris Brook. Drift diversity at the generic level
   was calculated using Brillouin's  formula and partitioned
    hierarchically following macroinvertebrate classifica-
   tions based on taxonomy (orders) and feeding strategies
   (functional groups or guilds).
                          4

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    The rate of movement of individuals and genera was
    significantly greater for those organisms leaving the
    acid-stressed area during the first five days than for
    those entering, whereas no  difference  between the
    rate of macroinvertebrates entering or leaving the acid-
    stressed area  was apparent for either numbers or
    genera over the remaining 25-day study period. For the
    acute period (first five days), the increased macro-
    invertebrate drift leaving the acidified area was signifi-
    cantly more diverse at the levels of aquatic  insect
    orders and functional groups but less diverse at the
    generic level than the drift entering. For the chronic
    period (25-day period) no significant differences were
    detected in  major taxa, functional group (with  the
    exception of collectors), or  generic diversity between
    the drift entering and leaving the  treatment  reach.
    Mayflies and chironomids  leaving the acid-stressed
    area during the acute period were generically more
    diverse than those drifting into the area. The overall
    change in  the normal pattern of spatial and temporal
    variation in drift rate and diversity provides quantitative
    evidence that H+ stress significantly altered the struc-
    ture and metabolism of the macrobenthic community.


5.  Hall, R. J.  and G. E. Likens. 1984. Effect of discharge
    rate on biotic and abiotic chemical flux in an acidified
    stream. Can. J. Fish. Aquat. Sci., 41, in press.

    Experimental manipulation of hydrogen ion concentra-
    tion was conducted in a mountain stream ecosystem to
    estimate the effects on biotic and abiotic chemical flux
    during high (spring) and  low (summer)  discharge
    periods. Dilute concentrations  of sulfuric  acid were
    added to Norris Brook, a stream in the Hubbard Brook
    Experimental Forest. The streamwater was maintained
    near pH 4.0 from April to September 1977.

    During acidification, Al, Ca, Mg, and K  were mobilized
    in the streamwater with the  concentrations progres-
    sively increasing downstream. Na and NOa were not
    affected. DOC concentrations decreased downstream
    during high discharge but did not change during low
    flow.  During storms these  elements increased in
    concentration below acid  addition as  well as  in  the
    reference section.

    The net flux of dissolved aluminum in the streamwater
    was significant during both the high and low discharge
    periods. The net flux of aluminum in  invertebrate
    biomass was significant only during high flow but was
    insignificant when compared to the  amount  in  the
    dissolved  state. On  the other hand, the net flux of
    nitrogen was significant only in the biomass of inverte-
    brates at high discharge. This latter amount, however,
    was totally overshadowed by the transport of dissolved
    nitrogen during storms.

6.  Hall, R. J.,  C. T. Driscoll, G. E. Likens, and J. M. Pratt.
    1984. Physical, chemical, and biological consequences
    of episodic aluminum additions to a stream ecosystem.
    Limno. and Oceanogr., in press.

    Experimental addition of AlClato a second-order stream
    was  conducted to simulate episodic increase*  in
    aluminum concentration during acidic snowmelt. With
    astepwise increase in aluminum in the stream water a
    significant decrease in pH and dissolved organic carbon
    (DOC) and an increase  in foam accumulations at the
    stream surface were observed. It was  hypothesized
    that foam production was visible evidence of reduced
    surface tension of the water. Controlled laboratory
    experiments with stream water corroborated the field
    results and  produced  a  20% reduction  in  surface
    tension. Electrophoretic mobility studies showed that
    increased aluminum concentration produced a surface
    charge reversal on suspended colloidal particles. The
    mechanism  for a change in  surface  tension,  it  is
    postulated,  is the binding of aluminum to functional
    groups (e.g.,  a carbonyl group) on DOC, thus rendering
    DOC more hydrophobic and less soluble. Some of these
    organo-aluminum complexes  subsequently accumu-
    late at the air-water  interface.

    Concomitant with physical-chemical changes in stream
    water following elevated aluminum levels were altera-
    tions  in terrestrial and  aquatic invertebrate  drift
    behavior in the field. These changes in drift behavior
    could be attributed to responses either to (1) chemical
    changes in the stream  related to potential aluminum
    and H+ toxicity or (2) a physical change in the surface
    film.


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                                                                       U. S. GOVERNMENT PRINTING OFFICE: 1985/559 111/10830

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