Genetic trends in parrot Bornavirus: a clinical analysis.

Parrot Bornavirus (PaBV) has been reported to cause indigestion and other wasting symptoms such as weight loss and lethargy. The pathogenesis of PaBV has yet to be fully elucidated. This study reports PaBV infections in South Korea and suggests a trend in the genetic information gathered from clinical cases. A total of 487 birds with or without clinical symptoms were tested for bornavirus. Twelve of 361 asymptomatic birds tested positive for bornavirus, while 15 of 126 birds with various symptoms tested positive. A segment of approximately 1,540 bps including the N, X, P and M proteins were obtained from 23 of the positive strains and analyzed with other strains found on GenBank that had clinical information. PaBV was type 2 and 4 in South Korea, and certain amino acid sequences showed a difference between symptom presenting animals and asymptomatic animals in the X protein and P protein. When considering that some asymptomatic cases may have been latent infections at the time of examination, it is plausible these trends may grow stronger with time. Majority of PaBV was type 4 in South Korea. If these trends are confirmed, diagnosis of potentially pathogenic PaBVs in a clinical manner will be possible during the early stages of infection.

Scarlet macaw (Ara macao) breeding at the Mimbres archaeological site of Old Town (early AD 1100s) in Southwestern New Mexico.

Examination of avian eggshell at the Old Town archaeological site in Southwestern New Mexico, United States of America, indicates that scarlet macaw (Ara macao) breeding occurred during the Classic Mimbres period (early AD 1100s). Current archaeological and archaeogenomic evidence from throughout the American Southwest/Mexican Northwest (SW/NW) suggests that Indigenous people bred scarlet macaws at an unknown location(s) between AD 900 and 1200 and likely again at the northwestern Mexico site of Paquimé post-AD 1275. However, there is a lack of direct evidence for breeding, or the location(s) of scarlet macaw breeding itself, within this area. This research, for the first time, provides evidence of scarlet macaw breeding using scanning electron microscopy of eggshells from Old Town.

Intralipid Emulsion Therapy for the Treatment of Suspected Toxicity in 2 Avian Species.

Intravenous lipid emulsion (ILE) therapy has shown promise as a treatment option for a variety of lipophilic toxins. Two birds presented for suspected ingestion of a toxic substance. A blue-and-gold macaw (Ara ararauna) presented after chewing a block of bromethalin rodenticide without overt clinical signs at the time of presentation. Additionally, a free-ranging bald eagle (Haliaeetus leucocephalus) was found weak and depressed near a municipal landfill after presumptive ingestion of pentobarbital. Both birds were treated with ILE therapy for potential intoxication without any adverse events. The macaw was clinically normal after 3 days of hospitalization and at a 1-week reevaluation. The eagle was transferred to a rehabilitation center after markedly improved mentation and strength and was released 7 days later. Clinicians should consider ILE therapy for the treatment of lipophilic toxicities; however, monitoring is recommended for persistent lipemia and other adverse effects that have been reported in the veterinary literature.

Pathogenic Idiopathic Extramedullary Hematopoiesis in a Yellow-Collared Macaw (Primolius auricollis).

1.5-year-old yellow-collared macaw (Primolius auricollis) was presented as a referral case for chronic breathing difficulties and coelomic distension. The bird was in poor body condition, and coelomic distension and green-colored urates were noted during the physical examination. Radiographic images revealed a large coelomic space-occupying soft-tissue lesion that was ultrasonographically confirmed to be hepatomegaly; the liver had a heterogeneous echogenic pattern. An ultrasound-guided fine needle aspirate of the liver was performed. The cytological results revealed immature hematopoietic cells with signs of dyserythropoiesis and were consistent with extramedullary hematopoiesis (EMH). The plasma biochemistry panel revealed a marked increase in aspartate aminotransferase and bile acids, consistent with severe hepatic disease. Following the results of the diagnostic tests, chemotherapy was initiated using hydroxyurea. Two weeks after the initial presentation and treatment, the bird died and a full postmortem examination was performed. Macroscopic examination confirmed severe hepatomegaly and severe splenomegaly. Histopathological examination of tissue samples confirmed severe EMH in the liver and spleen, splenic and renal hemosiderosis, and acute pulmonary congestion. The bone marrow was normal. The final diagnosis was pathogenic idiopathic EMH, and this case was unusual in both its presentation and severity. Extramedullary hematopoiesis is usually related to myeloid proliferative disorder, chronic blood loss, hemolytic disease, or chronic inflammatory disease. Mycobacteriosis and parasitic infection have been reported to be associated with EMH in birds; however, the inflammatory patterns seen in those cases were lacking in this case. Myeloproliferative neoplasia also appears an unlikely disease condition in this case considering that histopathology found normal architecture in the studied bone marrow; however, bone marrow abnormalities in locations other than the one sampled could not be excluded. A short review of homeostatic and pathogenic hematopoiesis in birds is provided to support the likely diagnosis of idiopathic EMH.

Uveal Malignant Melanoma in a Hybrid Macaw Parrot (Ara chloropterus × Ara militaris).

An adult male green-winged (Ara chloropterus) × military (Ara militaris) hybrid macaw of unknown age was presented to the Texas A&M Veterinary Medical Teaching Hospital for evaluation of left periocular swelling, blepharospasm, and ocular discharge. Complete ophthalmic examination and B mode ocular ultrasonography were performed to evaluate the affected eye. Ocular examination revealed a blind left globe with exophthalmos, periocular swelling, diffuse corneal edema, and severe anterior fibrinous uveitis obscuring visualization of the intraocular structures. An ultrasound examination revealed a hyperechoic mass that filled the posterior segment of the globe. Focal discontinuity of the posterior sclera suggested scleral rupture. Enucleation with histopathology was recommended and performed 1.5 months later. Intraoperatively, a heavily pigmented and friable soft tissue mass extruded through the posterior sclera and extended into the orbit. Histologically, the mass was consistent with a malignant melanoma. The patient died 18 hours after the surgical procedure because of unknown complications. Postmortem examination did not identify evidence of metastasis. This study described the clinical appearance and histopathologic findings of a rare ocular neoplasm with extrascleral extension in a hybrid macaw. Although uncommon in psittacine birds, primary ocular melanocytic neoplasms may display features of malignancy with scleral or orbital invasion, or both.

Comparison of Serum Thyroid Hormone Levels in Green-winged Macaws (Ara chloropterus) Using Radio and Chemiluminescent Immunoassays.

This study compared the use of radioimmunoassay (RIA) and chemiluminescent immunoassay (CLIA) to measure serum thyroid hormone levels in green-winged macaws (Ara chloropterus). The sample group comprised 20 male and 13 female (n = 33) healthy, captive green-winged macaws of reproductive age but outside of breeding season. Mean free triiodothyronine (T3), total T3, and free thyroxine (T4) measured by CLIA and RIA corresponded to 5.11 and 5.08 pmol/L (3.33 and 3.31 pg/mL), 1.59 and 1.56 nmol/L (103.5 and 101.5 ng/dL), and 8.25 and 8.82 pmol/L (0.64 and 0.69 ng/dL), respectively. Mean total T4 by RIA corresponded to 6.29 nmol/L (0.49 µg/dL). Mean free T4 and thyroid-stimulating hormone (TSH) levels differed according to immunoassay method, with higher values measured by RIA compared with CLIA. Sex influence was statistically significant in immunoassay results only on free T3 levels. Free T3 levels measured by RIA were higher than levels measured by CLIA in male birds, but this difference was not found with the female bird samples. Conversely, free T3 levels measured by CLIA were higher in the female macaws compared with male birds. Comparative analysis of thyroid hormone measurements in this study revealed that RIA and CLIA are equivalent methods to measure free T4 and total T3 levels but not TSH levels. These findings support the use of CLIA for free T4 and total T3 level determinations in green-winged macaws. However, the CLIA kit used in this study provided invalid total T4 level results for the macaws sampled. Radioimmunoassay and CLIA were equally ineffective for determining TSH levels in this species.

Impact of Dietary Fructose on the Lipid Profile in Six Macaws.

Atherosclerosis is frequently diagnosed in captive parrots. Recent studies have highlighted the potential role of high fructose intake in the development of human cardiovascular disease. The objective of this study was to investigate the effects of dietary fructose on the lipid profile in psittacine birds. Six macaws, composed of 3 green-winged macaws (Ara chloropterus, 2 females, 1 male), and 3 blue-and-gold macaws (Ara ararauna, 1 female, 2 males), ranging from 15 to 26 years of age, were used as subject animals for this dietary investigation. Initially, fruits were removed from the diet and replaced with vegetables lower in fructose for a month. Then, with a crossover study design, each bird was randomly selected to receive either 34 kcal of a fructose solution daily for 5 days then an equivalent volume of water for 5 days, or water in the first 5 days followed by the 34 kcal fructose solution. Lipid profiles were repeatedly performed over the course of the study at specific intervals. Serum cholesterol (3.01-5.55 mmol/L), triglycerides (0.27-2.70 mmol/L), high-density lipoprotein-cholesterol (1.43-4.79 mmol/L), low-density lipoprotein-cholesterol (0.02-0.85 mmol/L), and very low density lipoprotein-cholesterol (0.12-1.22 mmol/L) levels were not statistically significantly different over time and treatment. Although no significant effect could be demonstrated in this study, further studies are needed to investigate the effect of a high fructose intake in psittacine birds, because it could have a deleterious influence on their cardiovascular health if proven to be linked with dyslipidemia as reported in humans. Higher amounts of fructose and longer periods of administration should be considered for future studies.

Transsinus Pinning to Correct Lateral Deviation of the Upper Beak in Juvenile Macaws.

Lateral deviation of the upper beak ("scissor beak" or "wry beak") is a common malocclusion in many species of birds but appears to be a common presentation in macaws (Ara species). This article describes transsinus pinning, a procedure in which a pin is passed through the frontal sinuses, turned parallel to the upper beak, and attached to the tip of the beak with an orthodontic rubber band to provide constant tension on the beak as it grows. The tension of the rubber band is maintained until the beak is considered straight. The results of 16 cases in which this beak-straightening procedure was used are presented. The age of the chicks that had their beaks straightened ranged from 7 to 28 weeks, and they were placed into 2 groups: those younger than 12 weeks (12 chicks; 75%) and those older (4 chicks; 25%). Complete resolution was achieved in 87.5% (14 of 16) of the avian patients that were treated with this procedure. The 2 remaining cases (12.5%) failed to fully respond. The 12 younger birds (75%; age, 7-12 weeks; median, 10 weeks; range, 5 weeks) responded to treatment within 2 weeks (12-85 days; median, 14 days; range, 73 days); the remaining 4 older chicks (25%; age, 14-28 weeks; median, 17.5 weeks; range, 14 weeks) required a longer period (13-90 days; median, 25.5 days; range, 77 days) for the beak to straighten. Transsinus pinning is a simple, effective, and rapid technique for correcting this malocclusion in macaw chicks younger than 16 weeks old. With this approach, in most cases, excellent results can be expected in only 2-4 weeks.

Predicting oil content in ripe Macaw fruits (Acrocomia aculeata) from unripe ones by near infrared spectroscopy and PLS regression.

A method for early quantification of unripe macaw fruits oil content using near-infrared spectroscopy (NIR) and partial least squares (PLS) is presented. After harvest, the fruit takes about 30 days to reach its maximum oil accumulation. The oil content was quantified thirty days after harvest using Soxhlet extraction. PLS models were built using NIR spectra of shell obtained five days after harvest (Shell5). The Shell5 model was compared with models built using NIR spectra of the shell (Shell30) and mesocarp thirty days after harvest (Pulp30). Ordered predictors selection was used to select the most informative variables. The best models presented root mean square error of prediction and correlation coefficient of prediction of 4.87% and 0.89 for Shell5; 5.83% and 0.85 for Shell30; 4.76% and 0.92 for Pulp30. Thus, the anticipated prediction of oil content could reduce the time and costs of macaw palm quality control and storage.

Complete mitochondrial genome sequence of the northern red-shouldered macaw (Diopsittaca nobilis).

The northern red-shouldered macaw, Diopsittaca nobilis, is listed in the Convention on International Trade in Endangered Species of Wild Fauna and Flora Appendix II. Here, we report for the first time the complete mitochondrial genome of D. nobilis. This new sequence is 16,992 base pairs (bp) in length and includes 13 protein-coding genes, 22 transfer RNAs, two ribosomal RNAs, and a single non-coding control region. The overall nucleotide composition of this sequence consists of 30.40% A, 14.00% G, 23.60% T, and 32.00% C. The phylogenetic relationships suggested that the mitogenome of D. nobilis is close to that of three other macaw species. Our results provide useful mitogenomic information to support further studies on the phylogeny and taxonomy of Psittaciformes.

Characterization and phylogenetic analysis of the complete Green-winged macaw (Ara chloropterus) mitogenome.

In this study, we sequenced and characterized the complete mitochondrial genome of Green-winged macaw (Ara chloropterus) listed on CITES Appendix II. The circular double-stranded genome was 16,991 bp in size and included 22 transfer RNA (tRNA) genes, 13 protein-coding genes (PCGs), two ribosomal RNA genes (rrnL and rrnS), and one non-coding control region (D-loop). Both order and arrangement of genes were identical to those of other animal mitogenomes. The base content was 30.00% A, 14.38% G, 23.27% T, and 32.35% C, with an A + T content of 53.37%. Furthermore, phylogenetic analysis indicated that A. chloropterus is closely related to the Blue and yellow macaw (A. ararauna).

Genomic characterization of enteropathogenic Escherichia coli (EPEC) of avian origin and rabbit ileal loop response; a pet macaw (Ara chloropterus) as a possible zoonotic reservoir.

Enteropathogenic Escherichia coli (EPEC) constitutes one of the main causes of mortality in children in low- to medium-income countries. Diverse animal species have been linked as reservoirs, including birds. The aim of this study was to describe the genomic and phylogenetic features of an EPEC recovered from a pet macaw and further characterizing the macro and microscopic lesion in a rabbit ileal loop experimental model. The isolate was whole-genome sequenced (WGS) obtaining its genotypic and phenotypic in silico characteristics and inoculated in a rabbit experimental model with subsequently evaluating the strain's pathogenicity by scanning electron microscopy (SEM) and histopathology. The isolate was characterized as O109:H21-B1-ST40 typical EPEC, harboring several virulence factors of diarrheagenic E. coli. The macaw EPEC genome was located in a monophyletic clade of human and animal ST40 EPEC sequences. In vivo inoculation demonstrated severe hemorrhage with SEM and histopathological analysis confirming these lesions to be associated with intra-epithelial lymphocytes. Therefore, the isolate not only shared several genotypic and phylogenetic similarities with EPEC that affects humans and animals, but was able to induce severe tissue injury in a mammal model. These findings highlight the underrated role of pet birds as zoonotic reservoirs and the diversity in virulence factors being unraveled by new WGS studies.

Isolation and characterization of 15 new microsatellite markers for the globally endangered Lear's macaw Anodorhynchus leari.

A set of 16 microsatellite markers was characterized for Lear's macaw (Anodorhynchus leari) using DNA samples from captive individuals. Extending this molecular toolkit, including the use of samples from wild individuals, is expected to provide the required power of resolution for pedigree inference of both wild and captive individuals, and could support research on the genetic structure of wild populations. We characterize a set of 15 microsatellite markers optimized for the Lear's macaw, developed from a microsatellite-enriched library in a three-step procedure. Primer pairs were initially designed for 62 microsatellites with > 7 tandem repetitions. After amplification of DNA of five wild individuals from different localities, 22 loci seemed to be polymorphic and were further tested on 12 wild nestling samples. Fifteen unlinked loci showed unambiguous peaks and low to moderate polymorphism levels. The combination of the four most polymorphic markers allowed individual identification even of putative sibs.These markers complement previously described microsatellites developed for A. leari and constitute a fundamental genetic toolkit for the investigation of the genetics of both wild and captive populations, thus assisting integrated management plans for the conservation of this globally endangered species.

Experimental removal of invasive Africanized honey bees increased breeding population size of the endangered Lear's macaw.

BACKGROUND: Invasive Africanized honey bees potentially compete with cavity-nesting birds in South America. However, the impacts of this competition and its conservation consequences to threatened species are poorly known. We quantified the presence of these bees and assessed their competition for cliff cavities used by nesting Lear's macaws Anodorhynchus leari, a globally endangered parrot endemic to the Caatinga biome of Brazil. We treated beehives with permethrin by shooting them with a crossbow bolt that distributed the compound upon impact. When feasible, we removed the comb and applied an insecticide (fipronil) to deter bee recolonization. We subsequently surveyed the macaw breeding population to verify whether our treatment allowed for nest recruitment in cavities previously occupied by bees. RESULTS: We recorded > 100 beehives in the nesting cliffs. Hives outnumbered macaw nests tenfold in two areas recently recolonized by macaws. Cavities occupied by bees were significantly higher than those occupied by macaws, suggesting that macaws may be forced to breed in lower cavities. None of the untreated cavities (n = 50) were occupied by nesting macaws, whereas 15% of treated cavities (n = 52) were occupied within 2 years post treatment. Treated cavities occupied by macaws were significantly higher than those not occupied. Hive management increased macaw breeding population by 71% of the macaw breeding population increase. CONCLUSION: Experimental hive treatments were effective in restoring nesting resources lost due to bee infestation. An intensive and continued eradication program is recommended to enhance macaw habitat restoration, facilitating its expansion into historical areas. © 2020 Society of Chemical Industry.

INTESTINAL AND BLOOD PARASITES IN SCARLET (ARA MACAO) AND GREAT GREEN (ARA AMBIGUA) MACAWS IN WILDLIFE REHABILITATION CENTERS IN COSTA RICA.

Costa Rica undertakes continuous efforts to recover the native population of macaw species through rehabilitation programs for breeding and releasing birds in protected areas. In the summer of 2018, a total of 107 scarlet (Ara macao) and 93 great green (Ara ambigua) macaws were sampled in four wildlife rehabilitation centers in Costa Rica. Fecal samples representing 200 individuals were analyzed for intestinal parasites, and 23 individuals were sampled for hemoparasites. Ascaridia and Capillaria were found in fecal samples. No hemoparasites were found. The distribution of percentage of infection was analyzed by location, species, and housing type. As part of a health screening prior to release, parasitological examination is recommended.

A Novel Fungal Lipase With Methanol Tolerance and Preference for Macaw Palm Oil.

Macaw palm is a highly oil-producing plant, which presents high contents of free fatty acids, being a promising feedstock for biofuel production. The current chemical routes are costly and complex, involving highly harsh industrial conditions. Enzymatic processing is a potential alternative; however, it is hampered by the scarce knowledge on biocatalysts adapted to this acidic feedstock. This work describes a novel lipase isolated from the thermophilic fungus Rasamsonia emersonii (ReLip), which tolerates extreme conditions such as the presence of methanol, high temperatures, and acidic medium. Among the tested feedstocks, the enzyme showed the highest preference for macaw palm oil, producing a hydrolyzate with a final free fatty acid content of 92%. Crystallographic studies revealed a closed conformation of the helical amphipathic lid that typically undergoes conformational changes in a mechanism of interfacial activation. Such conformation of the lid is stabilized by a salt bridge, not observed in other structurally characterized homologs, which is likely involved in the tolerance to organic solvents. Moreover, the lack of conservation of the aromatic cluster IxxWxxxxxF in the lid of ReLip with the natural mutation of the phenylalanine by an alanine might be correlated with the preference of short acyl chains, although preserving catalytic activity on insoluble substrates. In addition, the presence of five acidic amino acids in the lid of ReLip, a rare property reported in other lipases, may have contributed to its ability to tolerate and be effective in acidic environments. Therefore, our work describes a new fungal biocatalyst capable of efficiently hydrolyzing macaw oil, an attractive feedstock for the production of "drop-in" biofuels, with high desirable feature for industrial conditions such as thermal and methanol tolerance, and optimum acidic pH. Moreover, the crystallographic structure was elucidated, providing a structural basis for the enzyme substrate preference and tolerance to organic solvents.

Modified Tail Amputation Technique in a Blue and Gold Macaw (Ara ararauna) With Uropygial Gland Adenocarcinoma.

A 33-year-old male blue and gold macaw (Ara ararauna) presented with a 5-month history of an ulcerated lesion and feather loss at the tail base. Two 4-mm biopsies obtained by the primary care veterinarian were consistent with uropygial gland adenocarcinoma. The bird was examined at the Veterinary Medical Teaching Hospital, University of California, and on physical evaluation, the dorsal and ventral surface of the tail base were devoid of feathers, ulcerated and crusted without an identifiable uropygial gland. Complete blood count, plasma biochemistry panel, whole-body radiographs, and an echocardiogram were performed before surgery. The bird was anesthetized, and a complete amputation of the tail was performed. The skin was incised with a radiofrequency electrosurgical system approximately 2 mm circumferentially cranial to the diseased tissue. The musculature was transected to the level of the vertebral column, disarticulating between the second and third caudal vertebrae and transecting the spinal cord with a no. 15 blade. Lateral vertebral processes of the second vertebra were removed with a rongeur. Coccygeus lateralis muscles and tensor fasciae latae muscles and skin were closed laterolaterally with 2 layers and 3-0 polydioxanone suture. The bird recovered uneventfully and was discharged after 6 days of hospitalization. The histopathological diagnosis was adenocarcinoma with squamous differentiation, marked scirrhous response, and superficial epithelial ulceration. It was determined that narrow margins of unaffected tissue were achieved from the pathological examination of submitted material. The bird was evaluated 24 days after surgery and again 3.5 months after surgery, without evidence of complications or recurrence. Approximately 10 days after the last reexamination, the bird was euthanatized after being found minimally responsive at home. A postmortem examination was not performed.

Necrotic Enteritis Caused by Clostridium perfringens in Blue and Gold Macaws (Ara ararauna).

Clostridium perfringens types A and C, which are gram-positive, anaerobic, spore-forming bacteria, can cause necrotic enteritis in birds. Although Clostridium perfringens is considered a commensal organism in the avian intestinal tract, in association with severe stress, other infectious agents, or immunosuppressive conditions, it can cause disease outbreaks. This report describes a disease occurrence of necrotic enteritis caused by C perfringens in macaws (Ara ararauna). Two adult male blue and gold macaws maintained in a zoo exhibit were presented for postmortem examinations after histories of sudden death. Based on the gross examinations and microscopic evaluation of submitted tissue from both birds, the cause of death was determined to be necrotic enteritis. Microbiologic assays followed by polymerase chain reaction analyses identified the isolated strains as C perfringens type A, indicated by only being positive for the cpa gene that encodes the α-toxin. The birds were maintained in an exhibit in which patrons can interact with the animals within their environment. Thus, organisms, such as this pathogen, may present a danger for other birds because visitors could disperse the bacterium to other parts of the zoo.

How much does it cost to save a species from extinction? Costs and rewards of conserving the Lear's macaw.

Although the limited resources available to save species from extinction necessitate the optimization of conservation actions, little is known about their costs and effectiveness. We developed a costs-rewards framework that integrates information on which sectors of society contribute to funding conservation, how much is contributed, how funds are distributed among conservation targets and how these investments drive not only conservation rewards but also the economic and ecosystem services that benefit society. We applied this framework to the Lear's macaw (Anodorhynchus leari), a species discovered in the wild in 1978 with only 60 individuals. Funds invested over the last 25 years reached US$3.66 million. The contribution of governments, non-governmental organizations and private funders varied over time, as did the funding targets. Funds were proportionally invested to mitigate the main causes of mortality, while no funds were devoted to protecting foraging habitats. Conservation rewards were satisfactory, with the cost and time needed to downlist the species from critically endangered to endangered being similar to those invested in other bird species. However, economic rewards (through ecotourism and handicrafts linked to the conservation of the species) were low and require promotion, while ecosystem services provided by Lear's macaws have yet to be quantified.

Key Ring Fixator: A Novel External Fixation Technique for Avian Long Bone Stabilization.

A 12-year-old blue-fronted Amazon parrot (Amazona aestiva) of unknown sex (case 1) and a 14-year-old female hybrid Catalina macaw (Ara ararauna × Ara macao) (case 2) were evaluated and treated for an open tarsometatarsal fracture and a tibiotarsal fracture, respectively. In case 1, 1 month of external coaptation resulted in a delayed union, significant osteolysis, and presumptive osteomyelitis, which led to the decision to treat with a key ring fixator. In case 2, a key ring fixator was chosen for fracture repair because of presumed resistance to destruction by the bird. In both cases, fractures were stabilized with makeshift circular external fixators composed of key rings, K-wires, orthopedic wire, and acrylic resin. After key ring fixator removal, radiographs confirmed complete bone healing. Both patients had acceptable function of the affected limbs 5 years (case 1) and 2 years (case 2) after their procedures. The key ring fixator described in this report is a viable option for fracture repair in pelvic limbs of moderately sized birds (300-1500 g).