Transfer of antibodies elicited by baculovirus-derived VP2 of a very virulent bursal disease virus strain to progeny of commercial breeder chickens.

A baculovirus-derived recombinant VP2 (rVP2) subunit vaccine elicited anti-infectious bursal disease virus (IBDV) antibodies in commercial flocks. The induced antibody levels were similar to those evoked against IBDV by a commercial vaccine. The levels remained higher than that of the negative controls for at least four and a half months in commercial chickens. The antibodies were also transferred to their offspring and were detected in the blood of the progeny for at least 20 days after hatching. These results, along with former data, that show that antibodies elicited by baculovirus rVP2 confer protection to chickens from IBDV [J. Pitcovski et al. (1996), Insect cell-derived VP2 of infectious bursal disease confers protection against the disease in chickens. Avian Diseases, 40, 753-761], imply that the baculovirus-derived rVP2 subunit may serve as a successful vaccine for commercial breeding flocks.

Insect cell-derived VP2 of infectious bursal disease virus confers protection against the disease in chickens.

Infectious bursal disease virus (IBDV) has become a major problem in recent years. Conventional vaccines make use of attenuated or inactivated viral strains, but these are gradually losing their effectiveness. We investigated the possibility of using purified VP2, a subunit of IBDV structural protein expressed in insect cells, as a vaccine. The VP2 gene was cloned into pAcYM1. The cloned gene was expressed in a baculovirus system, giving rise to a high quantity of recombinant VP2 (rVP2) protein. The length of the VP2 is 453 amino acids, and it contains two additional amino acids of the baculovirus at the carboxyl terminus. The molecular mass of the protein is about 48 kD. The rVP2 protein reacted with antibodies raised against viral VP2 and had a similar molecular weight. This protein was tested in a controlled vaccination experiment and compared with an inactivated commercial vaccine. High levels of antibodies were raised by the vaccinated birds. The vaccinated birds were challenged with a pathogenic viral strain. rVP2-vaccinated chickens exhibited high resistance to the virus. No mortality or weight changes in the bursa of Fabricius were observed in the vaccinated birds, whereas in the negative control birds, vaccinated with phosphate buffer, up to 50% mortality was found. Higher levels of antibodies were found by enzyme-linked immunosorbent assay in birds vaccinated with rVP2 compared with those vaccinated with the commercial vaccine. This study suggests the potential use of the isolated rVP2 as a subunit vaccine.

Coding region of segment A sequence of a very virulent isolate of IBDV--comparison with isolates from different countries and virulence.

We determined the sequence of the coding region of segment A, coding for the viral proteins (VPs) VP2, VP4, and VP3, of a very virulent (vv) infectious bursal disease virus (IBDV) isolated in Israel and named IBDVks. We compared the deduced amino acid sequences of the proteins of the new isolate with those of the same proteins from several IBDV isolates, as published in recent years. The amino acid sequences of VP3 and VP4 of the Israeli isolate were 1.9%-2.3% different from the sequences of their counterparts from classical strains. Thus, the stable region of VP2 of IBDVks was very similar (0-0.68% difference) to the same region of VP2 from vv strains from Europe and Japan but distinct from that of proteins from classical strains from Europe, the United States, and Australia (up to 9.42% divergence), showing that IBDVks is more closely related to the vv strains from Europe and Japan. We found that viruses isolated in recent years resemble each other more than isolates from the same areas isolated a few years earlier. Hence, IBDVks can be categorized in one group with vv new isolates from Europe and Japan. This group has been found to be distinct from new isolates in the United States and strains isolated before the IBDV epidemic during the late 1980s.

Kinetic Analysis of Resistance to Paraquat in Conyza: Evidence that Paraquat Transiently Inhibits Leaf Chloroplast Reactions in Resistant Plants.

Paraquat resistance has been claimed to be due to a sequestration of the herbicide before it reaches chloroplasts. This is based on the sensitivity of photosystem I in isolated thylakoids to paraquat, and autoradiographic analyses showing label from paraquat near veins 4 hours after treatment of a resistant biotype. Conversely, the enzymes of the superoxide detoxification pathway were found to be at constitutively elevated levels in intact class A chloroplasts of the resistant biotype of Conyza bonariensis (L.) Cronq. Evidence is presented here that physiologically active levels of paraquat rapidly inhibit chloroplast function in both the resistant and sensitive biotype, before the first sequestration was visualized. This inhibition is transient (completed in 2 hours) in the resistant biotype and irreversible in the sensitive type. Intact class A chloroplasts of the resistant biotype with or without paraquat are less susceptible to photoinduced membrane damage than the sensitive biotype without paraquat, as measured by ethane evolution. These data support a hypothesis that the ability to prevent superoxide damage keeps the resistant biotype viable while paraquat or its metabolites are being sequestered.

Increased Tolerance to Photoinhibitory Light in Paraquat-Resistant Conyza bonariensis Measured by Photoacoustic Spectroscopy and CO(2)-Fixation.

Tolerance to photoinhibition was compared between a paraquat-resistant and a sensitive biotype of Conyza bonariensis (L.). Cronq. Photoinhibitory damage was measured as a decrease in oxygen evolution or energy storage using photoacoustic spectroscopy, or as a decrease of (14)CO(2)-fixation. Prior to exposure to high fluence rates, both biotypes had similar quantum yields of oxygen evolution and energy storage. After exposure to high intensity light, the resistant biotype continued to evolve oxygen and to store energy with a high quantum yield while both energy storage and oxygen evolution were severely reduced in the sensitive biotype. CO(2)-fixation was less rapidly inhibited in the resistant biotype compared to the sensitive one. The data show that the paraquat resistant biotype with its high constitutive levels of the chloroplast localized enzymes of the oxygen detoxification pathway, is also partially protected from photoinhibition. This supports the theory that an enhanced radical scavenging system can give temporary protection against photooxidative damage from a variety of sources.