Previous Article | Next Article 
Journal of Virology, November 1998, p. 9298-9302, Vol. 72, No. 11
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Development of a Heterologous Model in Germfree
Suckling Rats for Studies of Rotavirus Diarrhea
C.
Guerin-Danan,1
J. C.
Meslin,1
F.
Lambre,1
A.
Charpilienne,2
M.
Serezat,1
C.
Bouley,3
J.
Cohen,2 and
C.
Andrieux1,*
Unité d'Ecologie et de Physiologie du
Système Digestif, Equipe Métabolites Bactériens et
Santé,1 and
Unité de
Virologie Immunologie Moléculaire,2 INRA,
78352 Jouy-en-Josas Cedex, and
Département Nutrition,
Danone, Le Plessis Robinson 92350,3 France
Received 19 February 1998/Accepted 24 July 1998
 |
ABSTRACT |
Germfree suckling rats were infected with an SA11 rotavirus strain.
Infected pups developed diarrhea associated with histopathological changes. The virus was detected in feces and in the small intestine. Cellular vacuolation was observed in the villi of the jejunum. These
results provide a new model for further investigations of group A
rotavirus infection.
| |
TEXT |
Group A rotavirus is the leading
cause of diarrhea among 6- to 24-month-old children worldwide. In
industrialized countries, the illness is usually self-limiting, with a
mean duration of 4 to 7 days (28). However, it is estimated
that 870,000 deaths/year are caused by rotavirus-associated diarrhea,
principally in developing countries (10). The primary
symptom is watery diarrhea, frequently associated with severe
dehydration (5). Limited investigation by mucosal biopsy of
infected infants has shown that rotavirus principally infects the cells
of the small intestine.
The physiopathology of rotavirus-associated diarrhea has been
experimentally studied in several animal species, including monkeys
(22), calves (19), dogs (14), pigs
(31), and rabbits (6). However, most of these
animals are costly and cumbersome to maintain (4).
To date, the murine model has been the most often used model for
studying rotavirus infection. However, difficulties in growing murine
rotavirus strains to high concentrations in tissue culture have limited
their use (21). The use of heterologous rotavirus, particularly the simian SA11, strain, has led to physiopathological symptoms in mice identical to those produced by the murine strain (3, 17, 18, 21) and may limit cross contamination among animals bred in the laboratory (6).
Surprisingly few studies of rotavirus infections have been conducted
with rats, even though this animal is commonly used in the experimental
field. To our knowledge, these experiments have described only
homologous models using rats infected with group B rotavirus (24,
27, 29, 30).
The purpose of the present study was to investigate a new simplified
model for rotavirus investigation. Clinical and histopathological symptoms of SA11 rotavirus strain-associated diarrhea in germfree suckling rats are described.
Animal inoculation.
The use of animals maintained in sterile
isolators prevents the exposure to extraneous rotavirus or other
enteric pathogens (31). Furthermore, intestinal microflora
can enhance the defense mechanisms of the host against pathogens
(25) and can influence rotavirus infection, as described
previously for mice (13). Under such conditions, the
germfree status makes it possible to study rotavirus infection without
any disturbing interference by other microorganisms. In our experiment,
rats (Fischer F344 strain) were delivered and maintained in sterile
isolators until the age of inoculation, and then they were housed in
separate isolators. Germfree pups were orally inoculated at 2, 5, or 8 days of age. Inoculation was performed with a graduated plastic Pasteur
pipette (Miniliquipette; Prolabo, Fontenay sous bois, France). After
inoculation, infant rats were returned to their dams and allowed to
suckle. Feng et al. (9) have demonstrated that diarrhea was
dose dependent according to the rotavirus strain. The minimal dose of
rotavirus provoking diarrhea in mice is 105-fold higher
with heterologous strains than with homologous ones. SA11-infected mice
developed diarrhea if the rotavirus dose was higher than 4 × 105 PFU/ml (17). Therefore, we chose to infect
the rats with a high concentration of SA11 rotavirus to ensure that
diarrhea would occur. Rats received a single 0.1-ml dose of either
virus suspension (1.6 × 109 PFU/ml), prepared as
described by Jourdan et al. (15), or modified Eagle medium
(MEM) as a control.
Clinical signs of diarrhea.
Diarrhea was assessed in 23 to 35 infected rats and 16 to 25 control rats per age group. Rats were
checked for diarrhea by gentle massage of the abdomen. Diarrhea was
diagnosed when poorly formed yellow-green feces occurred immediately
upon palpation. Control rats were treated identically to infected ones.
Individual stool specimens were carefully collected in sterile plastic
tubes on a weighed piece of plastic. Samples were stored at 
80°C
before analysis.
Our results demonstrate that diarrhea was induced in young germfree
rats by inoculation with 108 PFU of SA11 rotavirus. The
infection was self-limiting, and clinical signs were easy to interpret
since diarrheal feces occurred spontaneously in infected pups as
described previously for rat rotavirus-infected rats (24).
Although feces occurred in control pups upon gentle palpation as well,
samples were obtained from <20% of these animals (Fig.
1) except at 24 and 48 h
postinfection (hpi) for the control rats inoculated at 8 days.
Moreover, stools differed in color and weight. Control pups produced
dark and small feces. The weight of stool samples (mean ± standard deviation) collected from 5-day-old infected rats was
significantly greater (8 ± 4 mg) than the weight of stool samples
collected from control rats (4 ± 4 mg) (P = 0.04 [t test]). The percentage of diarrheal samples was
significantly higher from 24 to 192 hpi in pups infected at 2 days,
from 48 to 144 hpi in pups infected at 5 days, and from 72 to 96 hpi in pups infected at 8 days (Fig. 1). The susceptibility of germfree rats
was age dependent. The younger the rats were, the longer the period
during which they developed diarrhea was. We found that germfree rats
were susceptible to rotavirus infection for at least 8 days after
birth. Vonderfecht et al. (30) have demonstrated that
conventional rats were susceptible to a rotavirus-like agent up to the
age of 14 days. In these authors' experiments, clinical symptoms of
rotavirus diarrhea were defined as liquid or poorly formed content in
the distal colon associated with lesions of the perianal skin (29,
30). The relationship of age to the susceptibility to rotavirus
infection may be due to the maturation changes in the gut and to the
availability of rotavirus-specific receptors on the enterocytes
(28).
View larger version (14K):
[in this window]
[in a new window]
|
FIG. 1.
Percentage of rats delivering feces immediately upon
gentle palpation. Rats were inoculated with either SA11 rotavirus
(solid bars) or MEM as a control (open bars). An asterisk denotes a
percentage of samples significantly different from that of the control
group (P < 0.05 [ 2 test]). Rats were
infected at the age of 2 days (infected group, n = 35;
control group, n = 20) (A), 5 days (infected group,
n = 34; control group, n = 25) (B), or
8 days (infected group, n = 23; control group,
n = 16) (C).
|
|
Rotavirus shedding.
Virus antigen load was determined by
a double sandwich enzyme-linked immunosorbent assay (ELISA)
adapted from a previous study (12): each fecal sample was
diluted in 170 µl of phosphate-buffered saline (PBS) (pH 7.2).
Negative and positive control tests were included in each plate. The
baseline was determined with PBS. Samples obtained from noninfected
pups produced optical densities (OD) between 0.01 and 0.06. Positive
tests consisted of serial dilutions of the viral inoculum (from
1.6 × 108 to 1.9 × 105 PFU/ml).
Assays were performed in duplicate. The viral antigen load in samples
was determined relative to the standard curve and the dilution of
samples in each plate. Crude OD are not relevant to this study since
they are not really comparable. Indeed, the weights of feces were
different whereas all feces were diluted in the same buffer volume.
Therefore, dilutions and consequently OD were different from one sample
to another, although viral antigen loads could be similar. Using this
assay, we have been able to detect rotavirus antigen in virus stock
with a titer of 106 PFU/ml.
Rotavirus was detected in fecal samples for 192, 144, or 120 hpi in
pups infected at 2, 5, or 8 days, respectively (Fig.
2). Direct detection of rotavirus in the
intestinal segments of rats demonstrated that it remained in the small
intestine for at least 5 days postinfection (p.i.). Five-day-old pups
were inoculated with 0.1 ml of a mixture containing rotavirus (1.6 × 109 PFU/ml) and spores of Bacillus subtilis
(3 × 106 CFU/ml), usually used as an intestinal
transit marker (7, 8). At selected time intervals, two to
three pups were sacrificed. The entire intestinal tract of each pup was
removed and divided into stomach, small intestine, and colon segments.
The segments were separately diluted in sterile water (stomach and
small intestine, 1/10 [wt/vol]; colon, 1/100 [wt/vol]). Then, they
were homogenized in an Ultra-turrax homogenizer (Bioblock, Paris,
France) and tested for the presence of rotavirus and spores. Rotavirus
was tested by ELISA. B. subtilis spores were numerated on
agar medium (meat extract [8 g/liter], yeast extract [2 g/liter],
manganese sulfate [0.04 g/liter], glucose [1 g/liter], Bi-Tek agar
[28 g/liter]) after incubation at 53°C for 24 h under aerobic
conditions. The count was expressed as log10
CFU/milliliter. Rotavirus and bacterial spores were found in the
stomach immediately after inoculation but not at any time point
thereafter. Spores were found decreasingly from 12 to 120 hpi in the
colon but were not found in the small intestine during this period. By
contrast, rotavirus remained in the small intestine at 12 to 120 hpi
and was found in the colon (data not shown). Similar experiments with
mice have shown that murine rotavirus was detected in mucosal
homogenates from germfree mice at 8 days p.i. or from conventional mice
at 13 days p.i. (13). Rotavirus was present for 7 days p.i.
in the intestinal contents of conventional mice (16).
View larger version (9K):
[in this window]
[in a new window]
|
FIG. 2.
Relative viral antigen load/virus concentration ratio
determined in feces of infected pups. Rats were inoculated at the age
of 2 (A), 5 (B), or 8 (C) days. Feces were sampled and assayed for
rotavirus antigens by ELISA. A standard curve obtained from the viral
inoculum was determined for each plate. The viral antigen load in feces
was determined conventionally relative to the standard curve. Data
represent individual values obtained for fecal specimens.
|
|
Histological examination.
Infected and control pups were
inoculated at 5 days. Three pups were sacrificed 3 days p.i. Jejunal
segments were removed. The cell morphology was observed by light
microscopy. Fresh tissues were fixed in slightly modified Carnoy
fixative (absolute ethanol-30% formaldehyde-acetic acid [6:3:1;
vol/vol/vol]), dehydrated, and embedded in paraffin. Paraffin sections
(6 µm) were cut on a microtome (Leitz, Wetzlar, Germany) and were
polychromatically stained. Acid and neutral mucin were stained with
Alcian blue and periodic acid-Schiff stain, respectively. Cellular
nuclei were stained with Hansen ferric trioxyhematein, muscular fibers
were stained with picric acid, and collagen and basement membranes were
stained with indigo carmine. Care was taken that only longitudinal
sections cut perpendicularly to the muscular mucosae were studied.
Villus height, crypt depth, and the number of mucus-containing cells were determined for 10 different villi from the same intestinal section.
Morphologic changes associated with rotavirus infection were limited to
cellular vacuolation. The control section presented
cells characterized
by nuclei localized at their base and by a
large supranuclear area
occupying almost the whole apical cytoplasm.
Nuclear polarity and
cellular morphology of the villus basis were
not affected by the virus.
By contrast, large areas of cellular
vacuolation were observed in
infected intestinal sections at 3
days p.i. (Fig.
3). These lesions were revealed in the
enterocytes
localized at the upper one-third of the intestinal villi.
The
villus height/crypt depth and number of mucus-containing
cells/villus
height ratios of infected animals were not significantly
different
from those of control animals (data not shown). Mucus in
infected
villi was not completely released on the intestinal lumen, as
suggested by the presence of dark stained goblet cells. In rats
infected with a rotavirus-like agent, histological changes were
defined
in the distal small intestine by villus shortening, syncytial
cell
formation, and crypt elongation but limited vacuolation at
the tip of
the villus (
30). The damage intensities caused by
rotavirus
infection in the different parts of the small intestine
differ
according animals and rotavirus strains as reported by
different
authors. Some studies have reported greater changes
in the ileum than
in the jejunum in gnotobiotic dogs infected
with a canine rotavirus
(
14) or in suckling mice infected with
nonmurine rotavirus
strains (
23). By contrast, Kubelka et al.
(
17)
have observed more changes in the jejunum in SA11-infected
mice. Others
have demonstrated similar changes in the ileum and
jejunum in
SA11-infected mice (
18) or in group B rotavirus-infected
rats (
24). Heyman et al. (
13) have reported the
appearance
of vacuoles in enterocytes of the villi of the proximal
small
intestine associated with a release of mucus in the lumen of mice
infected with a murine strain.
View larger version (120K):
[in this window]
[in a new window]
|
FIG. 3.
Histological aspects of jejunum villi of germfree
suckling rats examined on day 3 p.i. Five-day-old rats were
inoculated with either SA11 rotavirus (A) or MEM as a control (B).
Intestinal sections were polychromatically stained and observed by
light microscopy. Magnification, ×300.
|
|
Rotavirus structural proteins in the cells were detected by
immunofluorescence. Paraffin sections were deparaffinized with
toluene
and rehydrated with decreasing concentrations of ethanol
(100, 95, 90, and 70%). The slides were washed in water and then
in PBS. After
drying at room temperature, the preparations were
incubated for 1 h at 37°C with a hyperimmune rabbit antirotavirus
serum. The serum
was diluted 1/100 in PBS containing 3% bovine
serum albumin. The
slides were again washed with PBS and incubated
for 30 min at 37°C
with a goat anti-rabbit immunoglobulin-fluorescein
isothiocyanate
conjugate (Boehringer SA, Meylan, France) diluted
1/100 in PBS
containing 3% bovine serum albumin. After final washings
with PBS,
fluorescence was examined by UV light microscopy. Greenberg
et al.
(
11) have reported that rotavirus has a tissue tropism
limited to the upper part of the villus. Our observations from
3 days
p.i. are in agreement with this result. Rotavirus antigen
was detected
by immunofluorescence in differentiated cells at
the tip of the villus,
although adjacent vacuolated cells closer
to the crypt did not show any
fluorescence (Fig.
4). Similar results
have been reported in infant rats infected with a rotavirus-like
agent
(
30): virus particles in the upper one-third of villi
of the
small intestine have been found to be associated with histological
changes.
View larger version (84K):
[in this window]
[in a new window]
|
FIG. 4.
Detection of rotavirus antigens in jejunum cells of
germfree suckling rats 3 days p.i. Five-day-old rats were inoculated
with either SA11 rotavirus (A) or MEM as a control (B). Rotavirus
structural proteins were labeled with a hyperimmune rabbit
antirotavirus serum and then a goat anti-rabbit
immunoglobulin-fluorescein isothiocyanate. Rotavirus antigens were
detected under UV light by immunofluorescence. Magnification, ×400.
|
|
Minimal replication and histological lesions have been associated with
diarrhea in animal models, suggesting a toxin-like
effect of rotavirus
(
1,
26). In our experiment, diarrhea
occurred whereas
enterocyte morphology presented mild modifications
and vacuolation was
observed in cells in which no rotavirus structural
antigens were
detected. This result might be explained by the
mechanism of infection
proposed by Ball et al. (
1). Rotavirus
particles bind some
cells, resulting in virus entry and gene expression
at the tip of the
villus. Then, the nonstructural protein NSP4,
expressed in infected
cells, is released into the lumen and interacts
with a specific
receptor on adjacent cells, increasing the endogenous
secretory
pathway.
Group A rotavirus is a significant human and veterinary pathogen in
terms of morbidity, mortality, and economic loss (
4).
Bass
et al. (
2) mentioned that natural group A rotavirus
receptors
had never been observed in rats. To our knowledge this is the
first report describing diarrhea in germfree rats infected with
a group
A rotavirus strain. The suckling rat model presents almost
the same
advantages as mouse or rabbit models in terms of cost,
diarrhea
intensity and duration, and histopathological changes.
However, one
advantage of the rat model over the other small-animal
models is found
with regard to the clinical signs. In rabbits,
both infected and
control animals normally excrete soft, moist
feces (
6). It
is of interest that our present model may further
the knowledge of
rotavirus infection. This report clarifies the
previously held concept
that group A rotavirus does not infect
rats (
2).
To date rotavirus pathogenesis remains poorly understood
(
1). The new animal model presented here may be useful for
further
investigations of group A rotavirus infection.
| |
ACKNOWLEDGMENTS |
We thank M. K. Estes (Baylor College of Medicine, Houston,
Tex.) for kindly providing the rotavirus strain.
This work was supported in part by the CIRDC, Danone, France.
| |
FOOTNOTES |
*
Corresponding author. Mailing address: UEPSD-MBS,
INRA-Domaine de Vilvert, 78352 Jouy-en-Josas Cedex, France. Phone: 33 1 34 65 24 68. Fax: 33 1 34 65 24 62. E-mail:
andrieux{at}biotec.jouy.inra.fr
| |
REFERENCES |
| 1.
|
Ball, J. M.,
P. Tian,
C. Q. Zeng,
A. P. Morris, and M. K. Estes.
1996.
Age-dependent diarrhea induced by a rotaviral nonstructural glycoprotein.
Science
272:101-104[Abstract].
|
| 2.
|
Bass, D. M.,
E. R. Mackow, and H. B. Greenberg.
1991.
Identification and partial characterization of a rhesus rotavirus binding glycoprotein on murine enterocytes.
Virology
183:602-610[Medline].
|
| 3.
|
Bell, L. M.,
H. F. Clark,
E. A. O'Brien,
M. J. Kornstein,
S. A. Plotkin, and P. A. Offit.
1987.
Gastroenteritis caused by human rotaviruses (serotype three) in a suckling mouse model.
Proc. Soc. Exp. Biol. Med.
184:127-132[Medline].
|
| 4.
|
Burns, J. W.,
A. A. Krishnaney,
P. T. Vo,
R. V. Rouse,
L. J. Anderson, and H. B. Greenberg.
1995.
Analyses of homologous rotavirus infection in the mouse model.
Virology
207:143-153[Medline].
|
| 5.
|
Burns, J. W., and H. B. Greenberg.
1994.
Viral gastroenteritis.
Infect. Dis. Clin. Pract.
3:411-417.
|
| 6.
|
Conner, M. E.,
M. K. Estes, and D. Y. Graham.
1988.
Rabbit model of rotavirus infection.
J. Virol.
62:1625-1633[Abstract/Free Full Text].
|
| 7.
|
Contrepois, M., and P. Gouet.
1969.
Utilisation d'une technique microbiologique pour la mesure de la vitesse de transit des microparticules dans le tractus digestif des ruminants.
C. R. Acad. Sci. Paris
268:1757-1759.
|
| 8.
|
Ducluzeau, R.,
M. Bellier, and P. Raibaud.
1970.
Transit through the digestive tract of the inocula of several bacterial strains inoculated "per os" into axenic and "holoxenic" mice. The antagonistic effect of the microflora of the gastrointestinal tract.
Zentralbl. Bakteriol. Abt. 1 Orig.
213:533-548.
|
| 9.
|
Feng, N.,
J. W. Burns,
L. Bracy, and H. B. Greenberg.
1994.
Comparison of mucosal and systemic humoral immune responses and subsequent protection in mice orally inoculated with a homologous or a heterologous rotavirus.
J. Virol.
68:7766-7773[Abstract/Free Full Text].
|
| 10.
|
Glass, R. I.,
J. R. Gentsch, and B. Ivanoff.
1996.
New lessons for rotavirus vaccines.
Science
272:46-48[Medline].
|
| 11.
|
Greenberg, H. B.,
H. F. Clark, and P. A. Offit.
1994.
Rotavirus pathology and pathophysiology.
Curr. Top. Microbiol. Immunol.
185:255-283[Medline].
|
| 12.
|
Guerin-Danan, C.,
C. Andrieux,
F. Popot,
A. Charpilienne,
P. Vaissade,
C. Gaudichon,
C. Pedone,
C. Bouley, and O. Szylit.
1997.
Pattern of metabolism and composition of the fecal microflora in infants 10 to 18 months old from day care centers.
J. Pediatr. Gastroenterol. Nutr.
25:281-289[Medline].
|
| 13.
|
Heyman, M.,
G. Corthier,
A. Petit,
J. C. Meslin,
C. Moreau, and J. F. Desjeux.
1987.
Intestinal absorption of macromolecules during viral enteritis: an experimental study on rotavirus-infected conventional and germ-free mice.
Pediatr. Res.
22:72-78[Medline].
|
| 14.
|
Johnson, C. A.,
T. G. Snider,
W. G. Henk, and R. W. Fulton.
1986.
A scanning and transmission electron microscopic study of rotavirus-induced intestinal lesions in neonatal gnotobiotic dogs.
Vet. Pathol.
23:443-453[Abstract].
|
| 15.
|
Jourdan, N.,
M. Maurice,
D. Delautier,
A. M. Quero,
A. L. Servin, and G. Trugnan.
1997.
Rotavirus is released from the apical surface of cultured human intestinal cells through nonconventional vesicular transport that bypasses the Golgi apparatus.
J. Virol.
71:8268-8278[Abstract/Free Full Text].
|
| 16.
|
Kanwar, S. S.,
V. Singh,
V. K. Vinayak,
A. K. Malik,
S. K. Mehta, and S. Mehta.
1994.
Differential tropism of EB rotavirus (serotype 3) to small intestine of homologous murine model.
Acta Virol.
38:269-276[Medline].
|
| 17.
|
Kubelka, C. F.,
R. S. Marchevsky,
P. R. Stephens,
H. P. Araujo, and A. V. Oliveira.
1993.
/1994. Murine experimental infection with rotavirus SA-11: clinical and immunohistological characteristics.
Exp. Toxicol. Pathol.
45:433-438.
|
| 18.
|
Majerowicz, S.,
C. F. Kubelka,
P. Stephens, and O. M. Barth.
1994.
Ultrastructural study on experimental infection of rotavirus in a murine heterologous model.
Mem. Inst. Oswaldo Cruz. (Rio de Janeiro)
89:395-402[Medline].
|
| 19.
|
Mebus, C. A., and L. E. Newman.
1977.
Scanning electron, light, and immunofluorescent microscopy of intestine of gnotobiotic calf infected with reovirus-like agent.
Am. J. Vet. Res.
38:553-558[Medline].
|
| 20.
|
Offit, P. A., and H. F. Clark.
1985.
Maternal antibody-mediated protection against gastroenteritis due to rotavirus in newborn mice is dependent on both serotype and titer of antibody.
J. Infect. Dis.
152:1152-1158[Medline].
|
| 21.
|
Offit, P. A.,
H. F. Clark,
M. J. Kornstein, and S. A. Plotkin.
1984.
A murine model for oral infection with a primate rotavirus (simian SA11).
J. Virol.
51:233-236[Abstract/Free Full Text].
|
| 22.
|
Petschow, B. W.,
R. E. Litov,
L. J. Young, and T. P. McGraw.
1992.
Response of colostrum-deprived cynomolgus monkeys to intragastric challenge exposure with simian rotavirus strain SA11.
Am. J. Vet. Res.
53:674-678[Medline].
|
| 23.
|
Ramig, R. F.
1988.
The effects of host age, virus dose, and virus strain on heterologous rotavirus infection of suckling mice.
Microb. Pathog.
4:189-202[Medline].
|
| 24.
|
Salim, A. F.,
A. D. Phillips,
J. A. Walker-Smith, and M. J. Farthing.
1995.
Sequential changes in small intestinal structure and function during rotavirus infection in neonatal rats.
Gut
36:231-238[Abstract/Free Full Text].
|
| 25.
| Salminen, S., E. Isolauri, and T. Onnela. 1995. Gut
flora in normal and disordered states. Chemotherapy
41(Suppl. 1):5-15.
|
| 26.
|
Shaw, R. D.,
S. J. Hempson, and E. R. Mackow.
1995.
Rotavirus diarrhea is caused by nonreplicating viral particles.
J. Virol.
69:5946-5950[Abstract/Free Full Text].
|
| 27.
|
Thillainayagam, A. V.,
J. A. Dias,
A. F. Salim,
F. H. Mourad,
M. L. Clark, and M. J. Farthing.
1994.
Glucose polymer in the fluid therapy of acute diarrhoea: studies in a model of rotavirus infection in neonatal rats.
Clin. Sci. Colch.
86:469-477[Medline].
|
| 28.
|
Uhnoo, I.,
T. Dharakul,
M. Riepenhoff-Talty, and P. L. Ogra.
1988.
Immunological aspects of interaction between rotavirus and the intestine in infancy.
Immun. Cell Biol.
66:135-145.
|
| 29.
|
Vonderfecht, S. L.,
J. J. Eiden,
R. L. Miskuff, and R. H. Yolken.
1988.
Kinetics of intestinal replication of group B rotavirus and relevance to diagnostic methods.
J. Clin. Microbiol.
26:216-221[Abstract/Free Full Text].
|
| 30.
|
Vonderfecht, S. L.,
A. C. Huber,
J. Eiden,
L. C. Mader, and R. H. Yolken.
1984.
Infectious diarrhea of infant rats produced by a rotavirus-like agent.
J. Virol.
52:94-98[Abstract/Free Full Text].
|
| 31.
|
Yuan, L.,
L. A. Ward,
B. I. Rosen,
T. L. To, and L. J. Saif.
1996.
Systemic and intestinal antibody-secreting cell responses and correlates of protective immunity to human rotavirus in a gnotobiotic pig model of disease.
J. Virol.
70:3075-3083[Abstract/Free Full Text].
|
Journal of Virology, November 1998, p. 9298-9302, Vol. 72, No. 11
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
This article has been cited by other articles:
-
McNeal, M. M., Sestak, K., Choi, A. H.-C., Basu, M., Cole, M. J., Aye, P. P., Bohm, R. P., Ward, R. L.
(2005). Development of a Rotavirus-Shedding Model in Rhesus Macaques, Using a Homologous Wild-Type Rotavirus of a New P Genotype. J. Virol.
79: 944-954
[Abstract]
[Full Text]
-
Boshuizen, J. A., Reimerink, J. H. J., Korteland-van Male, A. M., van Ham, V. J. J., Koopmans, M. P. G., Buller, H. A., Dekker, J., Einerhand, A. W. C.
(2003). Changes in Small Intestinal Homeostasis, Morphology, and Gene Expression during Rotavirus Infection of Infant Mice. J. Virol.
77: 13005-13016
[Abstract]
[Full Text]
-
Mori, Y., Borgan, M. A., Ito, N., Sugiyama, M., Minamoto, N.
(2002). Diarrhea-Inducing Activity of Avian Rotavirus NSP4 Glycoproteins, Which Differ Greatly from Mammalian Rotavirus NSP4 Glycoproteins in Deduced Amino Acid Sequence, in Suckling Mice. J. Virol.
76: 5829-5834
[Abstract]
[Full Text]
-
Ciarlet, M., Conner, M. E., Finegold, M. J., Estes, M. K.
(2002). Group A Rotavirus Infection and Age-Dependent Diarrheal Disease in Rats: a New Animal Model To Study the Pathophysiology of Rotavirus Infection. J. Virol.
76: 41-57
[Abstract]
[Full Text]
-
Guérin-Danan, C., Meslin, J.-C., Chambard, A., Charpilienne, A., Relano, P., Bouley, C., Cohen, J., Andrieux, C.
(2001). Food Supplementation with Milk Fermented by Lactobacillus casei DN-114 001 Protects Suckling Rats from Rotavirus-Associated Diarrhea. J. Nutr.
131: 111-117
[Abstract]
[Full Text]
Top
Abstract
Text
