Integrating in-situ (wild) and ex-situ (captive) conservation efforts can mitigate genetic diversity loss and help prevent extinction of endangered wild populations. The whooping crane (Grus americana) experienced severe population declines in the 18th century, culminating in a collapse to ~20 individuals by 1944. Legal protections and conservation actions have since increased the census population from a stock of 16 individuals to approximately 840 individuals, yet the impact on genomic diversity remains unclear. We analysed the temporal dynamics of genomic erosion by sequencing a high-quality reference genome, and re-sequencing 16 historical (years 1867-1893) and 37 modern (2007-2020) genomes, including wild individuals and four generations of captive-bred individuals. Genomic demographic reconstructions reveal a steady decline, accelerating over the past 300 years with the European settlement of North America. Temporal genomic analyses show that despite demographic recovery, the species has lost 70% of its historical genetic diversity and has increased its inbreeding. Although the modern population bottleneck reduced the ancestral genetic load, modern populations possess more realised load than masked load, possibly resulting in a chronic loss of fitness. Integrating pedigree and genomic data, we underscore the role of breeding management in reducing recent inbreeding. Yet ongoing heterozygosity loss, load accumulation, and persistent effects of historical inbreeding (i.e., background inbreeding) argue against the species' downlisting from its current Endangered status on the IUCN Red List and the Endangered Species Act. The presence of private genetic variation in wild and captive populations suggests that wild-captive crosses could enhance genetic diversity and reduce the realised load. Our findings emphasise the role of genomics in informing conservation management and policy.

The Whooping Crane (Grus americana; WHCR) is a large, long-lived bird endemic to North America. The remnant population migrates between Aransas National Wildlife Refuge, USA, and Wood Buffalo National Park, Canada (AWBP), and has recovered from a nadir of 15-16 birds in 1941 to ~540 birds in 2022. Two ongoing reintroduction efforts in Louisiana and the Eastern Flyway together total ~150 birds. Evidence indicates the U.S. Fish and Wildlife Service (USFWS) is strongly considering downlisting the species from an endangered to a threatened status under the Endangered Species Act (ESA). We examined the current status of the WHCR through the lens of ESA threat factors, the USFWS’s Species Status Assessment (SSA) framework, and other avian downlisting actions to determine if the action is biologically warranted. Our research indicates that WHCRs are facing an intensification of most threat drivers across populations and important ranges. The AWBP is still relatively small compared to other crane species and most birds of conservation concern. To date, only one avian species has been downlisted from an endangered status with an estimated population of <3,000 individuals. Representation in terms of WHCRs historic genetic, geographic, and life history variation remains limited. Also, the lack of spatial connectivity among populations, reliance of the reintroduced populations on supplementation, and continued habitat loss suggest that WHCR populations may not be resilient to large stochastic disturbances. Given that reintroduced populations are not self-sustaining, neither supplies true redundancy for the AWBP. Proposed downlisting before recovery plan population criteria have been met is objectively unwarranted and reflects USFWS inconsistency across ESA actions. Only by incorporating basic quantitative criteria and added oversight into ESA listing decisions can we avoid an action as misguided as downlisting the Whooping Crane without consideration of its recovery plan criteria or ostensibly its population ecology.

The objective of this study was to evaluate the pharmacokinetic properties of a single dose of ceftiofur crystalline-free acid (CCFA) in whooping cranes (Grus americana). Ceftiofur crystalline-free acid is a long-acting, injectable, third-generation cephalosporin antibiotic drug. A preliminary study evaluated CCFA administered intramuscularly in the pectoral or thigh muscle at 20 or 30 mg/kg IM to a single adult whooping crane for each dose. On the basis of these data, a dose of 30 mg/kg IM of CCFA was administered to five additional whooping cranes, and blood was collected at various time points from 0 to 288 h. Pharmacokinetic parameters for ceftiofur equivalents were determined and reached concentrations above minimum inhibitory concentrations of various bacteria in other avian species (>1 µg/ml) for at least 96 h in all birds, and for 144 h in two birds. From these findings, ceftiofur crystalline-free acid appears to be a long-acting antibiotic option for whooping cranes and may be dosed every 96 h; however, additional multidose studies are needed.

Pododermatitis and wing lesions are commonly reported issues in captive crane species. Regional limb perfusion has been used as a treatment for distal limb infections in several avian species, as systemic antibiotic therapy is often prolonged and unrewarding. A black-necked crane (Grus nigricollis), Siberian crane (Leucogeranus leucogeranus), and wattled crane (Bugeranus carunculatus) were treated with amikacin (5–10 mg/kg IV infusion) regional limb perfusion for cellulitis and osteomyelitis of hind limb digits and alular osteomyelitis and septic arthritis of the carpus, respectively, with a range of 1–3 treatments per case. Clinical signs of infection resolved within 10–40 days following regional limb perfusion combined with oral or parental antibiotic therapy. No side effects were observed following regional limb perfusion. This is the first report of regional limb perfusion in cranes and the first report of intravenous regional limb perfusion in a wing of an avian species.

A retrospective study of neoplasia was conducted from necropsy and histologic reports of 446 cranes representing all 15 extant species. Cases were obtained from the International Crane Foundation (ICF), Northwest ZooPath (NWZP), and six other zoologic institutions in the United States during 1993 to 2019. Only reports from ICF (n = 61) and NWZP (n = 374) were used for estimates of disease prevalence. Overall prevalence of neoplasia was 7.35% (32 of 435), with a metastasis rate of 31.8%. Seventeen types of neoplasms were identified. Geriatric cranes were the most common age class affected (60%). The digestive system was most frequently involved (n = 12; 27.3%), followed by urinary (n = 8; 18.2%) and integumentary systems (n = 6; 13.6%). Carcinoma was the most common tumor type across all species (n = 15; 34.0%) followed by benign epithelial tumors (adenomas; n = 11; 25.0%). Multiple neoplasms were observed in only one crane. Of the cases with metastasis, hematopoietic, reproductive, and respiratory tumors had 100% metastasis (2 of 2, 2 of 2, and 3 of 3, respectively), whereas tumors of the integumentary, nervous, and musculoskeletal systems had no evidence of metastasis (0 of 7, 0 of 3, and 0 of 1, respectively). Overall, Gruidae species were found to have a high prevalence of neoplasia but a low rate of metastasis compared with other avian species with the highest species-specific rates in Eurasian (Grus grus), demoiselle (Anthropoides virgo), and wattled cranes (Bugeranus carunculatus) (40.0%, 21.4%, and 19.0%, respectively). This is the first large-scale study of neoplasia in cranes.

CASE DESCRIPTION A 7-year-old female blue crane (Anthropoides paradiseus) was initially evaluated after it had suddenly developed signs of respiratory distress following aspiration of a rock. Emergency tracheotomy had been performed, and the rock had been removed from the proximal cervical portion of the trachea. Fifty-one days later, the clinical signs had returned and the crane was reevaluated. CLINICAL FINDINGS On reevaluation, no obvious external abnormalities were appreciated at the previous surgical site and no discharge was observed from the glottis. Computed tomography and tracheoscopy revealed marked tracheal stenosis and architectural collapse of the trachea at the previous surgery site. TREATMENT AND OUTCOME Tracheal resection and anastomosis was performed to remove the stenotic tracheal segment. Histologic examination of the resected tracheal segment revealed pyogranulomas with intralesional coccobacilli, fungal hyphae consistent with Aspergillus spp, possible parasitic ova, and features suggestive of mild to moderate heterophilic and lymphoplasmacytic tracheitis. The crane was treated with piroxicam, ceftiofur crystalline free acid, terbinafine, and itraconazole. At a follow-up examination 12 weeks later, no abnormalities were appreciated, and the surgical site had completely healed. CLINICAL RELEVANCE To the authors' knowledge, this is the first reported case of successful tracheal resection and anastomosis in a bird of the order Gruiformes. The surgical approach used for the blue crane may be useful for removal of tracheal foreign bodies in this and other long-necked avian species.

Endangered whooping cranes (Grus americana) have been produced in captivity for reintroduction programs since the 1980s, using techniques such as artificial insemination, multiple clutching, and captive-rearing to speed recovery efforts. Chicks are often hand-reared (HR) by caretakers in crane costumes, socialized into groups and released together, unlike parent-reared (PR) cranes that are raised individually by a male/female crane pair and released singly. HR cranes historically exhibit greater morbidity rates during development than PR cranes, involving musculoskeletal and respiratory system disease, among others. We hypothesized that HR crane chicks exhibit a higher baseline fecal glucocorticoid metabolite (FGM) concentrations during the development compared with PR chicks. Fecal samples were collected between 15 and 70 days of age from HR (n = 15) and PR (n = 8) chicks to test for differences in FGM concentrations using a radioimmunoassay technique following ethanol extraction for steroids. Linear mixed model analysis suggests increasing age of the chick was associated with an increase in FGM (p < .001). Analysis also supported the interaction between rearing strategy and sex of the crane chick (p < .01). Female PR chicks had greater FGM concentrations than all other groups (PR male,p < .01; HR female,p < .001; and HR male,p < .001). This result suggests that there may be an effect of rearing strategy on stress physiology of whooping crane chicks, especially among females. Further research is needed to investigate whether the FGM concentrations are reflective of true differences in stress physiology of young cranes and whether this may impact health and conservation success.

Abstract: We reviewed necropsy records of 124 Whooping Cranes (Grus americana) recovered following reintroduction of 268 individuals from 2001 to 2016 in the eastern US. Causes of death were determined in 62% (77/124) of cases facilitated by active monitoring that limited decomposition and scavenging artifact. The greatest proportions of mortality were caused by predation (0.468; 95% confidence interval 0.356–0.580; 36/77), collision with power lines or vehicles (0.260; 0.162–0.358; 20/77), and gunshot (0.169; 0.085–0.253; 13/77). Six deaths were attributed to infection (0.078; 0.018–0.138; 6/77), including bacterial and fungal etiologies. Lead analysis of 50 liver samples yielded two results with elevated concentrations (3.65 and 10.97 ppm wet weight), and 10 bone samples from partial carcasses lacking suitable liver tissue resulted in one elevated result (48.82 ppm dry weight). These data indicate that underlying subclinical or clinical lead toxicosis may be a factor in up to 5% of deaths attributed to predation or impact trauma. Brain cholinesterase activity testing indicated no exposure to organophosphate or carbamate pesticides (mean±SD=17.32±2.90 µmol/min/g, 31/71). The causes of death and potential underlying factors summarized in this study constitute the first definitive mortality survey of migratory Whooping Cranes based on a high carcass recovery rate. Causes of death by infectious etiologies remained comparatively rare in this study, and occurred as single cases with no evidence of sustained transmission among reintroduced Whooping Cranes.

In numerous animal clades, the evolutionary history of host species drives patterns of gut microbial community structure, resulting in more divergent microbiota with increasing phylogenetic distance between hosts. This phenomenon, termed phylosymbiosis, has been observed in diverse evolutionary lineages, but has been difficult to detect in birds. Previous tests of phylosymbiosis among birds have been conducted using wild individuals, and thus interspecific differences in diet and environment may have masked a phylogenetic signal. Therefore, we tested for phylosymbiosis among all 15 species of cranes (family Gruidae) housed in the same captive environment and maintained on identical diets. 16S rRNA sequencing revealed that crane species harbour distinct gut microbiota. Overall, we detected marginally significant patterns of phylosymbiosis, the strength of which was increased when including the estimates of absolute microbial abundance (rather than relative abundance) derived from microbial densities determined by flow cytometry. Using this approach, we detected the statistically significant signatures of phylosymbiosis only after removing male cranes from our analysis, suggesting that using mixed-sex animal cohorts may prevent the detection of phylosymbiosis. Though weak compared with mammals (and especially insects), these results provide evidence of phylosymbiosis in birds. We discuss the potential differences between birds and mammals, such as transmission routes and host filtering, that may underlie the differences in the strength of phylosymbiosis.

Juvenile whooping cranes (Grus americana) raised for wild release were found to have an increased incidence of rib fractures at fledging in 2017 compared with the previous 16 years. Serum analysis showed 30-day-old juveniles in 2017 (n = 12) had significantly lower 25-hydroxyvitamin D3 and significantly higher parathyroid hormone concentrations than juveniles in 2010 (n = 6) with no history of rib fractures. Increased serum parathyroid hormone concentrations in the 2017 juveniles persisted to fledging age. Review of dietary and environmental management revealed that juveniles in 2017 were provided a commercial diet with a lower, and perhaps suboptimal, calcium:phosphorus ratio and experienced reduced time outdoors in the first month after hatch, presumably resulting in less ultraviolet B radiation exposure. Mild hyperparathyroidism in precocial whooping cranes may result when dietary constraints and/or outdoor access is compromised and manifest as rib fractures in the absence of traumatic injury.

The central aim of conservation biology is to understand and mitigate the effects of human activities on biodiversity. To successfully achieve this objective, researchers must take an interdisciplinary approach that fully considers the effects, both direct and indirect, of anthropogenic disturbances on wildlife physiology and health. A recent surge in research has revealed that host-associated microbiota-the archaeal, bacterial, fungal and viral communities residing on and inside organisms-profoundly influence animal health, and that these microbial communities can be drastically altered by anthropogenic activities. Therefore, conservation practitioners should consider the disruption of host-associated microbial diversity as a serious threat to wildlife populations. Despite the tremendous potential for microbiome research to improve conservation outcomes, few efforts have been made to truly integrate these fields. In this review, we call for the microbial renaissance of conservation biology, where biodiversity of host-associated microbiota is recognized as an essential component of wildlife management practices. Using evidence from the existing literature, we will examine the known effects of anthropogenic activities on the diversity of host-associated microbial communities and integrate approaches for maintaining microbial diversity to successfully achieve conservation objectives.

Biochemical and trace element analyses of blood from wild Whooping Cranes ( Grus americana) were performed to assess the health of the only self-sustaining, migratory population in North America. Juvenile cranes ( n=31) approximately 49-70 d-old were sampled at Wood Buffalo National Park, Northwest Territories, Canada, in midsummer from 2010 to 2012. Archived serum ( n=24) and whole blood ( n=31) samples from captive juvenile cranes were selected as age-matched controls. Reference values were calculated for serum biochemical analytes and trace elements in whole blood from the captive juvenile Whooping Cranes reared under controlled conditions and with known health histories. Several statistical differences among blood biochemical and trace element values of the wild and captive juveniles were identified and were likely attributable to dietary differences between the populations.

For long-lived species with low fecundity rates, population growth rate can be sensitive to changes in annual survival. Understanding where, when, and why animals die provides useful information for prioritizing conservation practices designed to increase survival. As part of a satellite tracking study, we identified 19 confirmed and suspected deaths of Whooping Cranes of various ages from the Aransas-Wood Buffalo Population. Of these, more occurred during winter (∼45%), compared with summer (∼40%) or migration (∼15%). Summer mortalities occurred exclusively within Wood Buffalo National Park, and all winter mortalities occurred on the primary wintering grounds along the Texas Gulf Coast. Predation was the most common cause identified; proximate cause of mortality was not known for most of the birds. Previous assessments of mortality, based on assumptions and some supporting data, speculated that most mortality occurred during migration. Although preliminary, our results are based on stronger evidence and provide a different perspective. Migration may be less risky than previously assumed and of similar risk to other seasons; thus, conservation or management efforts to reduce mortality may have greater effect when focused during breeding and wintering periods, although feasibility and efficacy of these actions will need to be determined.

Brucellosis, or Bang's disease, caused by the bacterium Brucella abortus, is a potentially devastating cause of abortion and unthriftiness in cattle.

The central aim of conservation biology is to understand and mitigate the effects of human activities on biodiversity. To successfully achieve this objective, researchers must take an interdisciplinary approach that fully considers the effects, both direct and indirect, of anthropogenic disturbances on wildlife physiology and health. A recent surge in research has revealed that host-associated microbiota—the archaeal, bacterial, fungal and viral communities residing on and inside organisms—profoundly influence animal health, and that these microbial communities can be drastically altered by anthropogenic activities. Therefore, conservation practitioners should consider the disruption of host-associated microbial diversity as a serious threat to wildlife populations. Despite the tremendous potential for microbiome research to improve conservation outcomes, few efforts have been made to truly integrate these fields. In this review, we call for the microbial renaissance of conservation biology, where biodiversity of host-associated microbiota is recognized as an essential component of wildlife management practices. Using evidence from the existing literature, we will examine the known effects of anthropogenic activities on the diversity of host-associated microbial communities and integrate approaches for maintaining microbial diversity to successfully achieve conservation objectives.

The unicellular blood parasites in the order Haemosporida are highly diverse, infect many vertebrates, are responsible for a large disease burden among humans and animals, and have reemerged as an important model system to understand the evolutionary and ecological dynamics of host-parasite interactions. The phylogenetics and systematics of Haemosporida are limited by poor sampling of different vertebrate host taxa. We surveyed the Haemosporida of wild whooping cranes (Grus americana) and sandhill cranes (Grus canadensis) (Ayes: Gruiformes) using a combination of morphological and molecular approaches. We identified Haemoproteus antigonis in blood smears based on published morphological descriptions. Phylogenetic analysis based on partial cytochrome b (cytb) and cytochrome oxidase (coI) sequences placed H. antigonis parasites in a novel Glade, distinct from all avian Haemosporida genera for which cytb and/or col sequences are available. Molecular clock and divergence estimates suggest this crane Glade may represent a new genus. This is the first molecular description of H. antigonis and the first report of H. antigonis in wild whooping cranes, an endangered bird in North America. Further sampling of Haemosporida, especially from hosts of the Gruiformes and other poorly sampled orders, will help to resolve the relationship of the H. antigonis Glade to other avian Haemosporida genera. Our study highlights the potential of sampling neglected host species to discover novel lineages of diverse parasite groups. (C) 2016 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY-NC-ND license.