EXPERIMENTAL PARASITOLOGY 28, 30-36 (1970)

Anticoccidial Drugs: Lesion Scoring Techniques in Battery

and Floor-Pen Experiments with Chickens

Joyce Johnson and W. Malcolm Reid

Department of Poultry Science, University of Georgia, Athens, Georgia 30601

JOHNSOX, JOYCE, AKD REID, W. MALCOLM. 1970. Anticoccidial drugs: Lesion

scoring techniques in battery and floor-pen experiments with chickens. Experimental
Parasitology 28, 30-36. Although lesion scoring is a commonly accepted criterion
for determining the pathogenicity of coccidial species, no previous attempt has
been made to standardize and describe the scoring scale. In battery trials using
pure species, the six species Eimeria tenella, E. necatrix, acerdina, E. miuati,
E. maxima, and E. brunetti have been scored on a 0 to +4 scale with descrip-
tions of the gross pathologic changes for each score. Eimeria maxima and E.
brunetti were found to be the most difficult species to score since the gross patho-
logic picture has been difficult to correlate with weight gains or other indications of
pathogenicity. Birds from floor-pen trials infected with more than one species were
also graded on a 0 to +4 scale. Four sections of the intestine (upper, middle, lower,
and ceca) were each scored separately. No attempt was made to determine species,
but microscopic examination of scrapings was made to confirm the presence of
coccidia.
INDEX DESCRIPTORS: Pathogenicity; Coccidiosis, Eimeria tenella; E. necatrix; E.
aceroulina; E. micati; E. maxima; E. brunetti; Chemotherapy; Pathology; Techniques.

Although lesion scoring has frequently
been used as a technique to compare quan-
titatively the extent of gross lesions in
groups of birds on different coccidiosis con-
trol programs, few references give a full de-
scription of the different grades of patho-
genicity. A review of this literature has
been incorporated into the following de-
scriptions of the technique employed in our
laboratory. These include six species, al-
though the last two are so difficult to score
that it seems doubtful whether such a pro-
cedure should be attempted in any but
moderately severe infections. These are
Eimeria tenella, E. necatrix, E. acervulina,
E. mivati, E. maxima, and E. brunetti. Since
birds of the same breed and sex inoculated
with the same number of oocysts vary con-
siderably in their response to infection, ex-
amination of several birds on the same
treatment is highly desirable. For most pur-
poses 5 or 10 birds per treatment should
30
be compared with a similar number of sus-
ceptible unmedicated controls. Lesion scor-
ing of birds inoculated with mixed species
of oocysts or from birds grown in floor pens
where more than one species would be ex-
pected to occur, presents special problems.
We have abandoned all attempts to score
by species, and indicate only the degree of
coccidiosis damage in 4 areas of the intes-
tine. Lesion scoring is also helpful in de-
termining the coccidiostatic or coccidiocidal
nature of a drug (Reid and Johnson 1969).
RESULTS AND DISCUSSION
Eimeria tenella. Herrick et al. in 1942
first described a method of scoring E.
tenella lesions. They used a 0 to +4 scor-
ing system with +l for one or very few
lesions, +2 if lesions were “slight,” +3 if
many lesions and considerable blood were
present, and +4 if lesions were severe and
large amounts of blood were present. This
 ANTICOCCIDIAL DRUGS
scoring system has been followed by many
workers (Ripsom and Herrick 1945; Gar-
diner and Farr 1954; Cuckler 1957; Ban-
kowski et al. 1959; Horton-Smith et al.
1961; Lynch 1961; Britton et al. 1964; Turk
and Stephens 1967; Dunkley 1968).
Cuckler et al. (1958) added a +5 for
birds dying of the infection. Waletzky,
Hughes, and Brandt (1949-1950) gave a
+l for birds having less than 10 pinpoint
lesions, and included cecal cores in the
basic description, They included a +4 for
birds having ceca partially occluded with
cecal cores, and a +5 for birds with ceca
completely occluded with old cecal cores.
Eimeriu tenella has also been scored fre-
quently on a 0 to +3 basis as slight, mod-
erate, and severe (Ball 1959; Boney 1948;
Farr and Wehr 1945; Levine and Barber
1947; Waletzky and Hughes 1949-1950).
A 0 to +4 lesion scoring system was de-
vised for E. tenellu, based on this literature
review. A +5 score was not included since
birds dying of coccidiosis may have lesions
less severe in appearance than do survivors.
Lesions are scored on days 5 to 7 post-
inoculation in battery trials as follows:
0 No gross lesions.
+l Very few scattered petechiae on the
cecal wall; no thickening of the cecal
walls; normal cecal contents present.
+2 Lesions more numerous with notice-
able blood in the cecal contents;
cecal wall is somewhat thickened;
normal cecal contents present.
+3 Large amounts of blood or cecal
cores present; cecal walls greatly
thickened; little, if any, fecal con-
tents in the ceca.
+4 Cecal wall greatly distended with
blood or large caseous cores; fecal
debris lacking or included in cores.
Dead birds scored as +4.
Eimeria necatrix. The most complete de-
scription of E. necatrir lesions was pub-
lished bv Dunklev ( 1968). He used a + 1
31
for few hemorrhages with no balooning of
the intestine, +2 for numerous hemorrhages
with slight ballooning, +3 for extensive
hemorrhages with ballooning, and +4 score
for death from the infection. Few other au-
thors have described the lesions, although
Cuckler et al. ( 1956) and Turk and Steph-
ens (1966) have scored E. necatrix on 0 to
+4 scale.
The scoring of E. necatrix is described
below. Lesions are scored on days 5 to 7
postinoculation as follows :
0 No gross lesions.
+l Small scattered petechiae and white
spots easily seen from the serosal
side; little if any damage apparent
on the mucosal surface.
+2 Numerous petechiae on the serosal
surface; slight ballooning confined
to the midgut area may be present.
+3 Extensive hemorrhage into the lu-
men of the intestine; serosal surface
is covered with red petechiae and/or
white plaques. The serosal surface is
rough and thickened with many pin-
point hemorrhages. Normal intestinal
contents are lacking; ballooning ex-
tends over lower half of small intes-
tine.
+4 Extensive hemorrhage giving the in-
testine a dark color; intestinal con-
tents consist of red or brown mucus.
Ballooning may extend throughout
much of the length of the intestine.
Dead birds are scored as +4.
Eimeria acervulina. Lesion scoring has
been rather widely employed using a scale
of 0 to +4 ( Cuckler 1957). However, no
detailed description of the different grades
of lesions has appeared. Doran and Farr
(1965) used a 0 to +3 scale with lesions
described as follows: -I- 1 = bars rare, no
mucus; +2 = bars obvious but not coales-
cent, little or no mucus; +3 = bars coales-
cent, mucus present. Barber (1955) noted
32 JOHNSON
differences in the extent of lesions below
the duodenum ranging from 3 inches with
0.01% nicarbazin to 23 inches without
medication. Hein (1968) scored E. acervu-
Zina on a 0 to +4 scale of slight to very
severe lesions noting that the pathogenic
effects increased in severity and lesions ex-
tended further down the small intestine
with increased oocyst dosage. Aycardi
(1963) reported the typical ladder-like le-
sions of E. acwvulinu caused very little
pathologic disturbance, and should be con-
sidered coccidiasis and not coccidiosis. Le-
sions were scored on days 5 to 7.
A lesion scoring system was devised for
scoring E. acervulinu based on these de-
scriptions in the literature and by extended
pathogenicity trials with this species as
folIows :
0 No gross lesions.
+1 Scattered, white plaque-like lesions
containing developing oocysts are
confined to the duodenum. These
lesions are elongated with the longer
axis transversely oriented on the in-
testinal walls like the rungs of a
ladder. They may be seen from either
the serosal or mucosal intestinal sur-
faces. They may range up to a maxi-
mum of 5 lesions per square centi-
meter.
+2 Lesions are much closer together,
but not coalescent; Iesions may ex-
tend as far posterior as 20 cm below
the duodenum in 3-week-old birds.
The intestinal walls show no thicken-
ing. Digestive tract contents are nor-
mal.
+3 Lesions are numerous enough to
cause coalescence with reduction in
lesion size and give the intestine a
coated appearance. The intestinal
wall is thickened and the contents
are watery. Lesions may extend as
far posterior as the yolk sac di-
verticulum.
AND REID
+4 The mucosal wall is greyish with
colonies completely coalescent. Con-
gestion may be confined to small
petechiae or, in extremely heavy in-
fections, the entire mucosa may be
bright red in color (Morehouse and
McGuire 1958). Individual lesions
may be indistinguishable in the up-
per intestine. Typical ladder-like
lesions appear in the middle part of
the intestine. The intestinal wall is
very much thickened, and the intes-
tine is filled with a creamy exudate
which may bear large numbers of
oocysts. Birds dying of coccidiosis
are scored as +4.
Eimeria mivati. The species E. mivati
described by Edgar and Siebold (1964)
has not been scored on a separate scale as
reported in the literature. Since lesion ap-
pearance and location are similar in many
respects to E. acervulinu, descriptions given
above may be used but with the following
differences: E. mivati has a tendency to
move in a posterior direction with the pro-
gression of the infection. Thus late infec-
tions may extend further down the tract.
The shape of individual lesions is frequently
more rounded so that the ladder-like de-
scription is less likely to apply. Severe con-
gestion has not been noted with this spe-
cies. Although death is rare, if it occurs in
severely infected birds, it is scored as +4.
Eimeria maxima. This is a difficult spe-
cies to score since the degree of patho-
genicity does not always correlate with the
severity of the lesions. Since tissue damage
is produced late in the coccidial cycle, indi-
cations of pathogenicity are brief and ap-
pear largely on days 6 and 7. Rose (1967)
scored this species on a 0 to +4 scale, but
limited descriptions of her methods indi-
cate differences in degree of intestinal
thickening and extent of coloring and quan-
tity of orange red exudate.
E. maxima infections were scored on days
6 and 7 as follows:
 ANTICOCCIDW
0 No gross lesions.
+l Small red petechiae may appear on
the serosal side of the mid-intestine.
There is no ballooning or thickening
of the intestine, though small
amounts of orange mucus may be
present.
+2 Serosal surface may be speckled with
numerous red petechiae; intestine
may be filled with orange mucus;
little or no ballooning of the intes-
tine; thickening of the wall.
+3 Intestinal wall is ballooned and
thickened. The mucosal surface is
roughened; intestinal contents filled
with pinpoint blood clots and mu-
cus.
i-4 The intestinal wall may be ballooned
for most of its length; contains nu-
merous blood clots and digested red
blood cells giving a characteristic
color and putrid odor; the wall is
greatly thickened; dead birds are
recorded with this score.
Eimeria brunetti. In Levine’s original de-
scription of E. brunetii ( i942), light, mod-
erate, and severe lesions of the species were
described. He stated that light infections
produced no gross lesions, and since the
severe infections he described are seldom
seen in experimental infections, E. brunetti
was the most difficult of all the above spe-
cies to score. Boles and Becker (1954) did
not score the species, but described pro-
gressive daily changes in the pathologic
picture throughout the life cycle of the
parasite. The early stages of the heavy in-
fection are manifest in red pinpoint lesions
in the upper middle, and lower small intes-
tine. Rose (1967) scored E. brunetti on a
0 to +4 scale, but no description was given.
we have scored E. brunetti with a 0 to
+4 scale on days 6 and 7 following Levine’s
description as follows :
0 No gross lesions.
+1 No gross lesions. In the absence of
DRUGS 33
distinct lesions, presence of parasites
may go undetected unless scrapings
from suspicious areas are examined
microscopically.
+2 Intestinal wall may appear grey in
color. The lower portion may be
thickened and flecks of salmon-col-
ored material sloughed from the in-
testine are present.
+3 Intestinal wall thickened and a
blood-tinged catarrhal exudate pres-
ent. Transverse red streaks may be
present in the lower rectum and
lesions occur in the cecal tonsils.
Soft mucus plugs may be present in
this latter area.
+4 Extensive coagulation necrosis of the
mucosal surface of the lower intes-
tine may be present. In some birds a
dry necrotic membrane may line the
intestine and caseous cores may plug
the ceca. Lesions may extend into
the middle or upper intestine. Dead
birds are scored +4.
Other species. A lesion scoring system
for Eimeria hugani, E. praecox, and E. miti.s
is omitted since pure clones of these species
are unavailable.
Mixed species in battery experiments. At
the request of cooperating agencies, inocula
containing mixed species have sometimes
been administered in controlled battery ex-
periments. These are scored on the basis of
the degree of coccidiosis by intestinal re-
gion, without regard to species, using the
same system described below for floor pens.
Floor-pen experiments. In floor-pen ex-
periments a lesion score is determined to
indicate the degree of pathogenicity on a
0 to +4 scale. Scores are assigned to four
different areas of the intestine as follows:
the upper region including the duodenal
loop (U), the midgut area located above
and below the yolk sac diverticulum (NM),
the lower intestine, the cecal tonsil, and the
rectum (L), and the ceca ( C). A similar
34 JOHNSON
scoring method from three regions has been
described by Morrison et al. (1961).
Attempts to score individual species were
abandoned after unsuccessful attempts to
judge pathogenicity produced by two mix-
tures, each of three different species para-
sitizing different areas of the gut (Reid,
Womack, and Johnson 196S). Some indica-
tion of the species producing pathogenicity
may be recognized by a study of regions of
the intestine from which positive scores are
obtained. Lesions in the ceca are almost
certainly produced by E. tenella. Those in
the upper intestine are most often E. mivati
or E. acervrclina, although they could also
include any of the other intestinal species.
Lesions in the midgut are most often in-
duced by E. necatrir or E. maxima with the
possibility of species determination by pa-
thognomonic signs. In the lower intestine
E. brunetti is most commonly found, but
many of the other species may also be re-
covered. Since microscopic examinations of
unstained smears are required to determine
whether questionable lesions are due to
some cause other than coccidiosis, added
information on species may come from lo-
cation of pathognomonic parasite stages if
species considerations are of special inter-
est on special coccidiostats. However, diffi-
culties in separating coccidiosis from cocci-
diasis usually make such observations very
tentative. Coccidiasis has been used to de-
scribe a nonpathogenic low-grade infection
by coccidial organisms (Levine 1961) .
The overall lesion scoring technique re-
quires a minimum of two but better three
persons. The individual doing the scoring
examines the intestine under a strong light.
The serosal surface is examined for pete-
chiae, plaques, or other signs of lesions
throughout the length of the intestine. The
mucosal surface and intestinal contents are
similarly examined with shallow or deep
scrapings being prepared for microscopic
examination. The second and/or third oper-
AND REID
ator examines these preparations under low
and high power and keeps the records.
Presence of coccidial infection requires
demonstration of oocysts, schizonts, mero-
zoites, macrogametocytes, or microgameto-
cytes. Ideally a number of birds from each
pen should be examined since individual
birds show great variability in susceptibil-
ity. Five birds per pen (50/o of the total)
were examined in most trials reported in
the present studies (Brewer et al. 1969). A
much smaller percentage may be satisfac-
tory when larger floor pens are used (Gard
1969). Since gross lesion scoring requires
less time than microscopic examination, the
latter may be completed on only one or two
birds per pen. Scoring is best done “blindly”
by one person having no knowledge of the
treatment used. A third operator records
observations and suggests more extensive
studies of material from some birds should
coccidia be found in pens medicated with
experimental coccidiostats.
Lesion scoring is time consuming. Should
other disease conditions such as ulcerative
enteritis appear in the pens, more extensive
microscopic studies may be required to de-
cide whether lesions are induced by cocci-
diosis. The maximum number of birds that
may be examined in a day by a team of
three is 200 birds. Where infection is lim-
ited to one species following inoculation
scoring may be accomplished more rapidly.
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