Epizootic Gastritis Associated with Gastric Spiral Bacilli in Cheetahs (Acinonyx jubatus)
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Vet Path oI30:55-63 (1993)
Epizootic Gastritis Associated with
Gastric Spiral Bacilli in Cheetahs (Acinonyx jubatus)
K . A. EATON , M . J. RADIN , L. KRAMER , R . WACK,
R. SHERDI NG , S. KRAKOWKA , J. G. Fox, AND D. R. MORGAN
Departm ent of Veter inary Path obiology and Veterina ry Clinical Sciences, Th e O hio State Un ivers ity, Colum bus, O H
(KAE , MJR , LK, RW , RS, SK); Division of Com parative Medi cine, Massachu sett s Institute of Tec hno logy,
Ca mbridge, MA (JG F); and Procter and Ga mble Co., PO Box 398707, Cincinnati, O H (DRM)
Abstract. An outbrea k of vo m iting in a gro up of captive chee tahs (Acinonyx j ubat us) was inves tigated , and
histologic examinati on revea led chro nic gastritis characterized by infiltration o f lymphocytes and nume rous
pla sma cells and epithel ial erosio ns. Lymphoid follicles, globule leu kocytes, scatte red neutrophi ls, and (in on e
ani ma l) abscessed gastric glands were incon sistent finding s. In addi tio n, necrop sy of three cheetahs revea led
gastric mu cosal hyperplasia. T wo kinds of bacteria were identified in th e sto mac hs of infected cheetahs. Nu-
merou s long, tightl y coi led motile Gastrospi rillum -like organis ms were seen in gastric mu cu s and in Warthin-
Starry-stai ned sections of m ucosa . T hese bacteria could not be cultured but were transmi tted to conventional
mice in hom ogenates of gastric m ucosa from infected cheetahs. Ultrastructural exami natio n revea led helical
filam ents on some of these bacteria. In add itio n, a sma ller Helicobacter sp. was isolated . Th is organism co uld
be cultured in vitro und er m icroaeroph ilic co ndi tio ns. O ne or both of th ese bacterial species was proba bly
respon sible for th e gastritis in these cheetahs.
Key words: Bacteria; cheeta hs; gastritis; Gastrospirillum; Helicobacter pylori.
In 1983 , a ba ct eri al organism now known as H eli- and associa tio n with di sea se ha ve recei ved little a tte n-
cobacter py lori was first isolated fro m th e sto machs of tion . Althou gh th ey ha ve been incon sist entl y associ-
human pati ents with gastritis." This organism has since at ed with hi stologic ga stritis, th ey have not previ ou sly
be en shown to be a co m m o n cause of gastritis and ha s been associated with clinical disea se in anim als. G LO
be en associated with peptic ulcer di sease, gastric ca r- ha ve al so been described in an im al s and may be m ore
cin oma , and a number o f othe r co nd it io ns .'>' Because preva lent than in human beings. Their role in d isea se
of its association with peptic ulcer d isease, H. pylori is un cert ain , and they may be normal co m m ensa ls. P-"
has ev o ked a great deal of interest and stud y. Clinical Previous stud ies of naturally oc curring gastritis in cats
trials have shown tha t erad icatio n of infection is as- and dogs have not addressed the role of gastri c bac-
so ciated with healing of ulcers and that tr eatment for teria .1 3.21.27-31
ulcers is in effecti ve in th e absen ce of bacterial era d i- Pr eliminary in vestigation of a group of capti ve che e-
cation ," suggesting that H. pylori is a n important fac to r tahs (Acin onyx j uba tus) with chro nic vo m iting re-
in th e pathogenesis of human pepti c ulc er di sea se.' > ve a led Iymphoplasmacyti c gastritis as sociat ed with
In addition to H. pylori, other gas tric bact eria ha ve gas tric spira l ba ct eria. Thi s study was undertaken to
been described in human beings.v-' but th ey are un - in vestigate th e pr eval en ce of ga stritis in thi s chee ta h
co m mon and ha ve not been cult ured in vitro and thus popu lati on and to investigate th e rol e ofgastric ba ct eri a
cannot be taxonom icall y defin ed. They are m ost o fte n in th e pathogenesis of thi s disea se. Th ree catego ries of
referred to as Gastrospirillum hominis or Gastrospir il- findings a re presented: tissu es were ex a m ined grossly
lum-like ba cteria (GLO). Like H. pylori, they may be and microscopically and the character of th e gastritis
as sociated with gastriti s.s-" was co m pared to that of known d iseases , various tech-
Bact eri a l ga stritis is po orl y docum ented in an imals. niques were used to det ermine th e presence and iden -
R ecent rep orts, however , suggest that gastri c ba ct eri a tity o f gastri c bacteria, and th e role of these bacteria
are relati vely co m m on and that gast ritis ca used by th ese in th e pathogen esis o f gas tritis was addressed .
ba cteria ma y ha ve been co m m o nly overloo ked in th e
past. 12 H. pylori-like o rganis ms ha ve been isolat ed fro m Materia ls and Methods
ferrets with gastritis a nd gastric ulce rsv -? and from Ani ma ls
macaques' a nd pigs" with gastritis. These ba ct eria are Th e population studie d consisted of 25 cheeta hs at the
probably widespread in a nima ls, but their incidence Colum bus Zoo (Co lum bus, OH ) bet ween April 1988 and
55
Downloaded from vet.sagepub.com by guest on October 20, 201556 Eaton et al. Vet Patho l 30: I, 1993
Ju ne 1991 . T wenty-t wo o f th ese a nima ls (cheetah Nos. 1-1 9 Mouse inoculation
a nd 23-25) were in th e regula r co llectio n. A ll of these animals
Sa m ples ofgas tric tissue fro m th e th ree ana tomic sites were
were bo rn in captivity in the U ni ted Sta tes , exce pt for cheetah
co mbi ned and hom ogeni zed in 0.5- 1.0 ml of Brucella brot h
No . 24, whic h was wild caught. T hese ani ma ls were in co ntac t
(Difco Laboratories, Det roit, MI). Homogenat es were used
with each other eithe r d irectl y (in th e same cage) or indirectly
for oral inoc ulatio n of conventio na l BalbC m ice. T wo to fo ur
via com mo n uten sils an d hold ing areas. T hree ani ma ls (chee -
m ice per chee ta h were o rally gavaged wit h 0.2 ml of ho -
tah Nos. 20-22) were not in direct or indirect co ntac t wit h
m ogen at e. Fo ur mi ce were orally inocu lated with a bro th
any of th e othe r an ima ls. These three were acq uire d fro m a
cu lture of Helicobacter o f cheeta h o rigin. Mice were give n
ca ptive breeding facilit y overseas and were kept in qu ar antine
0.2 rnl of broth co ntai ning I x 10 9 cful ml. M ice were e u-
for th e durat ion of th eir sta y at th e Co lum bus Zoo . Gas-
th an at ized 4 to 16 weeks after inoc ulation, and th eir sto m -
tro scop y was perfo rm ed o n four different occasions. Chee ta h
ac hs were culture d an d exa mi ned his to logica lly and ult ra-
No . 23 was exa m ined endosco pica lly in April 1988 a nd Ma y
str uct ura lly for th e presen ce o f bacter ia.
1988. Bet ween Janua ry a nd April 1990, nin e chee ta hs were
exa mi ned endosco pica lly (ch eet ah Nos. 4, 6, 8, 10-13 , I S, Light microscopy
a nd 2 1). O ne chee ta h (N o. (0) was treat ed with a ntibiotics
2 weeks before endosco py. In A pril 1991 , 22 cheeta hs (N os. Sa m ples from eac h bi op sy site were exa m ined und er phase
1-22) were exa mi ned endoscopically. Non e of th ese a nima ls co ntras t illuminati on for the presen ce o f spiral bacteri a. Fou r
had been treat ed with a ntibiotics within 6 months o f ex- to five sa m ples from eac h site were fixed in 10% neutral
a m ina tio n. Three chee ta hs (N os. 23-2 5) d ied and were nec- buffered forma lin and em bedde d in paraffin . Six-micr om eter
ro psied . sec tions were sta ined with hem at oxylin and eos in a nd Wa r-
th in-Sta rry sta ins . Gastritis sco re was determined by ex-
Endoscopy a mi natio n of sectio ns stai ned with hem at oxylin a nd eos in.
Sectio ns were gra de d o n a sca le o f 0-3 as follows : 0 = no
For endoscopic exa mi na tion, all chee ta hs were a nes the- inflammat o ry cells in t he lami na pro pr ia; I = few lympho-
tized with 2 mg/lb int ramuscul a r telazol (A H Robbins, Ri ch - cytes a nd plasm a cells in the lam ina prop ria a nd in some
m ond , VA) a nd int ubated. Anes thesia was maint a ined with cases a mod erat e number of intraepit helia l leu kocytes; 2 =
isofl ura nc , Two-mi llime ter gas tric peroral fo rceps biop sies mo de ra te lym pho plas macytic gas tritis witho ut d istortion of
were ta ken wit h a videoendosco pe (Oly mpus G IF- PV 10 Vid- archi tect ure or epithelial cha nges; a nd 3 = severe Iym ph o-
eoe ndoscope with CV-I Videorecorder, Oly m pus Co rp., Lake plasmacytic or chronic suppurative gastritis with one or more
Success , NY) . Fifteen to 20 biopsies were ta ken fro m three of the foll owing con di tions: large numbers of intraepithelial
si tes from eac h chee ta h: ca rdia, fundus, and pylori c an tru m. leukocytes, disto rt ion of architecture, di latio n ofglands , gland
Sa m ples fro m eac h site were processed fo r bac terial cult ure , abscesses, or supe rficial epi thelial atte nua tio n, erosion, or
urease determinat ion , m ou se inocu lat ion , and histologic, ulcerat ion . Th e gas tritis score was based on th e m ost severely
phase co ntrast, and ult rast ructural exa m ina tions. in flame d sectio n fro m eac h ind ividu al. Sections were scored
witho ut know ledge of th e bacteriologic result s.
Bacterial culture
T wo sa m ples of gastr ic mu cosa fro m eac h site were in- Electron microscopy
oc ula ted o nto Brucella aga r co nta ining 10% rabbit blood and Sa m ples from eac h site were fixed in 3.0% gluta ra lde hyde
Skirrow's suppleme nt (O xoid , Co lum bia, MD). Plat es were (pH = 7.3), po st-fixed in 1.3% os m ium tetroxid e, a nd em-
incuba ted at 37 C in an atm osph ere o f 10% CO 2 for 4 to 5 bedded in Medc ast epoxy resin (T ed Pella, Inc ., Redding,
days. Fo llowing incubati on , a sterile swa b was rubbed over CA) . U ltra thin sections were stained wit h uran yl ace ta te a nd
th e surface of th e plat es and th en streaked o nto blood aga r lead citra te and were exa mi ned with a Ph illips 300 electron
plat es, whi ch were inc uba ted for a n add itiona l 3 days. T his mi croscop e.
technique faci lita ted detecti on of small co lonies o r light
grow th. Resulting co lon ies were ide nti fied as Helicobacter sp . Urease test
by co lony m orphology; Gram's stai n; ultrastructure; pro-
Sa mples fro m eac h site we re placed in 50 III of urease
d uct io n o f urease, oxidase, and ca ta lase ; o bligate mi cro aero-
indicato r bro t h, co ntai ning 330 mM urea, 10 m M sod ium
bic growt h; polyacrylam ide gel electro pho res is; and D NA
phospha te buffer (pH 6.5),0.02% phe no l red (w/v), and 0.02%
rest rict ion endo nuclease a na lysis. ' For mou se inoc ulation,
sod ium az ide (w/v) . A color cha nge from deep or a nge to
bacteria were cult ured in Brucella bro th wit h 10% fetal ca lf
cherry red within 2 ho urs was considered a positive rcac-
serum ." For en zy me- linked im m un oso rb ent assay (ELISA) ,
tion ."
bacter ia were grow n on trypticase soy aga r plat es (Re me l,
Lenexa, KS) at 37 C under micr oaero phili c co nd itio ns for 4
Se rology
to 5 days. In add itio n, Helicobacterfelis, ATCC Ty pe St rai n
49 179, was grown in Brucella broth co nta ining 5% fetal ca lf T o det ermin e a ntibod y titer agai ns t Helicobacter a ntigens ,
seru m. Antigens were prepared as described previ ou sly." th e ELISA was performed as described by Fox et al.' with
Bacteria were harvested into ph osph at e-buffered saline a nd th e follow ing modifi cati on s. Serum was seria lly d ilut ed 2-fo ld
washed . T he cells were di srupted so nica lly, an d cell deb ris a nd assayed at d ilut ion s of I : 64 th rou gh I : 2,000. Anti-ca t
was rem o ved by centri fugati on . T he protein co ntent of th e IgG conj ugated to alkal ine ph osph at ase (Kirkegaard and Per-
supe rna ta nt fluid was measu red by th e Low ry techniqu e." ry, Ga ithe rsburg, MD) was used at a d iluti on o f I : 400. Two
Downloaded from vet.sagepub.com by guest on October 20, 2015Vet Pathol 30: I. 1993 Bacterial Gastritis in Cheetahs 57
Table 1. T he type of path ologic lesion s present in cheetahs with gastritis.
Lesion Cheetah No .
Gr oss"
Rugal hypert roph y 6,8, 10- 13, 15,23-25
Eros ions 6,8, 10- 13, 15,23-25
Ulceration 10, 12, 25
No lesion s 21
Histologic
Lymphopl asm acytic gastritis]' 1-14, 16, 18, 20, 23-25
Intraep ith elial lymph ocytes 1-1 4, 16, 18- 20, 23-2 5
Epithelial atte nuatio n or eros ion 1, 3-6, 8-14,1 8, 23
Fibros is o f the lamina propria 1, 6, 8,1 0,1 2-14 ,1 7-1 9
Prominent globule leu kocytes 1-3,1 8
Lymphoid follicl es in the lamina propria 9, 13, 17,23
Neut rophil s in the mu cosa 5, 12, 23
Suppura tive gas tritis 23
No rrnalj 15, 21, 22
* Cheetah Nos. 6, 8, 10-1 3, 15, 21. and 23-2 5 were examined for gross lesions.
t Gas tritis sco re 2: 2.
*Ga stritis sco re 5 1, no other lesions.
negat ive con tro ls from SPF kittens free o f gast ric bacteria th ese except cheetah No. 2 1 had macroscopi c lesion s
were inclu ded on each plate. Posti ve controls were from a (Table I). The most consis tent findin g was mark ed ,
d og infec ted with H. pylori and a dog in fected with H. fe lis. widesp read th ickenin g of th e gastr ic rugae, which was
For these, anti-dog IgG co nj ugated to alka line phosph atase most pronounced in the fundu s. In addition, seve n
was used at a di lution of I : 400 (Kir kegaa rd an d Per ry). Ti ters anima ls had crateriform lesion s, int erp reted as epit he-
were deter mined as described by Correa et al.'
lial eros ions, and so me had multiple pun ctat e hem-
Re sults orrhages both withi n th e cra ters and sca tte red over th e
hypertroph ic rugae. T wo anima ls had frank ulcerati on
C linical findings
(chee ta h N os. 10 and I I).
Th e most prominent clinical sign seen in th ese chee - Three animals were necropsied (Ta ble I). In two of
tahs was vom iting (ch eetah Nos. 4, 8, 10, II , 12, 23, th ese (N os . 23 and 24), gastritis was seve re and wide-
and 24). Weight loss was observe d in three cheetah s spread . Both were eutha natizcd after a prolonged cou rse
(Nos. 10, 23, and 24). of vo m iting and weight loss, and at necropsy both were
seve rely emaciated. In th ese animals there was severe,
Gross lesion s
widespread gastric rugal hypertrophy. One cheetah (No.
Gross endoscopic findin gs were recorded in eight 23) had no other ma cros copic lesions. The other had
cheetahs (Nos. 6, 8, 10, II , 12, 13, 15,and21).All of mild telangi ectasis of th e liver , two 1-2-cm m yeloli-
pomas (one in the liver and one in th e spleen) , and
multi focal pancreati c du ctular ectasia (extent not re-
corde d). Th e third cheetah d ied naturally of unrelat ed
causes and had gastric rugal hypertrophy that was mod-
erate, co nfined to th e fundi c region , and surrounded a
gastric ulcer. Epith elial erosions, sim ilar to th ose seen
endosco pically, were present in th e areas of hypertro-
ph y (Fig. I).
Hi stologic lesion s
The most consis tent lesion in biop sies from a ffected
chee tahs was seve re Iymph opl asm acytic gastritis (Ta-
ble I), whi ch was cha racteri zed by marked , wides pread
infiltration ofly m phocytes and numerous pla sm a cells
Fig. 1. Sto mac h; chee ta h No. 25. Rugal folds ar e th ick- int o th e lamina propria and subm ucosa. In th e most
ened (arrow) and epithelial erosions are present (ar rowhead). seve re cases, the inflammatory infiltrate di storted or
Bar = I em . effaced th e normal architecture (Fig. 2). Intraepithelial
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Fig. 2. G as tric biop sy; cheeta h NO. 6. Th e normal architecture is di storted by a mark ed lymphopl asmacytic infiltrat e.
N ume ro us intraepithelial leuk ocytes (arrows), and super ficial epithelial atte nuation and eros ion (arrowhead) ar e present.
H E. Bar = 50 J.Lm .
Fig. 3. Gastric biopsy; chee ta h No . 6. Higher magnificat ion of Fig. 2. N umerous plasm a cells (arrow) are present.
Bar = 25 J.L m .
Fig. 4. Gastric m ucosa; cheetah No. 23 . Su ppurati ve gastritis is charac terized by a gland abscess and neutrop hils within
th e epithelium and lami na prop ria (arrows). Bar = 30J.L m
leukocytes; attenuation, ero sion , and ulceration of the In all three cheetahs necropsied , the most prominent
super ficial epithelium ; and numerous plasma cells (Fig. lesion was gastric muc osa l hyp erplasia (Tabl e I). In
3) were seen in all cheetahs with seve re gastritis (gas- th e mo st seve rely affected animal (No. 23), there was
tr itis score = 3). Prominent globule leukocytes and a seve re chronic suppurative gastritis . In thi s animal,
lymphoid follicles within the gastric mu cosa were seen neutrophilic inflammation was wides pread, as were
in a few ani mals (Table I). Fibrosis of the lamina pro- glands distended with neutrophils and cellular debris
pria was seen in ten animals but was mild . In all an- (gland ab scesses) (Fig. 4). Lymphoplasmacytic inflam-
imals, inflammation was multifocal and was most se- mation was moderate. In th e other two cheetahs nec-
ve re in the cardia and fundus and least severe in the ropsied, mucosal hyperplasia was accompanied by
pyloric antrum; but in many cases , no rm al mu cosa and lymphoplasmacytic gastritis typical of that seen in th e
seve rely inflam ed mu cosa could be found in th e same chee tahs exam ined endosco pically.
region of th e sto mach. Becau se of th e sma ll size of the Warth in-Starry-sta in ed sec t io ns reveal ed large
biop sies in those animals exa mined endoscopically, numbers of tightly coiled bacteria 7-10 /-Lm long (Gas-
gastric mucosal hyperp lasia could not be evaluated . In trospirillum-like organisms [G LO]). These were pres -
five cheetahs, the gastritis score was 1. In these animals, ent in 22/25 animals (Table 2). Animals from the reg-
Iymphoplasmacytic inflammation was mild and was ular collection and quarantined an imals were affected.
conside red to be within normal lim its. Two of these Th ese bacteria were present within glands, in surface
anima ls (chee ta h Nos. 19 and 17) had addi tio na l his- mucus, and in som e cases within gastr ic parietal cells.
tologic lesions (Table I). Cheeta h No s. 15, 21, and 22 No relationship between the presen ce of these bacteria
had no add itional lesion s. T wo of th ese an imal s (Nos . and th e seve rity of gastritis was apparent (Table 2). All
2 1 and 22) were in quarantine. three un infected animals had gastritis scores of 3.
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Table 2. Mi crobi ological a nd h istological findings in a group of ca ptive c hee ta hs with gas tritis.
Cheetah No.
Age
Sex
Helice- Gast ro- Gastritis Bacterial Species and Gastritis Score in
(years) bacter" spirillum" Scoret Previous Biopsy:j:
R eg u la r
co llecti on
I 12 F 3 2 3 N O§
2 10 F 3 2 3 NO
3 9 M 0 3 2 NO
4 8 F I 0 3 Helicobacter. Gastrosp irillum, gas tri tis sco re = 2
5 9 M 3 0 3 NO
6 7 M I 2 3 Helicobacter, Gastrospirillum , gas tritis sco re = 3
7 6 F I 3 2 NO
8 6 M 2 3 3 Helic obacter, Gastrospirillum , gas tritis sco re = 3
9 8 M 0 3 3 NO
10 4 M 0 2 2 N o ba ct eria, gastritis score = 3
II 4 F 3 2 3 Helicobacter, Gastrospirillum, gastritis score = 3
12 4 M 3 3 3 Helicobacter, Gastrospirillum, gas tritis score = 3
13 4 M 3 I 3 Helicobacter, Gastrospirillum , gas tritis sco re = 3
14 3 F 3 2 3 NO
15 4 M 0 2 I Helicobacter, Gastrospirillum , gas tri tis sco re = 3
16 2 M I 3 2 NO
17 2 M I 3 1 NO
18 2 F I 2 3 NO
19 2 F I 3 I NO
23 10 M 3 0 3 Gastrospirillum, lymphocyti c gas tri tis (sco re not giv en)
24 15 F NO 3 3 NO
25 6 F I 3 2 NO
Q uarant ine
20 3 F 0 3 2 NO
21 3 F 0 3 I Gastrospirillum , gas tritis score = 3
22 4 F 0 3 I NO
* 0 = no bacteria in histologic sections or culture, I = bacteria rare in Warth in-Star ry-stain ed sections but present in culture, 2 = bacteria
easily found in histologic sectio ns, 3 = numerous bacteria in histologic section.
t Gastritis was scored 0-3 (see text). Scores are given for the most recent endoscopy (cheetah Nos. 1-22) or for the necropsy examinations
(cheetah Nos. 23-25).
:j: Previous biopsy results are given for those animals examined more than once.
§ ND = not done.
I-Ielicobacter spp. were present in histologic sections seen by light microscopy. Of these 19 cheetahs, four
from cheetah Nos . 1,2,4,5 ,6 ,8 , 11-14, 17, and 23 . also had a second typ e of GLO. Th ese bacteria were
These bacteria were smaller than GLO (1-3 j.Im long), more loosely coiled, were 7-10 urn long, had paired
more loosely coiled, and difficult to identify in section, helical filaments closel y associated with the out er
particularly in animals with large numbers of GLO. membran e (Fig. 6), and were similar in size and overall
Thus, in som e an imals, I-Ielicobacter sp. could only be shape to the GLO without helical filam ents; th e two
identified by culture. I-Ielicobacter sp. were present in were not distinguishable by light mi croscopy.
17/ 25 chee tahs (Table 2). Som e relationship was sug- Short (1-3 j.Im) curved bacteria with no surface fea-
gested betw een the presence of I-Ielicobacter and gas- tures were identified within the gastric glands of four
tritis. Of the six animals in which I-Ielicobacter was chee ta hs (Nos. 5, 6, 13, and 23; Fig. 7). In the most
not identified , onl y one had a gastritis score greater seve rely affected animal (No. 23), these were the onl y
than 2. bacteria seen at necrop sy, alth ough this animal had
G LO when examined endosco pically some months
Ultras tructura l findings earlier (Table 2).
Gastric tissue from all 25 cheetahs was examined
ultrastructurally. Nineteen cheeta hs had tightl y coiled
Bacteriologic findings
bacteria that were long (7-10 j.Im) and had tufts of
flagella at one or both poles (Fig. 5). These bacteria Urease activity, indicative of the presence of bac-
had no external structures. They were interpreted as teria, was present in gastric tissue from 21/22 cheetahs
GLO, corresponding to the long, tightl y coiled bacteria examined endoscopically (all except cheetah No .4).
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."
Fi~. 5. Tra nsmission electro n micrograph . Gastric gland ; chee ta h No . 15. Gastrospirillum-like organis ms ar e present
within the gland . No te polar flagella (arrow) and lack or superficial structures . Bar = 0.75 j.Lm.
Fig. 6. T ran sm ission electro n micrograph . Gastric gland ; chee ta h No, 24 , Gastrospirillum -like organism with helical
filam ent s (ar row) is present. Bar = 250 nm .
Squash preparations of tissues from cheetah Nos. 4,6 , icobacter sp. recovered from infected cheetahs was not
8, 11- 13, 15, and 21 were exa m ined und er phase con- infectio us for mic e.
trast illumination. All of these cheetahs had acti vely
Serologic findings
motile pleomorphic sp iral bacter ia.
Helicobacter sp. were recovered fro m 15/2 2 cheeta hs T o determine th e durati on of infection in th ese chee-
exa mi ned endosco pically (Table 2). All 15 animals with tah s and to evaluate serologic exa m ina tion as a mean s
Helicobacter were from th e regular co llection. None of of diagn osis, sera from selected cheeta hs were tested
th e three cheetahs from outside the collection were for antibodies against two Helicobacter species. Three
infected with Helicobacter. Helicobacter sp. were re- of nin e cheeta hs tested had serum antibodies against
covered from one necropsied cheetah (No . 25). Cul- both the Hel icobacter isolated from the cheetahs and
tures from che etah No. 23 were overgrown with bac- a related organism , Helicobacter fe lis (Table 3). Pres-
terial conta minants. Chee tah No. 24 was not cultured. ence of an antibody titer was not correlat ed with se-
GLO co uld not be isolated in vitro, but were iden- verity of lesions or clinical signs. Titers of antibody
tified in the gastric mu cosa ofall m ice orally inoculat ed directed against H. fe lis were lower th an th ose of an-
with gastric mucosa from infected cheeta hs. T wo of tib od y di rected against autologous Helicobacter.
the seve n chee tah sto machs fed to mi ce had bacteria Discussion
both with and without helical filaments, but ultrastru c-
tural exa m ina tion revealed onl y smo oth bacteria in Character of gastritis
gastric mu cosa from mice. GLO were not pre sent in Gastritis in animals has received littl e attention, and
four control mice not given gastri c homogenates. Hel- in mo st cases gastritis in carnivores is considered id-
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Table 3. Serologic responses to Helicobacter sp , and IJ.
felis in cheeta hs,
Titer
Cheetah Helico-
No. Date bacter sp. H. felis
4 30 Oct 86 0* 0
4 10 Jan 90 0 0
6 2 Jan 87 0 0
6 28 Feb 90 0 0
8 2 Jan 87 0 0
8 7 Feb 90 0 0
10 27 Dec 89 955 64
10 31 Jan 90 III 64
II 21 Feb 90 416 169
12 12Dec88 0 0
12 24 Jan 90 0 0
15 14 Feb 90 0 0
21 6 Sep 89 0 0
23 25 Jan 89 724 64
* None. Titers less than 64 were considered negative.
postmortem examination; furthermore, protein-losing
enteropathy, typical of hypertrophic gastritis in Basenji
dogs, was not a clinical feature of this disease in chee-
tahs.
Parasitic gastritis has been described in cats with
Ollulanus tricuspis ." In spite of extensive gross and
histologic examination, O. tricuspis was not seen in
these cheetahs; furthermore , the histologic lesions as-
sociated with O. tricuspis are different from the lesions
seen in these cheetahs." Although epithelial hypertro-
phy and hyperplasia are prominent in both diseases,
fibrosis of the lamina propria, a striking feature of O.
Fig. 7. Transmission electron micrograph , Gastric gland; tricuspis infestation , was mild in these cheetahs. In
cheetah No, 23, The gland is packed with Heli cobacter-like addition, although both globule leukocytes and mu-
organisms, Bar = 2,5 /-Lm, cosallymphoid follicles were present in these cheetahs,
neither were prominent to the extent described in in -
iopathic, Reported causes of gastritis in carnivores in - festation by O. tricuspis.
clude idiopathic hypertrophy. v-" >' mycotic and viral Other causes of gastritis are unlikely in these chee-
infections,' duodenal reflux , 1 parasitic infections, 14 and tahs. Reflux and chemical gastritis are sporadic in na-
idiopathic pyloric hypertrophy in small-breed dogs. ' ? ture and are unlikely to occur as outbreaks. Uremia
In a recently published report, gastritis in dogs was causes gastric lesions that are dissimilar to those seen
classified histologically, but etiology of gastritis and in these animals. 1 No in fectio us agents other than gas-
the role of gastric bacteria were not addressed. v -'? tric spiral bacilli were identified in these cheetahs,
The gastritis in the cheetahs of this report does not Gastritis in these cheetahs was somewhat similar to
correspond to any type of previously described gastritis gastritis caused by Helicobacter pylori in human beings.
in carnivores, Hypertrophic gastritisv -" -" is most sim- In human beings, bacterial gastritis is characterized by
ilar to the gastritis in these cheetahs, Both are char- superficial lymphoplasmacytic gastritis, often with
acterized by marked mucosal hypertrophy, inflam- neutrophils and gland abscesses.v-" Such lesions are
mation , dilated glands, and intraepithelial leukocytes; very similar to those seen in these cheetahs. Lymphoid
furthermore , the clinical signs of vomiting and wasting fol licles" and intraepithelial leukocytes? have also been
are similar in dogs and cheetahs, In dogs, however, described in human beings (as in cheetahs), although
gastritis is associated with lym pho plasm acytic enteri- they are less prominent than lymphoplasmacytic in-
tis . T he small intestine in the cheetahs examined his - filtrates . Thus, in these cheetahs, gastric lesio ns were
to logically was not biopsied, but intestinal lesions were most similar to those found in human beings with H.
not seen in any of the three cheetahs examined by pylori,
Downloaded from vet.sagepub.com by guest on October 20, 201562 Eaton et al. Vet Pathol 30:I, 1993
Id entification of gas tr ic bact eri a o f th e bacteria ide ntified eithe r separa tely or in co m -
In human beings, H. pylori is most co m monly iden- binati on . In addi tio n, H elicobacter are difficult to d is-
ti fied on th e basis of hi stology, culture, biop sy ur ease tin guish fro m G LO und er th e light m icro scop e, par-
test, and sero logy.' These are all con sidered sensitive ticul arly when few bacteria are pr esent. Sim ilarly, th ey
and specific test s. Nonculturable ba cter ia (GLO) ha ve m ay not be identifiable by electro n m icroscopy. Thus,
been identified by light and electro n m icroscop y, ur e- culture is th e onl y reliable way to determine th e pr es-
ase test , an d m ou se inocul at ion .v'v-? In th e cheetahs ence of th ese ba cteria. The orga ni sms are fastidio us,
H elicobacter could be identi fied by culture, but wh en however , and infectio n may be spa rse . T hu s, a negat ive
G LO were present, H elicobacter co uld not be reliabl y culture ca nno t be tak en as pr oof tha t th e bacteria are
identified by histologic exami na tion. In th e pr esen ce ab sent. There are, however , so me indicat ion s th at He l-
of G LO, th e smaller H elicobacter were often not di s- icobacter may be th e more pathogen ic organi sm for
tinguish abl e. Thus, culture was th e only reliabl e way th ese chee ta hs . Infecti on by H elicobacter was restri cted
to ide nti fy H elicobacter. Biop sy urease test identifi ed to th ose animals in the regular co llection, as was severe
ba cter ia in all but one of th e chee tahs tested but did gas tritis . Non e of th e three chee tahs in quar antine had
not distinguish H elicobacter and GLO. Thus, th e bi- H elicobacter, non e had clini cal signs , and only one ha d
opsy urease test is useful for diagnosis of the pr esence m od erate gastritis. In add itio n, of th e two chee ta hs
of gastric bacteria in chee ta hs but is no t helpful in wit h H elicobacter in th e ab sen ce of G LO, both had
di st ingui shing bacterial ty pes . GLO co uld not be cul- severe gastritis, and one (chee ta h No. 23) was th e m ost
tured , but these bact eria co uld be iden tified by histo- severely affected animal in th e co llection. In co ntrast,
logic exami nation and m ou se inoculation. Bacteria with th ere was no direct corres ponde nce bet ween infecti on
and witho ut helical filam ents co uld be di stinguished with GLO and gastritis. Of th e seven chee tahs with
by electron m icroscop y. Thus, exa mi na tio ns by cult ure G LO alone (incl udi ng th e three in qu aran tine), only
and light an d electron mi cro scop y were all necessary one ha d severe gastritis . Becau se of the d iffi cult y of
to d iagnose an d ide ntify gas tric bacteria in th ese chee- cult uri ng H elicobacter, infec tio n by this organism can -
tahs. not be com pletely rul ed out in thi s case. Thus, if GLO
In human beings, sero logic exa mi na tio n is oft en used contribute to gastritis, th ey probably do so in co m bi-
as presumptive ev ide nce of infectio n by H. pylori.' it nati on with other ba cteria.
is used to identify infec ted peo ple and to determine All anima ls in thi s collectio n had at least one type
th e onse t of infectio n in population s. In chee ta hs, in of gastric bacteria. Thus, th e co ntribution of particular
contrast to human beings, sero logic exa mi na tion was ba cter ia to gastri tis cannot be det er m ined with cer-
not useful in dia gnosis; onl y three of nine infected chee- tainty. However, th e cha racte r of th e gastri tis and th e
tahs tested had sero logic titers to H elicobacter of chee - lack of other inciting causes pr ovid es suggestive ev i-
tah origin. den ce for a bacterial etio logy . Definitive de te rmi na tio n
of the ro les of th e various bacteria identified awa its
further study . It will be necessary to determ ine whether
Relationship of bact eria to gastritis th ese ba cteria are able to reproduce th e di sea se and
T here is some ev ide nce tha t gastric bacter ia cause fulfill Koch' s postulates. In addition , un in fected chee -
gastr itis in an imals." G LO have been associated wit h tah s must be ide nti fied. Altho ugh gastric bacteri a are
lym ph ofolli cul ar gastritis in dogs;" however , th ese or- th e most likely etio logic agents in this disease, the pres-
ganis ms m ay be normal co m me nsa ls, at lea st in some ence of ot her unidentified agen ts cannot be ruled out.
species. " H elicobacter sp. have been associated with T oxic agents, undet ect ed ba cteria or viruses, and a u-
gastritis in animals. H elicobacter m ustelae, a sho rt spi- toimmune or othe r ph en om ena must be co nsidered .
ral sim ilar to H. pylori. has been associated with chro n-
ic superficial gastritis and ulcerati on in ferrets.v'? and
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Req uest reprints from Dr. K. A. Eaton , Department of Veterinary Pathobiology, O hio State University, 1925 Coffey Road ,
Co lumbus, O H 43210 (USA) .
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