Application of 3D-printed prosthetic lower beak in an Oriental stork (Ciconia boyciana).

To our knowledge, ours is the first case of applying a 3D-printed prosthetic beak to an Oriental stork (Ciconia boyciana). A stork in captivity underwent several surgeries for beak fractures, but the lower-mandible fractures failed to be repaired. Therefore, we applied a patient-specific beak prosthesis of titanium alloy and nylon. Because the prosthetic beak could not be maintained due to mandible and soft-tissue inflammation, the stork was euthanized. Still, we confirmed typical behavior and feeding for ~3 months after surgery. This report highlights some of the challenges we encountered and identifies process improvements required for a more successful surgery.

Temporal Changes in Demography and Genetic Diversity of Oriental Storks at the Stage of Long-Term Captive Propagation and Reintroduction Initiation.

Captive propagation and reintroduction are the major steps in the ex-situ conservation of locally extirpated endangered species in a historical region. In a species restoration project conducted in South Korea, we examined temporal changes in demographics and genetic diversity of oriental storks (Ciconia boyciana). Demographic and genetic data from 1996-2018 were analyzed for 80% of all captive and recently reintroduced individuals. Founder establishment and pair formation induced increases in population size and genetic diversity during the early stage of captive propagation. The degree of genetic diversity was found to become saturated and stable with long-term captive propagation. However, this might be a concern for future genetic diversity of both captive and reintroduced populations simultaneously due to the extraction of captive populations at the early stage of reintroduction. Our findings suggest that periodic evaluation of genetic diversity and selection for releasing individuals, using effective genetic markers, would assist in balancing the genetic diversity of the captive and reintroduced oriental storks at the early stage of reintroduction.

Lysobacter ciconiae sp. nov., and Lysobacter avium sp. nov., isolated from the faeces of an Oriental stork.

Two Gram-stain-negative, mesophilic, strictly aerobic, nonspore forming, and yellow-pigmented strains with rod-shaped cells, designated H21R20T and H23M41T, were isolated from the faeces of an Oriental stork (Ciconia boyciana). Based on 16S rRNA gene sequences, both strains showed the highest similarity (98.3-98.4%) to the type strain of Lysobacter concretionis. Phylogenetic analysis based on the 16S rRNA genes and 92 bacterial core genes showed that strains H21R20T and H23M41T were robustly clustered with L. concretionis Ko07T. Whole genome sequencing revealed that the genomes of both strains were approximately 2.9 Mb in size. The DNA G + C contents of the H21R20T and H23M41T strains were 67.3 and 66.6%, respectively. The two strains showed 80.1-81.7% average nucleotide identity with L. concretionis Ko07T. Strain H21R20T grew optimally at 30°C and pH 8.0 and in the presence of 0.5-3% (wt/vol) NaCl, while strain H23M41T grew optimally at 30°C and pH 7.0-8.0 and in the presence of 0-3% (wt/vol) NaCl. Both strains possessed iso-C15:0, iso-C16:0 and summed feature 9 (iso-C17:1ω9c and/or C16:010-methyl) as the major cellular fatty acids, ubiquinone Q-8 as a predominant quinone, and diphosphatidylglycerol, phosphatidylglycerol, and phosphatidylethanolamine as the major polar lipids. A multifaceted investigation demonstrated that strains H21R20T and H23M41T represent novel species of the genus Lysobacter, for which we propose the names Lysobacter ciconiae sp. nov. and Lysobacter avium sp. nov. for strains H21R20T (= KCTC 82316T = JCM 34832T) and H23M41T (= KCTC 62676T = JCM 33223T), respectively.

Description of Ornithinimicrobium ciconiae sp. nov., and Ornithinimicrobium avium sp. nov., isolated from the faeces of the endangered and near-threatened birds.

Phenotypic and genomic analyses were performed to characterize two novel species, H23M54T and AMA3305T, isolated from the faeces of the Oriental stork (Ciconia boyciana) and the cinereous vulture (Aegypius monachus), respectively. Strains H23M54T and AMA3305T showed the highest similarities of 16S rRNA gene sequences and complete genome sequences with Ornithinimicrobium cavernae CFH 30183T (98.5% of 16S rRNA gene sequence similarity and 82.1% of average nucleotide identity, ANI) and O. pekingense DSM 21552T (98.5% of 16S rRNA gene sequence similarity and 82.3% of ANI), respectively. Both strains were Gram-stain-positive, obligate aerobes, non-motile, non-spore-forming, and coccoid- and rodshaped. Strain H23M54T grew optimally at 25-30°C and pH 8.0 and in the presence of 1.5-2% (wt/vol) NaCl, while strain AMA3305T grew optimally at 30°C and pH 7.0 and in the presence of 1-3% (wt/vol) NaCl. Both strains had iso-C15:0, iso-C16:0, and summed feature 9 (iso-C17:1ω9c and/or C16:0 10-methyl) as major cellular fatty acids. MK-8 (H4) was identified as the primary respiratory quinone in both strains. Strains H23M54T and AMA3305T possessed diphosphatidylglycerol and phosphatidylglycerol as major polar lipids. Moreover, strains H23M54T and AMA3305T commonly contained ribose and glucose as major sugars and L-ornithine, L-alanine, glycine, and aspartic acid as major amino acids. The polyphasic taxonomic data indicate that strains H23M54T and AMA3305T represent novel species of the genus Ornithinimicrobium. We propose the names Ornithinimicrobium ciconiae sp. nov. and Ornithinimicrobium avium sp. nov. for strains H23M54T (= KCTC 49151T = JCM 33221T) and AMA3305T (= KCTC 49180T = JCM 32873T), respectively.

Flaviflexus ciconiae sp. nov., isolated from the faeces of the oriental stork, Ciconia boyciana.

A novel Gram-stain-positive, non-motile, non-spore-forming, coccobacillus-shaped, strictly aerobic bacterium, designated strain H23T48T, was isolated from the faecal sample of an oriental stork collected from the Seoul Grand Park Zoo in Seoul, Republic of Korea. Optimal growth of strain H23T48T was observed at 30-37 °C, pH 8 and with 3 % (w/v) NaCl. 16S rRNA gene sequence-based phylogenetic analysis revealed that strain H23T48T was closely related to the genus Flaviflexus, with 97.0 and 96.7 % sequence similarities to Flaviflexus salsibiostraticola EBR4-1-2T and Flaviflexus huanghaiensis H5T, respectively. Strain H23T48T possessed MK-9(H4) as the major menaquinone and C16 : 0 (42.4 %), C18 : 1 ω9c (31.3 %) and C14 : 0 (17.7 %) as the major cellular fatty acids. The polar lipids included phosphatidylglycerol, two unidentified lipids, six unidentified phospholipids and two unidentified glycophospholipids. The amino acid composition of the cell-wall peptidoglycan was l-alanine, l-lysine, d-glutamic acid, l-aspartic acid and glycine. The genomic G+C content of strain H23T48T is 59.5 mol% and the average nucleotide identity value between H23T48T and F. salsibiostraticola KCT C33148T (=EBR4-1-2T) is 75.5 %. Based on the obtained data, strain H23T48T represents a novel species of the genus Flaviflexus, for which the name Flaviflexus ciconiae sp. nov. is proposed. The type strain is H23T48T (=KCTC 49253T=JCM 33282T).

Iodobacter ciconiae sp. nov., a bacterium isolated from feces of oriental stork, Ciconia boyciana.

A novel Gram-stain-negative, facultatively anaerobic, non-motile, non-violet-pigmented, rod-shaped bacterium, designated strain H11R3T, was isolated from the feces of Oriental stork, Ciconia boyciana, collected from Seoul Grand Park Zoo, Republic of Korea. Phylogenetic analysis of the 16S rRNA gene sequence revealed that H11R3T formed a monophyletic clade with Iodobacter fluviatilisDSM 3764T, Iodobacter arcticusDSM 100243T, and Iodobacter limnosediminisDSM 103822T, with sequence similarities of 98.8, 98.6 and 98.4 %, respectively. H11R3T grew optimally at 15 °C, pH 8, with 0.5 % (w/v) NaCl. The predominant isoprenoid quinone was ubiquinone-8 (Q-8), and polar lipids included phosphatidylethanolamine, three unidentified lipids, four unidentified phospholipids, and two unidentified aminophospholipids. The major fatty acids were summed feature 3 and C16 : 0, and the DNA G+C content of the genome is 48.0 mol%. The average nucleotide identity (ANI) value between strains H11R3T and I. fluviatilis NCTC 11159T (=DSM 3764T) is 83.7 %. On the basis of phenotypic, genotypic, phylogenetic and chemotaxonomic characteristics, strain H11R3T represents a novel species of the genus Iodobacter for which the name Iodobacterciconiae sp. nov. is proposed. The type strain is H11R3T (=KCTC 62666T=JCM 33283T).

A Case of Chaunocephalosis by Chaunocephalus ferox (Digenea: Echinostomatidae) in an Oriental White Stork, Ciconia boyciana, in Korea.

We intended to describe a case of chaunocephalosis and morphological characteristics of its causative agent, Chaunocephalus ferox, recovered from an oriental white stork, Ciconia boyciana, in the Republic of Korea. An oriental white stork was referred to the Wildlife Center of Chungbuk in Korea in February 2014 for severe depression with cachexia and it died the next day. At necropsy, the stomach was severely expanded and 7 thick-walled nodules were observed in the upper part of the intestine. Although the stomach was filled with full of foreign materials, the intestine was almost empty. The nodules were globular and total 9 flukes were recovered. They were 8,030-8,091 µm in length and 3,318-3,333 µm in maximum width. Because the flukes had bulbous forebody with short narrow subcylindrical hindbody, 27 collar spines, and vitelline follicles not reaching to the posterior end, the specimens were identified as being C. ferox. The cyst formation induced thickening of the intestinal wall with narrowing of the lumen that could have contributed to the gastric impaction to the death of the host. This is the first described case of chaunocephalosis and its causative agent C. ferox found from an oriental white stork in Korea.

Hematologic and serum biochemical reference intervals of the Oriental white stork (Ciconia boyciana) and the application of an automatic hematologic analyzer.

This study was conducted to establish accurate baseline values of clinical laboratory data with regard to age-related changes in the Oriental white stork (Ciconia boyciana). In addition, the availability of an automated hematological cell counter was evaluated. A total of 94 clinically normal storks, including 64 young storks (＜1 year old; 30 male and 34 female) and 30 adults (＞ 1 year old; 17 male and 13 female) were included. Hematological assays were performed using manual and automated cell counters and serum biochemistry profiles were examined using an automated analyzer. There were no significant differences in any parameters between male and female storks, while 16 parameters were significantly different between young and adult storks. Of these 16 parameters, total protein, albumin, aspartate aminotransferase, alanine aminotransferase, creatinine, triglyceride, total bilirubin, potassium, white blood cell count, packed cell volume, mean cell volume and hemoglobin levels were higher in adult storks than in young storks, while the latter showed higher glucose, uric acid and alkaline phosphatase levels, as well as a higher sodium/potassium ratio. The results presented herein will aid researchers who work for the conservation and rehabilitation of this endangered species.

Hematologic and serum biochemical reference intervals of the Oriental white stork (Ciconia boyciana) and the application of an automatic hematologic analyzer

This study was conducted to establish accurate baseline values of clinical laboratory data with regard to age-related changes in the Oriental white stork (Ciconia boyciana). In addition, the availability of an automated hematological cell counter was evaluated. A total of 94 clinically normal storks, including 64 young storks ( 1 year old; 17 male and 13 female) were included. Hematological assays were performed using manual and automated cell counters and serum biochemistry profiles were examined using an automated analyzer. There were no significant differences in any parameters between male and female storks, while 16 parameters were significantly different between young and adult storks. Of these 16 parameters, total protein, albumin, aspartate aminotransferase, alanine aminotransferase, creatinine, triglyceride, total bilirubin, potassium, white blood cell count, packed cell volume, mean cell volume and hemoglobin levels were higher in adult storks than in young storks, while the latter showed higher glucose, uric acid and alkaline phosphatase levels, as well as a higher sodium/potassium ratio. The results presented herein will aid researchers who work for the conservation and rehabilitation of this endangered species.

A Case of Chaunocephalosis by Chaunocephalus ferox (Digenea: Echinostomatidae) in an Oriental White Stork, Ciconia boyciana, in Korea

We intended to describe a case of chaunocephalosis and morphological characteristics of its causative agent, Chaunocephalus ferox, recovered from an oriental white stork, Ciconia boyciana, in the Republic of Korea. An oriental white stork was referred to the Wildlife Center of Chungbuk in Korea in February 2014 for severe depression with cachexia and it died the next day. At necropsy, the stomach was severely expanded and 7 thick-walled nodules were observed in the upper part of the intestine. Although the stomach was filled with full of foreign materials, the intestine was almost empty. The nodules were globular and total 9 flukes were recovered. They were 8,030–8,091 μm in length and 3,318–3,333 μm in maximum width. Because the flukes had bulbous forebody with short narrow subcylindrical hindbody, 27 collar spines, and vitelline follicles not reaching to the posterior end, the specimens were identified as being C. ferox. The cyst formation induced thickening of the intestinal wall with narrowing of the lumen that could have contributed to the gastric impaction to the death of the host. This is the first described case of chaunocephalosis and its causative agent C. ferox found from an oriental white stork in Korea.

Post-Mating Sexual Behaviors of Oriental Storks (Ciconia boyciana) in Captivity.

Parental behaviors that vary by sex and breeding stage facilitate the survival of offspring in birds. Females invest in reproducing according to the level of direct and/or indirect benefits provided by males. Males face trade-offs from defending territories or nests, providing for the young, and seeking additional mating opportunities. We examined whether post-mating sexual behaviors such as courtship display and nest-building were associated with reproductive investments made by males and females in the current study. The oriental stork (Ciconia boyciana) is known as a solitary breeder that nests in tall trees and provides biparental care with long post-hatching development. We filmed parental behaviors in seven stork pairs in captivity for 655 hours (245 hours during nest-building, 218 hours during incubation, 328 hours during nestling, and 192 hours during the fledgling period). We found that paired mating behaviors such as synchronous allopreening and bill-clattering were highly skewed to the pre-incubation period. Males participated in nest-building more than females although both sexes shared similar parental care during the incubation and nestling period. Male nest-building was negatively correlated with male nest attentiveness only during the nestling period. Our results suggest that male oriental storks with higher nest-building effort might spend more time taking precautions against conspecific intruders or nest predators near the nests with chicks while the females were attending the nests in the form of biparental care.