Methods of detection of Chlamydia psittaci in domesticated and wild birds.

OBJECTIVE: To study the occurrence of Chlamydia psittaci in domesticated and wild birds and compare the sensitivity of molecular detection with cell culture isolation. DESIGN: Study of cell culture isolation and PCR detection of C psittaci in avian samples. PROCEDURE: Samples were obtained from 485 birds. Domesticated birds were selected at random from pet shops, private aviaries and zoos, while wild birds were captured locally, sampled, and immediately released. Swabs were collected from choanal slit, conjunctiva and cloaca of each bird and pooled. Samples were divided into equal portions for use in PCR dot-blot and cell culture detection. PCR and dot-blot detection was based on the ompB gene. RESULTS: Prevalence of infection varied markedly between flocks of captive birds. It was highest where there were frequent changes in the flock members or where there were many birds confined in small areas. C psittaci was not detected in wild birds or water birds. The sensitivity of cell culture compared to PCR dot-blot detection was 68%. All samples positive by cell culture were also positive by PCR. CONCLUSIONS: PCR-dot blot detection of C psittaci in birds appears to be more sensitive than cell culture isolation in this study. C psittaci infection of birds may occur in clinically normal captive birds.

Screening for alcohol problems in the military: recommended tests.

This article suggests two new techniques for identifying alcohol problems in a military setting. The Alcohol Use Disorders Identification Test is a well validated self-report procedure and requires approximately 2 minutes for administration. Measurement of carbohydrate-deficient transferrin provides a useful biochemical index of recent heavy alcohol consumption. Employment of these tests could improve selection of individuals seeking entry to the military, aid recognition of current personnel in need of treatment, and assist in evaluating progress of patients in treatment.

A serologic survey for avian polyomavirus and Pacheco's disease virus in Australian cockatoos.

Sera collected from wild and captive Australian cockatoos and other psittacine species (n = 411) were tested for antibodies to avian polyomavirus (APV) and Pacheco's disease virus (PDV). Of 144 wild sulphur-crested cockatoos (Cacatua galerita) sampled at three regions in New South Wales (NSW) 96 (64.4%) birds had positive (>/= 1:32) neutralizing antibody titres to avian polyomavirus (range 1:32-1:2048). Two of 17 wild long-billed corellas (Cacatua tenuirostris) were also APV-antibody positive. However, no samples from 107 wild galahs (Eolophus roseicapillus) were positive for neutralizing antibody to APV. Sera were also collected from captive psittacine bird flocks from NSW, Tasmania, Victoria, and Western Australia. In a mixed aviary of cockatoos and lorikeets, APV antibody was detected in sera from sulphur-crested cockatoos, Major Mitchell's cockatoos (Cacatua leadbeateri), a white-tailed black cockatoo (Calyptorhynchus baudinii latirostris), a red-tailed black cockatoo (Calyptorhynchus magnificus) a single galah, a rainbow lorikeet (Trichoglossus haematodus), and a scaley-breasted lorikeet (Trichoglossus chlorolepidotus). All 411 wild and captive birds were negative for the presence of neutralizing antibody to PDV. These results indicate that wild sulphur-crested cockatoos in NSW are enzootically infected with avian polyomavirus and that the sampled populations are free of Pacheco's disease.

Epidemiological and clinical investigations into big liver and spleen disease of broiler breeder hens.

The epidemiological and clinical features of big liver and spleen disease (BLS) in flocks on two broiler breeder farms were investigated by serology and gross pathology. The most common necropsy findings on farm 1 were splenomegaly and hepatomegaly, with kidney enlargement in some birds. In one flock (farm 1), a decline in egg production began at 40 weeks of age and lasted for 9 weeks. Seroconversion to BLS antigen was first detected at 45 weeks (3.1% of birds) and increased to 72% at 50 weeks, which coincided with clinical recovery in the flock. Antigen was detected before antibody at 44 weeks and persisted at low incidence (< 15%). Farm egg production statistics and serology indicated that the disease affected all flocks on the farm. In three of eight flocks, seroconversion was detected in birds before peak production. The birds in the remaining sheds did not seroconvert or become sick until after peak production. On the second farm, sampling began within a flock already experiencing BLS. Clinical signs and pathology were similar to those seen in flocks on farm 1. However, the lesions that were seen in the pancreas in 15% of birds have not been reported previously. BLS antibody was detected in 78%, and circulating antigen in 14%, of sick birds.

The experimental production of big liver and spleen disease in broiler breeder hens.

Big liver and spleen disease (BLS) was reproduced experimentally by intravenous (IV) and oral (PO) administration of BLS inocula to susceptible broiler breeder hens 34 to 36 weeks of age. Serological and pathological signs of BLS similar to those seen in the natural disease occurred in inoculated and in-contact birds. Splenomegaly was the earliest and often the only necropsy finding, with hepatomegaly and kidney enlargement occurring in some birds later in the course of the disease. After IV administration, serum antigen was detected between 2 and 4 weeks, and antibody between 3 and 5 weeks. After PO administration, antigen was detected between 2 and 4 weeks, and antibody between 3 and 6 weeks. Antibody persisted in all birds to the end of the experiment (6 weeks), and horizontal transmission probably occurred since in-contact birds developed BLS. Liver probably contained the highest concentration of BLS agent because it had the highest infectivity.

The haemagglutination spectrum of psittacine beak and feather disease virus.

A simple method for concentrating psittacine beak and feather disease virus (PBFDV) from crude feather suspensions is described. The addition of 10% polyethylene glycol (MW 6000 to 9000) to feather suspensions facilitated the precipitation and pelleting of PBFDV by low speed centrifugation. Pellets were resuspended in one-twentieth of the original volume with caesium chloride (CsCl) buffer and subjected to isopycnic ultracentrifugation. Peak haemagglutination activity (HA) occurred at 1.35 g/ml in PBFDV CsCl gradients. CsCl purified virus agglutinated galah (Eolophus roseicapillus), eastern long-billed corella (Cacatua tenuirostris), sulphur-crested cockatoo (Cacatua galerita), Major Mitchell's cockatoo (Cacatua lead-beateri) and gang gang cockatoo (Callocephalon fimbriatum) erythrocytes, but not those of 19 other avian or five mammalian species. PBFDV agglutinated galah erythrocytes at 4 degrees C and 37 degrees C over a wide range of pH and no change in HA titre was observed when PBFDV was treated with chloroform. HA persisted in PBFDV suspensions heated to 80 degrees C for 30 min, but was not detected after incubation at higher temperatures. High HA titres were detected in the feathers, serum, liver and kidneys of PBFD-affected birds.

Carbohydrate-deficient transferrin as a measure of immoderate drinking: remaining issues.

A growing body of investigations demonstrate that elevated levels of carbohydrate-deficient transferrin (CDT) effectively distinguishes alcoholics recently consuming large amounts of alcohol from light social drinkers or teetotalers. Nevertheless, important questions still remain concerning the value of CDT as a more generalized marker of alcohol consumption. Most important, the nature of the drinking pattern, including quantity and frequency, necessary to raise levels of CDT significantly remains unclear. Neither has research convincingly demonstrated that CDT is as accurate a marker for women, young adults, or non-Caucasian ethnic groups as for White, middle-aged men. Whereas CDT might serve as a useful outcome measure in trials of alcoholism treatment effectiveness, current research suggests that CDT is of limited value in identifying problematic drinking in general medical or community settings in which a broad continuum of drinkers is represented. Combining CDT with other biochemical or self-report screening measures may, however, improve sensitivity in these contexts. At present, the most accurate laboratory technique to detect CDT seems to be isoelectric focusing. Additional research, however, is needed to resolve the issue of whether CDT is best quantitated as a simple value or if its ratio to total transferrin or non-CDT results in higher predictive validity.

CAGE as a predictor of hazardous alcohol consumption in U.S. Army personnel.

Sensitivities, specificities, and odds ratios for hazardous drinking for various cut scores on the CAGE are computed for a large sample active duty Army personnel. Contrasts on these properties are made between the standard CAGE and a "modified CAGE" consisting of standard CAGE items and two other items dealing with problematic drinking. The role of demographic variables--gender, ethnicity, marital status, rank category, and age in mediating relationships of both versions of the screening test to hazardous drinking--is also explored. At a cutoff score of one endorsed item, odds ratios were highest for female personnel and commissioned officers.

Vaccination and challenge studies with psittacine beak and feather disease virus.

Psittacine beak and feather disease virus (PBFDV) was administered to adult galahs (Eolophus roseicapillus) by mouth or by intramuscular injection. Concentration of PBFDV antibodies in serum and excretion of PBFDV were monitored by haemagglutination inhibition (HI) and haemagglutination (HA) respectively. After oral administration, 17 of 18 galahs remained clinically normal and a small rise in antibody titre was detected in 3 of 18 birds. After intramuscular administration, antibody was detected in all birds. PBFDV was not detected in the feather dander of birds in either group. One bird developed diarrhoea and high faecal HA titres within 4 days of oral administration and then died. Adult and nestling cockatoos were vaccinated with an experimental inactivated double-oil emulsion vaccine. PBFDV antibody responses are comparable to those induced by a primary-oil emulsion vaccination regimen using Freund's adjuvants. Both vaccines protected nestlings. Three sibling wild-caught sulphur-crested cockatoos were vaccinated but died of PBFD before experimental challenge despite antibody responses in all birds. Unvaccinated control chicks developed acute PBFD within 4 weeks of challenge, probably from PBFDV-induced hepatitis since high concentrations of PBFDV were detected in their livers.

Laboratory diagnosis of psittacine beak and feather disease by haemagglutination and haemagglutination inhibition.

Simple and sensitive haemagglutination and haemagglutination inhibition assays were developed for psittacine beak and feather disease (PBFD) virus and serum antibody, respectively. The assays were used in the examination of samples from 73 birds clinically affected with PBFD. High antigen titres (log2 9 to log2 12) were detected in feathers, faeces and cloacal contents of PBFD-affected birds. Antigen was not detected in either faecal or feather samples from 20 normal galahs (Eolophus roseicapillus) and 9 normal sulphur crested cockatoos (Cacatua galerita). After kaolin treatment and haemadsorption of serum, haemagglutination inhibition (HI) antibody titres could not be detected in serum from 42 PBFD-affected birds, whereas serum HI titres from 64 normal psittacine birds ranged from less than log2 1 to log2 8. Serum and yolk HI antibody responses of 6 PBFD virus-inoculated layer hens were measured. Pre-inoculation chicken sera contained high concentrations of non-specific haemagglutination inhibitors (not detected in chloroform-extracted yolk), which were removed by kaolin treatment and haemadsorption.

Seroprevalence of psittacine beak and feather disease in wild psittacine birds in New South Wales.

A haemagglutination inhibition assay was used to detect antibody to psittacine beak and feather disease virus in sera from wild sulphur crested cockatoos (Cacatua galerita), galahs (Eolophus roseicapillus), short-billed corellas (Cacatua sanguinea), eastern long-billed corellas (Cacatua tenuirostris) and other psittacine birds in New South Wales. The seroprevalence of psittacine beak and feather disease ranged from 41% to 94% in different flocks, indicating infection with the virus is widespread in wild populations.

Principles and practice of detoxification.

Detoxification is the medical care that safely carries the patient through withdrawal. It is normally not an end point but is a step toward rehabilitation. Protocols should be tailored to meet the needs of individual patients.

Newcastle disease.

Since 1926, there have been three epizootics of ND. The latter two have been directly linked with psittacine species and Racing Pigeons. The modern poultry industry is extremely vulnerable to the effects of NDV, once it gains entry to any facet of the industry. Consequently considerable expense and effort are expended to keep the virus at bay. The main threat continues to come from psittacine species and racing pigeons. The considerable international trade in these birds, together with rapid air transport, can allow virulent NDV to gain entry to a country while exotic birds are incubating the disease. It is hoped that quarantine barriers and requirements will prevent the virus from entering a country, but smuggling continues and constitutes the biggest risk. Domestic avian pets are also vulnerable to the virus. It is hoped that new in vitro testing procedures, such as monoclonal antibody and oligonucleotide fingerprinting techniques, may be used to identify rapidly and characterize emergent virulent strains, so that appropriate measures may be taken to prevent infection of commercial poultry and domestic pets.

Disorders of the avian integument.

The avian integument consists of highly specialized structures that show extreme variations between species. Feathers are the most obvious part of the integument, and their color and beauty forms an important basis for the attraction of birds to humans. Any disorder in the feathers can be distressing for a client and frustrating for the avian practitioner. The cause of some disorders may be apparent, but others have a complex pathogenesis involving management, human-bird interaction, malnutrition, psychologic factors, disease, or hormone imbalances.

The serological responses of chickens to mass vaccination with a live V4 Newcastle disease virus vaccine in the field and in the laboratory. 1. Meat chickens.

Meat chickens on commercial broiler farms were vaccinated once at 1 to 15 days of age with a live V4 Newcastle disease virus (NDV) vaccine administered by drinking water, aerosol or coarse spray. Hatchmates were housed and similarly vaccinated in laboratory isolation pens. Samples of birds were bled at weekly to fortnightly intervals and the serums tested for haemagglutination inhibiting antibody to NDV. Log2 mean titres of up to 6.26, and assumed protection levels (based on the percentage of birds with log2 titres of 4 or greater) of up to 89%, were obtained in field trials within 4 weeks of vaccination. Differences were observed between the results obtained from parallel field and laboratory trials. The presence of maternal NDV antibody reduced the response to vaccination. The results show that this V4 vaccine can produce an adequate serological response following mass administration to Australian meat chickens housed under commercial conditions.

Natural infection of broiler breeder chickens with endemic apathogenic Newcastle disease virus and their subsequent response to vaccination with a live V4 Newcastle disease virus vaccine.

Flocks of broiler breeder chickens housed on a commercial farm were monitored from 13 w of age for natural infection with endemic lentogenic Newcastle disease virus (NDV). Seroconversion was first detected at 17 w. By 24 w, all 8 flocks had achieved peak log2 mean haemagglutination inhibiting antibody titres of up to 4.8. Antibody titres then declined and rose again over several months, suggesting cyclic reinfection with NDV. A lentogenic NDV indistinguishable from V4 was isolated from the cloaca of one bird at 18 weeks of age. At 54 weeks of age, 6 of 8 flocks were vaccinated en masse with live V4 NDV vaccine, 3 flocks by drinking water and 3 flocks by aerosol. All flocks were serologically monitored for a further 8 w. Drinking water vaccination induced an anamnestic response in 3 flocks, showing that flocks with pre-existing active immunity to NDV may be successfully vaccinated with V4. However, in all aerosol vaccinated flocks, the procedures failed to induce a response different to that observed in unvaccinated flocks. The serological response to vaccination was greater in sires than in dams.

The serological responses of chickens to mass vaccination with a live V4 Newcastle disease virus vaccine in the field and in the laboratory. 2. Layer pullets.

Layer chickens on a commercial started pullet farm were vaccinated once at 31 to 52 days of age by drinking water or aerosol with live V4 Newcastle disease virus (NDV) vaccine. Flockmates which had been rehoused in laboratory isolation pens shortly beforehand were similarly vaccinated. Samples of birds were bled at intervals and the serums tested for haemagglutination inhibiting antibody to NDV. Log2 mean titres of up to 4.88 and assumed protection levels (based on the percentage of birds with log2 titres of 4 or greater) of up to 81%, were obtained in the field trials within 4 weeks of vaccination. A subsequent laboratory trial further compared the response of different breeds of chicken to different routes of vaccination. Differences were observed between breeds, routes of vaccination, and parallel field and laboratory trials. The results show that this V4 vaccine can produce an adequate serological response following mass vaccination of Australian layer pullets housed under commercial conditions, and that care should be exercised in extrapolating results obtained under laboratory conditions.

The resistance of meat chickens vaccinated by aerosol with a live V4 Newcastle disease virus vaccine in the field to challenge with a velogenic Newcastle disease virus.

Meat chickens housed on a commercial broiler farm in Australia were vaccinated once at 10 to 11 days-of-age by aerosol with live V4 Newcastle disease virus (NDV) vaccine. Groups of vaccinated and unvaccinated birds were flown to Malaysia, where they were challenged with a virulent strain of NDV. Survival rates in vaccinated chickens challenged 7, 14, 21 or 31 d after vaccination were 0.47, 0.77, 0.97 and 0.92, respectively. All unvaccinated chickens died due to Newcastle disease (ND) following challenge. Chickens in Australia and Malaysia were bled and the serums tested for haemagglutination-inhibiting (HI) antibody to NDV. Many vaccinated birds with no detectable antibody, and all birds with a log2 titre of 2 or greater, survived challenge. The results showed that this V4 vaccine induced protective immunity in a significant proportion of chickens within 7 d of mass aerosol vaccination. This early immunity occurred in the absence of detectable circulating HI antibody. Non-HI antibody mediated immunity continued to provide protection up to 31 d after vaccination. Almost all vaccinated birds were protected within 3 w of vaccination. It is concluded that the V4 vaccine is efficacious and could be useful during an outbreak of virulent ND in Australia.