Sex determination of three raptor species using morphology and molecular techniques

ABSTRACT.   Accurate identification of sex is important for many raptor studies, but may be difficult to determine in the field for some species. Because of size differences between males and females, morphological measurements have often been used to sex raptors. However, few investigators have examined the accuracy of using measurements of individuals made at one location to sex individuals of the same species at another location. Our objective was to develop more accurate region-specific methods for determining the sex of Red-tailed Hawks ( Buteo jamaicensis ), Red-shouldered Hawks ( B. lineatus ), and Cooper's Hawks ( Accipiter cooperii ) migrating through and wintering in California. We determined sex using a polymerase chain reaction (PCR)-based genetic test and grouped individuals based on sex, age, and geographic area. We did not combine groups due to differences in measurements between age classes and geographic areas. We then compared a suite of morphological measurements between males and females of each combination, and developed both a discriminant function and a flowchart to determine the sex of Red-tailed and Red-shouldered hawks in the field. The flowcharts were more accurate than the functions for both species. We also confirmed the accuracy of the current flowchart used to determine the sex of Cooper's Hawks migrating along the California coast. These region-specific methods for Red-tailed and Cooper's hawks were generally more accurate than published methods, possibly indicating different populations of these species and highlighting the importance of validating sexing methods when using them in different locations.     

Secrets in the eyes of Black Oystercatchers: a new sexing technique

ABSTRACT.   Sexing oystercatchers in the field is difficult because males and females have identical plumage and are similar in size. Although Black Oystercatchers ( Haematopus bachmani ) are sexually dimorphic, using morphology to determine sex requires either capturing both pair members for comparison or using discriminant analyses to assign sex probabilistically based on morphometric traits. All adult Black Oystercatchers have bright yellow eyes, but some of them have dark specks, or eye flecks, in their irides. We hypothesized that this easily observable trait was sex-linked and could be used as a novel diagnostic tool for identifying sex. To test this, we compared data for oystercatchers from genetic molecular markers (CHD-W/CHD-Z and HINT-W/HINT-Z), morphometric analyses, and eye-fleck category (full eye flecks, slight eye flecks, and no eye flecks). Compared to molecular markers, we found that discriminant analyses based on morphological characteristics yielded variable results that were confounded by geographical differences in morphology. However, we found that eye flecks were sex-linked. Using an eye-fleck model where all females have full eye flecks and males have either slight eye flecks or no eye flecks, we correctly assigned the sex of 117 of 125 (94%) oystercatchers. Using discriminant analysis based on morphological characteristics, we correctly assigned the sex of 105 of 119 (88%) birds. Using the eye-fleck technique for sexing Black Oystercatchers may be preferable for some investigators because it is as accurate as discriminant analysis based on morphology and does not require capturing the birds.     

Morphology versus molecules: sexing Red-winged Blackbird nestlings

ABSTRACT.   Past studies of offspring sex ratios in birds have often relied on sexually size dimorphic species where nestling sex could be determined based on weight at a given age. DNA-based sexing techniques allow us to assess the accuracy of those techniques and to refine them for use when costs or convenience make DNA methods impractical. Using nestling Red-winged Blackbirds ( Agelaius phoeniceus ) whose sex was determined using DNA, we compared sex ratios obtained using different morphological criteria. Conservative criteria from previous studies were completely accurate, but allowed sexing of few nestlings younger than 8 d old, and were more successful for sexing males than females. A new method was developed that allowed accurate sexing of nestlings beginning at day 6 posthatching and was less biased relative to known sex ratios. Using 11 years of data, the conservative method left an average of 55% of nestlings and 36% of fledglings unsexed, compared to 31% and 9% using the new method. Furthermore, the male bias in sex ratio estimates using the conservative method was greater, both absolutely and relative to estimates based on the new method, when the proportion of unsexed nestlings (because they were not weighed when older) was higher. Thus, estimates of population sex ratios will be more accurate as the number of nestlings measured on day 8 or older increases. However, if some nestlings that were not weighed past day 7 fledge, the new method allows more of those individuals to be sexed than the conservative method, and the population sex ratio estimate should be more reliable. Although our approach should apply to other sexually dimorphic species, the criteria used must be developed based on such species-specific attributes as growth patterns and degree of hatching asynchrony.     

Prey of White-throated Hawks (Buteo albigula) in the southern temperate forest of Argentina

ABSTRACT.   We examined the diet of White-throated Hawks ( Buteo albigula ) during the incubation and nestling periods in the southern temperate forest of Argentina. Pellets ( N = 74) and prey remains ( N = 59) were collected at 10 nests from 1998 to 2003, and preys delivered to two nests were monitored during the 2001–2002 breeding seasons. White-throated Hawks fed on small mammals, birds, lizards, and insects. The three methods of identifying prey (pellets, prey remains, and direct observation) produced different results. All types of prey except large birds were detected in pellets, and arthropods may have been over-represented in pellets due to secondary consumption. No remains of either arthropods or reptiles were identified among prey remains collected at nest sites and, during nest observations, we were unable to identify many of the prey items delivered by adults. Our results indicate that accurate determination of the diet of White-throated Hawks requires more than one method of identifying prey.     

Sex-determination of Continental European Robins Erithacus r. rubecula

Existing morphological sex-determination criteria for Continental European Robins proved inadequate when applied to birds captured in western Denmark. Only 49% of captured birds met with the wing-length criteria resulting in the classification of 2% males and 47% females respectively. This article presents a primer for correct sexing of Robins by amplification of a female-specific DNA element by a polymerase chain reaction (PCR) on genomic DNA. Further, two new criteria for sex-determination are proposed based on logistic regression and discriminant function analysis using morphological measures from birds with DNA-verified sex. Success rates of 80–81% for the new criteria were a considerable improvement compared with a success rate of only 36% for the old criteria.     

Hawk Eyes II: Diurnal Raptors Differ in Head Movement Strategies When Scanning from Perches

Background

Relatively little is known about the degree of inter-specific variability in visual scanning strategies in species with laterally placed eyes (e.g., birds). This is relevant because many species detect prey while perching; therefore, head movement behavior may be an indicator of prey detection rate, a central parameter in foraging models. We studied head movement strategies in three diurnal raptors belonging to the Accipitridae and Falconidae families.

Methodology/Principal Findings

We used behavioral recording of individuals under field and captive conditions to calculate the rate of two types of head movements and the interval between consecutive head movements. Cooper''s Hawks had the highest rate of regular head movements, which can facilitate tracking prey items in the visually cluttered environment they inhabit (e.g., forested habitats). On the other hand, Red-tailed Hawks showed long intervals between consecutive head movements, which is consistent with prey searching in less visually obstructed environments (e.g., open habitats) and with detecting prey movement from a distance with their central foveae. Finally, American Kestrels have the highest rates of translational head movements (vertical or frontal displacements of the head keeping the bill in the same direction), which have been associated with depth perception through motion parallax. Higher translational head movement rates may be a strategy to compensate for the reduced degree of eye movement of this species.

Conclusions

Cooper''s Hawks, Red-tailed Hawks, and American Kestrels use both regular and translational head movements, but to different extents. We conclude that these diurnal raptors have species-specific strategies to gather visual information while perching. These strategies may optimize prey search and detection with different visual systems in habitat types with different degrees of visual obstruction.     

Sex determination of African Penguins Spheniscus demersus using bill measurements: method comparisons and implications for use

African Penguins Spheniscus demersus are sexually dimorphic; on average, males are larger than females but measurements overlap making sex determination difficult through observations alone. We developed a discriminant function, using bill length and depth from a sample of birds sexed from gonad visualisation during post-mortem, which correctly classified 93% of the individuals. Cross-validation correctly assigned 90% of DNA-sexed birds and 91% of birds sexed by partner measurement comparisons. The use of discriminant function score cutpoints, while leaving 16% and 29% of birds unclassified, improved accuracy of birds sexed by DNA to 97% and of those sexed by partner comparison to 99%. Bill depth was found to be a discriminating variable. However, two techniques for measuring bill depth are currently in use for African Penguins. While these measurements are correlated (r = 0.85), they differ on average by 1.4?mm hindering accuracy of sex determination when using a discriminant function developed from the other bill depth measurement. Exploration of adult bill morphology of birds sexed from DNA at different colonies suggests the discriminant functions can be applied throughout the African Penguins’ South African range.     

Seabird nest counts: a test of monitoring metrics using Red-tailed Tropicbirds

ABSTRACT.   Counts of nesting birds are often used to monitor the abundance of breeding pairs at colonies. Mean incubation counts (MICs) are counts of nests with eggs at intervals that correspond to the mean incubation period of a species. The sum of all counts during the nesting season (MIC_total) and the highest single count during the season (MIC_max) are metrics that can be generated from this method. However, the utility of these metrics as measures of the number of breeding pairs has not been well tested. We used two approaches to evaluate the bias and precision of MIC metrics for quantifying annual variation in the number of breeding Red-tailed Tropicbirds ( Phaethon rubricauda ) nesting on two islands in the Papahānaumokuākea Marine National Monument in the northwest Hawaiian Islands. First, we used data from nest plots with individually marked birds to generate simulated MIC metrics that we compared to the known number of nesting individuals. The MIC_total overestimated the number of pairs by about 5%, whereas the MIC_max underestimated the number of pairs by about 60%. However, both metrics exhibited similar precision. Second, we used a 12-yr time series of island-wide MICs to compare estimates of temporal trend and annual variation using the MIC_max and MIC_total. The 95% confidence intervals for the trend estimates were overlapping and the residual standard errors for the two metrics were similar. Our results suggest that both metrics offered similar precision for indices of breeding pairs of Red-tailed Tropicbirds, but that MIC_total was more accurate.     

基于形态特征判定五种鹬类性别的可靠性

快速准确地鉴定两性同型鸟类个体性别在鸟类生态学研究中具有重要意义。本文选择2008年春季迁徙期在崇明东滩停歇的大滨鹬(Calidris tenuirostris)、红腹滨鹬(C.canutus)、红颈滨鹬(C.ruficollis)、尖尾滨鹬(C.acuminata)及翘嘴鹬(Xenus cinereus)5种两性同型的鹬类,用分子生物学方法进行性别鉴定,并基于个体的形态特征(体重、翅长、喙长、头喙长及跗跖长)采用判别分析方法对性别进行判定。结果表明,尖尾滨鹬雄性各形态特征均显著大于雌性,其他4种鹬类则相反。5种鹬类形态特征的性别差异指数在0.5%~25.3%之间,重叠度在29.4%~98.6%之间。5种鹬类判别分析判定性别的准确率在(0.69±0.06)~(0.96±0.01)之间,其中,尖尾滨鹬判别准确率(0.96)最高,翘嘴鹬判别准确率(0.69)最低。形态特征在两性间的差异程度影响性别的判别准确率。另外,两性性比对性别判别的准确率也有影响:性比偏雄性鸟类的雄性判别准确率高于雌性,而性比偏雌性鸟类的雌性判别准确率高于雄性。采用判别分析估测的性比与分子生物学鉴定结果相似,表明判别分析在判定种群的性比方面具有较高的可靠性。     

Bill length: a reliable method for sexing Purple Sandpipers

ABSTRACT.   Purple Sandpipers ( Calidris maritima ) are sexually dimorphic, with females larger than males. We tested the reliability of using bill length to sex individuals and estimate the sex ratio at a stopover site in Iceland in May 2003 and 2005. Feather samples from 65 of 324 captured birds were used for molecular sexing, and generalized linear models (GLMs) were used to discriminate the sexes from body measurements of the molecularly sexed birds. About 3% of the 324 individuals were misclassified by the Harding-Cassie method, and the proportion of males was 0.657 compared to 0.656 according to the best GLM. Our results showed that the reliability of determining the sex and sex ratio of Purple Sandpipers using a Harding-Cassie plot of bill length measurements was high, but that reliability was improved by including other variables in a GLM. The estimate of an uneven sex ratio in the population we studied was not due to a systematic error, and supports the conclusion of earlier studies that Purple Sandpipers exhibit an uneven sex ratio in favor of males.     

Sexing terns using biometrics: the advantage of within-pair comparisons

Capsule Within-pair comparisons substantially improve the accuracy of sexing from biometrics for two congeneric species of seabird with monomorphic plumage and soft-tissue colouration.

Aims To examine the extent to which statistical limitations of sexing birds from biometrics using sample-level analysis could be overcome by sexing Common and Arctic Terns (Sterna hirundo and S. paradisaea) using measurements obtained from breeding pairs.

Methods Incubating adults were caught at the nest using walk-in traps and wing, tarsus, head-plus-bill, tail length, tail fork, and body mass measured. Each bird was individually colour-ringed and dyed with picric acid, enabling subsequent sexing by behavioural observations of copulation and courtship feeding. Birds were sexed using biometrics and the proportion of birds sexed correctly this way at the sample level was compared with the accuracy achieved if, within a pair, the larger bird was classified as male.

Results Head-plus-bill length was the single most accurate measurement for sexing individuals of both species, and correctly classified 72% of Arctic Terns and 73% of Common Terns. Combinations of measurements derived from discriminant analysis achieved slightly higher accuracy (73% and 78% respectively). Within-pair comparisons were more accurate than sample-level analysis for both single measurements and discriminant functions, and were able to sex 84% of Arctic Terns and 86% of Common Terns correctly.

Conclusion Comparing birds within pairs improves accuracy and can eliminate the need to calculate cut points or discriminant functions from a sample of birds of known sex for each particular study. We strongly advocate such comparisons wherever possible to increase accuracy and simplify computational procedures for predicting sex, thus reducing associated sources of error.     

Morphological sexing of passerines: not valid over larger geographical scales

Sex determination of birds is important for many ecological studies but is often difficult in species with monomorphic plumage. Morphology often provides a possibility for sex determination, but the characters need to be verified. We tested whether five passerine species can be sexed according to standard morphological measurements applying a forward logistic regression with sex determined by molecular analysis as the dependent variable. Furthermore, we tested whether the results can be used on a larger geographic scale by applying morphological sexing methods gained by similar studies from other regions to our data set. Of the five species of this study only Garden Warblers Sylvia borin could not be sexed morphologically. In the Robin Erithacus rubecula, 87.2% of all individuals were sexed correctly. For Reed Warblers Acrocephalus scirpaceus, Willow Warblers Phylloscopus trochilus and Reed Buntings Emberiza schoeniclus, the respective values were 77.6, 89.4 and 86.4%. When the logistic regression functions from similar studies on Robins and Reed Buntings in Denmark and Scotland were applied to the birds from south-western Germany, they performed less well compared to the original dataset of these studies and compared to the logistic regression function of our own study. The same was the case for Willow Warblers when a wing length criterion used in Great Britain was applied to the birds of our study. These discrepancies may have several explanations: (1) the models are optimised for the dataset from which they were extracted, (2) inter-ringer variation in measurements, (3) the use of different age cohorts, (4) different morphology due to different habitat availability around the study site, or, most likely, (5) different morphology due to different migratory behaviour. We recommend that morphological sex differentiation methods similar to this study (1) be only used population specific, (2) only with one age cohort and (3) to adjust the extracted equations from time to time.     

A new method for sexing Oriental White Storks

ABSTRACT.   For species where males and females are monomorphic, or nearly so, determining the sex of individual birds generally requires either capturing birds or collecting samples, such as feathers, for DNA analysis. We developed a new method, involving the use of photographs, to determine the sex of endangered Oriental White Storks ( Ciconia boyciana ). Using photographs, we analyzed the lateral features of the heads of 25 captive storks of known sex (12 males and 13 females) and found differences between males and females in the distance from the bill tip to the nape and the distance from the bill tip to the commissural point. These differences were used to generate a discriminant function that was then tested on 22 captive storks at Hyogo Homeland Park (Toyooka, Japan), and we correctly determined the sex of 18 individuals (82%). In addition, the sex of two wild storks was correctly assigned. Our results suggest that good-quality photographs can be useful for determining the sex of both captive and wild Oriental White Storks and, further, that similar methods may prove useful for determining sex in other species of birds.     

Philopatry and natal dispersal in a sedentary population of western marsh harrier

Dispersal is a key life-history trait because it influences population dynamics and population genetic structure. From a behavioural perspective, the study of natal dispersal requires some understanding of the mechanisms that affect individual movements, because movements of an animal form a path that is continuous throughout its life. Our aim was to investigate juvenile dispersal strategies in the western marsh harrier Circus aeruginosus , between fledging and first breeding attempt (from 1 to 4 years later, depending on the sex and individual). Using radio tracking, we monitored dispersing juvenile harriers and their home-range size variations within a sedentary population in central western France from 2001 to 2007. Juvenile dispersal strategy was mainly characterized by a very high natal philopatry (i.e. birds that remained within the study area) and short-distance dispersal. All but one bird (out of 39) remained within the study area between their first winter and their first spring, and 96.0% during their first spring. The distance moved at 2 years of age was significantly larger for males than for females (3800± sd 3593 m and 935± sd 481 m for seven males and six females, respectively), in contrast to most bird species studied so far. Home-range size was not sex biased and significantly decreased with age. In addition, non-breeding birds had larger home ranges (1603± sd 2128 ha) than breeders (349± sd 185 ha). Using data obtained from other populations, juvenile marsh harrier dispersal strategies appeared to be determined by migratory status (migratory birds dispersing farther) and demographic parameters (juvenile survival or fecundity).     

Using interpubic distance for sexing manakins in the field

ABSTRACT Field methods for determining the sex of birds are often limited due to morphometric overlap between sexes, intermediate plumages, seasonality, and reliance on subjective age classification. Interpubic distance, characterized in birds as the distance between the distal ends of the pubic bones, has not been formally tested as a method for determining the sex of birds, despite references among parrot breeders and the frequent use of analogous measurements in mammals. We developed a harmless and easily performed field method for measuring interpubic distance in studies involving bird capture, and compared the interpubic distances of known sex White‐ruffed Manakins (Corapipo altera), Orange‐collared Manakins (Manacus aurantiacus), and Blue‐crowned Manakins (Lepidothrix coronata) to evaluate the possible use of this measurement to determine sex. Using interpubic distance ranges based on 85% confidence intervals where overlap existed between sexes, the sex of 92.8–100% of all manakins in our study was accurately determined with no misclassification. Interpubic distance performed better than plumage‐based methods that sexed 74.0% of all individuals and misclassified 1.5%. Using linear discriminant analysis, we developed classification equations that allowed us to accurately determine the sex of all individuals with 100% accuracy using mass and interpubic distance. Additionally, we compared the interpubic distances of female White‐ruffed Manakins to evaluate the potential to determine age and reproductive status. Despite an apparent relationship between interpubic distance, age and reproductive status, we concluded that interpubic distance has limited use for determining age and reproductive status due to extensive overlap (31.6–100%), but shows potential in other applications. Based on these results, we endorse the use of interpubic distance to determine the sex of manakins. We encourage further study to develop additional classification equations using different morphometric measurements and to test the efficacy of interpubic distance to determine sex in other bird species.     

Agricultural change and paddock tree loss: Implications for an endangered subspecies of Red-tailed Black-Cockatoo

Summary   Natural senescence and the intensification of agricultural practices are contributing to the continuing loss of paddock trees from agricultural regions in Australia. This is of particular concern in the southern Wimmera of western Victoria, where much of the endangered Buloke ( Allocasuarina luehmannii ) Woodland vegetation community is represented only by relict Buloke trees in paddocks, which also constitute critical feeding habitat for the endangered south-eastern subspecies of the Red-tailed Black-Cockatoo ( Calyptorhynchus banksii graptogyne ). I investigated the rate and correlates of loss of scattered Buloke trees in paddocks by examining aerial photographs taken over a period of 15 years in a region undergoing agricultural intensification. Tree loss over the period was measured using aerial photographs of five localities, covering a total of 7850 ha of agricultural land in the southern Wimmera. The average rate of loss (± 1 SE) was 25.8% ± 6.4% over the 15 years, or 1.7% per annum. The rate of tree loss was higher in areas under cultivation (32.5%) than areas under pasture (20.6%). A disproportionate number of trees was lost from locations where centre pivot irrigation systems were installed. Because of the slow growth rate of Buloke trees, revegetation efforts and offset planting are unlikely to compensate for losses of Red-tailed Black-Cockatoo habitat for approximately 100 years.     

Sexing birds using random amplified polymorphic DNA (RAPD) markers

We used random amplified polymorphic DNA (RAPD) markers to sex birds from small tissue (usually blood) samples. Arbitrarily chosen 10-mer PCR primers were screened with DNA from known-sex individuals for the production of a bright female-specific band. Suitable primers were found for seven bird species after screening about 30 primers (range 2–63), and no primer was found for three other species after screening about 50 primers for each species. Investigations into the reliability of RAPD markers for sexing great tits Parus major and oystercatchers Haematopus ostralegus show that: (i) when PCR reaction conditions for great tit DNA are varied, either the presence of the female-specific band correctly predicts the individual's sex or no DNA amplification occurs; (ii) the female-specific band in great tits can be sequenced, and subsequently amplified using specific PCR primers; (iii) null alleles of the female-specific fragment occur at an estimated frequency of 0% ( n = 241 females) in great tits and 0.6% ( n > 290 females) in oystercatchers; (iv) the female-specific fragment in great tits occurs in individuals from a wide geographical range encompassing two subspecies; and (v) the relative intensity of bands in great tit RAPD banding profiles is consistent across individual birds and scorers. The RAPD primers that we have identified are generally species specific, and the consequent time cost of screening for primers is the chief disadvantage of using RAPD markers to sex birds. However, with large sample sizes this disadvantage is outweighed by the relative technical simplicity and low cost of the technique.     

ON THE SOUTH AFRICAN RACE OF THE LITTLE SPOTTED WOODPECKER

Discriminant analysis functions have previously been determined for sexing Mediterranean Yellow-legged Gulls Larus michahellis michahellis from the western Mediterranean basin. However, data from eastern Mediterranean populations are lacking. In this work, we used morphometric data from a sample of 81 Mediterranean Yellow-legged Gulls (39 males and 42 females) breeding in the Gulf of Gabès in south-eastern Tunisia to (1) determine a discriminant function useful for sex discrimination, and (2) assess the accuracy of previously published functions in sexing Mediterranean Yellow-legged Gulls from our study area. Our results showed marked sexual differences in all morphological measurements, with males being significantly larger than females. The best discriminant function included head length, bill depth and wing length, and accurately classified 93% of sampled birds. We also found that Mediterranean Yellow-legged Gulls from the Gulf of Gabès could accurately be sexed using discriminant functions determined for another North African population, but not with a function determined for a South European population, although distances between sites are almost the same.     

A Comparison of Field and Molecular Techniques for Sexing Beavers

Abstract: The traditional method of sex identification in beavers (Castor canadensis) by external palpation can be inaccurate. We tested 2 genetic methods for determining sex in beavers, the zinc-finger DNA marker and the Y chromosome-specific sex determining region (SRY) marker. The SRY marker identified sex correctly in 57 of 67 (85%) beavers, whereas the zinc-finger technique was successful less often in only 48 of 67 (72%) animals. Sex was correctly assigned by palpation for 21 of 27 beavers (78%). Beaver studies in which accurate sex identification is critical may benefit by verifying the sex of individuals using one or both of these molecular markers.     

Sex determination in the Lesser Flamingo (Phoenicopterus minor) using morphological measurements

Morphological measurements and blood samples were taken from 154 Lesser Flamingos Phoenicopterus minor, including adults (>3 years old), immature sub-adults (2–3 years old) and first-year juvenile birds of both sexes, captured at Lake Bogoria, Kenya (0°11'–20' N, 036°06' E) during 2001 and 2002. PCR amplification of the CHD-Z and CHD-W genes using DNA extracted from the blood samples was used to determine the sex of each bird. There were significant differences in mass and tarsus length among the three age groups, indicating that Lesser Flamingos continue to grow in skeletal size and mass between fledging and the attainment of adult plumage at 3–4 years of age. On average, males were significantly larger than females in all age groups, although there was substantial overlap between the sexes in all morphological measurements. The element with the least amount of overlap was head-and-bill length. Discriminant functions utilising head-and-bill length that correctly predict the sex of juvenile and immature birds with approximately 93% accuracy are presented. By adding total tarsus length, the sex of wild adult Lesser Flamingos is correctly predicted with approximately 98% accuracy. The same discriminant function developed for wild adult birds predicted the sex of 19 captive adult Lesser Flamingos of known sex with 100% accuracy.