Analysis of Genetic Structure and Genetic Diversity in Japanese Black Kite Population Using mtDNA.

The black kite (Milvus migrans) is widespread in the "Old World" and is a common raptor species in Japan. However, examples of regional population dynamics and genetic research are limited. To reconsider the natural history and current status of the black kite in Japan, we analyzed the genetic diversity of the Japanese black kite population. We collected 59 black kite samples from 22 locations across Japan and extracted DNA; 1585 bp sequences of the cytochrome b and control region, which form part of the mitochondrial DNA, were determined for genetic analysis of the population. Consequently, six haplotypes were detected in 59 individuals, 50 of which had the same major haplotype, namely, Mm1. Moreover, the genetic analysis indicated that the Japanese black kite population would fit the population expansion model. Phylogenetic analysis using sequences obtained in this study or from a DNA database indicated that the Japanese black kite population can be divided into two groups: (1) Mm1 and its close haplotypes and (2) Mm5. The sequence of Mm1 was exactly the same as that of black kite in Pakistan, India, and Korea, suggesting that this haplotype is generally widespread in East Eurasia, and that the ancestral haplotype of the Japanese population likely migrated from continental East Asia and expanded its distribution throughout Japan. In summary, we found that the black kite population in East Eurasia, including Japan, is composed of at least two lineages.

A Comparison of Respiratory Variables in a Wild Population of Ventilated and Nonventilated Black Kites (Milvus migrans govinda).

Black kites (Milvus migrans govinda) often become injured due to kite-string injuries obtained during the International Kite Flying Festival that takes place during the celebration of Uttarayan, in Ahmedabad, India. The purpose of this study was to compare respiratory variables in ventilated and nonventilated black kites, to facilitate an evidence-based decision on whether to ventilate injured birds. A total of 56 juvenile and adult black kites of unknown sex were anesthetized and divided into a ventilated group (VG) and a nonventilated group (NVG). The variables recorded were end tidal carbon dioxide (ETCO2), peripheral capillary oxygen saturation, heart rate, respiratory rate, and maintenance percentage of isoflurane (%ISO). The ETCO2 values were higher in the NVG compared to the VG (72.9 ± 19.8 mm Hg compared to 16.0 ± 5.6 mm Hg, P < .01). The %ISO required to maintain adequate depth of anesthesia was higher in NVG compared to VG (3.5% ± 0.5% compared to 2.2% ± 0.4%, P < .001). Peripheral capillary oxygen saturation was not statistically different between groups (NVG 96.3% ± 3.5% compared to the VG 96.0% ± 5.1%, P = .867). Ventilation under anesthesia appears to lower the ETCO2 and the %ISO flow rate used to maintain a surgical plane of anesthesia. These findings should provide information that will be helpful in the anesthetic management of black kites and may apply to other avian species as well.

Assessment of the exposure to heavy metals and arsenic in captive and free-living black kites (Milvus migrans) nesting in Portugal.

Due to their high trophic level, raptor species may serve as important indicators of environmental contamination by heavy metals. This study was conducted to determine if the habitat of the black kite (Milvus migrans) is contaminated by heavy metals and arsenic and to assess the degree and type of exposure that may be present. For this purpose, this study was conducted on a group of captive birds (n = 12) and on a group of free-living birds admitted to two wildlife rehabilitation centers (n = 31). Blood samples were taken for analysis of arsenic (As), mercury (Hg) and lead (Pb) concentrations by inductively coupled plasma mass spectrometry (ICP-MS). Captive birds had the lowest blood concentrations for all toxic elements examined, but significant differences from the concentrations found in free-living birds were only observed for Hg and Pb (p < 0.01). Arsenic concentrations were almost three times higher in free-living birds (4.521 ±â€¯5.695 µg/dl) then in captive birds (1.566 ±â€¯0.753 µg/dl). In all the samples of captive birds' mercury was not detected, while in free-living birds we observed a concentration of 7.493 ±â€¯8.464 µg/dl (p < 0.01). Regarding lead, we observed a concentration almost four-fold higher in free-living birds (19.430 ±â€¯29.294 µg/dl) then in captive birds (4.449 ±â€¯1.987 µg/dl) (p < 0.01). Therefore, available sources of Pb and Hg seem to be present in the habitat of the black kite.

Lifetime variation in feather corticosterone levels in a long-lived raptor.

In long-lived animals, the challenges that threaten individual homeostasis, and the way they are dealt with, are expected to vary in an age-related manner, encompassing the progressive selection of superior phenotypes and the acquisition and improvement of key skills (e.g. foraging, breeding and fighting abilities). Since exposure to homeostatic challenges typically elevates circulating glucocorticoid (GC) levels in vertebrates (modulating the behavioural and physiological responses that mediate allostasis), we may expect concomitant age-related changes in these hormones. Here, we investigated whether the level of corticosterone (the main avian GC) deposited in feathers during regular moult reflected the expected lifelong progression of energetic challenges in a long-lived raptor, the black kite (Milvus migrans). Feather corticosterone values were highest in the youngest birds, gradually declined to reach minimum levels in prime age, 7- to 11-year-old birds, and then increased again slightly among the oldest, senescent birds (≥12 years old). This pattern mirrored the age-related changes in reproductive success and survival rates previously reported for this population, suggesting that feather corticosterone levels captured the most vulnerable and challenging periods experienced by these birds as they proceeded through life. Moreover, feather corticosterone levels were negatively related to body size, suggesting that larger birds either experienced fewer homeostatic challenges, or were better able to cope with them. Feather corticosterone measures thus provided a valuable snapshot of how allostatic loads vary along the life of individuals, supporting the idea of a tight, long-term link between cumulative physiological responses to ecological challenges and demographic performance.

Offal dumping sites influence the relative abundance and roosting site selection of Black Kites (Milvus migrans govinda) in urban landscape: a study from Kolkata metropolis, India.

Although Black Kites (Milvus migrans govinda) serve as major scavenging raptor in most of the urban areas, scientific studies on this important ecosystem service provider are almost non-existent in Indian context. The present study was carried out in a metropolis in eastern India to find out the factors influencing relative abundance and roosting site selection of Black Kites. Separate generalized linear models (GLMs) were performed considering encounter rate and roosting Black Kite abundance as response variables. The study conclusively indicated that encounter rates of Black Kites were significantly influenced by the presence of garbage dumps in its vicinity. Numbers of Black Kites were also higher in the roosting sites situated closer to garbage dumps and open spaces. In addition, expected counts of Black Kites significantly increased in roosting sites situated away from buildings and water bodies. However, built-up area and tree cover around the roosting sites had no influence on the abundance of Black Kites therein. With rapid urbanization and changing offal disposal patterns, our findings would be useful to ensure continued availability of food and roosting sites of Black Kites in urban areas.

Clinically relevant antibiotic resistance in Escherichia coli from black kites in southwestern Siberia: a genetic and phenotypic investigation.

Wild birds including raptors can act as vectors of clinically relevant bacteria with antibiotic resistance. The aim of this study was to investigate the occurrence of antibiotic-resistant Escherichia coli in black kites (Milvus migrans) inhabiting localities in proximity to human-influenced environments in southwestern Siberia and investigate their virulence and plasmid contents. A total of 51 E. coli isolates mostly with multidrug resistance (MDR) profiles were obtained from cloacal swabs of 35 (64%, n = 55) kites. Genomic analyses of 36 whole genome sequenced E. coli isolates showed: (i) high prevalence and diversity of their antibiotic resistance genes (ARGs) and common association with ESBL/AmpC production (27/36, 75%), (ii) carriage of mcr-1 for colistin resistance on IncI2 plasmids in kites residing in proximity of two large cities, (iii) frequent association with class one integrase (IntI1, 22/36, 61%), and (iv) presence of sequence types (STs) linked to avian-pathogenic (APEC) and extra-intestinal pathogenic E. coli (ExPEC). Notably, numerous isolates had significant virulence content. One E. coli with APEC-associated ST354 carried qnrE1 encoding fluoroquinolone resistance on IncHI2-ST3 plasmid, the first detection of such a gene in E. coli from wildlife. Our results implicate black kites in southwestern Siberia as reservoirs for antibiotic-resistant E. coli. It also highlights the existing link between proximity of wildlife to human activities and their carriage of MDR bacteria including pathogenic STs with significant and clinically relevant antibiotic resistance determinants. IMPORTANCE Migratory birds have the potential to acquire and disperse clinically relevant antibiotic-resistant bacteria (ARB) and their associated antibiotic resistance genes (ARGs) through vast geographical regions. The opportunistic feeding behavior associated with some raptors including black kites and the growing anthropogenic influence on their natural habitats increase the transmission risk of multidrug resistance (MDR) and pathogenic bacteria from human and agricultural sources into the environment and wildlife. Thus, monitoring studies investigating antibiotic resistance in raptors may provide essential data that facilitate understanding the fate and evolution of ARB and ARGs in the environment and possible health risks for humans and animals associated with the acquisition of these resistance determinants by wildlife.

Bilateral cataracts extraction by lens aspiration and foldable intraocular lens implantation in a black kite (Milvus migrans).

Background: Cataract is a common disease in the lenses of birds. Cataract surgery is the recommended treatment for birds with visual impairment. Nevertheless, reports of the results of cataract surgery in birds are limited. Case Description: An unknown-aged male black kite (Milvus migrans) was presented to Kasetsart University Veterinary Teaching Hospital to evaluate and treat bilateral ocular cloudiness. Bilateral hypermature cataracts were diagnosed. A-scan biometry was carried out to calculate the intraocular lens' (IOL) diopter power. Bilateral cataract extraction by lens aspiration with +17.5 diopter foldable IOL implantation was successfully completed. Two months after surgery, the black kite navigated well and could fly and catch his food. Retinoscopy at 4 months postoperative revealed -0.5 diopters OD, whereas OS could not be assessed due to a poor reflection. Follow-up at 3 years after surgery, the black kite's vision remained favorable. Conclusion: Phacoemulsification and IOL implantation were successful in this black kite because the black kite's vision was noticeably improved, and the black kite was able to navigate well. To the authors' knowledge, this is the first report of bilateral cataract surgery with IOL implantation using a calculated dioptric power (+17.5 D) IOL in a black kite.

The mitochondrial genome of Milvus migrans (Aves, Accipitriformes, Accipitridae), an endangered species from South Korea.

The complete mitochondrial genome sequence of black kite, Milvus migrans, one of the most common diurnal raptor, was characterized using next generation sequencing. The whole genome size was 18,016 bp and consisted of 13 protein-coding genes, 22 tRNAs, 2 rRNAs, a putative control region (CR), and a second control region (pseudo-CR). A frameshift mutation was found in the ND3 gene. Phylogenetic analysis illustrated monophyly of the subfamily Melieraxinae with high statistical support. The genetic resource obtained here will help to resolve taxonomic issues related to subspecies of M. migrans and will act as a starting point for conservation genetics.

Larval infestation in a common Pariah Kite Milvus migrans (Boddaert).

This paper describes a case of larval infestation in a common Pariah Kite Milvus migrans (Boddaert). Examination on the larvae revealed that the larvae were of the family Sarcophagidae (Diptera).

Sex determination in the wild: a field application of loop-mediated isothermal amplification successfully determines sex across three raptor species.

PCR-based methods are the most common technique for sex determination of birds. Although these methods are fast, easy and accurate, they still require special facilities that preclude their application outdoors. Consequently, there is a time lag between sampling and obtaining results that impedes researchers to take decisions in situ and in real time considering individuals' sex. We present an outdoor technique for sex determination of birds based on the amplification of the duplicated sex-chromosome-specific gene Chromo-Helicase-DNA binding protein using a loop-mediated isothermal amplification (LAMP). We tested our method on Griffon Vulture (Gyps fulvus), Egyptian Vulture (Neophron percnopterus) and Black Kite (Milvus migrans) (family Accipitridae). We introduce the first fieldwork procedure for sex determination of animals in the wild, successfully applied to raptor species of three different subfamilies using the same specific LAMP primers. This molecular technique can be deployed directly in sampling areas because it only needs a voltage inverter to adapt a thermo-block to a car lighter and results can be obtained by the unaided eye based on colour change within the reaction tubes. Primers and reagents are prepared in advance to facilitate their storage at room temperature. We provide detailed guidelines how to implement this procedure, which is simpler (no electrophoresis required), cheaper and faster (results in c. 90 min) than PCR-based laboratory methods. Our successful cross-species application across three different raptor subfamilies posits our set of markers as a promising tool for molecular sexing of other raptor families and our field protocol extensible to all bird species.