Psittacine Birds Research Articles

Chlamydia psittaci EN AVES PSITACIDAS EN DOS PARQUES ZOOLÓGICOS DE VENEZUELA

La determinación de Chlamydia psittaci (Cp) en aves psitácidas en parques zoológicos de Venezuela representa una estrategia de conservación y preservación para este grupo de aves, donde múltiples especies se encuentran amenazadas de extinción y otras han perdido su capacidad de reincorporación a su hábitat natural. A través de la reacción en cadena de la polimerasa anidada (PCR) fue amplificada la subunidad 16S del ADNr de Cp en 50 muestras de hisopado cloacal de aves psitácidas, reportando una frecuencia de 62 %. El trabajo fue realizado en el Parque Zoológico Las Delicias (PZD) 8 % y el Aquarium de Valencia (AV) 54 %. La elevada frecuencia fue asociada a un genotipo de baja concentración y virulencia debido a la ausencia de signos clínicos de clamidiosis aviar. Estos resultados demuestran la necesidad de promover la detección de Cp, principalmente para el AV que actúa como centro de recepción de ejemplares de decomiso, y, al igual que el PZD, poseen otras especies vulnerables a la extinción con riesgo de infección a Cp.

Temporal and spatial analysis of psittacosis in association with poultry farming in the Netherlands, 2000\u20132015

BackgroundHuman psittacosis is a highly under diagnosed zoonotic disease, commonly linked to psittacine birds. Psittacosis in birds, also known as avian chlamydiosis, is endemic in poultry, but the risk for people living close to poultry farms is unknown. Therefore, our study aimed to explore the temporal and spatial patterns of human psittacosis infections and identify possible associations with poultry farming in the Netherlands.MethodsWe analysed data on 700 human cases of psittacosis notified between 01-01-2000 and 01-09-2015. First, we studied the temporal behaviour of psittacosis notifications by applying wavelet analysis. Then, to identify possible spatial patterns, we applied spatial cluster analysis. Finally, we investigated the possible spatial association between psittacosis notifications and data on the Dutch poultry sector at municipality level using a multivariable model.ResultsWe found a large spatial cluster that covered a highly poultry-dense area but additional clusters were found in areas that had a low poultry density. There were marked geographical differences in the awareness of psittacosis and the amount and the type of laboratory diagnostics used for psittacosis, making it difficult to draw conclusions about the correlation between the large cluster and poultry density. The multivariable model showed that the presence of chicken processing plants and slaughter duck farms in a municipality was associated with a higher rate of human psittacosis notifications. The significance of the associations was influenced by the inclusion or exclusion of farm density in the model.ConclusionsOur temporal and spatial analyses showed weak associations between poultry-related variables and psittacosis notifications. Because of the low number of psittacosis notifications available for analysis, the power of our analysis was relative low. Because of the exploratory nature of this research, the associations found cannot be interpreted as evidence for airborne transmission of psittacosis from poultry to the general population. Further research is needed to determine the prevalence of C. psittaci in Dutch poultry. Also, efforts to promote PCR-based testing for C. psittaci and genotyping for source tracing are important to reduce the diagnostic deficit, and to provide better estimates of the human psittacosis burden, and the possible role of poultry.

Plasma protein, haematologic and blood chemistry changes in African grey parrots (Psittacus erithacus) experimentally infected with bornavirus

ABSTRACTBornaviruses are considered to be the causative agent of proventricular dilatation disease (PDD) in psittacine birds. In order to detect haematological and blood chemistry changes during the development of PDD and a possible correlation with clinical signs and the virological status, six African grey parrots (Psittacus erithacus) were experimentally infected with parrot bornavirus 4 (PaBV-4) by subcutaneous route. All six parrots developed clinical signs of varying extent and successful infection was confirmed in all the birds by seroconversion or detection of RNA of the PaBV-4 infection strain. Based on population-based and intra-individual reference ranges established during 12 months prior to experimental infection, only minor haematological changes were detected in individual birds after infection. Changes in blood chemistry were restricted to aspartate aminotransferase, creatine kinase, total protein, glucose and uric acid. Plasma protein electrophoresis revealed marked changes starting 10 weeks post infection characterized by an increase in the γ-globulin fraction and a gradual decrease to normal values during weeks 22–34. Indications of an acute-phase reaction at the initial stages of infection were not detected. While three birds suffered from clinical signs of PDD, which included weight loss and neurological disorders and died before development of haematological and plasma protein changes, recovery of clinical disease was paralleled in the remaining birds by an increase in γ-globulins and bornavirus-specific antibody titres.

A high prevalence of beak and feather disease virus in non-psittacine Australian birds.

Beak and feather disease virus (BFDV) is a circovirus and the cause of psittacine beak and feather disease (PBFD). This disease is characterized by feather and beak deformities and is a recognized threat to endangered Psittaciformes (parrots and cockatoos). The role that non-psittacine birds may play as reservoirs of infection is unclear. This study aimed to begin addressing this gap in our knowledge of PBFD. Liver samples were collected from birds presented to the Australian Wildlife Health Centre at Zoos Victoria's Healesville Sanctuary for veterinary care between December 2014 and December 2015, and tested for BFDV DNA using polymerase chain reaction coupled with sequencing and phylogenetic analyses.Results/Key findings. Overall BFDV was detected in 38.1 % of 210 birds. BFDV was detected at high prevalence (56.2 %) in psittacine birds, in the majority of cases without any observed clinical signs of PBFD. We also found that BFDV was more common in non-psittacine species than previously recognized, with BFDV detected at 20.0 % prevalence in the non-psittacine birds tested, including species with no clear ecological association with psittacines, and without showing any detectable clinical signs of BFDV infection. Further research to determine the infectivity and transmissibility of BFDV in non-psittacine species is indicated. Until such work is undertaken the findings from this study suggest that every bird should be considered a potential carrier of BFDV, regardless of species and clinical presentation. Veterinary clinics and wildlife rehabilitation facilities caring for birds that are susceptible to PBFD should reconsider biosecurity protocols aimed at controlling BFDV.

Survey of pathogens in threatened wild red-tailed Amazon parrot (Amazona brasiliensis) nestlings in Rasa Island, Brazil.

The red-tailed Amazon parrot (Amazona brasiliensis) is a threatened species of psittacine bird that inhabit coastal regions of Brazil. In view of the threat of this species, the aim of this study was to perform a health evaluation in wild nestlings in Rasa Island, determining the prevalence of enterobacteria and infectious agents according to type of nest. Blood samples were collected from 64 birds and evaluated for antibodies of Chlamydia psittaci by commercial dot-blot ELISA. Cloacal and oropharyngeal swabs samples were collected from 23 birds from artificial wooden nests, 15 birds from PVC nests and 2 birds from natural nests for microbiological analysis. Swab samples were collected from 58 parrots for C. psittaci detection by PCR and from 50 nestlings for Avian Influenza, Newcastle Disease and West Nile viruses’ detection analysis by real-time RT-PCR. Ten bacterial genera and 17 species were identified, and the most prevalent were Escherichia coli and Klebsiella oxytoca. There was no influence of the type of nest in the nestlings’ microbiota. All samples tested by ELISA and PCR were negative. There is currently insufficient information available about the health of A. brasiliensis and data of this study provide a reference point for future evaluations and aid in conservation plans.

Captive wild birds as reservoirs of enteropathogenic E. coli (EPEC) and Shiga-toxin producing E. coli (STEC).

Psittacine birds have been identified as reservoirs of diarrheagenic Escherichia coli, a subset of pathogens associated with mortality of children in tropical countries. The role of other orders of birds as source of infection is unclear. The aim of this study was to perform the molecular diagnosis of infection with diarrheagenic E. coli in 10 different orders of captive wild birds in the state of São Paulo, Brazil. Fecal samples were analyzed from 516 birds belonging to 10 orders: Accipitriformes, Anseriformes, Columbiformes, Falconiformes, Galliformes, Passeriformes, Pelecaniformes, Piciformes, Psittaciformes and Strigiformes. After isolation, 401 E. coli strains were subjected to multiplex PCR system with amplification of genes eae and bfp (EPEC), stx1 and stx2 for STEC. The results of these tests revealed 23/401 (5.74%) positive strains for eae gene, 16/401 positive strains for the bfp gene (3.99%) and 3/401 positive for stx2 gene (0.75%) distributed among the orders of Psittaciformes, Strigiformes and Columbiformes. None of strains were positive for stx1 gene. These data reveal the infection by STEC, typical and atypical EPEC in captive birds. The frequency of these pathotypes is low and restricted to few orders, but the data suggest the potential public health risk that these birds represent as reservoirs of diarrheagenic E. coli.

Are anti-ganglioside antibodies associated with proventricular dilatation disease in birds?

The identification of Parrot bornaviruses (PaBV) in psittacine birds with proventricular dilatation disease (PDD) has not been sufficient to explain the pathogenesis of this fatal disease, since not all infected birds develop clinical signs. Although the most accepted theory indicates that PaBV directly triggers an inflammatory response in this disease, another hypothesis suggests the disease is triggered by autoantibodies targeting neuronal gangliosides, and PDD might therefore resemble Guillain-Barré Syndrome (GBS) in its pathogenesis. Experimental inoculation of pure gangliosides and brain-derived ganglioside extracts were used in two different immunization studies. The first study was performed on 17 healthy chickens (Gallus gallus domesticus): 11 chickens were inoculated with a brain ganglioside extract in Freund’s complete adjuvant (FCA) and six chickens inoculated with phosphate-buffered saline. A second study was performed five healthy quaker parrots (Myiopsitta monachus) that were divided into three groups: Two quaker parrots received purified gangliosides in FCA, two received a crude brain extract in FCA, and one control quaker parrot received FCA alone. One chicken developed difficult in walking. Histologically, only a mild perivascular and perineural lymphocytic infiltrate in the proventriculus. Two quaker parrots (one from each treatment group) had mild lymphoplasmacytic encephalitis and myelitis. However, none of the quaker parrots developed myenteric ganglioneuritis, suggesting that autoantibodies against gangliosides in birds are not associated with a condition resembling PDD.

Development and evaluation of TaqMan real-time PCR assay for detection of beak and feather disease virus

Psittacine beak and feather disease (PBFD) is one of the most significant viral diseases in psittacine birds. The aim of the presented study was to develop a highly specific and sensitive TaqMan real-time PCR assay for universal detection of beak and feather disease virus (BFDV). Primers and a hydrolysis probe were selected on the highly conserved regions belonging to the ORF1 of the BFDV genome which were identified by aligning 814 genomic sequences downloaded from the GenBank database. The evaluation of the reaction parameters suggested a reaction efficiency of 97.1%, with consistent detection of 101 virus copies/μl of nucleic acid extract. The low values of standard deviation and coefficient of variation indicate a high degree of reproducibility and repeatability. The diagnostic applicability of the assay was proven on 36 BFDV positive and 107 negative specimens of psittacine origin representing 28 species. The assay showed a 100% ability to detect distinct genetic variants of the virus. Our data suggest that the presented TaqMan real-time PCR represents a specific, sensitive and reliable assay facilitating the molecular detection of BFDV.

Prevalence of Taxa of Pasteurellaceae Among Populations of Healthy Captive Psittacine Birds.

Sixty-two strains of Pasteurellaceae-like bacteria were isolated from the tracheas of 87 clinically healthy psittacine birds in two Danish zoos. The isolates were identified by a combination of rpoB and 16S rRNA gene sequencing and by matrix-assisted laser desorption-ionization time of flight. Twenty-eight strains belonged to the genus Volucribacter or were related to this genus and to the unnamed taxon 34 of Bisgaard, and 28 strains were related to the unnamed taxon 44 of Bisgaard. Four strains were identified as Pasteurella multocida , two isolates were classified with the related taxon 45 of Bisgaard, and a single isolate was classified as Pasteurella sp. The investigation documented an unrecognized reservoir of rarely reported and unclassified or unnamed species of Pasteurellaceae-like bacteria in psittacine birds. The results were in accordance with a recent report on isolation of Pasteurellaceae from diseased psittacine birds, and the investigation documented that the same taxa of Pasteurellaceae-like bacteria can be isolated from apparently healthy birds as well as from diseased birds.

The Influence of a Combined Butorphanol and Midazolam Premedication on Anesthesia in Psittacid Species

Premedication is considered routine for domestic animal and human anesthesia but is rarely applied to avian patients, and few controlled studies exist to document effects of premedication in avian species. To determine the effects of a butorphanol and midazolam premedication on general anesthesia and quality of induction and recovery phases in psittacid species, 17 clinically healthy birds undergoing anesthesia were randomly allocated into either a premedicated or control group. Anesthetic parameters were subsequently compared. Induction time and isoflurane concentration required for anesthetic maintenance were reduced in the premedicated group. Induction quality scores were improved in the premedicated group and no adverse effects on anesthesia and cardiovascular stability were observed. Use of a combined butorphanol and midazolam premedication in clinically healthy psittacine birds appears safe and effective. Premedication provides a beneficial effect at induction and enables maintenance levels of anesthetic gas to be reduced.

Viral vector vaccines expressing nucleoprotein and phosphoprotein genes of avian bornaviruses ameliorate homologous challenge infections in cockatiels and common canaries.

Avian bornaviruses are causative agents of proventricular dilatation disease (PDD), an often fatal disease of parrots and related species (order Psittaciformes) which is widely distributed in captive psittacine populations and may affect endangered species. Here, we established a vaccination strategy employing two different well described viral vectors, namely recombinant Newcastle disease virus (NDV) and modified vaccinia virus Ankara (MVA) that were engineered to express the phosphoprotein and nucleoprotein genes of two avian bornaviruses, parrot bornavirus 4 (PaBV-4) and canary bornavirus 2 (CnBV-2). When combined in a heterologous prime/boost vaccination regime, NDV and MVA vaccine viruses established self-limiting infections and induced a bornavirus-specific humoral immune response in cockatiels (Nymphicus hollandicus) and common canaries (Serinus canaria forma domestica). After challenge infection with a homologous bornavirus, shedding of bornavirus RNA and viral loads in tissue samples were significantly reduced in immunized birds, indicating that vaccination markedly delayed the course of infection. However, cockatiels still developed signs of PDD if the vaccine failed to prevent viral persistence. Our work demonstrates that avian bornavirus infections can be repressed by vaccine-induced immunity. It represents a first crucial step towards a protective vaccination strategy to combat PDD in psittacine birds.

External Beam Radiation Therapy of Squamous Cell Carcinoma in the Beak of an African Grey Parrot (Psittacus timneh)

Squamous cell carcinoma has been reported in a variety of bird species, most commonly psittacine and gallinaceous birds. The long-term prognosis in nongallinaceous birds is generally poor if complete surgical excision is not possible. Squamous cell carcinoma of the rhinotheca was diagnosed in a 34-year-old timneh African grey parrot (Psittacus timneh) with a 2-year history of beak abnormalities. No evidence of metastasis or local invasion were found on results of radiographs or computed tomography scan. The bird was treated with surgical debulking and palliative megavoltage radiation therapy. After 4 radiation treatments, the affected tissue was necrotic and was debrided to reveal healthy granulation tissue. The bird died approximately 7 months after diagnosis and 4 months after cessation of radiation treatment. At the time of death, a small scab lesion remained at the left oral commissure, but no visible tumor regrowth was evident. A postmortem examination was not performed, however, and tumor recurrence could not be ruled out in this bird.

A review of Sarcocystis spp. shed by opossums (Didelphis spp.) in Brazil

Os gambás Sul-americanos são os hospedeiros definitivos de Sarcocystis falcatula, Sarcocystis neurona, Sarcocystis speeri e Sarcocystis lindsayi. Estas espécies de Sarcocystis são morfologicamente similares, mas podem ser distinguidas por sua patogenicidade e infectividade em hospedeiros intermediários (aves e camundongos imunodeficientes) e técnicas moleculares. Os gambás são animais silvestres e sinantrópicos e amplamente distribuídos no território nacional. Estudos anteriores demonstraram uma alta contaminação ambiental com esporocistos de S. neurona em diversas regiões brasileiras. Este artigo revisa informações sobre Sarcocystis spp. excretados por gambás e sua ocorrência no Brasil.

A comparison of PCR assays for beak and feather disease virus and high resolution melt (HRM) curve analysis of replicase associated protein and capsid genes

Beak and feather disease virus (BFDV) threatens a wide range of endangered psittacine birds worldwide. In this study, we assessed a novel PCR assay and genetic screening method using high-resolution melt (HRM) curve analysis for BFDV targeting the capsid (Cap) gene (HRM-Cap) alongside conventional PCR detection as well as a PCR method that targets a much smaller fragment of the virus genome in the replicase initiator protein (Rep) gene (HRM-Rep). Limits of detection, sensitivity, specificity and discriminatory power for differentiating BFDV sequences were compared. HRM-Cap had a high positive predictive value and could readily differentiate between a reference genotype and 17 other diverse BFDV genomes with more discriminatory power (genotype confidence percentage) than HRM-Rep. Melt curve profiles generated by HRM-Cap correlated with unique DNA sequence profiles for each individual test genome. The limit of detection of HRM-Cap was lower (2×10−5ng/reaction or 48 viral copies) than that for both HRM-Rep and conventional BFDV PCR which had similar sensitivity (2×10−6ng or 13 viral copies/reaction). However, when used in a diagnostic setting with 348 clinical samples there was strong agreement between HRM-Cap and conventional PCR (kappa=0.87, P<0.01, 98% specificity) and HRM-Cap demonstrated higher specificity (99.9%) than HRM-Rep (80.3%).

Detection and Phylogeny of Beak and Feather Disease Virus and Avian Polyomavirus in Psittacine Pet Birds in Turkey

Beak and feather disease virus (BFDV) and avian polyomavirus (APV) infections cause various health problems in many psittacine species, and may ultimately cause death. The aims of this study were to investigate, for the first time, the presence of BFDV and APV in psittacine birds kept in Turkey by SYBR-Green real-time polymerase chain reaction (SYBR-Green real-time PCR) and to determine the phylogeny of any circulating strains. In this study, 62 samples consisting of feather samples from 35 Melopsittacus undulatus, 16 Psittacus erithacus, 5 Forpus coelestis, 4 Nymphicus hollandicus, 1 Agapornis roseicollis, and 1 Psephotus haematonotus were collected and analyzed for the presence of BFDV-DNA and APV-DNA using SYBR-Green real-time PCR. All birds had plumage defects and 50% were considered clinically depressed. BFDV-DNA and APV-DNA were detected in feather samples of 12 (19.3%) and 9 (14.5%) birds, respectively. Both BFDV-DNA and APV-DNA were detected in 23 (37%) feather samples, from which BFDV-Rep gene and APV-VP1 gene were partially sequenced and phylogenetic analyses were conducted. Birds suspected to be infected with BFDV and APV should be analyzed by real-time PCR for early diagnosis, and preventive measurements should be applied to reduce viral exposure to healthy birds. The results of this investigation indicate a high frequency of BFDV and APV infections in birds with plumage defects in Turkey. However, strains were similar to those previously reported in other areas of the world.

DEVELOPMENT OF REFERENCE RANGES FOR PLASMA TOTAL CHOLINESTERASE AND BRAIN ACETYLCHOLINESTERASE ACTIVITY IN FREE-RANGING CARNABY'S BLACK-COCKATOOS (CALYPTORHYNCHUS LATIROSTRIS).

Published avian reference ranges for plasma cholinesterase (ChE) and brain acetylcholinesterase (AChE) are numerous. However, a consistently reported recommendation is the need for species- and laboratory-specific reference ranges because of variables, including assay methods, sample storage conditions, season, and bird sex, age, and physiologic status. We developed normal reference ranges for brain AChE and plasma total ChE (tChE) activity for Carnaby's Black-Cockatoos (Calyptorhynchus latirostris) using a standardized protocol (substrate acetylthiocholine at 25 C). We report reference ranges for brain AChE (19-41 μmol/min per g, mean 21±6.38) and plasma tChE (0.41-0.53 μmol/min per mL, mean 0.47±0.11) (n=15). This information will be of use in the ongoing field investigation of a paresis-paralysis syndrome in the endangered Carnaby's Black-Cockatoos, suspected to be associated with exposure to anticholinesterase compounds and add to the paucity of reference ranges for plasma tChE and brain AChE in Australian psittacine birds.

Prevalence and genotype of Chlamydia psittaci in faecal samples of birds from zoos and pet markets in Kunming, Yunnan, China

Chlamydia psittaci is an important zoonotic pathogen in birds and may be transmitted to humans and result in severe respiratory disease. To assess the prevalence and genotype of C. psittaci in birds in Kunming, Yunnan, China, a total of 136 specimens of psittacine birds involving 8 species were collected from the city’s zoos (n=60) and pet markets (n=76). The frequency of C. psittaci infection was 19.9% (27/136) in the psittacine birds. The prevalence of C. psittaci was higher in pet birds (26.3%; 20/76) than in zoo birds (11.7%; 7/60) (P=0.034). In particular, among Agapornis fischeri, the C. psittaci infection (50%; 10/20) was significantly more frequent in the pet markets than in the zoos (P=0.006). In addition, the highest prevalence of 41.2% (7/17) was found in Ara ararauna. To determine the genotype of C. psittaci, 23 OmpA gene fragments (about 1.4 kb) in 27 positive samples were successfully amplified and sequenced. Phylogenetic analysis showed that all the 23 strains belonged to genotype A. Our results demonstrate the high prevalence of C. psittaci genotype A infection in psittacine birds in Yunnan Province, suggesting a potential threat to human health in this area. Therefore, it is necessary to take effective measures to prevent the spread of C. psittaci among psittacine birds, as well as among employees and customers.

Virulence and antimicrobial resistance of Klebsiella pneumoniae isolated from passerine and psittacine birds

ABSTRACTKlebsiella pneumoniae is considered one of the most important Gram-negative opportunistic pathogens. The contact between humans and birds poses health risks to both. The aim of this study was to investigate the resistance and virulence of K. pneumoniae isolates from psittacines and passerines, seized from illegal trade in Brazil. We analysed 32 strains isolated from birds of the orders Psittaciformes and Passeriformes by polymerase chain reaction (PCR) for virulence factor genes. Antibiotic resistance was assessed by disk diffusion assay and PCR. The results indicated that fimH (100%), uge (96.8%), kfu (81.2%) and irp-2 (68.7%) were the most common virulence genes, followed by kpn (46.8%), K2 (43.7%), mrkD (34.3%) and iroN (15.6%). The combination of virulence genes resulted in a great diversity of genotypes and the heterogeneity of the strains is also confirmed in the analysis by amplified fragment length polymorphism. The susceptibility profiles of the K. pneumoniae showed 25% of multiple antibiotic resistance strains. We identified seven strains that presented non-extended spectrum beta lactamase blaSHV variants SHV-1 and SHV-11 and one strain positive to the blaTEM-1 gene. Plasmid-mediated quinolone resistance was present in 10 strains (10/32). The data obtained in this study reveal the pathogenic potential of this pathogen and highlight the need for surveillance and monitoring.

Genetic Detection and Identification of Chlamydophila psittaci in Captive Psittacine Birds in Thailand

Genetic Detection and Identification of Chlamydophila psittaci in Captive Psittacine Birds in Thailand

Encephalitozoon hellem infection in aviary passerine and psittacine birds in Spain

A European goldfinch (Carduelis carduelis), a canary (Serinus canaria), and a lovebird (Agapornis roseicollis) captive-bred at three different private aviaries in Spain were submitted for necropsy with a history of weakness and ruffled feathers, weight loss associated with glossitis, and respiratory disease, respectively. Microscopically, enterocytes in the jejunum and ileum contained colonies of gram- and Stamp-positive, oval to elliptical microorganisms within parasitophorous vacuoles in the apical cytoplasm. Nested PCR using MSP primers that target microsporidian RNA genes produced amplicons of expected size for Encephalitozoon species, and analysis of forward and reverse DNA sequences confirmed the presence of Encephalitozoon hellem in all cases. The main cause of death of all three birds consisted of concurrent infections. However, intestinal encephalitozoonosis may have contributed to exacerbated catabolism. Encephalitozoonosis (or microsporidiosis) has been rarely described in passerine birds.