Flora, Vegetation and Bacteria in Relation to Patterns of Land Use StudyC~G~''i2~/ j'o
Property of t,~
1 r~ u h ~ b <~ ~~
rs®9~~~r~..~ ,,.3~~ .
/,'.
f 1
~~~. ~~~
Completion Report 72.4
.The Mill l~.iver and Its Floodplain
in Northampton and Williamsburg, Massachusetts:
A Study of The Vascular Plant Flora,
Vegetation, and The Presence of
.The Bacterial Fa~rnily Pseudomonadaceae
ix~, ILelation to Patterns of Land Use
Elizabeth D. Robinton
C. John Burk
~jATE~_
RESOURCES
RESEARCH
CENTER
UNIVERSITY OF MASSACHUSETTS AT AMHERST
,:.
THE MILL RIVER AND ITS FLOODPLAIN IN
NORTHAMPTON AND WILLIAMSBURG, MASSACHUSETTS:
A STUDY OF THE VASCULAR PLANT FLORA,
VEGETATION, AND THE PRESENCE OF THE BACTERIAL
FAMILY PSEUDOMONADACEAE IN RELATION TO ~ ,
PATTERNS OF LAND USE '
Elizabeth D. Robinton and C. John Burk
Principal Investigators
Smith College, Northampton, Massachusetts.
A cooperative project between Smith College and the
University of Massachusetts Water Resources Research
Center supported under Section 100, Water Resources
Research Act of 196+ (P.L. 88-379)
ABSTRACT
The Mill River, a stream about fifteen miles in length, located in
Northampton and Williamsburg, Massachusetts, has been subjected since the
settlement of the area in 1654 to disturbance by pollution, relocations
of the channel, and impoundment for industrial purposes. The structure of
vascular plant communities in marshes which have developed. behind impound-
ments reflects the effects of human activities. Downstream marshes were
shown to possess lower species richness and diversity and to be character-
ized by greater amounts of unoccupied substratum than upstream, less
disturbed marshes. The invasion of the lower marshes by an introduced
species, Marslea quadrifolia L., during. a period of extreme disruption
from pollution of several. types has been documented. A study. of the
invasion of drained impoundments by vascular plants indicated the.succes-
sional roles of several species .in the stream ecosystem. Leersia oryzoides
(L.) Sw., the only species found in all zones of all marshes studied, was
dominant in the early stages of secondary succession. A summer flora of
more than 275 species of vascular plants within the river and its flood-
plain is included.
Fluorescent strains of the genus Pseudomonas were found at all
sampling stations and may be considered a part of the indigenous microbial
flora of the stream. Pseudomonas aeruginosa (pigmented strains) were widely
,~ distributed. Coliform organisms, lacking at the source, occurred with high
densities in all downstream stations. A method of enrichment for obtaining.
members of the genus Pseudomonas was devised. Future studies of the vege-
tation and microbial flora will indicate the degree to which the stream
-11-
ecosystem responds to pollution abatement measures applied since the
4 field observations of this project.
key words:
pollution
marsh vegetation
land use
diversity
succession
Pseudomonas
coliformes
:~
-iii-
~~
ACKNOWLEDGEMENTS
The work upon which this report is based was supported in
large part by funds provided through the University of Massachusetts
Water Resources Research Center by the United States Department of
the Interior, Office of Water Resources Research, as authorized under
the Water Resources Research Act of 1g6~+.
The study could not have been conducted without the work, in
field and laboratory, of the following graduate and undergraduate
students: Wendy Brown, Lois,Davis, Paul Hosier, Arthur Lawry, Mary
Lincoln, Nick Lenz, Alexander Mesrobian; Deborah Reichert, Kenneth
Sherk, and Erica Swenson. We hope that their gain from participation
in the project was as great as our appreciation of their efforts. A
Sloan Foundation grant to Smith College supported essential.historical
research carried out by Darilyn Wilds.
We also wish to express our gratitude to Mr. Roble Hubley,
Director of Arcadia Wildlife Sanctuary, and to the staff of Forbes
Library, Northampton, for their co-operation, as well as to the staff
of Clark Science Center office for assistance with the preparation of
this manuscr_pt. The final versions of the figures were drawn by
Arthur Lawry.
- 1'V -
PREFACE
The scope of this .project required techniques of historical
research, plant taxonomy, ecology, and microbiology. It seemed wisest
to prepare the various chapters of this report in the separate formats
preferred by these disciplines. Chapters I and II were written by us
jointly; Chapters IIL, IV, and V were prepared by C. J. Burk and
Chapter VI by E. D. Robinton. We hope that segments of the report will
be of interest to specialists within our fields and that an overall con-
cept of the river as a system of inter-related physical, biotic, and
historical factors will emerge for the reader who peruses the entire
manuscript.
-v-
TABLE OF CONTENTS
Page
ABSTRACT i
~:
ACKNOWLEDGEMENTS
iii
PREFACE iv
LIST OF FIGURES vi
LIST OF TABLES vii
CHAPTER I
Objectives.. ... 1
Nature. and Scope of the Project 1
Degree of Achievement of Objectives 3
Literature Cited 5
CHAPTER II: THE RIVER AND ITS HISTORY 6'
CHAPTER III: THE EFFECTS OF DISTURBANCE ON THE STRUCTURE OF.
MARSH COMMUNITIES - -
Introduction 19
Study Sites 21
Methods and Results 23 -
Analysis and Discussion . 24
Observations of an Invading Species 35
Chemical Analyses 36
Conclusions and Summary 37
Literature Cited 39
CHAPTER IV: INVASION OF DRAINED IMPOUNDMENTS BY VASCULAR
PLANT SPECIES
Introduction 40
Methods .. 40
Results 42
Discussion and Conclusions 44
Literature Cited 47
CHAPTER V: A SUMMER FLORA OF THE MILL RIVER AND ITS FLOODPLAIN. 48
CHAPTER VI: .ISOLATION OF PSEUDOMONAD STRAINS FROM THE MILL .RIVER ~.
Introduction
. 57
Methods
Materials and 58
Results and .Discussion bO
Literature Cited 72
-V 1-
` LIST OF FIGURES
Page
1. The Mill River in Northampton prior to 1720 9
2. The First Diversion of the Mill River in Northampton: 1710-1723 10
3. The Second Diversion of the Mill River in Northampton: 1939 11
~_
-vii-
LIST OF TABLES
Page
1. Percent cover (C) and frequency (F) of species sampled in
' quadrats in high,. mid, and low zones of four Mill River
study sites 25
2. Richness, diversity,. and evenness of high, mid, and low-marsh
zones of four Mill River study sites 31
3. Total richness and mean diversity of four Mill River study sites,
computed from data obtained separately from high, mid, and low
marsh zones 31
4. Overlap of species between zones at each study site (species
shared by two zones/total number of species occurring in both
zones x 100) 34
5.~ Percentage of total substratum unoccupied by vascular plants in
quadrats of high, mid, and low zones of Mill River study sites 34
6. Frequency and density of species encountered in pioneer and
later seral stages on drained impoundments 43
1' Sampling points along the Mill River .. 62
2' Samples showing fluorescent pseudomonads 64
3' Total number of samples showing fluorescent pseudomonads 65
4' Individual samples showing Pseudomonas Pseudomonas (green- 67
pigmented strain)
5' Total number of samples yielding Pseudomonas aeruginosa 68
_1_
CHAPTER I
OBJECTIVES
The objectives of this project were to study the Mill River, a
stream which flows through five communities- (Northampton, Florence, Leeds,
Haydenville, and Williamsburg., Massachusetts) and. to determine the relation-
ship of land use in adjacent areas to the floras of vascular plants and
selected microorganisms growing within the river or its floodplain:
1. To inventory all species of vascular plants growing
within the river and its floodplain.
2. To determine the presence and abundance of each plant
species within segments of the river and its floodplain
adjacent to various patterns of land use.
3. To make quantitative ecological studies of the vascular
plant species within each land-use-related segment of
the river and its floodplain.
4. To determine the microbial flora of the river at selected
sites with regard to the presence of members of the family
Pseudomonadaceae.
NATURE AND.SCOPE OF THE PROJECT
The Mill River, located in the towns of Northampton and Williamsburg,
Massachusetts, is a relatively .short stream which passes through five
highly settled areas. The land bordering the river and its floodplain is
used for a number of purposes including industrial development, housing
of a variety of sorts, a city park, a college campus, pasture, cropland,,
-z-
and a Massachusetts Audubon wildlife sanctuary which may in time .become °
part of the Connecticut Valley National Recreation Area. At some points
along its course, the river is impounded by dams; at some points, the
channel has been straightened; in some segments the flow is essentially
unimpeded.
No comprehensive study of the vegetation of the river and its flood-
plain has been previously conducted, although it is known that the spread
from cultivation and establishment in disturbed areas below Smith College
of one aquatic vascular plant species, Marsilea quadrifolia, is somewhat
unique in the United States (Burk, 1966). The course of the river and the
condition of its floodplain have been extensively changed by Man's activi-
ties in the past, and it is likely that, in the future, they will be
manipulated even more. Among possible or likely sources of restructuring
of the environment at the time the project was initiated were the following:
(1) the abandonment of dams originally built for industrial
purposes. Most of these have been destroyed although a
few remain.
(2) the relocation of the Northampton City Dump which presently
abuts on the marsh at Arcadia Wildlife Sanctuary.
(3) the incorporation of at least a portion of the river and
its floodplain into a "greenbelt" within the city of
Northampton.
(4) the damming of the river at its entry into the Oxbow of the
Connecticut River in order to control the level of water in
Arcadia Wildlife Sanctuary.
(5) the enforcement of pollution-abatement measures.
-3-
DEGREE OF ACHIEVEMENT OF OBJECTIVES
' As the research for this project progressed, certain aspects of
the study appeared to yield particularly significant or interesting data.
A recent history of man`s relation to the river was prepared, partially
through funds provided by a Sloan Foundation grant to Smith College.
The vegetation in marshes developed behind impoundments along the stream,
proved most responsive to land-use (and hence water quality) within the
watershed ,and extensive, quantitative, ecological studies were conducted
in these marshes. The draining of one impoundment provided a unique
opportunity to observe the earliest stages of secondary .succession in
the floodplain, while the presence of older, partially drained impound-
ment beds allowed these stages to be contrasted with later portions of
the sere, No direct relationship of the presence of any given species
(with the exception of Marsilea quadrifolia, to be discussed in Chapter
IV) with overall patterns of land-use. within large segments of the stream
or floodplain were noted; a summer flora was, however, prepared and the
general relative abundance of each species within the river ecosystem
noted. A wide range of species within the Pseudomonadaceae was encoun-
tered in the stream. Fluorescent strains of the genus Pseudomonas were
found at all sampling stations while pigmented strains of Pseudomonas
aeru~inosa were widely distributed; a method of enrichment for obtaining
~'' members of this genus was, devised.
Although the use of larger aquatic vegetation as an indicator of
general biological productivity must be employed with caution (Welch,
1952), general observations prior to the initiation of this study
-4-
suggested that these forms might well be used to indicate environmental
deterioration or improvement. The family Pseudomonadaceae is comprised
of a heterogenous group of genera found in water, soil, and on or in
plants and animals; classified in the order Pseudomonadales, some of
its genera are closely related to Eubacteriales (Bergey). The pseudo-
monads possess a high degree of nutritional versatility and could be
expected to be present under numerous and diverse conditions. The
distribution and ecological relationships of both vascular plants and
micro-organisms in the river and its floodplain hence were expected to
reflect water quality in any given segment of the stream, providing
information useful in planning for proper use of the watershed and in
interpreting environmental changes which might occur there.
-5-
LITERATURE CITED
Burk, C. J. 1966. Marsilea quadrifolia L. in Western Massachusetts.
Amer. Fern. Jour. 53: 140-41.
Welch, P. S. 1952: Limnology, Ed. 2. McGraw-Hi11, New York
-6-
CHAPTER II
THE RIVER AND ITS HISTORY
The Mill River in Northampton and Williamsburg, Massachusetts, is a
stream about fifteen miles in length which arises from the convergence of
two branches, the West Branch originating in the western uplands of Goshen
Township, and the East Branch, formed by a converging of brooks in Williams-
burg. The two branches unite in the town of Williamsburg, about seven miles
above Northampton; the stream then follows a south-southeasterly course
through Haydenville, Leeds, Florence, and Northampton to a confluence with
the Connecticut River Oxbow. The Mi11 River drains about 37,760 acres (59
square miles). While the greater portion of this watershed is situated in
the eastern. part of the .Berkshire Hills i.n the New England upland, the
lowermost portion of the drainage area lies within the Connecticut Valley
lowland. Elevations range from 1700 feet to about 100 feet above mean sea
level. At present, 74% of the watershed is forested, 9% is cropland, 5%
is pasture, 7% is urban, and 5% is used for miscellaneous purposes. The
greater part of the stream bed runs through urbanized areas.
The first settlers arrived in the vicinity of Northampton in 1654; an
understanding of subsequent human activities is essential to an understanding
of the present river ecosystem. These activities, while inter-related and
closely tied to the slow but relatively steady growth of population along
the stream, may be categorized as follows: changing farming practices,
relocations of the channel for flood control, the construction of mill dams
and the subsequent abandonment of many of these structures, and the release
of various pollutants into the stream flow. Each of these factors will be
separately discussed.
-7-
changing farming practices - The earliest settlers were attracted to the
region by the flat, almost treeless meadowlands along the floodplain of the
Connecticut, and their first habitations were at the foot of a hill at the
border of the meadows and the forest, close to the Mill River, then a small
but fast-flowing stream. During the pioneer period, a self-sufficient
economy was maintained,principally through the abundance of fertile virgin
soil. Corn, wheat, and hay were the most important crops; apples, rye,
kidney beans, and barley were also grown. The success of corn growing and
the abundance of timber stimulated the building of gristmills and sawmills;
in time hemp and flax were raised and cloth mills and presses .for linseed
oil were established. Livestock included cattle, pigs, horses, and sheep;
wool from the latter was first processed locally at a fulling mill con-
structed in 1702. ,An abortive silk industry during the period 1836-1840
resulted in the planting of 100 acres or more along the banks of the river
with white mulberries, the leaves of which were intended as food for the
silkworms.
As industrialization increased along the river, agriculture became
less important, the greater part of the raw materials used in the mills being
imported. By the end of the 18th century, moreover, the soil was exhausted,
crop rotation being unknown at that time, and planting failed repeatedly. The
cultivation of wheat was largely abandoned and the worn-out fields were seeded
with broom corn, oats, grasses for hay, and tobacco. In 1845, according to
'j official reports prepared for the Secretary of the Commonwealth of Massachu-
setts, corn, hay, and fruit were the principal crops., followed by oats, rye;
wheat, and barley in decreasing order of importance. Nearly twenty thousand
pounds of tobacco were also produced yearly during this period.
-8-
Much. of the town of Williamsburg originally was low .swampy ground on
which grew an abundance of hemlock and alder bushes. The first resident of
the area, arriving in 1735, subsequently built a cider mill along the river.
General settlement of the area occurred about 1780; the land, after drainage,
supported chiefly pasturage and fruit trees.
At present, agriculture in the Mill River watershed is declining
although the average farm size, about 116 acres in 1964, has increased 10%
since 1959. Approximately 55 farms are operative with dairying the most
important enterprise. One farm is used for growing shade tobacco, and it
has been predicted that commercial agriculture along the stream will have
been abandoned by about 1980 (Preliminary Investigation of the Mill River
Watershed, May, 1971).
relocation of the channel - The early settlers soon learned that the Mill
River tended to flood adjacent grounds, particularly the meadows, yearly
during spring. In 1699 (see Figure 1) embankments were raised to control
the course of the lower portion of the river, to "Stop the mouths of the
gutters that carry the water... into the greate Swamp and so into the
meadows whereby much dammag is done Both to the highways And in the meadow"
(in Trumbull, 1898). This procedure proved ineffective, and during the early
years of the 18th Century (see Figure 2), the colonists, in an elaborate feat
of engineering for that time, diverted the stream across the meadows directly,
shortening and straightening the river's course while turning it into the
Connecticut north of the Oxbow through the tract of land known as the Great
Swamp.
Although the river was to follow this diversion for more than 200 years,
flooding was a frequent problem as the river attempted to shift its course
-9-
CONNECTICUT RIVE R
Mill River
Manhan
River
k'igure 1
The Mi.11 River in Northampton prior to 1720
(after Cestre, 1963)
-10-
Figure 2
The kirst Diversion of the Mill River in Northampton
1710-1723 (after Cestre, 1963)
CONNECTICUT RIVFR
-11-
Mill Rive'
Manhan
River
?figure 3
'1'he Second lliversion of-the Mill River in Northampton
1939 (after Cestre, ].963)
-12-
back to its old bed. A dike was built in 1856 to prevent water from in-
updating the lower portions of the city during high spring floods. This
dike was strengthened and heightened later but, in 1936, during a period of
extremely high water, it proved ineffective and, when the floods receded,
was deliberately broached to avoid insanitary conditions resulting from
stagnation of water trapped behind it. The hurricane of September 21, 1938,
also caused extensive damage within the floodplain, and in February of 1939,
a federally-funded flood control and diversion project was begun (see
Figure 3). The diversion channel built in the early 1700's was closed off
by a reinforced levee and a new diversion channel was created, taking the
river into the Oxbow of the Connecticut along a route rather similar to that
it had followed at the time of the first settlements.
construction of mill dams - The first mill on the banks of the Mill River
was a small gristmill constructed in 1658; the river was dammed to provide
a source of water for this enterprise. During the next two centuries, in-
dustrialization along the stream increased until, by 1850, there were 74
establishments there, all dependent on water power. Statistics prepared
five years earlier for the Secretary of the Commonwealth (in Wright, 1949),
indicate that Northampton alone supported "one woolen mill, two paper mills,
three silk manufacturing plants, and manufactures of hollow-ware, iron cast-
ings, jewelry, gold and silverware, saddlery, hats, caps, coaches and other
carriages, soap, candles, chairs, tin and cabinet-wares, leather, boots,
shoes, straw hats, bricks, books, corn .... broad-cloth and cassimere."
The use of steam for power was introduced in the late 1850's; nonethe-
less, in 1865, a group of industrialists formed the Williamsburg and Mill
River Reservoir Company for the purpose of constructing a new, large dam to
-13-
provide a reserve supply of water for the mills and factories already present
along the river and to attract new industry to the area. This dam, constructed
at the head of a narrow gorge above the town of Williamsburg, maintained a
reservoir which covered 124 acres to an average depth of 24 feet. In May of
1874, the structure collapsed, loosing a wall of water which destroyed or
damaged a number of the downstream enterprises. The transition from the use
of water power to the use of steam was accelerated by this incident, and the
total industrialization of the river began a decline which has continued to
the present.
By 1935, steam and electricity had largely supplanted water, power,
although water was still used as a supplementary source of .energy. At
present, no industries depend on the river as a power source. Of_the Mill _
River dams, several remain; one on the Smith College campus and a second near
the ~'rophylactic Brush Company are maintained to provide impoundments used
for recreation. Other dams situated in the villages of Leeds and Haydenville
are in various states of disrepair. One of these, the "Starbuck" dam, lost
its capstones in 1969 and gave way almost completely the following year.
Once-trapped sediments loosed by the failure of these dams carry downstream
and cause damages of various sorts along the river and its floodplain.
release of pollutants - While the rise of industry and the diversions of
the channel of the stream are fully chronicled, the history of pollution is
rather hard to document. Without question, the Mill River is at present
,>
subject to pollution from a number of sources. .The bulk of Northampton's
sewage entered the river until shortly after the beginning of the 20th
century and a main sewer line, which on occasion has broken, discharging
quantities. of raw sewage into the stream, still runs along the river bed.
-14-
At the time this study was initiated, raw sewage entered the river directly
from the town of Williamsburg, a situation to be remedied in the near
future.
During the period of heavy industrialization, Mill River water was
kept in good condition for paper manufacture, and all leases of water
rights in the neighborhood of paper mills contained clauses protecting
these industries against pollution of the stream. Nonetheless the various
industries still in operation discharge or have discharged chemicals,
including heavy metals and synthetic organic compounds, in varying amounts
and concentrations; it is difficult to believe these discharges were not
greater in the past.
Nutrients enter the stream, principally with runoff from dairy farms
and agricultural properties along Roberts Meadow Brook, a main tributary,
and other chemicals enter with runoff from streets which have been salted
and sanded during winter months.
Particularly devastating in their effects on the river ecosystem
are accidental incidents of pollution which may often never be recorded.
In 1963, as one example of these incidents, oil originating from the
Northampton City dump (which verged, at that time, on the stream above
Arcadia Wildlife Sanctuary near the convergence of Mill River with the
Oxbow) resulted in the destruction of much marsh vegetation. During the
period of the present study, similar episodes occurred. During the spring
of 1969, several thousand gallons of oil burner sludge, emptied on afield
below the State Mental Hospital, entered the river north of Paradise Pond.
This oil, in conjunction with sediments released by failing dams, necessitated
the drainage and deepening of Paradise Pond that summer. A second spillage
-15-
occurred within a year when a thousand gallons of fuel oil, channeled '
into the wrong inlet at Northampton High school, was then released into
the stream.
In conclusion, the stream ecosystem has been, for nearly three
centuries, highly disturbed by man's activities. It is not surprising
that the communities of organisms found within the river and its flood-
plain reflect the effects of these disruptions. The following partially
annotated bibliography contains the source of the information in this
chapter, as well as other data of interest to the total study.
-16-
BIBLIOGRAPHY
A Fu11 and Graphic Account of the Terrible Mill River Disaster in
Hampshire County, Massachusetts. (Contents acknowledged by the
author as having been borrowed from the Springfield Republican.)
Forbes Library, Call number F844Wi3.F.
Cestre, Gilbert. 1954. Northampton's Growth. (Typewritten, 13 pages).
Forbes Library, Call no. F844N+C338n.
1963. Northampton, Mass.: Evolution Urbaine. Paris:
Societe d'Ed~.tion d'Enseignement Superieur. Forbes Library, Reference
Room and Circulation.
Dean, Charles J. 1935. The Mills of Mi11 River. (Unedited work, 160 pages).
Northampton, Mass. Forbes Library, Kingsley Room Collection, Call
no. HE844N+D34.
Deming, Phyllis Baker. 1946. History of Williamsburg in Massachusetts.
Northampton: The Hampshire Bookshop. Available at Forbes Library,
Reference Room and Circulation.
Early Northampton. Northampton, Mass.: Betty Allen Chapter of the Daughters
of the American Revolution, 1914. Available at Forbes Library.
Everts, L. H. 1879. History of the Connecticut Valley. (2 vols.)
Philadelphia: J. B. Lippincott. Forbes Library.
Flooded Area in and around Northampton, March 19-22, 1936. (Map; statistics
on 1927 and 1936 floods). Compiled by Sea Scouts of Ship no. 113,
Boy Scouts of America. Forbes Library.
Hannay, Agnes. 1935-36. Chronology of Industry on the Mill River. Smith
College Studies in History, vol. 21. Northampton, Mass. Available
at Forbes and Neilson (Smith College) Libraries.
Hifny, Mohamed and Niildred Danklefsen. 1948. i~~.storical Geography of
Northampton, Massachusetts. (A Special Project Report). Worcester,
Mass.: Graduate School of Geography, Clark University. Forbes Library,
Kingsley Room Collection, Ca11 no. G844N+H533h. (Includes information
about the physical geography of Northampton, advantages and disadvan-
tages of location, etc. Summarizes briefly some information, about
the geology of Northampton and surrounding areas, which may be found
in much more detail as The Flow of Time in the Connecticut Valley.
This latter work, by Bain and Meyerhoff, is available at the Clark
Science Center Library, Smith College.)
Historical Localities of Northampton. Northampton, Mass.: Gazette Printing
Company, 1904. (Published in celebration of the 250th anniversary of
the toc~m's settlement.) Forbes Library, Reference Room and Circulation.
-17-
Hampshire History: Three Hundred Years of Hampshire County, Mass.
" Tercentenary Editorial Committee, eds. Northampton, Mass.: Hampshire
County Commissioners, 1964. Forbes Library, Ca11 no. F844H17+W64h.
Index to Local News in the Hampshire Gazette; 1786-1937. Boston: The
_-_
Historical Records Survey, Works Progress Administration, 939. Forbes
Library, Reference Room.
Mill River Flood, 1874. (Scrapbook of articles from the New York Daily Graphic,
May 20, 1874, and Frank Leslie's Illustrated Newspaper, May 30, 1874.)
Forbes Library, Kingsley Room Collection, Call no. F844Wi3//M6.
Mill River k'1ood, 1874. .(Copies of N.Y. Daily Graphic, May 18-20, 1874.)
Forbes Library, Kingsley Room, Ca11 no. F844Wi3//D.
Northampton Dike. (Scrapbook of Hampshire Gazette clippings, 1927-1931, on
the Maple Street-Fruit Street Dike. Effects of floods, moves to improve
structure of dike, etc.) Forbes Library, Kingsley Room, Call no. F847N+F74d.
Northampton of Today. 'Charles .F. Warner, editor. Northampton, Mass.:
Picturesque Publishing Company, 1902. (Includes information about prob-
lems of sewage disposal in the town, controversy involving local govern-
ment and the State Board of Public Health regarding pollution of Mill
River water, etc.) Forbes Library; Call no: G844N+9N+2.
Picturesque Hampshire: Su lement to the Quarter-Centennial Journal.
Charles F. Warner, ed. Northampton: Wade, Warner and Co:, 1890.
Forbes Library.
P'roprietors' Records, Town of Northampton, Mass., 1653-1680. .City Clerk's
Office, City Hall, Northampton..
Report of Preliminary Investigation of the Mill River Watershed by the United
r States Department of Agriculture Soil Conservation Service, Amherst,
Massachusetts. May, 1971. Published under authority of the Flood
Prevention and Watershed Protection Act, Public Law 566.
Trumbull, J. R. 1898 and 1902.. The History of Northampton. (2 vols.)
Northampton, Mass. Available at Forbes Library, Reference Room and
Circulation.
Wegmann, Edward. 1901. The Design and Construction of Dams. (4th edition,
enlarged and revised.) New York: John Wiley and Sons. Forbes Library,
Call no. SDID//W42.
Western Massachusetts: A History, 1636-1925. (J. H. Lockwood et al., editors).
New York: Lewis Historical Publishing Company, Inc., 1926. Forbes
Library, Reference Room and Circulation.
Williamsburg and Mill River Disaster. (Collection of clippings from the
Springfield Union and the Hampshire Gazette, dated 1921-1934. Forbes
Library, Kingsley Room.
-18-
Wright, Harry Andrew. 1849. The Story of Western Massachusetts. New
York: Lewis Historical Publishing Company, Inc. (Includes statistics,
compiled in 1845, which give a comparative view of industry in towns
of four Western Mass. counties, including Hampshire.)
Wright, W. H. 1886. Scrapbook of Industries on Mill River with Article
from the Gazette and Courier on the Burning of riaynard's Hoe Shop.
Forbes Library,. Kingsley Room Collection, Call no. HE844N.W93.
Wright, William Henry. 1938. "Industries on Mill River...". Article from
the .January 2, 1938 issue of the Springfield Republican. (History of
industrial development which follows very closely the work of Hannay
already cited.) Forbes Library, Kingsley Room. HE844N.W93 (combined
with Wright, 1886).
-19-
CHAPTER III
THE EFFECTS OF DISTURBANCE ON THE STRUCTURE OF MARSH COMMUNITIES
INTRODUCTION
Several sites on the Mill River are particularly suitable for. quanti-
tative ecological studies of the effects of disturbance on communities of
vascular plants in marshes. The diversity of Marsh vegetation in relation
to pollution and other disruptions of the marsh environment is a topic on
which surprisingly little research has been done. Iri 1908, Kolkwitz and
Marsson (in Hynes ,. 1963) described a polluted German river in which three
zones were maintained by a sewage outfall: a polysaprobic zone in which
no vascular plants were present, a mesosaprobc zone where some rooted
aquatics survived, and an oligosaprobic zone which supported a wide range
of plant life. In England,.-the Royal Commission on Sewage Disposal reported
in 1913 on the abundance of "water weeds" in waters ranging from "very clean"
to "bad". At best this report indicated only a vague relationship between
clean water and plant diversity .(Klein, 1966). Butcher (summarized with
additional observations in Hynes, 1963) categorized English vascular plants
according to their tolerance to siltation, defining five principal plant
communities as follows: (1) torrential on rock or heavy shingle, character-
ized by river mosses, (2) non-silted on shingle, characterized by Ranunculus
fluitans, Myriophyllum spicatum, and Berula in harder water,. (3) partly
silted on gravel and sand, with Ranunculus .fluitans accompanied by various
species of Potamo~eton, (4) silted on sand, with Callitriche, Potamogeton sp.
and Elodea canadensis, and (5) littoral, with a varied group of species
-20-
including Glyceria aquatica and Spar anium erectum.
In North America, within an area floristically similar to that of the
present study, several researchers have noted deteriorative changes in marsh °
vegetation which may be attributed to disturbances of various kinds. On
the Oswegatchie and Black River systems in New York State, Farrell (1931)
found no vascular plant species growing in the septic zone of Black River
Bay. Myriophyllum exalbescens, Potamogeton gramineus var. graminifolius,
Najas flexilis, and Potamogeton epihydrus were growing in the zone of
recovery, and Potamogeton pectinatus, P. crispus, Radicula aquatica, and
Myriophyllum heterophylum, excluded from the zone of recovery, were abundant
in nearby unpolluted waters. On the upper Hudson watershed (Muenscher, 1932),
only Potamogeton epihydrus and Eleocharis acicularis were found in areas
heavily polluted by pulp, sewage, shavings, chemicals, cinders, oil, grease,
and other wastes, while near bake Ontario, pollution was considered (Clausen,
1940) to be the major cause of a depauperate flora in certain bays.
The effects of a n~imber of specific environmental factors on aquatic
vascular plants are discussed in Hynes (1963) and elsewhere. Several examples
may be cited. Drainage from lead and zinc mines tends to destroy all higher
plant growth. Oil prevents diffusion of gases into or out of leaves. The
toxicity of most pollutants doubles with every 10°C increase in temperature,
halving the survival times of plants growing in the heated waters. As oxygen
concentration lowers, the toxicities of cyanides, cresols, alkyl aryl sulpho-
nate detergents, and ammonia increase. even extremely small amounts of
detergents inhibit oxygen diffusion through. the surface of water, and Roberts
(in Yapp, 1959) has shown that 2.5 ppm alkyl benzene sulphonate detergent is
-21-
'fatal to Ranunculus aquatilis. In zones of gross pollution, rooted plants
are often 'smothered although Potamo eton pectinatus, the setaceous leaves
of which shed depositing silt, may frequently survive and thrive. Organic
effluents have been shown to be toxic to Callitriche sp. and Potamogeton
densus while the emergent Glyceria maxima is adversely affected by the
accumulation near its roots of heavy metal ions released with sewage. Hence,
as Wilhm and Dorris (1968) conclude, chemical substances affecting water
quality are numerous, are effective in a great range of concentrations, and,
within the stream ecosystem, vary continuously and erratically in concentra-
tion. Because of the large number of potentially toxic materials, of biotic
species which may be affected, of interactions among compounds, and of varia-.
tions in physical and chemical conditions, diversity indexes derived from
information theory and applied to biological data gathered within the stream
are often the most useful criteria of water quality. These indexes, as well
as more traditional methods of analysis, have been utilized in the present
study.
STUDY SITES
A reconnaissance of the Mill River was conducted in June of 1969 to
determine areas in which marsh vegetation had developed at the edges of
impoundments. Of the four such areas present along the stream, one, on
Paradise Pond on the Smith College campus, was unsuitable for investigation
because the pond was drained and scraped during the study period to remove
sediments and oil deposits which had accumulated in large quantities the
preceding spring (see historical .section of this report). Four sites were
chosen from the remaining three areas as follows:
-22-
1. Haydenville site: a marsh developed in an impoundment identified `
as "Brass Mill Pond'' on the Williamsburg Quadrangle of the 1964 U. S. Dept.
of the Interior Geological Survey topographic map. The Brass Mill Pond is
the first impoundment imposed on the river from its source and the major
contaminant of the stream to this point is raw sewage released by the town
of Williamsburg. The maximum width of the impoundment, as measured from the
topographic map, is about 25 meters.
2. Leeds site: a marsh developed at an area identified as "Leeds
Substation" on the Easthampton Quadrangle of the 1964 USDI Geological Survey
topographic map. Between this site and the Haydenville site, the river passes
over several dams, some faulty, others in good condition. A few industries
currently operate above this point; Roberts Meadow Brook, a major tributary
stream of the Mill River, also enters between this site and the Haydenville
site. The maximum width of the impoundment, as measured from the topographic
map, is about 50 meters.
3. Duck Pond site: a marsh within Arcadia Wildlife Sanctuary, north-
west of the impoundment identified as "Hulbert's Pond" on the Easthampton
Quadrangle of the 1964 USDI Geological Survey topographic map. The history
of the sanctuary area i5 complex, the river having flowed through the marsh
until the early 18th Century and from 1939 to the present. The water level
at this and the following site is maintained in large part by the Holyoke
Water Power Company dam on the Connecticut River downstream; the Holyoke
dam, hence, impounds the lower portion of the Mi11 River as well as the -
Oxbow of the Connecticut and portions of the Connecticut. During the period
of this study, water level in the Arcadia Wildlife Sanctuary marshes fluctu-
ated markedly as the Holyoke Dam was raised or lowered. The maximum width
-23-
of the Duck Pond impoundment is 100 meters, as measured from the topographic
map. The Duck Pond referred to as the "Wood-duck Pond" by the wildlife
sanctuary staff) is separated from Hulbert's Pond by the embankment of an
old trolley line.
4. Arcadia marsh site:. the largest marsh within the wildlife sanctuary
marsh complex, identified as "Hulbert's Pond" on the Easthampton Quadrangle
of the 1964 USDI Geological Survey topographic map. The history of this site
is summarized above. The maximum width of the impoundment is approximately
225 meters, measured from the topographic map.
METHODS AND RESULTS
In three of the four sites studied.(Haydenville, Leeds, and Arcadia
marsh) three distinct zones of vegetation were evident, high marsh, in which
the soil was moist but where no standing water was present, mid-marsh, the
zone of emergent vegetation with water standing among plant stems, and low
marsh, the zone of predominantly floating or submersed vegetation.
A line transect was run across each site from high marsh on one bank
to high marsh on an opposite side in such a way that low marsh and two bands
of mid marsh were intersected. Although vegetation touching or crossing any.
segment of the line was recorded, .these data were considered less useful than
data derived from quadrats and are hence not included here. In each of the
~• three main zones in each of the three marshes, ten .25m2 quadrats were evenly
spaced on a 40 foot base line perpendicular to the transect line.
At the Duck Pond site, high marsh and low marsh were present and were
sampled as in the other marshes. A zone of open, still water was lacking on
-24-
this transect, a wet mud bar dominated by Pontederia cordata being situated
here between the high and low marsh areas on either side. This area was
sampled with ten .25m2 quadrats and this data substituted for low marsh data
from this site.
Percentage cover was recorded for each species of vascular plant in
each quadrat. In some quadrats, total cover surpassed 100% because of the
layering of the vegetation. Mean percent cover and frequency of each species
within each group of quadrats was determined and is summarized in Table 1.
ANALYSIS AND DISCUSSION
Various criteria have been used to evaluate the structure of communi-
ties. Richness, diversity, and evenness were computed in the manner described
below. Succession and the amount of unoccupied substratum are separately
discussed.
richnesst Richness, the total number of species recorded in each zone of
each marsh is included in Table 2. Table 3 contains the total number of
species occurring in all zones of each marsh. Since many species in each
marsh occurred in more than one zone within that marsh, the total richness
of each marsh is less than the sum of the high, mid, and low marsh figures.
The high marsh zone possessed the greatest richness in every marsh except
the Duck Pond site where high and mid-marsh were equal in richness. The
low marsh showed least richness in every instance. The total richness was J
greatest at the Leeds site, which possessed the greatest richness for each
respective zone, and least at the Duck Pond site. Generally, the two
upstream marshes possessed a markedly higher richness than the two downstream
marshes.
-25-
3
a~ o
~ ~
• ~ `f:
o o
N ~ O r-I ~t M
,.d
~ ~ ~
x ~ O O O
,~ rl r-I O r--I O r-I
~ 3 ~
+~ o • o
cd .-1 O r--I
O
r-1O
tf'1
O
N
O
M
O
~ O
~ O
O' •rl 41 ~ V rl O rl r1 ~' O N O '-1 rl rl
[A N ,
a
`O 0 0 ~ ~ ° "~ OO
b ~ ~ ~o~--io •o 00 .0 ~0 0 0 •o o •o
N {.1 ,L O ri V r-I O N r1 N ri M r--I ~-I V ri I~ r-I O M r1 r-i rl *--~ O M
~ ~
N O
to r-i
~ ~
~
~ O 'C)
. p.~ .
U ~ ~
N •ri ..~
~
~ ~ ~
~+ A b0 ~ O
0 ~n o0
0 .~.
r-I 4-I
u, a~ w w 3
N rl ~/ O O
i .a ~
ni ~ ~
H N O
•i-I
~ N b ~
C7' Cd
~
~
4-1 r-1 ~ b0 M O
'
''d b ~" rl ~ 00
~
G ~+
ro ro
,-.
~~
~
~w
~w
~w
~w
~w
~w
c~w
~w
~w
~w
~w ~w ~w vw w
a~
~~
o cn
U •rl
~ ~
~i ~'+ ~ ~ ~ r-i
N •ri cd
U ~
~
N ~
~
y
P-+
'~ ~F-I ~ ~ ~U
'o ~ ro .,~
~
~ ,-I
0 N U
~ y
a 4-I ~ O
''-~ a a o ~ ~
~ ~ .~ ~ .a p .
N v ~ cn m o cv a ,a •~ +~
cd ..a v ~ m a cd ~-i f1. n. m ~ u ~ ~
,~ ~ cd •rl m •rl
p. N m +~ cd rn ~ ~ r%I ~ G ~
~ ?C N
~ ~+ ~+ o m ~ ~ ~ ~
~ ~
~ a~ ~
~ a a~
m .[
a
v v v ~ ~ ~ ~ ~ a
i a
i ~ c
v ~ i
a
~ ~ ~ ~ ~ ~ ~ as as as Pa v ~ ~ v
-26-
oVNO oMOO ~o o.
~ ~
•~ v ~o `~'
o ~o
•~oo
•o o~oriooo ~ ~
' O
O
O ~
.
ri rl O N M ~O V M r-i N ~
~
V O
Cp
x bDcr u1 O ~1 u1
r. O~ MOr-ION 000 ~O '~
O
. ri u1 N N d' rl N ~t O~ V~ O
«'1 rl
Q O Oo0O~tO
~ N r-i r-I r-I r-I N N ~O
W
~
~ b
~ ul ~ u'1 u1
O O O O n
OI~O N N
O O
ra \O ~ O N O r-I r-I r--I O d' V r--I ONO t11
.C
O
~ ~ O rl 1` N
V
V rl V N
• O O
O ~ O r-I N
O O O
O rl ~' M rl O ~p V r1O M O M
7r O
.~ ~ r• N
>-'+
O
W
zi ~
.~'
•~ •
M ~
U r-i ~
A
on ,,~
''~
~ O
~O N
O O
r-i ~
RS
.r{
cti rd
•~ ~
O 00 NO
~O
U M n N~ N n O ,-I
C' ~ N -
''~
G r-I O r-i O rl O ~ O ~ O
.. r-I CO V r-I V rl O N O r1
U w U W V F~ U FL+ U W V W V W C.) w U W U (~ U W U W U W U W U W U W V W U W
~ ~
•r•I .~' U1 .~ ~i
~ N •ri ~ d) ~ +~
+J P+ p r1 cd ++ +~ cd rl O U
c~U r~I '17 U N ~ •rl R} rl O •r~~i •}~.~ ~
cd ,.C ~ U ,.O td rl ~ td N rl ~ d-~
~ a +~ ro ~d o a cd ~ q a ~ ~
G ~ u o •r-I o 0 0 'd •~I cd ro •,~ u a ~, v
•~ o v ~ ~ ~ r°1-I r~i ~ ~ °ui ~ ~ ~ ~ ~ ~ ~
~ ~ `-' ~ A r~ w w w w w w w o c7 ~ x x
-a~-
~ ~:o ~o~o .Ho
,~ T° o M v ~ v ~ v, ~-+
.~,
~ .~ `"•o .-to o ~o O o •o
b ~ O rl V r-I r--I I~ V r-I ONO ~t O N
ro
x' ~b0 ^ O~OmO~ONO ~OrlO x-10
•rl O M O r-I O rl O r-i N ~' O rl V M V rl
`~ ~ O• m O r-I O r-I O
r-Oi O r-I O 01 V rl V rl
~ .~ ~ O r-I O r-I O ~ O ~-I O ~ O
N ~ O~ V r-I V r-I rl U1 V N O .-I
a
~i r--I I~ ~ ~ cV
•bD OONO~OOO V ~ O'-IOON
O H O '
'T7 ,--~ V M
G
~ o
v ~
~ ~~
°
~
~ O O
N ~' ~O I~•
~ O N O H O
N N O r1 r-I I~
~O M
n
00 H O O+-I O rl O
r-I ~' V M rl ~t V N N rl
O
pa
.b
6~
~t O
.~ O ~O O~
,x ~ o ~o
U ~
~
A r-I
M ~1
~ O Q O
•rl N O~ rl M 00 M
..C
g ~ o M o
O V rl N I~
•t3 r-I
.
M
~' 1
O 0
O
~b ~+ O M O r-1 00 M
U O
N
~'
~
rl.O rl O r-i O
O O
•~ V ri V' rl V N ~ v1
,.G
r-I
• O
O N
• O
O O
r-1 rl
H ~
r-I
• O
~ O
U (srUf~+CJF~UFTiUfzrUF~±U(z+UF=•~ U fs•IUf~+UfsaUF~UFLtUf=aCJw UFT~ UFT•+U FTa
ro
.~
H
O
4-I
O
D"
~ •
~ r~ D
•`
b0 N O tp •ri •rl cA ~' ro ,.o H W
3-r p, rl ~ O ~ ~ N b0 •r-1 H H ~ td
•rl Cd O ~ N `~ r-I bA ~ ~
(A Cd
•,~ . ~,-~ ~ o a. a •~ ~+ a~ a a •~ ,~
~ ro w o ~ ro cn w ~+ ro ro a ~ ~ v G
G ~ N •~I •rl ro ~ ~ m O O
U N ~ cd q G •r-I cn ~ ro ~ ~ w
•~ •~ P ro •~ ~ cw ~ ro ~ N a~ ~ ~
!~ +~ ~ cn ro ~ •~ p. ~ r-I ro ~ •~I v v ,~ ro
~ ro ~ u ~+ b S o •u ~ ,.ro v ~ •~ •,~ +~ ro
•
ro ~ rd v ~ ~ a o ro a G ~ ~--~
~ ~ ~ ~ i
a a~ •~ ~ ~ a~ •~ ~ G 5C ro ro ro •~
x H H h a a a a a ~ ~ z o o w w a w
-28-
M
N
O
~
O .~ O `r.' O
~
~ O
M M . O
~7 M rl d• r-I N
•~
'~ "d
•
u•1
u1 M 1~
~
~
O~
O
O
~ r{ O
O
~ a1N
O
M
M V rI~OM
~ "
ctf ,.C
'~' ~ ~O r-I M ul
•
C O M O O O O
• O N ~ N r-I M O ~-I O rl
~
'~ al
• O
O
-I O O M ir1
~ N N O ri
u1 b .-I
't7 •rl O O O r-I O ul O H O ~ O
~ ~' O N N N V rI ~- r~ M N
a
~ ~
on ~ o
`r:
~
`°
~ o ~o •o ~o
o
o ~o •o
,.G O N N M ~t ~t N ~1 O r-I O rl V r-I rl N
ul
O M
O O O
'd r-1 ~O 01 ,-~ ~y-
G.
O
P~ b r-I M ,~
~' ~ O
Or1 O
ON O
U ~, ~O~t
~i
A .•~
bA ~O
•,~ O
..4 r-I ~
3 ~n
o •o
rl rl M
•ri b 00 61
' ~
0.i ~ ~' O
C' .C
b0 ul ~
O
~-+ rl
N O ~ M ~
Uf=+ Ufs+ C~G4Ufx+C7f~U(=a Ufa Ufs+UW VFs.iUwC..)Fs.iU(=aUFsa UF~Uf3.~Ufst
~ ~
G
ro +~ ~ O U7 U ro W U
~
~
•
aJ *t} b •ri CA G • •rl ro G' S-+
• • A •
bD O O C N N rl cd 1-I O ,G ~ ~ ~
~ v .u ~, ro ~ •,~ a~ a~ ~, .u v ro ro
~ ,-~ ~ a G v ~ u a. ,a •~ ~ ~ a
ro a~ o ~ •~ ro ro ~+ •~ ~, ~ ~ a o co m G
~ •~ •v .u ~ ,.~ •~ ~ cw ~+ v ~ m ~ b o
~ ~+ a~ a. ~+ on •~ ~ o o •~
o a~ ~ on ~ ~, ro ~ ~ ~ w ua m •
o v ~ o ~ .u +~ ~ ~ ~ ~ ~ ~ ~ ro
on a~ ~ ~ v .u ~, ~ ~ a a w a a b b o0
T--i ~ a +~ a~ o ao ~ ,~ ~ •~ •~ •~ •~ r--i ~ ~ ro
0 0 0 0 ~ ,.G m ro ro u u v v o 0 o a
w P-+ w w d w ~ ~ cn ~ cn ~ ~ cn rn cn ~
-29-
3
~ O
r-I rl
•rl
'J b r-I
•
~ ~ O r1
'tJ
a1 .~ N
x oo ,-i o • o
,~ V rl ,-I c+~
,C
3
0
U1 b ~-i ~ M
b •rl O O
~ ~ O r--I.O rl
a
~
bn rt o
•rl V N
3
0
G
o v
w •~
~
x
U
~ ~
A d0
.,~
'C
3
0
r-{
N b
.,~ .,~
b ~ .
c~
U
~+ ~
~ ~
M
• O
~ ~
N M ~7
N~ Otf~r--I~
00 N
•o ~o •o
O N V ~t rl ~O
N
~ o ~ o • o
v~nv~~~
W
• O .-I O
M ~O V rl
~o
V N
Uf~UFT~fJFT~Uf=+Uf=+Uf~ UFO
c[3
N
td cd •rl
•r-I ~ ,-G
rl O ~ ~d
O +.~ '~ •ri pa
4-1 ~ ~ ~ W
,.
o a'
~
.-I +.~ cd +~ !~
~ as ro a
~
ro
~
~ p
i
a a
i b ~ 3
m cd o ro u ro o
•~+ •~+ •~ a ~ o ,.~
a a a ~, +~ •~+ G
v] [n v~ H '~~ D r~-
-30-
diversity: An index of diversity was calculated for each zone of each marsh
using Shannon's formula (Shannon and Weaver, 1964),
d = - j~ niI Log2 (niJ
where d represents diversity, n the total average per cent cover of all
species in the quadrats for each zone, and ni the average total per cent
cover of species i in the quadrats. Because of unoccupied space within the
quadrats n may be less than 100; n may exceed 100 because of overlapping of
plants. Shannon's formula was used in preference to Brillouin's formula
(Brillouin, 1962) for reasons elaborated in Pielou (1966).
The indexes of diversity for the three zones of each marsh are shown
in Table 2. An average index for each marsh, computed by averaging the
indexes of each zone, is shown in Table 3. Other studies utilizing communi-
ties of benthic macroinvertebrates (summarized in Wilhm and Dorris, 1968)
have yielded values of less than 1 in areas of heavy pollution, values from
1-3 in areas of moderate pollution, and values exceeding 3 in clean water
areas. If these generalizations may be extended to communities of vascular
aquaties, then the open water communities in the two upstream marshes show
effects of low-moderate pollution and the high and mid-marshes in these
areas show no pollution effects. Similarly the high and mid-marshes at the
Arcadia site show low-moderate pollution effects whi7;e the open water commu-
nities possess an index of diversity indicative of high-moderate pollution.
At the Duck Pond site, that portion of the wildlife sanctuary closest to a
city dump which operated until two weeks before the actual study period, the
open water community shows the effects of severe pollution and the high and
mid-marsh indicate high-moderate pollution. Since the area sampled as low
marsh at the Duck Pond site was a mud-bar dominated by Pontederia, the index
-31-
Table 2 .
Richness, diversity, and evenness of high, mid, and low-marsh zones
of four Mill River study sites
Haydenville -
Leeds
Duck Pond
Arcadia
Marsh
high marsh richness
32 diversity
3.7 evenness
.7
mid marsh 31 3.8 .7
low marsh 14 2.6 .7
high marsh 44 4.2 .7
$rid marsh 33 3.9 .7
low marsh 15 2.6 .7
high marsh 7 1.7 .6
mid marsh 7 1.3 .5
low marsh 4 .4 .2
high marsh 15 2.5 .8
mid marsh 10 2.4 .7
low marsh 4 1.3 .7
Table 3
Total richness and mean diversity of four Mi11 River study sites,
computed from data obtained separately from high, mid, and low marsh zones
total
richness. mean
diversity
Haydenville 41 3.4
Leeds 64 3.6
Duck Pond 10 1.1
Arcadia Marsh 18 2.1
-321
of diversity for that site may not reflect conditions comparable to those
measured by the low marsh indexes in the other marshes. High and mid-marsh
indexes for the Duck Pond site would be, however, valid.
evenness: Evenness (also known as equitability), a measure of pattern
diversity, was calculated as proposed by Pielou (1966) and Wilhm and Dorris
(1968) as follows;
__ a
J Log2S
where S is species richness. The range of J is from 0 to 1, J equalling 1
in a situation of perfect evenness when diversity is theoretically infinite
and all individuals belong to separate species. Evenness is, hence, an
inverse measure of the degree of dominance in a community.
Evenness as shown in Table 2 is relatively constant in all zones of
the various marshes studied, excluding the low marsh of the Duck Pond site,
although the abundance of a few species stands in the downstream marshes
(Cephalanthus occidentalis covering 43.5% of Arcadia high marsh, Acer
saccharinum 57.0% of the Duck Pond high mash, Nuphar varie~atum 23.0% of
Arcadia low marsh and 64.0% of Duck Pond high marsh), coupled with the
greater size of the wildlife sanctuary marsh complex, create a strong visual
impression that large single-species stands are characteristic there.
Succession and unoccupied substratum: Any consideration of the structure
of zones of vegetation within a marsh must take into account the .process of
hydrarch succession. Almost certainly if the flow of the river into and out ,
of the impounded areas were to cease, mid-marsh would succeed low marsh, high
marsh would replace mid-marsh, and more terrestrial communities would eventu-
-33-
ally replace the entire marsh complex. A measurement of the overlap of
species between zones may be obtained as follows:
Overla ~ number of species shared by two zones x 100
p total number of species occurring in
both zones
Table 4 contains the measure of overlap calculated by this means.
Species overlap is greatest between high and mid-marsh zones at all sites
except at the Duck Pond where true low marsh is lacking. No species occurs
in high and low marsh but not in mid-marsh at any site. Only Leersia
oryzoides occurred in every zone of every study site. The zones of the
Leeds site are least similar of the four study areas, while those of the
Haydenville site are most similar, suggesting that the differences in
species. richness and diversity between the two upstream marshes and the
Arcadia Wildlife Sanctuary marsh ..complex cannot be explained as simple
differences between seral states.
Table 5 lists the per cent of substratum unoccupied within each zone.
Unoccupied substratum may be interpreted (1) as the result of the exclusion
of species for some reason from otherwise suitable niches or (2) as areas
which species have not yet had time to colonize. Unoccupied space is least
in all zones at the Haydenville site and greatest in all zones at Arcadia
marsh. The similar percentages of unoccupied space at the Leeds site
(which, being least homogenous in species composition between zones, is
hence perhaps at the earliest seral stage of the marshes studied) and at
the Duck Pond site, undoubtedly one of the most disturbed marshes, suggest
~~ that explanations (1) and (2) may both be operative within the river eco-
system.
-34-
Table 4
Overlap of species between zones at each study site (species shared by
two zones /total number of species occurring in both zones X 100)
Haydenville Leeds Duck Pond Arcadia
high and mid-marsh 65.8 30.5 27.3 47.1
mid and low marsh 32.4 26.3 57.1 27.3
Table 5
Percentage of total substratum unoccupied by vascular plants
in quadrats of high, mid, and low zones of Mill River study sites
Haydenville Leeds Duck Pond Arcadia
high marsh 2.9 0.0 12.5 19.0
mid-marsh 0.0 29.6 21,2 58,2
low marsh 23.0 46.3 31.0 66.4
a
-35-
OBSERVATIONS OF AN INVADING SPECIES
Prior to the time of this project, a study (Burk, 1966) was conducted
" of the invasion of Paradise Pond, on the Smith College campus, and the down-
stream portions of the Mill River by Marsilea quadrifolia, the water clover,
an Old World plant now found occasionally throughout the northeastern United
States and adjacent Ontario. Marsilea quadrifolia has been found in several
lakes and rivers in eastern Massachusetts (Churchill et a1, 1933); it first
appeared in western Massachusetts in Paradise Pond, having escaped from the
college botanical gardens sometime. between 1941 and 1945. By summer of 1965,
the water clover had spread along the shore of the 25-acre pond and downstream
a distance of two miles along the Mill River to the large marsh at Arcadia
Wildlife Sanctuary, the mode of dispersal being transport of fragmented
rhizomes and possibly sporocarps by running water. In the pond, M, quadri-
folic occupies areas. of deeper water than Sagittaria and other emergent
aquatic vegetation and, when the pond is drained, it quickly invades exposed
mud areas. Downstream, the most abundant growth of the water clover seemed
to be correlated with man-made disturbance at the edge of the river. In 1965
near Arcadia Wildlife Sanctuary, M. quadrifolia occurred with Nuphar and other
aquatics at .the foot of a bank of rubble pushed onto the marsh from the city
dump. At that time, it was absent from undisturbed sites within the wildlife
sanctuary itself. In summer of 1966, a small colony of water clover became
established near the site of the old trolley bridge which separates the main
" marsh from the Duck Pond area; this colony did not survive the winter of
1966-67.
During the summer of 1969 while the present study was being conducted,
a colony of M. quadrifolia became established over several hundred square
-36-
feet of substratum in the lower portion of the main Arcadia marsh near the
outlet of the Mill River into the Oxbow of the Connecticut. This colony
spread, and by early fall of 1970, the water clover had become generally
abundant throughout the marsh, including the site where the transect had
been run in 1969. The recent success of M, quadrifolia is believed to be
related to disturbance, including the extensive "letting down" of the marsh
caused when the height of the Holyoke Water Power Company dam is lowered and
the series of accidental upstream incidents which released pollutants which
were carried downstream.
CHEMICAL ANALYSES
Certain chemical analyses were conducted to derive data which might
be correlated. with the biological assessments which were the major objectives
of the study. Water samples were taken at each of the study sites and at
other sites along the river during five periods from July to December, 1969,
and tested for various substances with a Hach Chemical Co. portable water
chemistry kit (DR-EL). Samples of sediments were taken from each of the
sites on five dates and tested for oil as well as other substances. While
detailed methods and results of these tests are beyond the intended scope
of the present treatment, several trends may be summarized. During periods
of high water, the samples showed a high degree of similarity except for
slight increases in some substances downstream. The pH tended to increase
from Haydenville to Arcadia Wildlife Sanctuary, probably as a result of the
addition of raw sewage to the stream. Iron increased downstream and was
concentrated in the marsh areas where flushing is less complete. The pH
-37-
tended to be depressed in portions of these marshes, perhaps as a result
of the accumulation of iron humates. Iron was continually released over a
9 period of several weeks when the sediments were steeped in the laboratory,
suggesting that iron may be released continuously from marsh ooze and con-
centrated in the overlying water during periods of stagnation. Oxidized
forms of nitrogen were entirely absent from the sediments.
Oil residues within the sediments increased downstream, a set of
determinations in samples collected at the Haydenville site on September 15
being .42 , .28, and .16 mg./gm. dry weight as contrasted with samples of
1.20, 1.30, and 1.02 mg./gm. dry weight recorded from Arcadia marsh on the
same date.
Hence, while the environmental factors affecting the marsh communities
are undoubtedly complex and difficult, if not impossible, to assess separately,
the accumulation of certain harmful substances in the downstream areas cam. be
demonstrated by standard laboratory procedures. A full analysis of the
chemistry of the.Mill River remains to be completed.
CONCLUSIONS AND .SUMMARY
The structure of vascular plant communities occurring in marshes which
have developed behind impoundments on the Mill River .reflects the effects of
man's activities in the area. The downstream marshes possess lower species
richness, a lower diversity, and are characterized by greater amounts of
r unoccupied substratum than the upstream, less disturbed marshes.- The invasion
of the lower marshes by an introduced plant species has been documented.
The variations in marsh composition are correlated with pollution, rerouting
-38-
of the stream bed, and abrupt raising and lowering of the water level.
It is likely that in other stream systems the structure of marsh vegetation
would also be a useful indicator of pollution and other disruptions of the
river ecosystem.
-39-
Literature Cited
Brillouin, L. 1962. Science and Information Theor 2nd Edition.
~> Academic Press, Inc., New York.
Burk, C. J. 1966. Marsilea quadrifolia L. in Western Massachusetts.
Amer. kern Jour. 53: 140-141.
Churchill, J. R., et al. 1933. Reports of the Flora of Massachusetts, II.
Rhodora 35: 351-359.
Clausen, R. T. 1940. Aquatic vegetation of the Lake Ontario watershed in
"A Biological Survey of the Lake Ontario Watershed". State of N. Y.
Conservation Dept., Supplemental to 29th Annual Report, 1939.
J. B. Lyon Co., Albany, N. Y.
karrell, M. A. 1932. Pollution studies in "A Biological Survey of the
Oswegatchie and Black River Systems". State of N. Y. Conservation
Dept., Supplemental to 21st Annual Report, 1931. Burland Printing
Company, Inc., N. Y.
Hynes, H. B. N. .1960. The Biology of Polluted Waters. Liverpool U. Press,
- - _.
Liverpool.
Klein, Louis. 196E. River Pollution III. Control. Butterworth Pub.,
Washington, D: C:
Muenscher, W. C. 1933. Aquatic vegetation of the Upper Hudson watershed
in "A Biological Survey of the Upper Hudson Watershed". Supplemental
to 22nd Annual Report, 1932. J. B. Lyon Co., Albany, N. Y.
Pielou, E. C. 1966. The measurement of diversity in different types of
biological collections. Jour. .Theoretical Biology 13: 131-144.
Sharinor~, C. E. and W. Weaver. 1964. The Mathematical Theory of Communica-
tion. U. of Illinois Press, Urbana, Illinois.
Wilhm, J. L, and T. C. Dorris. 1968. Biological parameters for water
quality criteria. Bioscience 18: 477-481.
Yapp, W. B. (ed.) 1959. The Effects of Pollution on Livin Material.
Sy~posium of the Institute of Biology 8, London.
-40-
CHAPTER IV
INVASION OF DRAINED IMPOUNDMENTS
BY VASCULAR PLANT SPECIES
INTRODUCTION
The invasion of drained impoundments by vascular plant species has
been described by various workers in various areas of North America (see
De Selm and Shanks, 1967, for a tabulation of these studies); none of these
investigations, to my knowledge, have been carried out in New England nor
have the earliest phases of any invasion been described quantitatively.
The draining of Paradise Pond on the Smith College campus during the early
phases of this project presented an excellent opportunity to observe the
initial establishment of vegetation on the exposed substrata while the
presence of a site in the rived ecosystem on which invasions had been pro-
ceeding over a period of years permitted comparisons of the pioneer stage
with older seral states. Both the pioneer and later seral stages, examples
of secondary succession, were then compared with hydrarch primary succession
as evidenced by the zonation of the river marshes.
ME'~HOD S
The Paradise Pond impoundment was released on June 9, 1969; although
the Mill River continued to flow through a portion of the former pond bed,
nearly 25 acres of substratum was exposed. No vascular plants were rooted
in these sediments at the time of the release; on June 11, seedlings of
-41-
Leersia oryzoides were observed on the portions of the surface on which
drainage was complete. On July 23 and 24, 1969, quadrats were placed on
the exposed pond bed to determine species composition, frequency, and den-
sity of the invading seedlings. A 500 foot base line was established along
the length of the impoundment beginning near the dam; this line was divided
into five 100 foot segments; five randomly placed .25 m2 (0.5 x 0.5 m) quad-
rats were placed within each segment. This method of quadrat placement
assured randomness in site selection as well as a distribution of the 25
quadrats along the length of the base line. Any plant rooted within a
quadrat was considered to be present in that quadrat.
In addition, two .5 m2 (0.5 x 1.0 m) quadrats were placed on selected
sites on the Paradise Pond bed. These sites were representative of several
areas which exhibited a high density of seedlings; one.site was dominated
by Leersia oryzoides and the other by Polygonum ~ensylvanicum.
A second study site was established directly north of the Northampton
Corporate Boundary in .the town of Haydenville behind a dam which has, over
~.
a period of years, in the recent past, been partially ruptured by the river.
At this site, three distinct zones of vegetation occur on the floodplain
inward from the area of open water. It was assumed that these zones repre-
sent distinct seral stages, the zone farthest from the river having developed
first as the dam began to fail. Three separate 100 foot base lines were
established parallel to the river, perpendicular to the dam and directed
through the center of each zone of vegetation at distances of 6 feet, 20 feet,.
and 35 feet from the stream respectively. Five 0.25 m2 quadrats were randomly
placed along each base line, and species composition, frequency, and density.
were determined for each vascular plant rooted in each quadrat.
-42-
RESULTS
Table 6 summarizes frequency and density of each species recorded at
the study sites. The recently-exposed sediments of Paradise Pond supported
an invading flora of 30 species. Leersia oryzoides, the most abundant of
these, exhibited 100% frequency and represented nearly half the total number
of individual plants recorded in all quadrats. The mean number of individual
plants per square meter was 559.6 with a range of from 260 to 1928 plants/m2.
In the quadrat selected for a dense Polygonum pensylvanicum population,
3438 individuals were counted in an area of .5 m2; 2636 of these were P.
pensylvanicum. In the quadrat selected for a dense Leersia oryzoides popula-
tion, 1494 individuals were counted in an area of .5 m2, 1238 of these being
L. oryzoides and other grasses.
At the older site, the set of quadrats nearest the river possessed the
lowest number of species (11) and the lowest mean number of plants/m2 (373.6)
of the three zones sampled. Leersia oryzoides was the dominant plant, com-
prising 82.2% of the individuals sampled. Species richness and the total
number of individuals, 16 and 1474.4/m2 respectively, were greater in the
middle zone, which also was dominated by L. oryzoides (70.1% of the indivi-
dual plants counted with a frequency of 100%). Polygonum pensylvanicum,
15.7% of the individuals recorded, was second in importance, with a frequency,
of 80%. In the zone farthest from the stream and hence assumedly in the
most advanced seral state of the three zones, the greatest number of species
(25) was recorded; the mean number of plants/m2 was 1222.4, somewhat lower
than the mean density of the middle zone. An increase in biomass from the
zone closest to the stream to the zone farthest from open water was obvious,
-4:i
Table 6
Frequency and density of species
encountered in pioneer and later seral stages on drained impoundments
PIONEER
STAGES LATER SERAL STAGES
Total zone middle zone
Total
"~ near est zone farthest
stream from stream
freq. density fr X fr X fr X fr X
m2 % m2 % m2 % m2 % m2
Acer saccharinum 20 0.8 0 0 20 1.6 0 0 6.7 0.5
Agrostis stolonifera 0 0 0 0 60 36.8 20 27.2 ' 26.7 2"1.3
Alisma subcordatum 40 2.0 0 0 0 0 0 0 0 0
Ambrosia sp. 4 0.2 20 0.5 0 0 60 40.8 26.7 13.8.
Barbarea vulgaris 0 0 0 0 0 0 40 5.6 13.3 1.9
Bidens sp. 16 1.0 0 0 100 41.6 80 22.4 60.0 21.3
Cardamine pratensis I2 0.6 0 'O 0 0 20 1.6 6.7 0.5
Chelidonium majus 0 0 0 0 0 0 20 3.2 6.7 1.1
Chenopodium sp. 4 0.2 0 0 0 0 0 0 0 0
Cruciferae* 8 0.4 0 0 20 3.2 0 0 6.7 1.1
Cyperus sp. 92 33.6 0 0 20 0.8 80 9.6 33.3 3.5
Eleocharis sp. 88 11.6 40 5.6 0 0 60 28.0 33.3 11.2
Eupatorium maculatum 0 0 20 0.8 20 1.6 20 1.6 20 1.3
Eupatorium perfoliatum 0. _ 0 20 1.6 0 0 0 0 6.7 0.5
Gnaphalium sp. 8 0.4 0' 0 0 0 0 0 0 0
Hydrocotyle.americana 4 0.2 0 0 0 0 20 3.2 6.7 1.1
Hypericum sp. 20 1.2. 20 1.6 0 0 60 15.2 26.7 5.6
Impatiens capensis 0 0 0 0 20 1.6 80 57.6 33.3 19.7
Juncus sp. 80 11.4 20 2.4 0 0 0 0 6.7 0.8
Labiatae* 100 39.2 0 0 0 0 0 0 0 0
Leersia oryzoides 100 278.4 100 307.2 100 1031.2 100 635.2 100 657.9
Lindernia dubia 92 20.7 0 0 0 0 0 0 0 0
Ludwigia sp. 76 20.3 20 0.8 0 0 0 0 6.7 0.3
Mimulus ringens 12 0.6 0 0 0 0 60 32.0 20 10.7
Mollugo verticillata 24 1.4 0 0 0 0 0 0 0 0
Oxalis sp. 32 2.0 0 0 20 3.2 60 16.8 26.7 6.7
"
Panicum sp. 92 14.8 40 2.4 40 4.0 80 26.4 53.3 10.9
Pi1ea fontana 48 3.4 0 0 0 0 80 129.6 26.7. 43.2
Plantago sp. 0 0 0 0 0 0 20 3.2 6.7 1.1
Poa palustris 0 0 0 0 0 0 20 4.0 6.7 1.3
Poaceae * 100 74.2 0 0 0 0 0 0 0 0
Polygonum sagittatum 0 0 0 0 20 28.8 80 12.0 33.3 .13.6
Polygonum sp. 60 8.0 60 2.4 80 231.2 80 17.6 73.3 83.7
Populus deltoides 92 20.3 0 0 0 0 0 0. 0 0
Portulaca sp. 12 0.8 0 0 0 0 0 0 0 0
Potentilla sp. 8 0.5 0 0 0 0 0 0 0 0
Rubus sp. 0 0 0 0 20 1.6 0 0 6.7 0.5
Setaria glauca 0 0 0 0 0 -0 20 0.8 6.7 0.3
Solanum sp. 28 1.2 0 0 0
0 0
0 0
0 0
0 0
6.7 0
0.5
Spergularia rubia 0 0 20
0 1.6
0 0 0 80 19.2 26.7 6.4
Stellaria sp. 16
0 1.6
0 0 0 20 0.8 40 4.8 20 1.9
Tr ifolium sp.
Ulmus sp. 4 0.2 0 0 20 1.6 0 0 6.7 0.5
3
2
Viola sp. 0 0 0 0 20 0.8 60 8.8 26.7 .
Unidentified 20 7.2 - 46.4 - 84.0 - 96.0 - 75.5
*unidentified beyond family, possibly more than one species
-44-
with large specimens of Ambrosia, Bidens, Eupatorium, and Impatiens showing
aspect dominance. Leersia oryzoides, with an average density of 635.2
individuals/m2 and a frequency of 100%, was nonetheless the most abundant
species in this zone.
DISCUSSION AND CONCLUSIONS
Erosion, deposition, the release of impounded waters, relocations of
the stream bed, and other changing physical features of the river ecosystem
result in sites on which both hydrarch and secondary successional processes
may occur alternately. A comparison of the data obtained in this phase of
the study with that compiled in other sections lends insight into the role
of several species within the river ecosystem, as well as into means by which
plant communities are adapted to changing seral conditions.
Some weedy species, including Mollugo verticillata and Chenopodium sp.
were encountered in the pioneer stages of succession on drained impoundments
only. Others, including Amb, rosia sp., Barbarea vulgaris, Chelidonium majus,
Eupatorium maculatum, E. perfoliatum, Poa palustris, and Setaria lauca were
found only in the later stages of this sere.
Certain species typical of low and mid-marsh participated in the
pioneer stage of secondary succession but do pot persist into later seral
stages. These include Lindernia dubia and Alisma subcordatum; their growth
is undoubtedly encouraged by the moist substratum left after release of the
impoundment; as soil conditions become more dry, they are probably excluded.
Other mid-marsh species including Pilea fontana and Mimulus ringens, however,
persist through the perennial herb stage.
Rice Cutgrass, Leersia oryzoides, occurs in all zones of the four
-45-
marsh sites studied. Although, in each of the sampled areas, it comprised
only a small portion of the total vegetative cover, its frequency was as
high as 90% in the high marsh zone of the Duck Pond. Leersia oryzoides is
the dominant pioneer species on drained impoundments, persisting past the
initial stages of invasion and within the marsh habitat, might well occupy
niches only recently cleared of other vegetation.
Acer saccharinum, a typicalspecies of the upper marsh zones, occurred
in all the stages of secondary succession studied, while Agrostis stoloni-
fera, Impatiens capensis, Polygonum sagittatum, characteristic species of
the high marsh zone, occurred in the later secondary seral stages.
Populus deltoides, a very common plant on the river's floodplain (see
flora) was present at a density of 20.3 plants/m2 on the bed of Paradise
Pond six weeks after the sediments were drained, but lacking in the-later
seral stages at the upstream site. At the time of the draining'of Paradise
Pond, seeds of P. deltoides were being released and .carried in great abundance
by the wind. Hence, the presence or absence of the cottonwood. in seral stages
of the floodplain may be to a. large degree dependent on the, time of year that
the substratum was open to invasion.
A major unsolved problem in ecology centers about the questions of how
much stress an ecosystem can tolerate and what mechanisms ecosystems use to
withstand stress. In the Mill River ecosystem, where extents of substratum
may be either rapidly exposed for colonization and the earlier stages of
secondary succession or, equally rapidly, be inundated for a sufficient time
that primary hydrarch succession may proceed, the presence of species within
the seral communities common to both successional types may be a major stabl-
izing force, assuring a rapid vegetative cover of the substratum when the
-46-
environment is disrupted. The study of the river marshes showed that pollu-
tion and other disturbance lowered the diversity of species in the downstream
marshes; if those species common to marsh and to secondary seral communities
were to be among those eliminated by these factors, the total ecosystem
might well be severely damaged.
-47-
LITERATURE CITED
De Selm, H. R. and R. E. Shanks. 1967. Vegetation and floristic
changes on a portion of White Oak Lake bed. Ecology 48: 419-425.
-48-
CHAPTER V
A SUMMER FLORA OF THE MILL RIVER AND ITS FLOODPLAIN
An inventory of the flora of vascular plants occurring on the Mill
River and its floodplain was conducted from June 3 to August 28, 1969. All
specimens were taken at or below the spring high water level of the river;
this level could be easily determined by observation of flood-bourne debris,
either entangled in tree or shrub branches br as a strand deposited along
the bank. A11 perennial species collected (except for the few which may
have become established since the spring floods of 1969) can therefore be
assumed to be sufficiently hydrophytic to withstand a period of inundation.
A total of more than 500 specimens were collected, representing about 275
species and varieties, 165 genera, and 75 families. A few plants which
flower in early spring and a larger number of fall-blooming species,
including some Poaceae and Compositae which may be important elements of
the flora are not included in this listing, although fall collections have
been made and deposited in the herbarium of Smith College. Vouchers for
all species in the following list have been deposited in the herbarium of
Smith College. Nomenclature follows Fernald (1950).
A scale of relative abundance was established with the following
categories: very common, common, uncommon, rare, and local. Species
categorized as local were restricted to a small area along the course of
the river but relatively common in that area. The greatest number of
species collected were common or very common in abundance.
-49-
EQUISETACEAE
Equisetum arvense
E. arvense forma pseudo-sylvaticum
E. fluviatile
E. hyemale
E. palustre
E. sylvaticum
OSMUNDACEAE
Osmunda cinnamomea
0. regalis
POLYPODIACEAE
Athyrium filix-femina
Dryopteris marginalis
D. thelypteris
Onoclea sensibilis
Pteretis pensylvanica
MARSILEACEAE
Marsilea quadrifolia
PINACEAE
Pinus strobus
TYPHACEAE
Typha latifolia
SPARGANIACEAE
Sparganium americanum
ZOSTERACEAE
Potamoge. t on crispus
P. epihydrus
ALISMATACEAE
Alisma subcordatum
Sagittaria latifolia
S. latifolia forma gracilis
HYDROCHARITACEA
Elodea canadensis
E. nuttallii
POACEAE
Agrostis perennans
A. stolonifera
Anthoxanthum odoratum
Bromus ciliatus
Cinna arundinacea
Dactylis glomerata
Echinochlora crusgalli
E. pungens
Elymus riparius
Festuca elatior
F. octoflora
Glyceria canadensis
G. grandis
G. striata
Leersia virginica
L. oryzoides
Panicum clandestinum
P. lanuginosum var. imp licatum
common
common
very common
common
common
uncommon
common
common
common.
local
very common
very common
common.
very common,
Paradise Pond & downstream
local
very common
very common
common
very common
very common
very common
common
uncommon
very common
common
very common
common
common
common
common
common
common
very common
very common
common
common
common
common
common
very common
very common
very common
-50-
POACEAE, CONTINUED
Phalaris arundinacea
very common
Phleum pratense
very common
Poa palustris
very common
Setaria lauca
Aizania aquatica common
local
CYPERACEAE
Carex angustior
C. annectens common
C. crinita common
very common
C. intumescens
~ uncommon
C.
laevivaginata
C. laxiflora very common
common
C. lupulina
very common
C. lurida
very common
C. scoparia common
C. tenera uncommon
C. tribuloides
common
Cyperus strigosus
common
Dulichium arundinaceum common
Eleocharis acicularis very common
E. obtusa
very common
E. palustris
very common
E. tenuis uncommon
Scirpus atrovirens very common
S. cyperinus common
S. expansus common
S. pedicellatus very common
S, polyphyllus
uncommon
S. rubrotinctus very common
S. smithii local
S. validus uncommon
ARACEAE
Arisaema atrorubens common
A. stewardsonii
common
Peltandra virginica comm
Symplocarpus foetidus on
common
LEMNACEAE
Lemma minor very common
Spirodela polyrhiza uncommon
COMMELINACEAE
Commelina communis very common
Tradescantia virginiana uncommon
PONTEDERIACEAE
Pontederia cordata common
JUNCACEAE
Juncus acuminatus common
J. bufonius common
J. effusus very common
J. effusus var. compactus uncommon
J. marginatus uncommon
J. militaris uncommon
J. tenuis common
-51-
JUNCACEAE, CONTINUED
Luzula campestris local
LILIACEAE
Aspara~ s officinalis common
Hemerocallis fulya common
Polygonatum canaliculatum common
Smilacina racemosa very common
Smilax herbacea very common
IRIDACEAE
Iris pseudacorus common
I. versicolor very common
ORCHIDACEAE
Epipactis helleborine uncommon
SALICACEAE
Populus deltoides very common
P. tremuloides common
Sa? ix nigra very common
S. sericea uncommon
JUGLANDACEAE '
rya ovata
Ca common
,
CORYLACEAE
Alnus ru osa very common
A. serrulata very common
Betula papyrifera common
Carpinus caroliniana very common
k'AGACEAE .
uercus ~alustris common
Q. rubra very common
ULMACEAE
Ulmus americans very common
URTICACEAE
Boehmeria cylindrica very common
Laportea canadensis very common
Pilea fontana ~ very common
P, u~ mila common
Urtica dioica very common
POLYGONACEAE
Poly~onum aviculare very common
P. cespitosum common
P. lapathifolium very common
P. pensylvanicum very common
P. gensylvanicum var. laevigatum common
P. persicaria very common
P. sagittatum very common
Rumex acetosella very common
R. cris~us very common
R. obtusifolius very common
Tovara virginiana common
PHYTOLACCACEAE
Phytolacca americana common
AIZOACEAE
Mollugo verticillata very common
CARYOPHYLLACEAE
Cerastium vulgatum common
-52-
CARYOPHYLLACEAE, CONTINUED
Lychnis flos-cuculi
Saponaria officinalis common
Spergularia rubra very common
Stellaria media common
NYMPHAEACEAE common
Nuphar variegatum very common
RANUNCULACEAE
Ranunculus abortivus
R
a
i common
.
cr
s very common
R. bulbosus
R. recurvatus common
R. tricophyllus common
local
BERBERIDACEAE
Berberis chunbergii v
PAPAVERACEAE ery common
Chelidonium majus very common
CRUCIFERAE
Barbarea vulgaris ver
y common
Capsella bursa-pastoris very common
Cardamine pratensis ver
y common
Hesperis inatronalis very common
Lepidium campestre very common
L. virginicum very common
SAXIFRAGACEAE
Penthorum sedoides
uncommon
PLATANACEAE
Platanus occidentalis very common
ROSACEAE
Amelanchier laevis
un common
Agrimonia striata common
Geum canadense
common
Potentilla canadensis
common
P. noryegica uncommon
Prunus pensylvanica common
P. serotina common
P. virginiana uncommon
Rosa multiflora
common
R. palustzis com
n
R. virginana mo
common
Rubus alleghenienis very common
Spiraea latifolia very common
S. tomentosa very common
LEGUMINOSAE
Apios americana common
Amorpha fruticosa very common
Robinia pseudo-acacia very common
Trifolium pratense common
T. repens common
OXALIDACEAE
Oxalis europaea ~ very common
0, stricta very common
GERANIACEAE
Geranium maculatum un common
-53-
GERANIACEAE, CONTINUED
Geranium sibiricum uncommon
EUPHORBIACEAE
Acalypha rhomboidea common
Euphorbia maculata common
CALLITRICHACEAE
Callitriche heterophylla very common
ANACARDIACEAE
Rhus radicans very common
R. typhina very common
AQUIFOLIACEAE
Ilex verticillata common
CELASTRACEAE
Celastrus scandens very common
ACERA.CEAE
Acer rubrum very common
A. saccharinum very common
A. spicatum local
BALSAMINACEAE
Impatiens capensis very common
VITACEAE
Parthenocissus quinquefolia common
Vitis labrusca very common
V. riparia very common
TILIACEAE
Tilia americana _ very common
GUTTIFERAE
Hyp_ericum ellipticum very common
H. mutilum very common
H. perforatum very common
H. ~unctatum very common
H. virginicum very common
VIOLACEAE
Viola papilionacea common
LYTHRACEAE
Lythrum salicaria uncommon
ONAGRACEAE
Circaea ~uadrisulcata common
Epilobium glandulosum var. adenocaulon uncommon
E. leptophyllum uncommon
Ludwigia palustris very common
Oenothera biennis common
UMBELLIFERAE
Cicuta bulbifera common
C. maculata common
Cryptotaenia canadensis very common
Dancus carota common
Hydocotyle americana very . common
Sium suave common
Zizia aurea common
CORNACEAE
Cornus racemosa very common
C. stolonifera, very common
-54-
ERICACEAE
Lyonia ligustrina local
PRIMULACEAE
Lysimachia ciliata m
L. quadrifolia very co
mon
L. nummularia common
OLEACEAE common
Fraxinus americana very commgn
F. pennsylvanica very commgn
APOCYNACEAE
Apocynum cannabinum very common
ASCLEPIADACEAE
Asclepias incarnata
very common
A. syriaca very common
Cynanchum ni rum local
CONVOLVULACEAE
Cuscuta cephalanthi
BORAGINACEAE common
Myosotis,laxa
uncommon
M. scorpioides common
VERBENACEAE
Verbena hastata very common
V. urticifolia
LABIATAE common
Galeo sis tetrahit
Glechoma hederacea common
very common
Lycopus americanus very common
L. uniflorus
common
Mentha arvensis common
M. p~erita uncommon
Prunella vulgaris very common
Scutellaria lateriflora very common
SOLANACEAE
Solanum dulcamara very common
SCROPHULARIACEAE
Gratiola neglecta common
Lindernia dubia very common
Linaria vulgaris common
Mimulus ringens very .common
Scrophularia nodosa uncommon
PLATAGINACEAE
Plantago major very common
E ~. r~u .Lelii very, common
RTTB TACEAE
Cephalanthus occidentalis very .common
Galium asprellum common
G. palustre common
G. ~klnctarium very common
Houstonia caerulsa common
CAPRIFOLIACEAE
Lonicera canadensis uncommon
L. morrowi local
Sambucus canadensis common
Vibernum acerifolium uncommon
CAPRIFOLIACEAE, CONTINUED
Vibernum dentatum
Weigela florida
CUCURBITACEAE
.~ Echinocystis lobata
Sicyos angulatus
CAMPANULACEAE
<, Campanula aparinoides
COML'OS ITAE
Achillea millefolium
Erigeron philadelphicus
E. strigosus
Eupatorium maculatum
E. perfoliatum
Galinsoga ciliata
Lactuca biennis
Solidago graminifolia
S. gigantea
=55-
common
local
common
common
common
very common
very common
common
very common
very common
very common
common
very common
common
-56-
LITERATURE CITED
Fernald, M. L. 1950. GrayTs Manual of Botany, $th Edition. American
Book Co., New York, N. Y.
-57-
CHAPTER VI
ISOLATION OF PSEUDOMONAD STRAINS FROM THE MILL RIVER
INTRODUCTION
The aerobic pseudomonads were among the earliest microorganisms to
be studied. They are ubiquitous, easy to grow, although taxonomically
difficult to identify. Their significance as plant and animal pathogens,
as indicators of. pollution, as digestors of complex organic materials,
and as food spoilage organisms has been widely discussed and debated.
The present study represents a simple and direct approach to the
question of the presence and .isolation of the green pigmented strain of
Pseudomonas aeruginosa and fluorescent strains of pseudomonads from the
waters of the Mill River. This fifteen mile stream presents a variety of
ecological .sampling points, ranging from areas free from human and Indus-
trial wastes, to those portions of the river directly receiving raw human
wastes and/or some industrial discharges.- Ten sampling points were chosen
in connection with the botanical survey of the stream. Laboratory proce-
dures involved a comparison of an enriched isolation medium (Trypticase Soy
broth) and a highly selective primary medium (Drake's ~~10 medium) which was
designed to encourage the growth of Pseudomonas aeruginosa. For isolation
of pure cultures, streak plates were made from the Trypticase Soy broth
comparing the merits of MacConkey`s agar and the selective Pseudosel agar,
and further selecting strains growing in Drake's medium by passage through
Acetamide broth before streaking positive cultures on MacConkey's agar.
No review of the literature of the genus Pseudomonas will, be included
-58-
here, for excellent papers have been published by Tanner (1918), Haynes
(1951), Shewan et al (1960), Lysenko (1961), Colwell (1961 and 1964),
Jessen (1965), Stanier et al (1966), Hoadley and McCoy (1966), and Ballard
et al (1970). It is evident from all of these studies that the taxonomic
status of the genus at present is not clear cut, nor does there appear to
be any appropriate media or methods of isolation and identification of
species.
The present study makes no attempt to clarify this situation which
may be well described as "chaotic", but merely offers information gleaned
from the isolation and preliminary identification of strains of Pseudomonas
aeruginosa which produced definite water-soluble green pigment, and of
strains of pseudomonads which were shown to :be fluox'escent on special
medium under ultra-violet light.
MATERIALS AND METHODS
Collection of samples. All samples were collected in sterile water
sampling bottles and returned immediately to the laboratory within an hour's
interval. All. procedures were carried out at once.
Culture media. The following dehydrated media, each representix~~ a single
lot number, were used: MacConkey's agar (BBL), Trypticase Soy broth (BBL),
Trypticase Soy agar (BBL), Pseudosel agar (BBL), Simmon's Citrate agar (BBL),
Lief son's 0/F medium (BBL), Flo agar (BBL), Tech agar (BBL), Moeller's agar
(BBL), Motility broth (BBL), D-Nase agar (Difco).
-5 9-
The following media were made from the basic ingredients following
the directions of the authors: Drake's ~~10 medium (HLS 3:10-19, 1966) and
Acetamide broth (Rhodes, General Microbiol. 21:221-263, 1959).
Isolation of organisms. Method A.
Trypticase Soy broth was used as an enrichment medium in single
and double strength. Three 10-m1 portions were inoculated in five
sets of double strength and three 1-ml portions into five sets of
single strength tubes. Sets of each were incubated at 5,20, 30, 35,
and 42 C. for 16-18 hours. Growth occurred in all tubes, each of
which was then streaked so as to insure isolated colonies on
MacConkey's agar and Pseudosel agar plates and incubated at the
temperature of initial growth. The 5 C plates were checked daily
for growth for a period of eight days before being discarded and
recorded as "no growth". All other plates were read at 16-18 hours.
Representative types of non-lactose fermenting colonies were fished
from MacConkey's plates to Trypticase Soy agar slants and repre-
sentative types developing on Pseudosel agar were similarly isolated.
All were held for further study.
Method B.
Drake's ~~10 medium was used as a presumptive broth and multiple
tubes were inoculated as described under Method A. All tubes showing
growth were transferred to Acetamide broth for confirmation and
following incubation all tubes showing a positive. reaction were
streaked to MacConkey's agar plates and representatives of each
non-fermenting colony were picked to Trypticase Soy agar slants and
following growth were held for furthex study
-6 0-
RESULTS AND DISCUSSION
The present study was designed to indicate the presence of fluores-
cent pseudomonads in the Mill River and hence every non-lactose fermenting
colonial type which developed in MacConkey's agar or Pseudosel agar plates
was obtained in pure culture on trypticase soy agar slants. This resulted
in the isolation of over a 1000 strains of organisms. Following Gram
staining and the elimination of the oxidase negative strains a total of 387
cultures were held for further study. A11 of these strains grew readily on
Simmons citrate agar. Motility tests were carried out and 294 cultures
were found to be actively motile while 87 were non-motile at all times
examined.
Leif son's 0/F glucose medium was used to further screen the organisms
and a total of 197 strains were able to both oxidize and ferment glucose.
Although these organisms were considered to be non-pseudomonads they were
also grown on Tech and Flo agar slants and of these 68 showed fluorescence
under ultra-violet light. The colors varied: 30 strains were greenish-
yellow, 15 were a brownish-yellow, 5 showed a slight or pale yellow tinge,
9 were distinctly brown, and 9 were a bright orange. These fermenting
organisms had been isolated at all five temperatures. Twenty-one had grown
at 5 C, nine of which showed fluorescence. Nine strains were obtained at
42 C, one of which showed a soluble green pigment on, primary isolatianand
one showed fluorescence only. All other strains were isolated at 20,20
or 35 C. These organisms represent a heterogenous group and further
studies indicated that they represented species of Aeromonas, Achromobacter,
Vibrio, and Flavobacterium.
-61-
selected as pseudomonads were the organisms which oxidized but did
` not ferment glucose in Leif son's medium and were lactose negative. These
wee°e obtained from all five temperatures used, 5, 20, 30, 35, and 42 C.
The majority were isolated in the range of 20-35 C as was to be expected.
Plates of MacConkey`s agar streaked from Trypticase Soy broth (herein after
designated as T-M) yielded the greatest number of cultures at 5 C, while
from the sequence of Drake's medium to Acetamide broth to MacConkey`s plates
(herein after designated as D-A-~M) developed the fewest. At 42 C, the
highest .yield was obtained from plates streaked from Trypticase Soy broth.
to Pseudosel agar (herein after designated T-P), while with D-A M and T M
the same number were positive. Further analysis of these results will be
discussed later..
Parallel studies for the.prese~.ce of col.iform organisms (MPN)-were
carried out on all water samples and where the densities of these organisms _
were low or absent, no pseudomonads were isolated. (Samples ~~2, 6, and 10).
The ten sampling points, as previously mentioned, represented eco-
logically different environments (Table l~. At the source of the river
(sample ~~10) there are no habitations and although no coliforms were ever
found to be present, fluorescent strains of Pseudomonas were isolated at
all temperatures. Thus, while these organisms are most frequently found
in the presence of coliforms the absence of coliforms does not indicate
the absence of pseudomonads. Green pigmented strains of Ps. aeruginosa
were present from this source only at the higher temperatures of 35 and 42
and were obtained only from the selective method of D-A-M. Downstream
(samples ~~9, 8, 7) the coliform densities, increased and pseudomonads were
obtained at all temperatures of isolation. Here the most productive methods
-62-
Table 1'
Sampling Points Along the Mill River
Sample ~~ Location
1 Smith College Boat House, Northampton
2 North end of Paradise Pond, Smith College Campus, Northampton
3 Railroad bridge, south of Route 66 cross over, Northampton
4 Mouth of Mill River at the Ox-Bow of the Connecticut,. Northampton
5 North of Prophylactic Brush Shop, Florence
6 Leeds above the dam -North of Northampton Country Club
7 Adjacent to the road leading to the Hampshire County
Sanitarium, Leeds
8 Second dam in Haydenville
9 South of Kellog Road, off Route 5, Williamsburg
10 Source of River just South of Mi11 Street, Williamsburg
-63-
was T-M, while the green pigmented strains of Ps. aeruginosa were present
at the higher temperatures. Raw sewage is received into the stream just
above the ~~9 sampling point.
Sample ~~6 was low in coliform numbers and only a few pseudomonads
were isolated, none of which showed green pigment at any time or temperature.
No pollution is received into the river between sampling points 4~7 .and ~~6.
Sample 4~5 was taken from an area receiving wastes and the coliform
and pseudomonad densities were again high..
Samples ~~4, 3, and 1 were all obtained below the city dump and the
river at these points received the leachings from this area. Hence, the
coliform count was high and pseudomonads were also present.
Thus, from the ten sampling points, although the coliform group was
absent from the source of the stream and present thereafter,"fluorescent
pseudomonads were isolated from all the samples taken, and green-pigmented
strains were obtained from seven of the ten waters, indicating that members
of this genus are commonly occurring organisms.
An analysis of the isolation procedures is most interesting. Method A
used Trypticase Soy broth as the primary medium and both MacConkey's and
Pseudosel agar was used for isolation procedures. Method B used,Dr.ake's ~~10
medium for a primary broth and Acetamide for confirmatory purposes and
MacConkey's agar plates for isolation. The individual samples yielding
fluorescent pseudomonads from the various media and at the different tempera-
tures are shown in Table 2: A summary of the numbers of these samples is
given in Table 3: The enriching effect of preliminary growth in Trypticase
Soy broth is clear, for the greater number of samples yielded psuedomonads
from this medium. Except at the lower temperature of 5 C, there appears
-64-
Table 2'
Samples Showing Fluorescent Pseudomonads
Sample 5 C 20 C 30 C 35 C 42 C
Number T-M T-P D-A-M T M T-P D-A-M T-M T-P D-A-M T-M T-P D-A-M T-M T-P D-A-M
1 0 0 0 + + + + + + + + + + + +
2 + 0 + 0 0 0 0 0 + 0 + + 0 0 0
3 0 0 0 + 0 0 + + 0 + + 0 + 0 +
4 + 0 0 + + 0 + + + 0 + + + + 0
5 + 0 + + + 0 + 0 + + 0 + 0 0 +
6 + + 0 0 0 0 0 0 0 0 0 + 0 0 0
7 + + 0 0 + 0 + + 0 0 0 0 + + 0
8 + 0 0 0 0 0 0 + 0 0 0 + + 0 0
9 + 0 0 0 + 0 + + 0 + + 0 + + 0'
10 + + 0 + 0 0 + + 0 + + 0 + + 0
T-M = Trypticase soy broth streaked to MacConkey's agar
T-P = Trypticase soy broth streaked to Pseudosel agar
D-A-M = Drake's medium to Acetamide broth streaked to
MacConkey's agar
+ Recovery of organisms
0 = No recovery of organisms
-65-
Table 3'
Total Number of Samples Showing Fluorescent Pseudomonads
5 20 30 35 42
T-M 8 5 7 5 7
T-P 3 5 7 6 5
D-A-M 2 1 4 6 3
T-M = Trypticase soy broth streaked to MacConkey's agar
T-P = Trypticase soy broth streaked to Pseudosel agar
D-A-M = Drake's medium to Acetamide streaked to MacConkey's
agar
-66-
to be little choice between the use of MacConkey's or Pseudosel agar for
isolation of the organisms. No single medium yielded pseudomonads from
every sample, at every temperature. The temperature of incubation was a
factor in the isolation of the various strains and the relative value of
42 C for obtaining pigmented strains is evident. Strains were isolated
from all samples at one temperature or another showing the value of using
more than one temperature for primary recovery purposes. It is apparent
that 42 C enhances the opportunities to isolate the green pigmented strains
from any medium and that the T-M methods at this temperature is very produc-
tive in the early identification of these strains as is seen in Table 4'and
Table 5:
Cultures which .showed fluorescence, but no distinct pigment were
common from all sampling points as shown.in Table 2~. No one combination of
media yields 100 per cent recovery although T-M procedure is the most eco-
nomical and highest yielding of the methods used.
On MacConkey's agar, streaked directly from the Trypticase Soy broth
five types of non-lactose fermenting colonies were observed which were later
all identified as pseudomonads. There were two types which produced pin-
point colonies, one a pale blue and the other a shiny gxay. The most
commonly occurring type measured 1 mm in diameter and was an opaque blue.
A flat, spreading blue colony, 2 mm in diameter and a 3 mm creamy, gray-blue
type were also observed aad isolatedo No correlation could be made between
these types and the final identification of the cultures.
There were also five types of colonies developing on Pseudosel agar,
none of which were pinpoint. Two types measured 1 mm in diameter and one
was a cream color, while the other was blue in appearance. A 2 mm colony
-67- I!
i
~i
Table 4'
Individual Samples Showing Pseudomas Pseudomonas (green-pigmented strains)
Sample 5 G 20 C 30 C 35 C 42 C
Number
T-M T-P D-A-M T-M T-P D-A-M T-M T-P D A-M T-M T-P D-A-M T-M T-P D-A=M ~I
1 0 0 0 + 0 + + 0 + + 0 + + + +
2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 ''
3 0 0 0 0 0 0 0 + 0 0 0 0 + 0 +
i
4 + 0 0 + 0 0 0 0 0 0 0 0 0 0 0
5 + 0 0 0 0 0 0 0 0 0 0 0 0 0 0
6 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
7 0 0 0 0 0 0 0 + 0 ~0 0 0 + + 0' '.
8 0 0 0 0. 0 0 + 0 0 0 0 0 + 0 0
i
9 0 0 0 0 0 0 0 0 0 + 0 0 + 0 0 ~;'
10 0 0 0 0 0 0 0 0 0 0 -0 0 0 0 +
~_.___
T-M = Trypticase Soy broth streaked to MacConkey' s agar
T-P Trypticase Soy broth streaked to Pseudosel agar
D-A-M = Drake's medium to Acetamide broth streaked to MacConkey's agar
0 = Organisms not isolated
. + = Organisms isolated
-68-
Table 5' a
Total number of samples yielding Pseudomonas aeruginosa
5C 20C 30C 35C 42C
T M 2 2 2 2 5
T-P 0 0 2 0 2
D-A-M 0 1 1 1 3
T-M = Trypticase Soy broth streaked to MacConkey's agar
T-P = Trypticase Soy broth streaked to Pseudosel agar
D-A-M = Drake's medium to Acetamide broth streaked to
MacConkey's agar
0 = Organisms not isolated
+ = Organisms isolated
-69-
with a cream colored center and blue periphery and an all cream colored
3 mm colony were also present. The predominating colony was 5 mm in dia-
meter and cream colored. The smaller colonies all proved to be fermenters
of glucose in Lief son's medium, while the 5 mm types were all oxidizers.
From the MacConkey's agar plates streaked from the Acetamide broth, -
all colonies were small and again five-types could be distinguished. A
pinpoint blue colony proved to be a motile,' non-fermenter of glucose. A
1 mm bluish colony which predominated, was isolated in numbers. All
strains proved to be motile, glucose fermenters. Some brown pigmented
strains were among this group. A l mm and a 1.5 mm creamy type of colony
was characteristic of many strains isolated which were motile and which
oxidized glucose in Lief son's medium.
Confirming the work of previous investigators, this diversity of
colonial types appears to. be characteristic of the pseudomonads and of no
value in identifying any of them.
Some comments are in order concerning the organisms isolated which
were not pseudomonads. Aeromonas species appeared to be the most common
and were obtained at all five temperatures, from all media used and from
all ten samples. However, they were found in greatest frequency from the
Trypticase Soy broth. Alcaligenes species were found in seven of the
samples and from all of the temperatures. These two genera are commonly
found in fresh waters. Six samples yielded Vibrio species and from two
samples, strains of Xanathomonas were obtained. All of these genera were
among the oxidase positive strains selected for study. All oxidase negative
strains were discarded and thus it must~be emphasized that many types of
organisms which were lactose negative were omitted from this study, even
-~o-
though they represent part of the total bacterial flora of the water
samples.
A year after the initial study was made, further samples were
obtained from the various points, and using only Trypticase Soy 'broth and
MacConkey's agar plates it was shown that green pigmented strains of
Pseudomonas aeruginosa were present i~ all of the samples previously showing
its presence and that fluorescent, non-pigmented strains were again isolated
from all ten sampling sites. The conclusion may be drawn that these organ-
isma, indeed, are a part of the indigenous flora of this stream.
Hoadley (1968) reported that sewage was a major source of Pseudomonas
aeruginosa~ with storm drainage contributing the next. greatest numbers.. He
further reported that the numbers decreased downstream from sewage outfalls.
The Mill River receives both raw sewage and storm drainage which probably
accounts for the consistency of the presence of these organisms. Bonde
(1966) stated that other Pseudomonas species as well as Ps. aeruginosa may
indicate pollution. The present study clearly demonstrated that fluorescent
pseudomonads were present in large numbers at all times, from all points
along the stream including its source which was free of coliform organisms.
k'uture studies fqr these orgaulsms will be of great value, for all habita-
tions now contributing pollution to the stream are being required by law to
desist and within a five year period this river should be free from sewage
and industrial wastes.
SITMMARY
The Mill River which is free of coliform organisms at its source, but
which at all other sampling points reveals high densities of this group,
-71-
was shown to have as part of its indigenous microbial flora fluorescent
" strains of the genus Pseudomonas in all waters tested. Pseudomonas
aeruginosa (pigmented strains) were shown to be widely spread throughout
the river having been isolated from seven of the ten sampling points.
The isolation of members of the genus Pseudomonas may be readily
carried out by inoculating water samples directly into Trypticase Soy
broth and incubating at 5, 20-35, and 42 C for 16 hours. Streaking
directly to MacConkey~s agar plates will yield a maximum number of poten-
tial pseudomonads. The use of a highly selective medium such as Drake's
~~10, although quite specific, is apparently toxic enough to greatly reduce
the opportunity to recover a maximum number of strains.
-72-
LITERATURE CITED
L
Ballard, R. W. Taxonomy of the aerobic pseudomonads. J. Gen. Micro-
biol. 60; 199, 1970.
Bonde, G. Bacteriological Methods for Examination of Water Pollution.
Health Laboratory Science 3: 124, 1966.
Colwell, R. R. A Study of Features Used in the diagnosis of Pseudomonas
aeruginosa. J. Gen. Microbiol. 37: 181, 1964.
Colwell, R. R. and J. Liston. Taxonomic Relationships among the pseudo-
monads. J. Bact. 82: 1, 1961.
Drake, C. H. Evaluation of Culture Media for the Isolation and Enumeration
of Pseudomonas aeruginosa. Healtk Laboratory 3: 10, 196b.
Haynes, W. C. Pseudomonas aeruginosa - its characterization and identi-
fication. J. Gen. Microbiol. 5: 939, 1951.
Hoadley, A. W. On the Significance. of Pseudomonas aeru inosa in Surface
Waters. N.E.J.W.W. Assoc. LXXXII, 99, 1968.
Jessen, 0. Pseudomonas aeruginosa and other green fluorescent pseudo-
monads. Munkgaard, Copenhagen. 1965.
Lysenko, 0. Pseudomonas - an attempt at a general classification.
J. Gen. Microbiol. 25: 379, 1961.
Rhodes, M. E. The Characterisation of Pseudomonas fluorescens. J. Gen.
Microbiol. 21: 221, AJ 9.
Shewan, J. M., G. Hobbs, and W. Hodgkins. A Determinative Schema for the
identification of certain Gram negative bacteria with special refer-
ence to the Pseudomonadaceae. J. Appl. Bact. 23: 379, 1960.
Stanier, R. Y., N. J. Palleroni, and M. Doudoroff. The Aerobic Pseudo-
monads: A Taxonomic Study. J. Gen. Microbiol. 43: 159, 1966.
Tanner, F. W. A Study of the green fluorescent bacteria from water.
J. Bact. 3: 63, 1918.