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strain has been of interest, because it is apparently immune to leaf rust, a factor
which may explain why it is more persistent than commercial strains. Although
meadow fescue is apparently unsatisfactory when grown alone with Ladino clover,
there are indications that five to eight pounds might be seeded along with four to
five pounds of a leafy strain of orchard grass or eight to ten pounds of smooth
brome grass, with excellent results. Meadow fescue is a fast-growing grass which
establishes itself quicklj- and would retard weed growth and yield considerable
feed while the slow-growing brome grass or orchard grass was becoming estab-
lished.

Strains of the so-called tall meadow fescue, such as Alta tall fescue, have not
performed satisfactorily. They are coarse and unpalatable in comparison with
several of the other grasses and offer too much competition for Ladino clover.



DEPARTMENT OF ANIMAL HUSBANDRY
Victor A. Rice in Charge

A Study of the Mineral Elements of Cow's Milk. (J. G. Archibald, C. H. Par-
sons, and H. G. Lindquist.) For two reasons the work with manganese reported
last year was repeated during the winter of 1941-42: (1) the findings were at
variance with some earlier work at another station, so it was thought advisable
to confirm them; (2) it was desired to investigate the possible effect of metabolized
manganese on the development of oxidized flavor in milk.

The results of this second season's work confirmed the finding reported last
year that the amount of manganese in milk can be doubled by adding manganese
to the ration fed. This additional metabolized manganese did not retard or in-
hibit the development of oxidized flavor either in ordinary milk or in milk to
which copper had been purposely added to accentuate the effect.

The element zinc is being studied this \ear.

Investigation of the Merits of Legume and Grass Silage for Massachusetts
Agriculture. (J. G. Archibald and C. H. Parsons.) War econom\ has resulted in
a definite shortage and a corresponding increase in price of the two most common
preservatives for grass silage, molasses and phosphoric acid. Our efforts this
year have, therefore, been devoted to a study of other materials which might
possibly be used as preservatives, and to an improvement of methods in general
so that smaller amounts of preservative or none at all, may suffice.

It was not possible to conduct any feeding trials with the molasses silage stored
in the large experimental silo in June 1941. The excellent quality of this silage



14 MASS. EXPERIMENT STATION BULLETIN 398

was, however, very evident; the odor was mild and sweet, and the cows ate it
much more readily than they did a similar lot preserved with phosphoric acid in
the same silo the previous year.

A second silo containing 75 tons of mixed grass was inoculated at five different
levels at filling time in June 1941, with a pure culture of Bacillus hulgaricus. No
other preservative was used. This silo was opened in January 1942; there was
considerable spoilage at the top and sides and the odor at first was very objec-
tionable, indicating formation of excessive amounts of butyric acid. The odor
improved after the top layers were fed off, but continued more sharply acid than
the odor of the molasses silage referred to above, although the pH was 4.4 in con-
trast to 3.9 for the molasses silage. The station bacteriologist was unable to recover
B. hulgaricus from the silage, which suggests that the inoculation may have
played little if any part in the fermentation.

The silage was fed to young cattle; they ate it readilj' but made somewhat
smaller gains than when fed corn silage. This may have been due, however, to
inherent feeding value of the original material rather than to method of preserva-
tion. It would have been desirable to include another siloful of similar material
not inoculated, as a control. This has been done this year, but at this date
(Nov. 30) the silos have not been opened.

In addition to the three large silos filled this year, using respectively molasses,
bacterial inoculation, and no preservative (control), sixteen miniature silos have
been filled, each containing approximately one and a half bushels of chopped
grass or alfalfa, and weighted with concrete blocks to produce a pressure of about
120 pounds to the square foot. Two crops, mixed grass and alfalfa, at two dif-
ferent moisture levels, have been ensiled in these miniature silos with the follow-
ing treatments: (1) no preservative, (2) inoculation with B. hulgaricus, (3) salt,
(4) B. hulgaricus plus salt. At date of writing (Nov. 30) six of these small silos
have been opened and their contents studied from a biochemical, and to a lesser
extent from a bacteriological standpoint. The outstanding facts thus far are:
(1) the excessive amount of butyric acid which has developed regardless of the
preservative used, and (2) the complete absence of lactic acid from these silages.

A Study of Urea as a Partial Substitute for Protein in the Rations of Dairy
Cows. (J. G. Archibald.) This project has recently been discontinued for the
duration of the war. Final conclusions are not being drawn at this time, but
results of feeding trials extendmg over three years indicate that while the urea
has been utilized by the cows to some extent it has not proved equal to the stand-
ard protein concentrates as a source of nitrogen for milking cows.

The Effect of Feeding Irradiated Dry Yeast on Reproduction and General
Health in Dairy Cows. (J. G. Archibald and J. D. Neville.) Two years' results
with sixty cows in the Gardner State Hospital herd show no differences of any
significance between the cows getting irradiated yeast and those on the control
ration. This lack of response may be due to the fact that this herd has always
been maintained at a rather high nutritional level. The level of vitamin D fed
was approximately 10,000 international units daily per cow.

Determination of the phosphatase level in the blood of twenty of these cows
(ten in each group) at regular intervals during January-, February, and March
of this year showed no significant differences between the groups in this constit-
uent. Since a rise in blood phosphatase is considered to be a sensitive indicator
of vitamin D deficiency, it seems evident from these results that the cows in this
herd were not suffering from even a slight deficiency of this vitamin; therefore,
the lack of response to the feeding of irradiated 3'east is quite understandable.
The project in its present form and as involving the use of the Gardner herd has
been terminated.



ANNUAL REPORT, 1942 15

DEPARTMENT OF BACTERIOLOGY
Leon A. Bradley in Charge

Nitrification in Soils Containing Plant Residues of Varying Lignin Content.

(James E. Fuller, cooperating with the Agronomy Department.) This investi-
gation, so far as the field work is concerned, is obviously seasonal and consequently
proceeds slowly. A preliminary' statement was made in the 1941 report from this
Department (Mass. Agr. Expt. Sta. Bui. 388: 19, 1942). The crops employed
were millet, rye, wheat, sudan grass, sorghum, corn, oats, buckwheat, barley,
rape, artichoke, tobacco, and sunflower. There were six plots with all of the crops
in each, except the sunflower, which was alternated with fallow strips. The
crops were grown and then plowed under. The following season tobacco was
grown on the plots, and during that season three series of soil samples were col-
lected — one early in the season before the tobacco was planted, another in mid-
summer, and the third in the early autumn after the tobacco had been h?rvested.
These samples were tested for their ability to nitrify dried blood and ammonium
sulfate.

The general tendency seemed to be that the capacity of the soil to nitrify dried
blood declined in mid-summer as compared to the spring, and then recovered to
some extent later in the season. There was considerable variation among the
different plots, and no definite relationship could be noted between the variety
of crop and the nitrifying capacity of the soil of the plot. The ability of the
soils to nitrify ammonium sulfate declined markedly, in general, when the early
and mid-season samples were compared, and there was little recovery of activity
when the late-season samples were analyzed.

The nitrification results are being compared with the production and the grade
and crop indices of tobacco grown on the plots. No relationship has been noted,
but further study of the data is being made.

The Determination of the Sanitary Quality of Drinking Utensils. (Ralph L.
France and James E. Fuller.) This project had been carried on b}' France, and
after he entered the armed service of the country the work was taken over by
Fuller, who completed some unfinished work and prepared a report. The results
may be summarized as follows: A wet swab with excess moisture squeezed out
was more effective than a dr\- or a moist swab for removing bacteria from used
drinking glasses. A diluting fluid containing a phosphate (Butterfield's phosphate)
was more effective than distilled water or physiological salt solution for preserving
the viability of bacteria on swabs when they had to be held for some time before
plates were made. When swabs had to be held for several hours before plates
were made, it was found that chilling the containers with ice helped materially in
preventing loss of viability of the bacteria. A yeast-dextrose medium smiilar to
that used for the Standard-Methods procedure for plating milk samples gave
good recovery of bacteria for swabs. Practical examinations were made of local
establishments serving food and drink.

Bacteriological Study of Chocolate Milk. (James E. Fuller and R. W. Swan-
son, in cooperation with W. S. Mueller of the Department of Dairy Industry.)
This study has been completed and the results published. They may be sum-
marized as follows: The addition of chocolate syrups or cocoa powders inhibited
the growth of bacteria as compared with growth in the milk without the powders
or syrups. Even though the bacterial content of some of the powders and syrups
was high, the resultant growth of bacteria in chocolate milk made from these
powders or syrups was not nearly as great as was anticipated. Experiments
indicated that the tannic substances in the chocolate or cocoa were responsible for
the inhibition of bacterial growth.



16 MASS. EXPERIMENT STATION BULLETIN 398

Bacteriological Studies of Rural Water Supplies. (James E. Fuller.) This
project has been started within the year. Some preliminary work along this
line has already been reported (James E. Fuller and Sonnia Levine, Mass. Agr.
Exp. Sta. Bui. 378, 1941). Present experiments are being done on an amplified
scale. The study aims to attempt a differentiation of the intermediate members
of the coliform group of bacteria, so frequently encountered in rural water sup-
plies (wells or springs), to learn how many of them indicate serious pollution and
how many indicate merely surface wash. The differential reactions (Imvic tests)
of these bacteria are being studied at several incubation temperatures, from room
temperature to 46°C. (Eijkmann-test temperature), to determine the proportion
that are related to the fecal Escherichia coli. Present indications are that the
use of the Eijkmann-test temperature for incubating the Imvic tests may be
useful in evaluating the sanitary quality of rural water supplies.

Laboratory Service. (R. L. France, until Julv 1942 ; James E. Fuller, beginning
July).

Milk (bacteria counts) 392

Ice cream (bacteria counts) 91

Milk (butter fat) 27

Water 107

Miscellaneous 3



DEPARTMENT OF BOTANY
A. Vincent Osmun in Charge

Diseases of Trees in Massachusetts. (M. A. McKenzie and A. Vincent Osmun.)

The Dutch Elm Disease Problem. The Dutch elm disease caused by the fungus,
Ceratostomella itlmi (Schwarz) Buisman, and spread by bark beetles was found
in Massachusetts in 1941. During 1942, investigation of this disease in the State
has had three main objectives: the discovery and eradication of diseased trees;
the evaluation of the importance of woodpiles as sources of the causal fungus and
carrier beetles; and the elimination of conditions favorable to the spread of the
disease.

Results of the inoculation of several species of elm with the causal fungus in
early summer under controlled conditions in the greenhouse indicate that all
species of elm are susceptible; but American elm ( Ulmus americana L.) showed
symptoms of the disease most promptly and succumbed most rapidh- of all
species, potted plants of the species being completely killed within 10 days after
inoculation. Death presumably is due to a toxin produced b}- the fungus, but the
word "toxin" is here employed only in a nonspecific sense. Studies previously
reported bj' others, as well as the writers' investigations, indicate that micro-
scopic symptoms indistinguishable from those resulting from fungus infection
may occur in plants variously treated with sterilized extracts from fungus cultures.
If the symptoms are associated with a specific toxin, a neutralizing agent may be
found, although no consistently reliable agent has been found by the writers.

Considerable interest in the work has been shown by municipal tree depart-
ments and private citizens, and many specimens for laboratory study were re-
ceived during the past year. Through the cooperation of the United States
Bureau of Entomology and Plant Quarantine, valuable information relative to
the general distribution of the disease has been obtained regularly. Seven dis-
eased trees have been eradicated in Massachusetts: in 1941, 1 in Alford; in 1942,
3 in Egremont, 1 in Great Barrington, 1 in Westfield, and 1 in Sheffield. The



ANNUAL REPORT, 1942 17

eradication of diseased trees is under the direction of the State Department of
Agriculture which has cooperated also in the entire program. The infection of
trees in the southwestern part of Berkshire County and Westfield, Massachusetts,
apparently does not indicate direct spread of the disease between these two regions
within the State, but rather the introduction of the disease into the State at two
separate points, presumably from a commcn source in the extension of the in-
festation radiating from the area around New York City.

In order to protect disease-free 'elms, concerted effort has been directed toward
the elimination of freshly cut elm wood. At present this procedure is the most
practical control that can be employed effectively in checking the spread of the
disease. Since the disease fungus is virtually a prisoner within an affected tree,
it cannot spread significantly except as it is carried from a diseased to an unin-
fected tree by a vector. The best evidence indicates that the smaller European
elm bark beetle, Scolytus multistriatus Marsh., is the principal carrier Insect.
This insect invades the bark of weakened trees or freshly cut elm wood, where
eggs are laid in galleries engraved between the bark and wood. Upon emergence,
the beetles feed on healthy elm twigs and in this manner may facilitate fungus
infect'on of healthy trees if the feeding beetles come from galleries in diseased
wood. The beetle is now known to occur in Springfield, West Springfield, and
Westfield in Hampden County, and widel> in Berkshire County, in addition to
the originally known eastern Massachusetts section of infestation which has
been enlarged to include most of the area east of Worcester County.

The prompt destruction by burning of all freshly cut elm wood which is well
suited for infestation by bark beetles is urged upon all agencies and Individuals
whose work brings them into contact with it, unless there is complete assurance
that the bark will be removed immediately after the wood is cut or other provision
is made for the consumption of any fungus- free wood as fuel in cooperative agree-
ments. Tree wardens, foresters, arborists, fire wardens, highway departments,
state departments, and public utilities have cooperated generously in aiding in
this practical method of protecting disease-free elms. As in the case of all diseases
of plants, however, unbroken continuity of the program is most essential.

Other Tree Problems. Sixty-three diseases of thirty-four species of trees, in-
cluding nine diseases of elm, were identified from approximately 300 specimens
and inquiries received during the year. The Cephalosporium wilt of elm was re-
ported from 4 municipalities in which no previous cases of the disease had been
reported, making a total of 177 cities and towns in which the disease has been
found in Massachusetts. The fungus, Verticillium sp., was isolated from several
species of woody plants, but no specimens were received from municipalities not
included in the total of 96 reported for 1941.

Following an extended period of wet weather early in the growing season, leaf-
inhabiting fungi caused considerable damage to foliage. Trees throughout
Massachusetts were affected.

A nonparasitic disease of white pine, commonly known as needle-blight, In
which needles of the current season discolor to varying degrees beginning near
the tips, was rather prevalent both on ornamental trees and in plantations.

At the request of the Massachusetts Tree Wardens' and Foresters' Association,
a report on wartime municipal tree programs was prepared, and subsequently a
survey of tree diseases and other defects was made, with a view to outlining a
program for tree protection and the prevention of damage by defective trees .to
persons and property. Current miscellaneous activities included the preparation
of parts of the program of the annual Five-day Short Course for Tree Wardens,
the compilation of a progress report, revision and publication of a manuscript
on a Peridermium of northern hard pines, the discussion of the control of wood-



18 MASS. EXPERIMENT STATION BULLETIN 398

destroying fungi at the Eastern Pest Control Operators' Conference, the prepara-
tion of a mimeographed circular on "Trees in War," and the writing of newspaper
press releases. Publications referred to are found listed at the end'of this bulletin.

Damping-off and Growth of Seedlings and Cuttings of Woody Plants as
Affected by Soil Treatments and Modification of Environment. (W. L. Doran.)

As a result of increasing interest in the beach plum, a native plant heretofore
relatively neglected, work is now being done in cooperation with J. S. Bailey of
the Department of Pomology on its vegetative propagation, and an article on
the subject was recently published. There was no rooting of hardwood stem
cuttings, but softwood cuttings rooted fairh well and best (67 percent in 25 days)
when taken here in mid-June and treated with indolebutyric acid (50 mg./ 1.,
24 hr.). Work with root cuttings taken in the fall is now in progress.

The vegetative propagation of garden sage, a plant not now available to spice
manufacturers from the usual European sources, was investigated in cooperation
with A, M. Davis of the Division of Horticulture, and an article published. Un-
treated cuttings taken in winter rooted well in s?nd-peat or sandy soil, better
than in sand; but their rooting was hastened or improved by treatment for 24
hours with rapthaleneacetic acid or indolebut>Tic acid 25 mg./l

None of the treatments (red cuprous oxide, Barbak D, zinc oxide, Spergon, and
Semesan) which were applied to seeds of trees sown in a cold frame in March gave
wholly satisfactory protection against infection of seedlings by soil fungi. So far
as there were differences in final stands, they were in favor of the use of red cuprous
oxide with blue spruce, Barbak D with white pine, and zinc oxide with hemlock,
arbor-vitae, Douglas fir, and sycamore.

In propagation by cuttings, it is important to know which species need no
treatnients with root-inducing substances. It is noteworthy, therefore, that
August cuttings of Actinidia arguta and February cuttings of Dtcaisnea Fargesii,
both of which bear edible fruit, also February cuttings of Viburnum rhytidophyl-
lum, rooted 100 percent without treatment.

Cuttings of Norway spruce are not very responsive to treatment with the com-
monly used root-inducing acids. But the rooting of February cuttings of two
varieties, pygmaea and nana, was hastened and improved by treatments with
solutions of manganese sulfate, 1.0 percent and 5 hours for the former, 0.5 per-
cent and 24 hours for the latter. Rooting of late fall or early winter cuttings of
Norway spruce, variety nana, as well as cuttings of black spruce, yew, and
Hinoki cypress was also im.proved or hastened by treatment with monobasic
potassium phosphate 0.5 percent, 20 hours. Rooting of the last named was more
improved by this treatment than b> indolebutyric acid.

A mixture of sand and European sphagnum peat has been favored as a rooting
medium, but an American sedge peat, similarly used, may give better results.
January cuttings of Lonicera syringantha, treated with indolebutyric acid 50 mg.
/I., 24 hr., rooted 92 percent in a mixture (2:1) of sand and Florida sedge peat,
decidedly less well in a sim.ilar mixture of sand and European sphagnum peat.

To determine the possible effect of a powder-dip treatment in combination with
and immediately after solution-immersion treatments, cuttings of Chinese juniper
(the variety torulosa) taken in February were variously treated. Untreated
cuttings rooted only 1 1 percent. There was 83 percent rootmg of cuttings treated
with Hormodin No. 3 after treatment for 20 hours with indolebutyric acid 50 m.g.
/L or naphthaleneacetic acid 25mg./l., while cuttings given onK the solution
treatm.ents rooted not more than 50 percent.

As compared with our knowledge of soil disinfection prior to seeding, there
has been relatively little information available about the dismfection of rooting
media for cuttings. When a rooting medium, sand-peat, was disinfected by



ANNUAL REPORT, 1942 19

steaming, by formaldehyde, or by vinegrr within 24 hours before the insertion of
winter cuttings, there was injury to arbor-vitae, red cedar, Hinoki cypress, yew,
and two species of Ilex. Similar treatment of the rooting medium with steam or
formaldehyde four days before the insertion of cuttings was also harmful or of no
benefit. Such treatments are apparently more tox'c to cuttings, even dormant
cuttings, than to some seeds and it is evident that there should be considerable
delay between the disinfection of a rooting medium and the planting of cuttings.
Cuttings of white pme, made of the previous year's growth, were taken in late
wnter. They rooted neither more nor less well in sand-peat previously used for
this purpose, or in sand-peat to which soil from under pine trees had been added,
than they did in new sand-peat, never before used, or in new sand-peat steam-
sterilized fifteen days previously. Fungi in the rooting medium are, it appears
from this, without effect on the rooting of cuttings of white pine. Cuttings from
difi^erent white pine trees differ in rooting capacity, and treatments which improve
the rooting of cuttings from some trees are without much or any effect on the
rooting cf cuttings from others. Rooting of cuttings from some trees was im-
proved by relatively short solution-immersion treatments in relatively concen-
trated indolebutyric acid or napthaleneacetic acid solutions, by powder-dip
treatments with Hormodins No. 2 and No. 3, and by combinations of solution-
immersion and powder-dip treatments.

Effect of Chromated Zinc Chloride on Plants. (L. H. Jones.) The scarcity of
lumber which is naturally decay-resistant has led to the use of a chemically treated
substitute. This has been successful in the case of telephone poles, railroad ties,
building supports, etc. However, when such chemically treated lumber was used
in the construction of benches in which plants are grown in the greenhouse, re-
sults have indicated that the wood preservative may be toxic to the plants, in-
stead of only to the fungi and bacteria which cause decay.

In tests made here, where chromated zinc chloride was used as the preservative,
injury to growing plants was very common and was most pronounced during the
warmer months and with the more acid soils.

Study of Diseases of Plants Caused by Soil-Infesting Organisms, with Par-
ticular Attention to Control Measures. (W. L. Doran.) Seed treatment with a
mixture of oxyquinoline sulfate and talc (1:1) markedly improved stands, but
seed treatment with a mixture of potassium dichromate and graphite (1:1) gave
even better results; as good results, in fact, as did Semesan, Spergon, or red copper



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