SPECULUM 2

THE SURVIVAL OF OOCYSTS OF AVIAN COCCIDIA IN THE SOIL

F. R. KOUTZ, D.V.M., M.Sc.
Department of Veterinary Parasitology

It is rather common to read state- E. tenella and E. maxima disappeared

ments in farm journals that oocysts of in all plots in less than a year's time
avian coccidia, especially Eimeria te- and the greatest length of time infec-
nella, will not survive a severe winter. tive oocysts of E. acervulina were re-
During the past four years experiments
have been continuously conducted at covered from plots was 86 weeks.
the Animal Disease Research Labora-
tories, Ohio Agricultural Experiment MATERIALS AND METHODS
Station, Reynoldsburg, Ohio, on the New Hampshire day-old chicks were
viability of oocysts of avian coccidia reared coccidia-free in wire-bottomed
under various climatic conditions. battery brooders and held in the bat-
Warner1 was one of the first to test teries until ready for use in the experi-
the survival of coccidia of chickens in ments. The chicks used in each of the
the soil. He fed material, taken from experiments were from the same hatch
poultry ranges and houses, to chickens and divided evenly into groups on the
in which infections were produced at basis of live weights and sex.
intervals up to 49 days but not at 81 Weekly fecal examinations, using the
days or 370 days after exposure. Other sodium nitrate flotation method, were
chickens fed soil from experimental made on the chicks in the battery brood-
plots heavily seeded with oocysts pro- ers to assure they remained coccidia-
duced infection in chicks at all periods free. Fecal examinations were made
up to 197 days after such seeding, but before the chicks were placed in the ex-
failed after 217 and 231 days. Warner perimental pens or cages. In addition,
made no atempt to identify the species several chicks from each selected group
of Eimeria used in his experiments. were killed and autopsied and the entire
Delaplane and Stuart 2 found that intestinal tract examined for any
mixed Eimeria species survived in the pathological changes or any forms of
soil from experimental plots for a period coccidia. Fecal examinations were also
of from four to nine months following made periodically from the chicks on
the removal of chickens from the range. experiment.
Viable oocysts were found to survive in Two types of outdoor wire-enclosed
soil of a wooded range at 15 to 18 pens were used. The larger pens (A
months. The soil from the experimental and B) were 14 feet square, completely
plots were fed to chickens. enclosed sides and top, with one inch
Patterson3 found that Eimeria tenella square mesh wire nettings, with the
caused infection of chickens for 21 same type of wire buried two feet. The
weeks in soil under natural conditions smaller pens were 25 inches square, also
of weathering and shaded from direct completely enclosed with the one inch
sunlight and ten weeks under natural square wire netting and the same type
conditions and exposed to direct sun- of wire buried around the sides of the
light. ground. Both types of pens were rat
Farr and Wehr 4 reported that the and bird proof.
length of time oocysts survive on soil Cultures of either E. tenella or mixed
in outdoor plots depended on sunlight, (Continued on Page 3)
shade and vegetation. They reported
SPECULUM 3

COCCIDIOSIS autopsied. Sixty-four additional three-

(Continued from Page 2) week-old chicks were placed in the pen
cultures of E. tenella, E. acervulina, E. to test the infectivity of the ground.
mitis, and E. maxima were given, by These chicks developed clinical symp-
pipette, to susceptible coccidia-free toms of acute cecal coccidiosis with a
chicks before they were placed in the death loss of 20 per cent from the di-
various pens to develop coccidiosis and sease. The remaining chicks were al-
contaminate the ground with oocysts. lowed to stay in the pen until Septem-
The number of birds used to contami- ber, a total of 50 days. Weekly com-
nate the ground, the species of coccidia posite fecal examinations showed the
used, and the length of time the ground presence of E. tenella.
was left idle will be given in each ex- Pen A was sealed and remained idle
periment. from September 2, 1947 until June 1,
Two methods were used to test the 1948 (272 days) . The highest air tem-
viability of oocysts in the contaminated perature recorded during this period
pens. In one method, samples of the was 89° F. (September 1947 and April
soil from five different areas of the pen 1948) ; the lowest temperature was
were collected and mixed in the feed minus 13° F. (January 1948). The
given to susceptible coccidia-free chicks. weeds in the pen had grown about three
By the other method, susceptible coc- feet during the spring. These were cut
cidia-free chicks were placed in the pen down before any chicks were placed in
under natural conditions for sufficient the pen or any soil removed for the ex-
time to ingest any available oocysts. periments. Care was taken so that no
Both of these methods were applied in outside contamination was introduced.
experiments 1 and 2 ; the latter method
only was applied in experiment 3.
The feed was handled in a sterile
mariner, kept in sterile containers and
the feed from the same sack or lot was
fed to all experimental chicks, controls
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oocysts by the pipette method. One HOG CHOLERA VIRUS
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contamination with oocysts. At the end
of this period, all chicks were killed and
SPECULUM 4

PERIOD OF SURVIVAL OF OOCYSTS OF AVIAN Cheesecloth was wrapped around the

COCCIDIA OF THE SOIL cages to keep flies and other flying in-
Species Experiment 1 Experiment 2 Experiment 3
272 Days 322 Days 365 Days sects from contaminating the feed.
Eimeria Twelve days after the start of the soil
tenella Viable Nonviable Nonviable feeding, the experimental and control
Eimeria .. .. Viable Viable
chicks were killed and autopsied. E.
acervulina tenella oocysts were found in the ceca
Eimeria .. .. Viable Viable of the experimental chicks, while the
mitis controls were negative. The mucosa and
Eimeria .. .. Viable Nonviable contents of the ceca were scraped into
maxima
sterile petri dishes, covered with two
Soil from five different areas, six percent potassium dichromate and al-
inches square and aproximately 1/9 inch lowed to sporulate for two days. The
deep was removed from Pen A, mixed culture was washed and given, by pi-
together and this mixture added to the pette, to nine six-week-old coccidia-free
feed. The entire mixture of soil and chicks. Seven days later these chicks
feed was fed for a period of six days to were killed and autopsied ; all showed
ten three-week-old coccidia-free chicks typical lesions of acute coccidiosis. The
kept in small wire-bottom cages. At ten controls were negative for oocysts
the same time, ten controls, the same or lesions.
age and from the same hatch as those At the same time the chicks in the
fed the soil and feed, were fed portions small cages were fed mixtures of soil
of feed alone from the same source. (Continued on Page 5)

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SPECULUM 5

COCCIDIOSIS until September, a total of 54 days.

(Continued from Page 4) Fecal examination periodically showed
and feed, 70 three-week-old chicks were the presence of numerous oocysts of the
placed in pen A (from which samples four species.
of soil had been taken) to determine
whether oocysts that might have sur- Pen B was sealed and allowed to re-
vived over winter would be ingested main idle from September 2, 1947 to
under natural conditions. Control chicks July 20, 1948 (322 days). The highest
from the same hatch were placed in air temperature during this period was
wide-bottomed brooder cages and fed 96° F. (June 1948) ; and the lowest
the same feed. Eighteen days after the temperature was minus 13° F. (Jan-
chicks were placed in the pen, 18 were uary 1948). Two small trees close to
killed and autopsied. Numerous oocysts the pen gave partial shade during part
were found in the ceca of all chicks au- of the day. The weeds in the pen had
topsied. On the twenty-first day, the grown very tall and thick during the
remaining chicks were killed and au- spring and early summer, protruding
topsied. All showed typical lesions of through the top of the six-foot pens.
cecal coccidiosis. The controls remained These were cut before the chicks were
negative. placed in the pen, care being taken to
Seventeen six-week-old coccidia-free avoid outside contamination.
chicks were placed in pen A to deter- In July 1948, soil scraped from five
mine the build-up of oocysts from the different areas (six inches square and
first group of chicks. All of these showed approximately 1/2 inch deep) was re-
severe clinical cecal coccidiosis, but onlymoved, mixed with ordinary feed and
one chick died from the disease. fed to ten three-week-old coccidia-free
Experiment 2: Pen B—To determine chicks for a period of five days. On the
the survival, over a winter, of various fifth and sixth day after feeding the
contaminated soil, oocysts were found in
species of coccidia, cultures of E. tenella,
E. acervulina, E. mitis, and E. maxima a fecal flotation, while examination of
were used in this experiment. To con- the controls showed no oocysts. Re-
taminate the soil in pen B, 80 four- peated checks showed oocysts in those
week-old chicks were given, by pipette, fed soil and feed, but no oocysts were
a mixed culture of the above four found at any time in the feces from
species. One week after the inoculation, the controls. Seventeen days after the
28 chicks were dead from acute cecal start of the soil feeding, four chicks in
coccidiosis with the remainder of the the cages were killed and examined.
chicks showing clinical evidence of the Oocysts of E. acervulina and E. mitis
disease. The remaining chicks were left were found in the anterior parts of the
in the pen for 34 days to spread all intestine, while E. maxima were found
in the middle intestine. Twenty-two
species of oocysts. Periodic checks days later the remaining six chicks were
showed the chicks passing numerous killed and examined. Each showed
oocysts of all four species. Two weeks oocysts of E. mitis, E. acervulina, and
later, 60 three-week-old coccidia-free
chicks were placed in the pen to test E. maxima.
infectivity. All chicks showed typical At the same time the soil was fed to
clinical symptoms of acute cecal coc- the chicks in the cages, 25 three-week-
cidiosis with a mortality of 22 per- old coccidia-free chicks were placed in
cent from the disease. The remainder
were allowed to stay in the infected pen
 6SPRING 1950

pen B to test whether oocysts had sur- deepshade under low trees where there
vived and whether chicks would pick was little or no sun.
up oocysts under natural conditions. The ground in the pens was contami-
Five days later oocysts were found in nated during a three-week period in
the fecal samples from the chicks. Fifteen September 1948 by placing various
days later all chicks were killed and groups of young chicks in the pens that
autopsied. Oocysts of E. acervulina, E. had been given cultures of E. tenella,
mitis and E. maxima were found in the E. acervulina, E. mitis, and E. maxima.
small intestine. In each pen, the inoculated chicks
Ten additional chicks were later showed clinical evidence of cecal coc-
placed in pen B to check the build-up of cidiosis. Fecal examination showed the
oocysts. There was no evidence of cecal passage of numerous oocysts of each
coccidiosis in this latter group, but all species. After the three-week period,
developed chronic coccidiosis from E. four coccidia-free chicks were placed
acervulina, E. mitis and E. maxima. in each pen to test the degree of con-
Experiment 3—In this experiment tamination. The ground was sufficiently
four small pens, 25 by 25 inches were contaminated for in each pen all chicks
used. Pens 1 and 2 were placed so the showed clinical evidence of coccidiosis,
area of ground was always in the sun, with deaths from acute cecal coccidiosis.
pen 3 was placed so that there was Post mortem revealed infection with all
partial shade on the ground during part four species of coccidia.
of the day ; pen 4 was placed in The pens were sealed and left idle for

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SPECULUM 7

one year following the seeding of the young chicks placed on the soil. If suc-
ground with the four species of coccidia. ceeding groups of young chicks are
During this year period between Sep- placed on the same ground, following
tember 1948 and September 1949, the the first group, then severe cases of
highest air temperature was 96° F. acute cecal coccidiosis will develop from
(July 1949) ; the lowest temperature the build-up of oocysts. It is also likely
was 6° F. (January 1949) . that oocysts protected by sticks, stones,
In September 1949, seven three-week- leaves, grass, and weeds in the yard will
old coccidia-free chicks were placed in have a better chance of surviving the
each of the pens. Seven coccidia-free winter.
controls the same age and from the same While some oocysts of E. tenella will
hatch were placed in separate wire-bot- survive a severe winter, these survivors
tomed cages. Another group of three- will usually perish during the hot
week-old coccidia-free controls were put months of summer. Even though the
in a large outdoor pen recently contami- oocysts are protected by weeds and
nated with the four species of coccidia. grass that might have grown during
After six days one of the chicks was the spring, the oocysts do not survive
taken from each pen, killed and au- the heat.
topsied. The results were as follows: It is well known that other Eimeria
Pens 1 and 2 were negative, pens 3 and species are usually present along with
4 were positive for E. acervulina and E. E. tenella. The data showed that mixed
mitis. Single chicks were taken from species such as E. tenella, E. acervulina,
each of the pens every other day and E. mitis, and E. maxima survived a
examined for oocysts. All chicks in pen severe winter, but that E. acervulina
1 were negative to all types of coccidia, and E. mitis were the only species still
the first four chicks from pen 2 were infective after a period of one year.
negative, but the last three showed Sunlight does have a deleterious effect
slight infection with E. acervulina and on all types of oocysts as shown by the
E. mitis. In pens 3 and 4 all chicks were fact that E. acervulina and E. mitis did
slightly infected with E. acervulina and not survive in a bare area with no pro-
E. mitis. The controls in the wire-bot- tection from grass or weeds. Grass and
tomed cages examined before, during, weeds offer some protection even though
and after the experiment were all nega- there is no other shade. Deep shade pro-
tive. Controls placed in the known in- tected the oocysts so that they were able
fected outdoor pens were all positive for to survive long periods.
the four species of coccidia with several The oocysts of E. tenella were found
deaths from acute cecal coccidiosis. to be viable after 272 days through fall,
After the chicks were removed from winter, spring ; but not infective after
the pens, seven coccidia-free chicks were 322 days through fall, winter, spring,
again put in each pen to determine the and part of the summer.
build-up of oocysts. The chicks in pen The oocysts of E. maxima were found
1 remained negative, while those in to be still infective after 322 days during
pens 2, 3 and 4 all became heavily in- fall, winter, spring, and part of the
fected with E. acervulina and E. mitis. summer ; but did not survive a full year
DISCUSSION of exposure.
The data presented shows the oocysts
of E. tenella will survive a severe winter
such as occurs in Ohio and will be viru-
lent enough to cause the disease in
SPECULUM 8

The oocysts of E. acervulina and E. vulina and E. mitis survived a year's

mitis were found to be infective after a exposure. In one pen left in the direct
full year of exposure, but in most cases sunlight and not protected by grass or
were few in number. weeds no oocysts of any species sur-
vived. In another pen protected from
SU M M AR Y the sun by some grass, there was light
survival of E. acervulina, and E. mitis.
Experiment 1: E. tenella oocysts were In pens in partial and deep shade the
found infective after 272 days exposure oocysts of E. acervulina and E. mitis
from September to June. They were were found in great numbers.
found infective to chicks both by feeding
portions of soil from contaminated REFERENCES
ground and from ingestion of oocysts
under natural conditions. 1 WARNER, DAVID E.: Survival of Coccidia of

Experiment 2: A mixed culture of E. the Chicken in Soil and on the Surface of

Eggs. Poultry Science, 12, (1933) : 343-348.
tenella, E. acervulina, E. mitis, and E. 2 DELAPLANE, JOHN P. AND STUART, HOMER 0.:
maxima was used in this experiment. The Survival of Avian Coccidia in Soil. Poultry
Science, 14, (1935) : 67-69.
E. tenella did not survive a severe winter 3 PATTERSON, F. D.: Studies on the Viability of
and partial summer up to 322 days, even Eimeria tenella in Soil. Cornell Vet., 23,
though heavy weeds had grown during 4 (1933) : 232-349.
FARR, MARION M. AND WEHR, EVERETT E.:
the spring to afford some protection. Survival of Eimeria Acervulina, E. tenella,
The oocysts of E. maxima, E. and . E. Maxima Oocysts in Soil under Various
Field Conditions. Annals New York Academy
aeervulina, and E. mitis did survive. Science, 52, (1949) : 468-472.
Experiment 3: A mixed culture of
four species, E. tenella, E. maxima, E.
aeervulina, and E. mitis was used in
this experiment. Two species, E. acer-