Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving virus or bacteriophage
Reexamination Certificate
2001-06-05
2002-06-18
Park, Hankyel T. (Department: 1648)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving virus or bacteriophage
C424S184100, C424S204100, C435S235100
Reexamination Certificate
active
06406843
ABSTRACT:
The present invention is directed to chicken virus isolates or strains (Texas RB 3, Texas RB 4, HBS, F57-7, W/L 39, GAR 1) believed to be a new strain or strains of infectious bursal disease virus (IBDV) which infects the proventriculus, a vaccine made from these isolates, a method of treatment, vaccination scheme, and the like.
In accordance with the present invention, numerous experiments were conducted to determine the role of infectious bursal disease (IBD) virus in the induction of lesions associated with proventriculitis syndrome in chickens. Parameters examined included age of the chicken at IBD virus exposure, concentration of IBD virus at exposure the strain of IBD virus, dietary influence in the presence of IBD virus, mixed IBD virus infection, autoimmune mediated IBD reactions associated with lesion production, viral induced apoptotic tissue injury and isolation and characterization of the causative agent.
The experiments were carried out in SPF white leghorns, with the experimental birds being examined for the presence of gross and microscopic lesion at 4 and 11 days post challenge. Tissue homogenates were analyzed for the presence of IBDV at 4 and 11 days post challenge with Antigen Capture ELISA (AC-ELISA). Determinations of neutralizing antibody levels and IgG antibody responses were monitored as well as depletion of serum complement following IBDV infection. Physical parameters were also considered utilizing body weights and organ to body weight ratios to determine IBDV effects following experimental challenge.
Physical parameters indicated that the primary viral response is predominantly in the bursa of Fabricius, but changes were also noted in the proventriculus. Physical changes in the proventriculus occurred primarily during the acute stage of the IBD virus infection.
Gross and microscopic lesions in the proventriculus were exacerbated by the presence of copper sulfate supplementation in the feed. There were also very evident negative effects on the weight gain of SPF white leghorn chickens given copper sulfate feed supplementation.
Infectious bursal disease virus strains show different affinities for producing lesions in the proventriculus following challenge. Standard challenge IBD virus USDA/STC stain produced the most consistent lesions in the proventriculus often accompanied by hemorrhage. This correlates well with antibody enhanced pathology as observed with antigen-antibody complex activation of serum complement. Evidence of yet another mechanism of IBDV induced pathology, was demonstrated through the use of TUNEL apoptosis staining of infected tissues. The degree of apoptotic involvement was also reflected in the pathogenicity of the IBD virus strain used for challenge.
Infectious bursal disease viral inclusions were demonstrated by thin section electron microscopy in the proventriculus at 4 days post challenge. Viral inclusions resemble previously documented inclusions produced in the bursa following IBD infection.
In accordance with the present invention, infectious bursal disease viruses were isolated from broiler chickens experiencing proventriculitis in, for example, Oklahoma, Texas, West Virginia, and California.
IBDV Antigen Capture (AC) Elisa tested positive for Texas RB 3, Texas RB 4, HBS, F57-7, W/L 39, and GAR 1. Virus isolation in SPF embryonated eggs was completed and 3rd and final passage for the isolates. SPF chickens were inoculated with egg harvest for the isolates. Tissues from inoculated birds and histopathology confirmed lesions. AC-ELISA of tissue and serological confirmation of the isolates was performed.
Proventriculitis is a major problem to the broiler industry. These viruses may be one of the major causes of this condition and if this proves to be true, could be utilized as or in a vaccine to prevent the disease condition. The viruses are believed to be a significant finding in the search for a causative agent for proventriculitis in broiler chickens and as such may be utilized in the development of a vaccine or vaccines. The viruses can be attenuated to be used as a modified live vaccine or utilized in an inactivated form in a killed vaccine.
TABLE OF CONTENTS
INTRODUCTION
BRIEF DESCRIPTION OF THE DRAWINGS
I. SECTION 1
D
ETERMINATION OF THE
R
OLE OF
I
NFECTIOUS
B
URSAL
D
ISEASE
V
IRUS
S
TRAIN
V
ARIANT
1084-E
IN THE
I
NDUCTION OF
P
ROVENTRICULITIS IN
SPF C
HICKENS
: A
GE
R
ELATED
E
FFECTS
II. SECTION 2
D
ETERMINATION OF THE
R
OLE OF
I
NFECTIOUS
B
URSAL
D
ISEASE
V
IRUS
S
TRAINS
USDA/STC
AND
V
ARIANT
1084-E
ON THE
I
NDUCTION OF
L
ESIONS
A
SSOCIATED WITH
P
ROVENTRICULITIS
: I
NFLUENCE OF
V
IRUS
C
ONCENTRATION AND
T
IME
III. SECTION 3
T
HE
I
NTERACTION OF
I
NFECTIOUS
B
URSAL
D
ISEASE
V
IRUS
, REO V
IRUS, AND
C
OPPER
S
ULFATE IN THE
P
RODUCTION OF
L
ESIONS
A
SSOCIATED WITH
P
ROVENTRICULITIS IN
SPF W
HITE
L
EGHORN
C
HICKENS
IV. SECTION 4
I
NFECTIOUS
B
URSAL
D
ISEASE
V
IRUS
I
NDUCED
A
POPTOSIS
A
SSOCIATED WITH
P
RODUCTION OF
L
ESIONS IN THE
P
ROVENTRICULUS AND
B
URSA
V. SECTION 5
M
EASUREMENT OF
H
EMOLYTIC
C
OMPLEMENT
L
EVELS AND
V
IRUS
N
EUTRALIZING
A
NTIBODY
T
ITERS
4, 6, 8
AND
11 D
AYS
P
OST
C
HALLENGE WITH
I
NFECTIOUS
B
URSAL
D
ISEASE
V
IRUS
S
TRAINS
USDA/STC, V
ARIANT
E/DEL,
AND
V
ARIANT
E/1084
VI. SECTION 6
T
HE
I
NTERACTION OF
I
NFECTIOUS
B
URSAL
D
ISEASE
V
IRUS
(USDA/STC)
AND
C
OPPER
S
ULFATE IN THE
P
RODUCTION OF
L
ESIONS
A
SSOCIATED WITH
P
ROVENTRICULITIS IN
B
ROILER
C
HICKENS
VII. SECTION 7
I
SOLATION AND
C
HARACTERIZATION OF
I
NFECTIOUS
B
URSAL
D
ISEASE
V
IRUS
F
ROM
B
ROILER
C
HICKENS
E
XHIBITING
S
YMPTOMS OF
P
ROVENTRICULITIS
VIII. FURTHER DISCUSSION
IX. APPENDIX CLAIMS ABSTRACT OF THE DISCLOSURE
INTRODUCTION
The primary impetus for this invention came as a result of a request for help from a number of commercial poultry processors. Proventriculitis has been described in broiler chickens from a number of geographical locations. This syndrome is characterized by the enlargement of the proventriculus, with feed impaction, and structural weakness. Affected birds can be either normal or underweight at processing, with either a high or normal feed conversion efficiency. Problems generally manifest at the processing plant with breakage of the proventriculus during mechanical evisceration, resulting in an increased number of washouts, slowed or stopped processing lines, excessive trims, and higher than normal levels of condemnation. This proventriculitis syndrome has been linked to a number of environmental, nutritional, genetic, and infectious causes.
Preliminary studies indicate that infectious bursal disease virus is capable of producing lesions in the proventricular mucosa. While proventriculitis induced by IBDV alone does not exactly mimic what is observed in field situations, it is contented that IBDV plays the role of a facilitator, and thus presents an opportunity for other viral, bacterial, fungal or chemical pathogens to exert an effect.
Some objects of this invention are to: (1) attempt to establish infectious bursal disease virus as a causative agent in the induction of lesions associated with proventriculitis, (2) determine the pathogenic mechanism by which IBDV exerts an effect on the proventriculus, and (3) to isolate and characterize IBDV virus from field cases of proventriculitis.
Infectious Bursal Disease Virus
Infectious bursal disease (IBD) is an acute viral infection which predominantly affects domestic chickens. This viral disease has also been described in commercial turkeys (22, 60, 81, 82, 119), and represents an occasional problem in ducks (81). Infectious bursal disease virus infection and replication occurs primarily in lymphoid tissues throughout the body. This results in induction of significant pathological lesions within the follicles of the bursa of Fabricius, which contains the highest concentration of lymphoid cells. When infection occurs at an early age (<3 weeks), the disease syndrome may be further complicated by an accompanying immunosuppression (33, 76, 95).
In 1962, Cosgro
Newberry Lisa A.
Skeeles John K.
Head Johnson & Kachigian
Park Hankyel T.
University of Arkansas
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