The Papovaviridae contain two very different virus families, the papillomaviruses and the polyomaviruses. With further investigation, it was determined that the Budgerigar Fledgling Disease Virus is a polyomavirus. Subsequently, the virus was found to infect many different species of psittacine birds (parrots) and thus it is generally the convention to call it the avian polyomavirus (APV).
APV is wide spread and can be found in most countries of the world where psittacine birds are raised. As will become clearly apparent, generalizations about this virus cannot be made and over simplification about the issues of infection and disease, while convenient, are often misleading.
Avian Polyomavirus Disease
Budgerigars
Nonbudgerigar Parrots
Nonbudgerigar parrots are also susceptible to avian polyomavirus infection. Some are highly susceptible to disease, while others rarely if ever develop disease. APV- disease in these birds occurs at different ages in different birds. In conures, deaths typically occur in birds less than six weeks of age. Deaths in macaws and Eclectus Parrots occur in birds 14 weeks and younger. Most, possibly all, of the nestlings lost are being hand fed. Infected nestlings appear healthy, show few premonitory signs, and then die suddenly. When signs do occur, they proceed death by only a few hours. Observant owners may notice delayed crop emptying, weakness, a generalized pallor, or bruising under the skin in the preceding hours before death. Yellow discoloration of the urates is another rare observation. Necropsy findings commonly include generalized pallor with subcutaneous and subsersoal hemorrhage and enlargement of the spleen and liver. Less commonly, acites and cardial effusion may be present. Microscopic examination of the tissues reveals extensive areas of necrosis (cell death) in the liver. Virus inclusion bodies are found in the spleen, mesangial cells of the kidney, and Kupffer cells of the liver. Necrosis of splenic cells is often massive. Less commonly, virus inclusions are found in other organ systems including the feather follicles. An immune complex glomerulopathy occurs in a significant percentage of the birds with this disease. These complexes contain antibody and APV proteins.
Lovebirds
APV disease in lovebirds is distinct enough to merit special attention. Like the Budgerigar, this disease occurs in nestling birds and inclusion bodies can be found in multiple organs. Unlike the Budgerigar, birds up to one year of age can also be affected. This unusual age susceptibility has not been fully explained. However, in at least some of these older birds, concurrent infection with PBFDV is also occurring and may permit APV disease in a bird that would otherwise be resistant to it.
Infection Versus Disease
It has become evident that infection and disease are not synonymous. Many birds are infected by the virus but only a certain, and sometimes small, percentage of these birds will develop disease. Whether disease will develop is dependent on the species of bird infected, the age of the bird infected, whether that bird is also infected with the PBFDV, and other factors that remain unclear. Birds that are infected and do not develop disease still have virus replication within their bodies and shed virus in their droppings for a period of time. The length of time that virus shedding occurs, again, depends on the age of the bird at the time of infection and the species of the bird.
Infection in Budgerigars
In the United States, the English variety of Budgerigar appears to have some resistance to APV infection. The most devastating outbreaks of disease occur in large commercial aviaries of the American variety of Budgerigar where birds are bred in rooms containing tens or hundreds of free-flighted birds. Both nestlings and adult Budgerigars are susceptible to infection. Deaths, however, is confined to young birds between the ages of 10 and 25 days. The nestling mortality (death) rate is often high and may approach 100% when the virus is first introduced to an aviary. If there is no intervention in subsequent breeding seasons, mortality rates will decline but production will always remain depressed. Birds that survive infection may have abnormal feathering or appear completely healthy. Survivors shed virus in their droppings and probably from their skin and feather dander for up to six months after infection. Virus shedding stops with the onset of sexual maturity or during the first breeding cycle. The infection cycle is then maintained through the shedding of virus by nestlings and young adult birds. Thus birds are exposed to the virus immediately after hatch and have virus circulating in their blood by the time they are seven to 10 days old. Fledglings and young adult birds are also important sources of virus exposure for other birds when they are taken to bird shows, bird marts, and sold to pet stores. It has been suggested that egg transmission of APV occurs in the Budgerigar. This conclusion is based on two observations. First, intra-nuclear inclusion bodies were reported in day old nestlings suggesting that these birds had virus growing in them before they hatched. Secondly, in a clinical trial, eggs were removed from a flock of Budgerigars experiencing an outbreak of disease and placed under the hens of a clean flock. The young from these eggs subsequently developed disease. In another study inclusion bodies are not seen in birds less than a week old. Also, there is another interpretation for the results of the clinical trial. If the transferred eggs were contaminated with virus, then the chicks could have been exposed at hatch. Additionally, Budgerigar hens eat the egg shells. Thus they could have become infected and then passed the infection on to their young. There is only very limited and circumstantial evidence that egg transmission occurs in any parrot species.
Infection in Nonbudgerigar Parrots
Susceptible birds infected with APV infection will die. Rarely, a susceptible bird will have transient signs and survive. In birds resistant to disease, infection is unapparent. In these birds,, viral DNA can be first detected in blood after which it is detected in the cloaca. Cloacal samples may intermittently be negative, but generally the blood will remain positive. When the bird is about to stop shedding, the blood will become negative and within a week or two, cloacal swabs will also become negative. The length of time that birds are blood and cloacal positive is dependent on the species of bird and the age that it was infected. It appears, for the most part, that the older the bird is at the time of infection, the shorter the duration of shedding.
Conures
Many, possibly most, conure nestlings exposed to APV at six weeks of age or younger will develop disease and die. In birds older than six weeks, APV causes an unapparent infection. In conures, unapparent infections are best detected by examining the blood for virus DNA.Virus shedding can be expected for four to six weeks inmost birds, but up to 16 weeks in the rare individual.
Macaws
Macaws are susceptible to APV infection and disease up to approximately 14 weeks of age, after which infection is unapparent. Peak mortality in macaws occurs from four to eight weeks of age. Unapparently infected birds will become blood positive and cloaca positive. In a recently completed study, two Blue and Gold nestlings that survived infection shed virus 14 weeks. Two fledgling Red-fronted Macaws shed virus for 10 weeks. Adult Blue and Gold Macaws and Hyacinth Macaws shed for six weeks or less. Unlike most other birds examined, the Blue and Gold nestlings were occasionally blood negative, but cloacal positive.
Eclectus Parrots
Infection of otherwise healthy nestling Eclectus Parrots will cause their death if they are less than 14 weeks old. Specific studies on the length of virus shedding in these birds have not been done.
Cockatoos
As a general statement, cockatoos of any age are highly susceptible to infection with APV, but are extremely resistant to disease. Healthy adult cockatoos are not expected to ever develop APV disease and the same is true for nestling cockatoos under most circumstances. In a recent study, it was found that Goffin's and Ducorp's nestlings exposed to the virus at less than three weeks of age developed abnormal feathers. These birds showed transient signs of a systemic illness, then recovered with supportive care. Older birds and other cockatoo species remained healthy, although nearly all of them became infected. Virus shedding, as determined by cloacal swab, lasted eight to 10 weeks. Virus could be detected in the blood consistently until just before shedding stopped. In this group of birds, cloacal swabs were not consistently positive and several birds that were originally cloacal positive became negative and then positive again.
APV Infection and Disease in Adult Parrots; the PBFDV Connection
APV readily infects adult parrots. Most infections, probably greater than 99.9% of them, are completely asymptomatic. These birds become infected, shed virus for a period of time, and never look or act ill. APV disease, however, has been documented in adult birds. So why do these few adult birds develop APV disease? The answer in most cases is that they are immunosuppressed with a concurrent infection of PBFDV. In a documented outbreak of APV disease in adult Eclectus Parrots, all birds had PBFDV. Disease has also seen identified in adult cockatoos, again when these birds were tested for PBFDV they have been found to be positive. Young adult lovebirds can die with APV disease. Again, concurrent infections with PBFDV may be the explanation for why. On every occasion that APV outbreaks have occurred in lovebirds, PBFDV could also be found in the aviary. PBFDV-infected birds are a common source for APV in an aviary.PBFDV-infected birds, like AIDS patients, have a poorly functioning immune system, Therefore, if they become infected with APV they cannot clear the virus. Some of these birds will develop full blown APV disease and die. Most will become persistently infected. These persistently infected birds will then shed virus continuously from their skin and in their feather dander. This constant virus shedding contaminates the environment and makes it likely that it will be tracked into the nursery.
Are Caiques and Eclectus Parrots More Susceptible to APV Infection, Even as Adults, Than Other Parrots?
One of the first reports of APV disease in adult birds documented an outbreak where an Eclectus, a Painted Conure, and three White-bellied Caiques died. These birds clearly had APV-disease. They were not, however, tested for the PBFDV. So we do not know if this means that they were normal birds that have a predilection for APV disease, or were birds infected with PBFDV and were immunosuppressed. Since that time, the author has heard of a number of deaths in adult caiques. However, none of these birds were tested for PBFDV. Thus, the answer to this question remains elusive and requires further investigation. Because PBFDV may not cause histologic evidence of disease, It is felt that it is essential that when APV disease occurs in adult birds or in species where it is not normally a problem, that they be tested by DNA probes for the PBFDV. Numerous asymptomatic APV infections in adult Eclectus Parrots has been documented. Yet, recently I became aware of four adult Eclectus Parrots, originating from one source, that died with APV-disease. These birds were tested negative for PBFDV. What does this mean? Was this a particularly hot strain of APV or was there another factor involved? At this point we do not know.
Testing
Currently there are three types of tests available for detecting APV infection in birds; serology, examination of blood for virus DNA, and examination of cloacal swabs for virus DNA.
Serology
Serology is the examination of the liquid portion of blood (plasma or serum) for antibodies that are made specifically against a virus, bacteria, or fungus. If a bird is infected with APV and survives, it will develop antibody to the virus. Antibody can be detected in the Budgerigar by nine days after infection, in most other birds antibody is not present in the blood until two to three weeks after infection. Antibody concentrations rise very quickly and by four to six weeks after infection they reach their peak. Antibody to APV can be detected in the blood for months to many years after infection depending on the species. Budgerigars maintain an antibody titer for life. Cockatiels probably only maintain antibody titers for about six months. However, for most parrot species, antibody can be detected for at least two to three years following infection and possibly for many more years. So what does APV serology tell us? In the Budgerigar, it tells us that the bird was infected with APV. If the bird is a young adult it is probably still shedding virus. If the bird is an older experienced breeder it is not shedding virus and most likely will not. A positive antibody titer in a Cockatiel means that the Cockatiel has been infected within the last six months and this bird may be shedding virus. In other parrots, it tells us very little. If the bird has antibody, then we know that it has been infected with virus but we do not know whether the bird is shedding virus. If the bird was infected recently, then it probably is shedding virus. If the bird was infected over 16 weeks ago, then it is probably not shedding virus, unless it is also infected with PBFDV. Therefore, with the exception of the Budgerigar, serology is generally not very helpful in detecting virus shedding birds. Unfortunately, this test has been inappropriately used in the past.
PCR Assay of Cloacal Swabs and Blood
The polymerase chain reaction, or PCR, is an assay that has become an incredibly important tool for the diagnosis and control of infectious diseases. This assay takes a low concentration of the APV DNA and amplifies it to a concentration that can be detected. Therefore, as few a 10 copies Of the virus can be detected if the test is properly preformed. The sensitivity of this test is one of its greatest strengths as well as one of its greatest weaknesses. The potential problem with this assay is that even the smallest contamination of the sample, either at the collection site or in the laboratory will result in a negative sample becoming positive. Therefore, if one is testing multiple birds, it becomes very easy to get the sample from a negative bird contaminated with the feather dust or dried feces from a positive bird. The original discovery that APV could be detected in the live bird was made by Dr. Frank Niagro at the University of Georgia. He and his collaborators found that APV could be detected in cloacal swabs of unapparently infected birds. This technology was licensed to the Research Associate Laboratories (Drs. Dalhausen and Radabaugh) and has been offered by them for five years. During this time, these scientists have modified and improved this assay and have discovered that APV DNA can also be detected in the blood of birds recently infected with APV. The bottom line is that neither test will pick up app APV infected birds that are shedding or will be shedding the virus. On the other hand both tests will pick up most birds shedding virus. So which one will you choose to screen birds? In a recently completed study, the cloacal virus PCR, blood PCR and serologic assay were compared. Of all the birds ( > 50) that were examined with multiple tests, both tests picked up all but one of the birds that seroconverted. Not all birds were positive on both tests each time. In cockatoos and conures it was found that birds stayed consistently positive with the blood PCR, while several were intermittently positive on the cloacal swab. As virus was cleared from the bird, the blood test became negative first and the cloacal swab became negative to two to four weeks later. Therefore, for these species, I recommend that the blood PCR be used as a screening tool. if it is positive, the bird should be retested in two to three months, if negative, the bird should be quarantined for four additional weeks and then will he considered free of virus shedding. In macaws, we found that in most situations both tests were positive. Virus shedding and viremia stopped almost simultaneously. In a few rare occasions, however, the blood PCR test was negative when the cloacal swab was positive. Thus in the limited macaw species examined, a cloacal swab may be more sensitive than the blood PCR. Research Associates is now offering a PCR assay that screens both blood and cloacal samples from the same bird in the same reaction. This should be the most sensitive assay of all. It has been said that blood PCR testing of live birds following vaccination or swabs of tissue in recently vaccinated birds that die, will detect fragments of DNA from the vaccine. Recent work by Drs. Dalhausen and Radabaugh has shown that viral DNA is never present in the blood of nestlings vaccinated for APV. The veterinarian and bird owner must therefore conclude that if a bird is blood PCR positive, vaccinated or not, that it is infected with APV and is most likely shedding virus.
The APV Vaccine: Possibly a Tool, Not a Panacea
In the past few years an APV vaccine (Biomune, Lenexa, KS) has been on the market. The developers of this vaccine are advocating its use in essentially all parrots, and suggest that if adequate numbers of birds are vaccinated that we can essentially eliminate APV as a problem. Not all investigators, however, share this opinion. While we all share in the desire to control this terrible virus, I believe that the widespread and indiscriminate immunization of pet and aviary birds, is often unnecessary and for the aviculturist may not make economic sense. In somewhat reverse order, consider the following two points. First, if the vaccine is effective, which birds can it be expected to protect from infection and disease.? Secondly, do we have significant and substantial data to suggest that the vaccine does work?
APV immunization to protect from disease
Adults: If our goal is to prevent APV disease by immunization, then it is essential to understand basic APV biology. As has been discussed, healthy adult parrots rarely if ever develop disease. Thus, vaccinating adult birds to protect them from APV disease in most cases is unnecessary (see discussion on caiques and Eclectus Parrots). Nestlings: it is the nestling that when infected with APV will die. Recall, however, that only certain nestlings of certain species are susceptible to disease. To protect these nestings, according to the vaccination manufacturer, nestlings should be vaccinated at five weeks or older and then again two to three weeks later. They are said to be protected four weeks after the first immunization. Thus the vaccine has the potential to protect susceptible chicks from infection and disease in the window of nine to 14 weeks. We cannot immunized conures at an early enough age to protect
