Phylogeny and phylogeography of critically endangered Gyps species based on nuclear and mitochondrial markers

Populations of Oriental White-backed Vulture (Gyps bengalensis) and Long-billed Vulture (Gyps indicus) dramatically declined by 95–100% on the Indian subcontinent in mid-1990s. The present study was conducted to discover the phylogeny and phylogeography of Gyps species based on nuclear (recombination activating gene, RAG-1) and mitochondrial (cytochrome b, cytb) markers. Gyps species showed monophyly and no geographic partition was observed within the three groups of Gyps species (G. bengalensis, G. indicus and G. fulvus) despite the large sample size available (n = 149). Our study supports the treatment of G. indicus and G. tenuirostris as separate species. In all analyses, the earliest divergence separated G. bengalensis from all other Gyps taxa while a sister relationship was supported between G. fulvus and G. rueppellii, and these two taxa together were sister group to a clade consisting of G. indicus, G. tenuirostris and G. coprotheres. Molecular clock estimates of both nuclear and mitochondrial (RAG-1, cytb) genes indicated a rapid and recent diversification within the Gyps species. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Systematics within Gyps vultures: a clade at risk

Background  

Populations of the Oriental White-backed Vulture (Gyps bengalensis) have declined by over 95% within the past decade. This decline is largely due to incidental consumption of the non-steroidal anti-inflammatory veterinary pharmaceutical diclofenac, commonly used to treat domestic livestock. The conservation status of other Gyps vultures in southern Asia is also of immediate concern, given the lack of knowledge regarding status of their populations and the continuing existence of taxonomic uncertainties. In this study, we assess phylogenetic relationships for all recognized species and the majority of subspecies within the genus Gyps. The continuing veterinary use of diclofenac is an unknown but potential risk to related species with similar feeding habits to Gyps bengalensis. Therefore, an accurate assessment of the phylogenetic relationships among Gyps vultures should aid in their conservation by clarifying taxonomic uncertainties, and enabling inference of their respective relatedness to susceptible G. bengalensis.     

Cape Vulture Gyps coprotheres breeding status in southern Africa: monitoring results from 2010–2014

Monitoring of eight Cape Vulture Gyps coprotheres breeding colonies in South Africa and Botswana took place between 2010 and 2014 using standardised counting methods. Counts of breeding pairs, nestlings and fledglings were used to calculate breeding success and general population trends. While our data suggests multiple colonies are stable or increasing, the extinction of peripheral colonies and contraction of the species’ range is alarming and gives evidence for unsustainable population declines. Monitoring efforts chronicled the extinction of the previously large core colony at Roberts’ Farm, which was abandoned as a breeding site in 2013. Standardised monitoring is urgently needed across the entire species range to better understand current population dynamics.     

The Population Decline of Gyps Vultures in India and Nepal Has Slowed since Veterinary Use of Diclofenac was Banned

Populations of oriental white-backed vulture (Gyps bengalensis), long-billed vulture (Gyps indicus) and slender-billed vulture (Gyps tenuirostris) crashed during the mid-1990s throughout the Indian subcontinent. Surveys in India, initially conducted in 1991–1993 and repeated in 2000, 2002, 2003 and 2007, revealed that the population of Gyps bengalensis had fallen by 2007 to 0.1% of its numbers in the early 1990s, with the population of Gyps indicus and G. tenuirostris combined having fallen to 3.2% of its earlier level. A survey of G. bengalensis in western Nepal indicated that the size of the population in 2009 was 25% of that in 2002. In this paper, repeat surveys conducted in 2011 were analysed to estimate recent population trends. Populations of all three species of vulture remained at a low level, but the decline had slowed and may even have reversed for G. bengalensis, both in India and Nepal. However, estimates of the most recent population trends are imprecise, so it is possible that declines may be continuing, though at a significantly slower rate. The degree to which the decline of G. bengalensis in India has slowed is consistent with the expected effects on population trend of a measured change in the level of contamination of ungulate carcasses with the drug diclofenac, which is toxic to vultures, following a ban on its veterinary use in 2006. The most recent available information indicates that the elimination of diclofenac from the vultures’ food supply is incomplete, so further efforts are required to fully implement the ban.     

AN AFTERNOON WITH THE WADERS

Houston, D. C. 1974. Mortality of the Cape Vulture. Ostrich 45: 57–62.

An attempt was made to analyse the ringing returns of the Cape Vulture Gyps coprotheres from Transvaal colonies. Although it is not yet possible to determine annual mortality rates, some conclusions can be deduced from the results. Mortality during the early age classes is high and is over 500; during the first year. Mortality must decline rapidly with age, but it seems probable that recruitment to the breeding population at these colonies is insufficient to balance adult mortality and the population may be declining.     

The Cape Vulture at Colleywobbles: 1977–1997

Vernon, C.J. 1998. The Cape Vulture at Colleywobbles: 1977–1997. Ostrich 70 (3&4): 200–202.

In the late 1970s there were about 180 breeding pairs of Cape Vulture Gyps coprotheres at Colleywobbles colony. In the 1980s the colony increased to over 300 pairs, presumably as a result of birds immigrating to the Transkei. In the 1990s the colony declined to about 60 pairs, either through emigration or mortality. The decrease from the 1980s to the 1990s was probably related to a reduction in food supply (carcasses of domestic livestock) as a result of changes in human social activities.     

BREEDING OF THE INDIAN WHITEBACKED VULTURE AT JODHPUR

Vulture populations have declined globally as well as regionally within Africa. Little is known about the status of the African White-backed Vulture Gyps africanus in Kenya, but ongoing studies indicate that its population has declined over the last two decades. A total of 32 African White-backed Vulture nests were monitored in the Masai Mara National Reserve over a five-year period between 2003 and 2007. Mean nesting success was 59%, which is comparable to that of populations from southern Africa. Nearest neighbour distances were significantly closer in wooded habitats (‘trees and shrubs savanna’) than in more open grassland habitats (‘open low shrubs’). Based on nearest neighbour distances, the estimated total breeding population within the Masai Mara National Reserve is 1 106 pairs, a figure that may be an overestimate and requires ground-truthing. Collecting baseline data on numbers of breeding pairs and regular nest monitoring are essential in order to assess the impact of various threats to vultures in Kenya, which include growing threats (elephant-mediated habitat disturbance and fire) as well as emerging threats (such as poisoning with the carbamate-based pesticide Furadan?).     

Effects of superabundant food on breeding success and behavior of the red-winged blackbird

Summary The effects of food on breeding success and behavior of the red-winged blackbird (Icteridae: Agelaius phoeniceus) were investigated during 3 successive breeding seasons. In the second season, a 4-week pulse of abundant food in the form of a periodical cicada emergence (Homoptera: Cicadidae: Magicicada spp.) occurred in the forest adjacent to the marsh where the birds were breeding.During the cicada period, the bird population showed: 1) an increase in foraging trips to the forest and a decrease in trips per h, 2) increased biomass of nestlings, 3) increased nestling survival caused by decreased starvation, 4) increased fledging success, and 5) bimodal weight distributions of older nestlings (reflective of the sexual dimorphism in this species). These data suggest the temporary removal of food limitations on the breeding population when the pulse of food was available.     

High interannual variation in the hatching sex ratio of Tengmalm’s owl broods during a vole cycle

The sex ratio at hatching in broods of Tengmalms owl (Aegolius funereus) in northern Sweden was investigated for 3 years characterized by different phases of the vole and owl cycle. Previous work showed the sex ratio in this species to be male-biased for 1 year with a favourable food supply, and that in feeding experiments male nestlings (but not females) suffered higher mortality when food was limited, but not otherwise. Here we present data from a complete 3-year owl cycle, showing that mean brood sex ratio varied significantly among years, being male-biased (65% males) in the first year of high owl breeding density, unbiased (49%) in the second year of high owl breeding density, and female-biased (33% males) in the owls low year. Brood sex ratio did not vary significantly within years with laying date or parental age. Vole availability, and therefore the owls food supply, declined during the 3 years studied. Tengmalms owl parents thus appear to adaptively adjust the sex ratio of their broods according to the expected annual mortality risk of sons.     

Sex-biases in the hatching sequence of cooperatively breeding apostlebirds <Emphasis Type="Italic">Struthidea cinerea</Emphasis>

In the cooperatively breeding apostlebird (Struthidea cinerea, Corcoracidae) both sexes are philopatric and help to raise offspring. However, male helpers provision nestlings more often than females, an activity associated with reduced nestling starvation and enhanced fledgling production. Presuming that males are the more helpful sex, we examined the helper repayment hypothesis by testing the predictions that offspring sex ratio should be skewed toward the production of males (a) among breeding groups with relatively few helpers, and (b) in the population as a whole. The relationship between sex and hatching order was examined as a potential mechanism of biasing sex allocation. The sex ratio of all sexed offspring was male biased (57.9%; n = 171) as was the mean brood sex ratio (0.579; n = 70 broods). These biases were less pronounced in the subset of clutches/broods in which all offspring were sexed. This overall bias appeared to result from two distinct patterns of skew in the hatching order. First, mothers in small breeding groups produced significantly more males among the first-hatching pair. This is consistent with the helper repayment hypothesis given that later hatching chicks were less likely to survive, particularly in small groups. Second, almost all fourth-hatching chicks, usually the last in the brood, were male (91.7%, n = 12). This bias is difficult to interpret but demonstrates the value of examining hatching sequences when evaluating specific predictions of sex allocation theory in birds.     

Status and structure of the griffon vulture (Gyps fulvus) population in Crete

Griffon vulture (Gyps fulvus) population surveys were conducted during 1996–2002 in the island of Crete (Greece) to document population status and structure. Fieldwork was carried out during the breeding period when birds could be monitored in their colonies. Total population size was estimated at 379 individuals (range = 341–417) with adult birds comprising 63%. The breeding population was estimated at 141 pairs, which were distributed on an average in 23 colonies per year (range = 16–30) while the mean number of breeding pairs that laid eggs was 98 (range= 64–126). Crete thus supports the largest insular population of the species in the world and hosts 70–80% of the breeding population of the species in Greece. Population density was estimated at 6.9 individuals/100 km², 2.6 breeding pairs/100 km² and 1.8 nesting pairs/100 km². The average home range of an occupied colony (i.e., breeding group) was estimated at ca. 204 km² producing a theoretical foraging range of 8 km radius around the breeding cliff. No trends in the total number of individuals and breeding pairs appeared to exist, although significant differences in population size of individual colonies occurred between the years. The majority of the population was concentrated in small-sized colonies, which showed a low occupancy rate. The number of abandoned sites and the colonization of new ones could represent a shift of breeding pairs to alternative colonies provoked by local food abundance and conspesific attraction.     

Temporal changes in food resources,parental feeding and breeding success of Heron Island silvereyes,Zosterops lateralis chlorocephala

The influences of the temporal change in food supply on the parental feeding effort and breeding success of silvereyes,Zosterops lateralis chlorocephala, was investigated on Heron Island, Australia. Food supply (arthropods and figs) declined as the breeding season progressed. The parental feeding rate and growth of nestlings were lower when food supply was poor. When available, dominant pairs fed their young more figs and fewer arthropods than lower ranking pairs. Dominant pairs raised heavier young than lower ranking pairs when food supply was poor, while there were no significant differences between them when food supply was rich. When food supply was rich, pairs delivering greater amounts of arthropods reared nestlings better, whereas feeding more figs did not improve growth of nestlings. When food supply was poor, pairs spending a longer time at the nest reared nestlings better.     

Reproductive success and nestling diet in the Lesser Spotted Woodpecker ( Picoides minor ): the early bird gets the caterpillar

Populations of Lesser Spotted Woodpecker (Picoides minor) are decreasing in size, necessitating management strategies. However, data on the reproductive biology of this species are scarce. The aim of this study was to investigate the influence of timing of breeding and nestling diet on the reproductive success of the Lesser Spotted Woodpecker and thereby contribute to an understanding of the determinants influencing its reproductive success. During 6 study years between 1996–2003, we investigated various variables of reproductive success in a Lesser Spotted Woodpecker population in the Taunus low mountain range, Germany. We observed nestling feeding at breeding holes to assess the composition of nestling diet and how it changes during the season. Clutch size, number of fledglings as well as body mass of the nestlings declined with the start of egg laying. Pairs composed of individuals that had breed together in previous years started egg laying earlier than newly established pairs. Nestling diet consisted mainly of aphids, caterpillars, craneflies and wood-living larvae. The composition changed considerably within the breeding season in both early and late broods. However, early broods were provided with caterpillars more frequently, whereas late broods received wood-dwelling larvae more often. Our results suggest that Lesser Spotted Woodpeckers time their breeding so as to coincide with the early nestling stage with the highest availability of caterpillars, since this is the time of highest energy demand. Moreover, food availability declines during the breeding season, and the decline in reproductive performance seems to be an effect of this development.     

Seasonal decline in cell-mediated immunity of collared flycatcher Ficedula albicollis nestlings: does the sex of offspring matter?

The seasonal decline in offspring performance is a frequently reported phenomenon in species breeding in a temperate zone, but the potential effect of brood sex ratio on such declines has not been studied. Here, we predicted that this decline may occur if the sex that exhibits the lower immune response or lower survival rate tends to be more frequent among late broods. The seasonal patterns of four performance parameters of collared flycatcher Ficedula albicollis nestlings have been examined during 4 years. Sex was assigned to all studied individuals using molecular techniques. We found significant seasonal decline in cell-mediated immune response, tarsus length and survival of the chicks. The lack of interactions between gender and hatching date revealed that both sexes contributed equally to the observed decline. The brood sex ratio did not vary with the laying date. On the basis of available data, we suggest that the breeding date may only exceptionally induce female-driven sex allocation in species with only slight sexual size dimorphism. In consequence, we suggest that seasonal sex ratio shifts do not account for seasonally declining fitness of nestlings in passerines.     

Characterization of plasma cholinesterase activity in the Eurasian Griffon Vulture Gyps fulvus and its in vitro inhibition by carbamate pesticides

The legal and illegal use of organophosphorus and carbamate pesticides represents one of many threats to birds. The activity of the cholinesterase enzyme in plasma is used as a non‐destructive biomarker to diagnose the exposure of birds to these pesticides. Scavengers are one of the most important bird groups threatened by the use of baits poisoned with anticholinesterase pesticides. Knowledge of the characteristics of this enzyme in each bird species is crucial, as several studies indicate that more than one cholinesterase form may be present in the plasma of birds. In this study, cholinesterase activity was characterized in the plasma of the Eurasian Griffon Vulture Gyps fulvus by using several substrates and inhibitors of the enzyme, and its normal activity value was also determined. The in vitro sensitivity of Gyps fulvus plasma cholinesterase to carbamate insecticides (aldicarb, carbaryl and methomyl) was also investigated. The results indicated that propionylthiocholine iodide was the preferred substrate to determine plasma cholinesterase activity, followed by acetylcholine iodide and S‐butyrylcholine iodide, and acetylcholinesterase was the predominant enzymatic activity in Gyps fulvus plasma. Aldicarb was the most potent in vitro inhibitor of plasma cholinesterase activity in this species. However, cholinesterase enzymatic activity was significantly inhibited by all tested carbamates, providing further evidence that this biomarker is a suitable tool to monitor the exposure to these poisons in the field, highlighting its utility in conservation programmes.     

Increased detrimental effects of ectoparasites on their bird hosts during adverse environmental conditions

Parasites are usually assumed to negatively affect fitness components of their hosts, particularly during adverse environmental conditions. We experimentally tested whether the ectoparasitic house martin bug (Oeciacus hirundinis) had larger negative effects on its house martin (Delichon urbica) host during second clutches. Conditions for reproduction decreased during the season, being less favourable for the second clutch than the first clutch. This was demonstrated by smaller clutch sizes, lower breeding success and smaller and lighter nestlings during the second clutch, as compared to the first, in the absence of parasites. Experimental manipulation of the intensity of parasitism resulted in the reproductive success of the host being reduced with increasing numbers of house martin bugs. This effect was much stronger during the second, as compared to the first, clutch. The house martin bug therefore had a greater detrimental effect on the reproduction of its host when conditions for reproduction were poor.     

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.     

Mortality over 16 years of cacti in a burnt desert grassland

Mortality of cacti after grassland fires is usually <25% within 2 years. Little, however, is known about long-term mortality. This study followed the fate of 50 marked plants of each of four species of small cacti (Coryphantha vivipara, Echinocereus pectinatus, Echinomastus intertextus and Mammillaria heyderi) on burnt and unburnt desert-grassland in Arizona. All marked plants were dead within 16 years. The first three species suffered increased mortality after being burnt while M. gummifera survived better after fire. The average rate of mortality of burnt plants over unburnt plants was 10.2 times higher in E. pectinatus, 3.7 times in C. vivipara and 1.5 times in E. intertextus. In M. heyderi, however, the mortality of burnt plants was 0.25 that of control plants. Fire more rapidly removed the breeding populations of the first three species, reducing seed availability in the occasional years favourable for establishment, increasing the risk of local extinction. Cacti did establish on the study areas and by the end were of a similar size to the dead cohort giving the superficial impression that the site was unchanged except for a reduction on unburnt sites in total cactus density (burnt: 445/ha in 1987, 401/ha in 2003; control: 2235/ha and 675/ha, respectively). Evidence of fire and dead plants rapidly disappeared, so without plants being marked the death of one population and establishment of another would have been easily missed. Estimates of growth rates support the assertion that the final plants established during the study.     

Microflora and internal parasites of the digestive tract of Eurasian griffon vultures (Gyps fulvus) in Croatia

Faecal and vomit samples were collected from 12 griffon vultures (Gyps fulvus) in May 2002 and examined for bacteria, fungi and parasites. Bacteria present included Escherichia coli, Aeromonas salmonicidia, Proteus sp., Escherichia fergusonii, Providencia alcalifaciens, Micrococcus sp. and Streptococcus sp. Mycological examination revealed the presence of Candida sp., yeasts and in one case, Rhodotorula rubra. Strongyle eggs were found in several samples. In most faecal samples, ascarid eggs of Toxocara type were found. Tapeworm eggs of Moniezia type were present in one faecal sample. Both microbiologic and parasitic findings are most likely the result of the feeding habits of griffon vultures.     

High occurrence of extra-pair partnerships and homosexuality in a captive Cape Vulture Gyps coprotheres colony§

The Cape Vulture Gyps coprotheres is an Endangered colonial cliff-nesting species that is typically cited as monogamous. Observations of wild Cape Vulture colonies note extra-pair breeding activities but homosexual activity has never been confirmed. Observations of breeding behaviours within a captive colony were conducted at VulPro’s breeding facilities in South Africa The captive breeding programme included 10 breeding pairs and a total of 41 non-releasable, individually recognisable, and genetically sexed subadults or adults. Observations were conducted over the 2014 breeding season as part of a study investigating the social and rearing conditions of chicks intended for release in a population supplementation programme. A high percentage of total colony copulation attempts (22%) were extra-pair copulations but only 23 interactions resulted in cloacal contact or possible sperm transfer. Four paired males formed extra-pair relationships, two of which were homosexual.