Isolation and amplification of shrew DNA from barn owl pellets

The last decade has seen a number of studies reporting the extraction of DNA from ancient sources, such as fossil bones. Owl pellets, which contain an excellent skeletal record of small mammals consumed, can be used in a non-invasive sampling method for genetic studies of free-ranging animals without the need for direct capture or even observation. Such a non-invasive sampling method will allow us to address questions that cannot be answered using conventional methods and will lead to a more integrated study of micromammals. In the present study, various protocols used for ancient DNA extraction were investigated, in order to determine the applicability of owl pellets as a source of DNA for phylogenetic and phylogeographical studies of micromammals. Of the 12 bone samples used in this study, 11 gave sequences of expected species ( Crocidura , Rattus and Mus ) and size (around 300 bp), using the pair of primers L14841/H15149, which target a highly conserved region of the cytochrome b gene. The results obtained demonstrate that mitochondrial DNA can be isolated and amplified using bones of micromammals found in barn owl ( Tyto alba ) pellets. The recovery of genetic data from owl pellets will enable the identification of prey species for several phylogenetic and phylogeographical studies of small mammals (Muridae and Insectivora) appearing in the owl's diet. The main advantages of this new approach are that (a) the pellets are very easy to find and collect, (b) the pellets can potentially provide a large number of individuals of small mammals, and (c) the method can cover a wide geographical area complementary to the range of owls of this type.  © 2005 The Linnean Society of London, Biological Journal of the Linnean Society , 2005, 85 , 331–340.     

Impacts of Agriculture on the Diet and Productivity of Mackinder's Eagle Owls (Bubo capensis mackinderi ) in Kenya

Land conversion for agriculture is an increasing threat to biodiversity conservation, but its ecological effects on African birds is practically unknown. We investigated the impacts of agriculture on the diet and productivity of a small, disjunct population of Mackinder's eagle owls ( Bubo capensis mackinderi  ) in central Kenya. Owl diet was determined by analysis of pellets and other remains and compared to small mammal populations estimated by live trapping in two habitats. Small mammal abundance was low and averaged 7.4 small mammals/ha in farms and 0.5 small mammals/ha in grassland. Owls consumed a wide diversity of prey. The majority were mammals (87%) followed by birds (7%) and insects (5%). The percentage of small mammals in owl diet correlated positively with the relative abundance of small mammals during monthly trapping sessions. Diet composition did not influence owl breeding success. Farming activities affected owl diet composition through crop production. The amount of maize, peas, and carrots growing in farms was correlated with the abundance of Mastomys sp. and Procavia sp. in the owl's diet. Agricultural activities had a large effect on Mackinder's eagle owl diet by increasing the abundance of certain small-mammal prey and attracting owl prey to farms, though farming practices harmful to owls were observed.     

巴基斯坦旁遮普中部仓鸮食性的季节性变化

在巴基斯坦对仓鸮食性的季节变化进行了研究。通过分析连续3年在6个地区搜集的2 360个仓鸮回吐食物团,发现其食物主要是小型哺乳动物(95.6 %)。其中,家(Suncus murinus)有最高的比例,达65.6%(冬季最多78%,夏季最少27%)。就生物量而言,小型哺乳动物占仓鸮食物总生物量的99% 。     

Application of molecular methods to identify food resources of short-eared owl (Asio flammeus) in wetland community

The short-eared owl (Asia flammeus), which lives in open wetland habitats, has been threatened and become endangered in South Korea due to the destruction and loss of its habitats. However, the diet of the short-eared owl remains to be studied. The prey of the short-eared owl was monitored using molecular methods as an alternative tool. A total of 121 pellets of the species were collected in the Dalsung wetland. Of these pellets, about 30% (n = 33) were selected for prey identification in 2002 (n = 12), 2003 (n = 6) and 2006 (n = 15). Two hundred and thirty-two bone fragments were successfully analyzed and 9 prey species were detected. In an expanded analysis, excluding 4 pellets used for the preliminary analysis, small mammals constituted the highest percentage (89.1%): 47.6% Eurasian field mouse (Apodemus agrarius); 27.2% European harvest mouse (Micromys minutus); 6.1% Japanese wild house mouse (Mus musculus); 5.4% Asian lesser white-toothed shrew (Crocidura shantungensis); and 2.7% Norway rat (Battus norvegicus). Not only small mammals, but birds and amphibians were also identified: 4.1% Japanese quail (Cotumix japonica); 3.4% vinous-throated parrotbill (Paradoxornis webbianus); 0.7% eastern turtle dove (Streptopelia orientalis); and 2.7% Kori salamander (Hynobius yangi). This result would be valuable in understanding the feeding ecology of the short-eared owl in its wetland habitats.     

Using scats of a generalist carnivore as a tool to monitor small mammal communities in Mediterranean habitats

Owl pellets have long been used to analyze communities of small mammals, while analogous analyses of faeces of mammal carnivores are not available. We demonstrate that common genet (Genetta genetta) scats can be used as a reliable method to sample small mammal communities and to monitor their variations. We have compiled data on 6350 small mammal remains of 18 species found in scats from 51 different latrines in a 1200 km² area of northeastern Spain. Genet scats sampled effectively 95.6% of the small mammal species ranging in size from 2.7 to 385 g. Spatial patterns of diet composition along environmental gradients of elevation, climate and land-use matched expected changes in small mammal communities along these gradients according to ecological requirements of prey species. Frequencies of occurrence of prey in genet scats were strongly correlated with frequencies of occurrence in barn owl (Tyto alba) pellets. Genet scats included two forest species not preyed upon by owls, whereas only one species was not preyed upon by genets. Forests species were more frequent in genet than in barn owl diets after correcting for environmental effects, whereas the opposite was true for open-habitat and synanthropic species. Scats of generalist carnivores can be used to estimate the spatial patterns of distribution and abundance of small mammal communities. Genet scats in fact overcome some of the limitations of more traditional sampling methods (live-trapping and owl diets), as genets were less selective and their diets reflect more accurately changes in community composition.     

Effects of vole fluctuations on the population dynamics of the barn owl <Emphasis Type="Italic">Tyto alba</Emphasis>

Many predator species feed on prey that fluctuates in abundance from year to year. Birds of prey can face large fluctuations in food abundance i.e. small mammals, especially voles. These annual changes in prey abundance strongly affect the reproductive success and mortality of the individual predators and thus can be expected to influence their population dynamics and persistence. The barn owl, for example, shows large fluctuations in breeding success that correlate with the dynamics in voles, their main prey species. Analysis of the impact of fluctuations in vole abundance (their amplitude, peaks and lows, cycle length and regularity) with a simple predator prey model parameterized with literature data indicates population persistence is especially affected by years with low vole abundance. In these years the population can decline to low owl numbers such that the ensuing peak vole years cannot be exploited. This result is independent of the length and regularity of vole fluctuations. The relevance of this result for conservation of the barn owl and other birds of prey that show a numerical response to fluctuating prey species is discussed.     

The influence of meal size on prey DNA detectability in piscivorous birds

Molecular methods allow noninvasive assessment of vertebrate predator–prey systems at high taxonomic resolution by examining dietary samples such as faeces and pellets. To facilitate the interpretation of field‐derived data, feeding trials, investigating the impacts of biological, methodological and environmental factors on prey DNA detection, have been conducted. The effect of meal size, however, has not yet been explicitly considered for vertebrate consumers. Moreover, different noninvasively obtained sample types remain to be compared in such experiments. Here, we present a feeding trial on abundant piscivorous birds, Great Cormorants (Phalacrocorax carbo), to assess meal size effects on postfeeding prey DNA detection success. Faeces and pellets were sampled twice a day after the feed of large (350–540 g), medium (190–345 g) and small (15–170 g) fish meals contributing either a large (>79%) or small (<38%) share to the daily consumption. Samples were examined for prey DNA and fish hard parts. Molecular analysis of faeces revealed that both large meal size and share had a significantly positive effect on prey DNA detection rate postfeeding. Furthermore, large meals were detectable for a significantly longer time span with a detection limit at ~76 hr and a 50% detection probability at ~32 hr postfeeding. In pellets, molecular methods reliably identified the meal consumed the previous day, which was not possible via morphological analysis or when examining individual faeces. The less reliable prey DNA detection of small meals or meal shares in faeces signifies the importance of large numbers of dietary samples to obtain reliable trophic data.     

巴基斯坦旁遮普中部横斑腹小鸮的食性

横斑腹小鸮(Athene brama)是巴基斯坦旁遮普中部农业生态系统中最常见但研究匮乏的猛禽。我们通过分析吐弃块(n=498)以了解其季节性和地域性食性变化。结果显示:小型哺乳类占所有食物生物量的98.4%,而昆虫类在数量上占据绝对优势(56.4%)。在小型哺乳类食物中,鼠类(38.8%)是稳定的食物成分。而在昆虫食物中,甲虫最多(42.8%)。横斑腹小亦捕食鼩鼱类(2.3%)和鸟类(1.7%)。昆虫类食物还有蟑螂(6.4%)、臭虫(3.3%)、蝗虫(6.7%)和蚂蚁(3.2%)。小家鼠(Mus musculus)和柔毛鼠(Rattus meltada)在鼠类食物中出现频次最多。不同地区中,Hafizabad的食物多样性最高(2.07),而秋季的食物多样性最高(1.99)。     

Molecular scatology as a tool to study diet: analysis of prey DNA in scats from captive Steller sea lions

The DNA of prey present in animal scats may provide a valuable source of information for dietary studies. We conducted a captive feeding trial to test whether prey DNA could be reliably detected in scat samples from Steller sea lions (Eumetopias jubatus). Two sea lions were fed a diet of fish (five species) and squid (one species), and DNA was extracted from the soft component of collected scats. Most of the DNA obtained came from the predator, but prey DNA could be amplified using prey-specific primers. The four prey species fed in consistent daily proportions throughout the trial were detected in more than 90% of the scat DNA extractions. Squid and sockeye salmon, which were fed as a relatively small percentage of the daily diet, were detected as reliably as the more abundant diet items. Prey detection was erratic in scats collected when the daily diet was fed in two meals that differed in prey composition, suggesting that prey DNA is passed in meal specific pulses. Prey items that were removed from the diet following one day of feeding were only detected in scats collected within 48 h of ingestion. Proportions of fish DNA present in eight scat samples (evaluated through the screening of clone libraries) were roughly proportional to the mass of prey items consumed, raising the possibility that DNA quantification methods could provide semi-quantitative diet composition data. This study should be of broad interest to researchers studying diet since it highlights an approach that can accurately identify prey species and is not dependent on prey hard parts surviving digestion.     

High efficiency protocol of DNA extraction from Micromys minutus mandibles from owl pellets: a tool for molecular research of cryptic mammal species

Owl pellets have high potential as a source of DNA. However, this noninvasive method of collecting DNA is rarely used, and its methodological aspects are poorly understood. We investigated the methodology for DNA extraction and amplification from owl pellets containing the smallest European rodent—the Harvest mouse Micromys minutus—as an example. We used mandibles identified in owl pellets for mitochondrial and nuclear DNA amplification. For DNA extraction, we tested two commercial protocols and utilized a protocol being a combination of two commercial kits which ensured high efficiency of DNA extraction. Additionally, we recorded that the amount of DNA was five times higher in extracts from teeth as compared to DNA extracts from jawbones derived from the same mandible. The quantity of DNA was significantly positively correlated with biological sample weight; however, the age of the pellet remains had an impact on the level of inhibition. We recorded inhibition in 40 % of mtDNA extracts derived from pellets older than 150 months, whereas in DNA extracts from pellets younger than 80 months, we did not observe a negative impact of inhibition on PCR efficiency. The amplification success rate was 89.9 % for the mitochondrial fragment and 39.4 % in the case of the nuclear fragment. We observed partial degradation of DNA evidenced by the fact that the longest fragments that we were able to amplify in the case of mtDNA were 450 and 200 bp for nuDNA. The study shows that pellets can be considered as a source of DNA and have high potential for molecular research in the case of threatened species and species that are difficult to study using standard field techniques.     

南充高坪机场短耳 鸮 越冬期的食性分析和习性观察

2006年3～5月,收集南充高坪机场草地和周围松林越冬期短耳鸮(Asio flammeus)的食团,进行食性分析。在805个完整食团和一些零碎的食团中,小型兽类是短耳食物组成的主要成分,占98.4%,其中四川短尾(Anourosorex squamipes)所占比例最高,为97.2%。通过调查短耳鸮的食团在南充高坪机场草地里的分布情况,分析短耳v在机场草地的栖息地选择,发现短耳鸮并没有选择草高和密集的区域作为栖息场所,而高20cm左右的白茅(Imperata cylindrical)丛是短耳最喜欢栖息的场所。     

Food habits of genetically identified pine marten (<Emphasis Type="Italic">Martes martes</Emphasis>) expanding in agricultural lowlands (NW Italy)

We assessed the diet of pine marten (Martes martes) expanding in the heavily human-altered agricultural plain of the River Po, northern Italy. Between February 2008 and November 2009, surveys were carried out twice a week during seasonal sampling sessions of three to four consecutive weeks. To distinguish the faecal samples from those of sympatric carnivores, a polymerase chain reaction-restriction fragment length polymorphism method was applied. The availability of small mammals was assessed by the analysis of 59 barn owl (Tyto alba) pellets. A total of 109 pine marten faeces were analysed. Its diet consisted of fruit, rodents, lagomorphs and birds. Seasonal variation occurred for fruit, which prevailed in summer, and rodents, which were more preyed upon in autumn. In winter, the diet of the pine marten was almost totally based on vertebrates, and lagomorphs were the main source of protein in summer. Use of small mammals differed significantly from their availability, voles, particularly bank vole Myodes glareolus, being preferred to mice (Apodemus sp.) and rats (Rattus sp.). Medium-size mammals formed about 18% of pine marten diet, a value generally reported for high-latitude habitats. In agricultural areas, the pine marten proved to be an opportunistic predator, able to face the reduced availability of small mammals by preying upon medium-size prey and fruit.     

On the diet of the Pharaoh Eagle Owl,Bubo ascalaphus (Savigny, 1809), in Qatar,with an overview of its feeding habits

The diet of Bubo ascalaphus in Qatar was assessed based on pellets collected from the first known nesting site of the species in the country. The pellets contained a total of 68 prey items, representing 9 different species: 4 mammals, 1 bird, 1 reptile, and at least 3 scorpions. Mammals clearly comprised the major food source (89.7% and 97.7% in frequency and biomass respectively). Our data suggest that Pharaoh Eagle Owls are opportunistic predators that feed on a variety of prey depending on their temporal/spatial availability, which is consistent with previous studies. A literature review clearly suggests that Eagle Owls in arid to semi-arid environments are opportunistic predators with small mammals being their main prey. Predation on migrating Blue-cheeked Bee-eaters Merops persicus supports this hypothesis.     

Vulnerability of small and medium-sized prey mammals in relation to their habitat preferences,age classes and locomotion types in the temperate Monte Desert,Argentina

In this study we examined predation by small felids upon small and medium-sized prey mammals in the central region of the Monte Desert. We analyzed the degree of vulnerability of prey mammals in relation to their habitat use patterns, modes of locomotion and age classes. Medium and small rodents were the predominant prey in the diet of small cats. The most consumed small mammal was Akodon molinae, which inhabits areas of dense cover, while species occurring in open habitats such as Eligmodontia typus were less consumed. Among medium-sized mammals, Galea leucoblephara that depends on patches with high plant cover for refuge was more consumed than Microcavia australis that lives in colonies and displays a complex set of antipredator strategies. Prey selectivity on small rodents showed that E. typus, which uses bipedal locomotion and an erratic escape behaviour, was consumed in greater proportion than its availability. We suggest that although its specialized locomotory mode would diminish predator attacks, its preference for open habitats would increase the probability of predator–prey encounter. Consumption of small mammals focuses mostly on adult individuals. However, we found a significant consumption of senile individuals of A. molinae, which points out that vulnerability tends to be high towards the age of maturity in this species.     

乌鲁木齐市区越冬期长耳鸮的食性分析

2009～2011年间,利用食团分析法对乌鲁木齐市越冬长耳鸮(Asio otus)的食性进行分析。3年累计收集长耳鸮食团683份,辨认出1 132只猎物。分析结果表明,长耳鸮在冬季共捕食小型哺乳类6种,鸟类2种。小家鼠(Mus musculus)是最常见的食物,占总捕食量的53.45%。小型哺乳类是长耳鸮的主要食物,它在食物组成中出现的总频率为88.16%,以生物量计,小型哺乳类占食物构成的95.13%。长耳鸮的食物组成年度间差异显著,与当地猎物资源多样性和可获得性密切相关,表明长耳鸮可能采用机会主义者的捕食策略。     

Food web structure and climate effects on the dynamics of small mammals and owls in semi-arid Chile

Population dynamics of small mammals and predators in semi-arid Chile is positively correlated with rainfall associated with incursions of El Niño (El Niño Southern Oscillation: ENSO). However, the causal relationships between small mammal fluctuations, predator oscillations, and climatic disturbances are poorly understood. Here, we report time series models for three species of small mammal prey and two species of owl predators. The large differences in population fluctuations between the three small mammal species are related to differences in their respective feedback structures. The analyses reveal that per capita growth rate of the leaf-eared mouse is a decreasing function of log density and of log barn owl abundance together with a positive rainfall effect. In turn, per capita population growth rate ( R -function) of the barn owl is a negative function of log barn owl abundance and a positive function of leaf-eared mouse abundance, suggesting a predator–prey interaction. The dramatic population fluctuations exhibited by leaf-eared mouse ( Phyllotis darwini ) are caused by climate effects coupled with a complex food web architecture.     

The relationship between intra–guild diet overlap and breeding in owls in Israel

Even though intra-guild predators frequently prey on the same species, it is unclear whether diet overlap between two predators is a source of interspecific competition or whether predators simply use the same abundant prey resource. We measured the extent to which the diets of barn owls (Tyto alba) and long-eared owls (Asio otus) in Israel overlap and examined whether yearly differences in diet overlap correlate with barn owl breeding success. Pianka’s index of niche overlap was positively related to barn owl population size but not to its breeding success. The number of breeding barn owls was higher when long-eared owls consumed more rodents, suggesting that diet overlap most likely increased when rodents became more abundant. Therefore, in Israel, when these two owl species prey more often on rodents, their diets are more similar and interspecific competition is reduced. Unlike sympatric populations in Europe, in years when rodents are less abundant in Israel long-eared owls switch to hunting alternative prey (e.g., birds), perhaps to avoid competition with barn owls.     

Effect of snow cover on seasonal changes in diet, habitat, and regional distribution of raptors that prey on small mammals in boreal zones of Fennoscandia

Seasonal changes in spatial distribution of search effort of birds that prey on small mammals were studied in two structurally different coniferous forest habitats in the northern boreal zone in SE Norway. During the season with snow cover both the proportion of Microtus relative to that of Clethrionomys in the predators' diet, and their use of a clear-cut relative to that of older forest were lower than during the snow-free season. This was related to a lower relative availability of prey ( Microtus agrestis and M. oeconomus ) in the clear-cut when the ground was snow-covered than when it was snow-free. Based on this local pattern I suggest the following explanation for differences in migratory strategy between raptors that prey on small mammals in Fennoscandian boreal zones: species that migrate to snow-free areas in winter are either adapted to hunt by the energetically expensive method of quartering in open grassland habitats, where prey ( Microtus ) availability is relatively low during periods with snow cover (hen harrier Circus cyaneus , short-eared owl Asio flammeus , and longeared owl A. otus ), or by sit-and-wait in open grassland and forest habitats, the latter with relatively high prey availability during periods with snow cover, but unable to locate concealed prey (kestrel Falco tinnuculus , common buzzard Buteo huteo , and roughlegged buzzard B. lagopus ). In contrast, species that remain in areas with permanent snow cover during winter use the energetically cheap sit-and-wait tactic, and are able to hunt in closed forest habitat and localize concealed prey (the remaining owl species). Interspecific differences in prey availability as determined by hunting habitat and hunting mode is probably more important in shaping the migration patterns of Fennoscandian owls than is nest site availability.     

What determines prey selection in owls? Roles of prey traits,prey class,environmental variables,and taxonomic specialization

Ecological theory suggests that prey size should increase with predator size, but this trend may be masked by other factors affecting prey selection, such as environmental constraints or specific prey preferences of predator species. Owls are an ideal case study for exploring how predator body size affects prey selection in the presence of other factors due to the ease of analyzing their diets from owl pellets and their widespread distributions, allowing interspecific comparisons between variable habitats. Here, we analyze various dimensions of prey resource selection among owls, including prey size, taxonomy (i.e., whether or not particular taxa are favored regardless of their size), and prey traits (movement type, social structure, activity pattern, and diet). We collected pellets of five sympatric owl species (Athene noctua, Tyto alba, Asio otus, Strix aluco, and Bubo bubo) from 78 sites across the Mediterranean Levant. Prey intake was compared between sites, with various environmental variables and owl species as predictors of abundance. Despite significant environmental impacts on prey intake, some key patterns emerge among owl species studied. Owls select prey by predator body size: Larger owls tend to feed on wider ranges of prey sizes, leading to higher means. In addition, guild members show both specialization and generalism in terms of prey taxa, sometimes in contrast with the expectations of the predator–prey body size hypothesis. Our results suggest that while predator body size is an important factor in prey selection, taxon specialization by predator species also has considerable impact.