Selection of Anti-Sulfadimidine Specific ScFvs from a Hybridoma Cell by Eukaryotic Ribosome Display

BACKGROUND:Ribosome display technology has provided an alternative platform technology for the development of novel low-cost antibody based on evaluating antibiotics derived residues in food matrixes. METHODOLOGY/PRINCIPAL FINDINGS:In our current studies, the single chain variable fragments (scFvs) were selected from hybridoma cell lines against sulfadimidine (SM(2)) by using a ribosome library technology. A DNA library of scFv antibody fragments was constructed for ribosome display, and then mRNA-ribosome-antibody (MRA) complexes were produced by a rabbit reticulocyte lysate system. The synthetic sulfadimidine-ovalbumin (SM(2)-OVA) was used as an antigen to pan MRA complexes and putative scFv-encoding genes were recovered by RT-PCR in situ following each panning. After four rounds of ribosome display, the expression vector pCANTAB5E containing the selected specific scFv DNA was constructed and transformed into Escherichia coli HB2151. Three positive clones (SAS14, SAS68 and SAS71) were screened from 100 clones and had higher antibody activity and specificity to SM(2) by indirect ELISA. The three specific soluble scFvs were identified to be the same molecular weight (approximately 30 kDa) by Western-blotting analysis using anti-E tag antibodies, but they had different amino acids sequence by sequence analysis. CONCLUSIONS/SIGNIFICANCE:The selection of anti-SM(2) specific scFv by in vitro ribosome display technology will have an important significance for the development of novel immunodetection strategies for residual veterinary drugs

Mesopredator Release by an Emergent Superpredator: A Natural Experiment of Predation in a Three Level Guild

Chakarov N, Krüger O. Mesopredator Release by an Emergent Superpredator: A Natural Experiment of Predation in a Three Level Guild. PLoS ONE. 2010;5(12): e15229.Background: Intraguild predation (IGP) is widespread but it is often neglected that guilds commonly include many layers of dominance within. This could obscure the effects of IGP making unclear whether the intermediate or the bottom mesopredator will bear higher costs from the emergence of a new top predator. Methodology/Principal Findings: In one of the most extensive datasets of avian IGP, we analyse the impact of recolonization of a superpredator, the eagle owl Bubo bubo on breeding success, territorial dynamics and population densities of two mesopredators, the northern goshawk Accipiter gentilis and its IG prey, the common buzzard Buteo buteo. The data covers more than two decades and encompass three adjacent plots. Eagle owls only recolonized the central plot during the second decade, thereby providing a natural experiment. Both species showed a decrease in standardized reproductive success and an increase in brood failure within 1.5 km of the superpredator. During the second decade, territory dynamics of goshawks was significantly higher in the central plot compared to both other plots. No such pattern existed in buzzards. Goshawk density in the second decade decreased in the central plot, while it increased in both other plots. Buzzard density in the second decade rapidly increased in the north, remained unchanged in the south and increased moderately in the center in a probable case of mesopredator release. Conclusions/Significance: Our study finds support for top-down control on the intermediate mesopredator and both top-down and bottom-up control of the bottom mesopredator. In the face of considerable costs of IGP, both species probably compete to breed in predator-free refugia, which get mostly occupied by the dominant raptor. Therefore for mesopredators the outcome of IGP might depend directly on the number of dominance levels which supersede them

Interaction of climate change with effects of conspecific and heterospecific density on reproduction

We studied the relationship between temperature and the coexistence of great titParus majorand blue titCyanistes caeruleus, breeding in 75 study plots across Europe and North Africa. We expected an advance in laying date and a reduction in clutch size during warmer springs as a general response to climate warming and a delay in laying date and a reduction in clutch size during warmer winters due to density-dependent effects. As expected, as spring temperature increases laying date advances and as winter temperature increases clutch size is reduced in both species. Density of great tit affected the relationship between winter temperature and laying date in great and blue tit. Specifically, as density of great tit increased and temperature in winter increased both species started to reproduce later. Density of blue tit affected the relationship between spring temperature and blue and great tit laying date. Thus, both species start to reproduce earlier with increasing spring temperature as density of blue tit increases, which was not an expected outcome, since we expected that increasing spring temperature should advance laying date, while increasing density should delay it cancelling each other out. Climate warming and its interaction with density affects clutch size of great tits but not of blue tits. As predicted, great tit clutch size is reduced more with density of blue tits as temperature in winter increases. The relationship between spring temperature and density on clutch size of great tits depends on whether the increase is in density of great tit or blue tit. Therefore, an increase in temperature negatively affected the coexistence of blue and great tits differently in both species. Thus, blue tit clutch size was unaffected by the interaction effect of density with temperature, while great tit clutch size was affected in multiple ways by these interactions terms.Peer reviewe

Landscape homogenization due to agricultural intensification disrupts the relationship between reproductive success and main prey abundance in an avian predator

Selecting high-quality habitat and the optimal time to reproduce can increase individual fitness and is a strong evolutionary factor shaping animal populations. However, few studies have investigated the interplay between land cover heterogeneity, limitation in food resources, individual quality and spatial variation in fitness parameters. Here, we explore how individuals of different quality respond to possible mismatches between a cue for prey availability (land cover heterogeneity) and the actual fluctuating prey abundance.Peer reviewe

Interspecific variation in the relationship between clutch size, laying date and intensity of urbanization in four species of hole-nesting birds

The increase in size of human populations in urban and agricultural areas has resulted in considerable habitat conversion globally. Such anthropogenic areas have specific environmental characteristics, which influence the physiology, life history, and population dynamics of plants and animals. For example, the date of bud burst is advanced in urban compared to nearby natural areas. In some birds, breeding success is determined by synchrony between timing of breeding and peak food abundance. Pertinently, caterpillars are an important food source for the nestlings of many bird species, and their abundance is influenced by environmental factors such as temperature and date of bud burst. Higher temperatures and advanced date of bud burst in urban areas could advance peak caterpillar abundance and thus affect breeding phenology of birds. In order to test whether laying date advance and clutch sizes decrease with the intensity of urbanization, we analyzed the timing of breeding and clutch size in relation to intensity of urbanization as a measure of human impact in 199 nest box plots across Europe, North Africa, and the Middle East (i.e., the Western Palearctic) for four species of hole-nesters: blue tits (Cyanistes caeruleus), great tits (Parus major), collared flycatchers (Ficedula albicollis), and pied flycatchers (Ficedula hypoleuca). Meanwhile, we estimated the intensity of urbanization as the density of buildings surrounding study plots measured on orthophotographs. For the four study species, the intensity of urbanization was not correlated with laying date. Clutch size in blue and great tits does not seem affected by the intensity of urbanization, while in collared and pied flycatchers it decreased with increasing intensity of urbanization. This is the first large-scale study showing a species-specific major correlation between intensity of urbanization and the ecology of breeding. The underlying mechanisms for the relationships between life history and urbanization remain to be determined. We propose that effects of food abundance or quality, temperature, noise, pollution, or disturbance by humans may on their own or in combination affect laying date and/or clutch size

Age and sex differences in numerical responses, dietary shifts, and total responses of a generalist predator to population dynamics of main prey

Fluctuations in the abundance of main prey species might shape animal communities, by inducing numerical responses and dietary shifts in predators. Whether numerical responses and dietary shifts differ among individuals of different age and sex has so far gained little attention. These differences could affect how much predators consume main and alternative prey, thus causing variation in predation pressure on main and alternative prey species. We studied the effect of fluctuating main prey abundance (voles) in autumn on the age and sex composition of a food-hoarding population of Eurasian pygmy owls Glaucidium passerinum (327 individuals), and on the species composition of their food stores in western Finland during 2003–2017 (629 food stores). Numbers of yearlings (< 1-year old) of both sexes and adult (+ 1-year old) females increased with increasing vole abundance. During low vole abundance, adult owls stored more small birds and less small mammals than yearlings. Females stored more small mammals than males and showed a tendency to store less birds. The amount of consumed birds (the most important alternative prey), and in particular of crested, willow, great, and blue tits, increased with low vole densities. Our results show that numerical, functional, and total responses of pygmy owls, and probably also other vertebrate predators, to the availability of the main prey in winter are shaped by the age and sex composition of the predator population, which both show large spatio-temporal variation in boreal forests

Climate change and perishable food hoards of an avian predator: Is the freezer still working?

Changing climate can modify predator–prey interactions and induce declines or local extinctions of species due to reductions in food availability. Species hoarding perishable food for overwinter survival, like predators, are predicted to be particularly susceptible to increasing temperatures. We analysed the influence of autumn and winter weather, and abundance of main prey (voles), on the food-hoarding behaviour of a generalist predator, the Eurasian pygmy owl (Glaucidium passerinum), across 16 years in Finland. Fewer freeze–thaw events in early autumn delayed the initiation of food hoarding. Pygmy owls consumed more hoarded food with more frequent freeze–thaw events and deeper snow cover in autumn and in winter, and lower precipitation in winter. In autumn, the rotting of food hoards increased with precipitation. Hoards already present in early autumn were much more likely to rot than the ones initiated in late autumn. Rotten food hoards were used more in years of low food abundance than in years of high food abundance. Having rotten food hoards in autumn resulted in a lower future recapture probability of female owls. These results indicate that pygmy owls might be partly able to adapt to climate change by delaying food hoarding, but changes in the snow cover, precipitation and frequency of freeze–thaw events might impair their foraging and ultimately decrease local overwinter survival. Long-term trends and future predictions, therefore, suggest that impacts of climate change on wintering food-hoarding species could be substantial, because their ‘freezers’ may no longer work properly. Altered usability and poorer quality of hoarded food may further modify the foraging needs of food-hoarding predators and thus their overall predation pressure on prey species. This raises concerns about the impacts of climate change on boreal food webs, in which ecological interactions have evolved under cold winter conditions

Food hoarding of an avian predator: sex- and age-related differences under fluctuating food conditions

Abstract: Hoarding behaviour (storing food for a later use) has evolved to reduce starvation risk when resources are scarce. Different age and sex classes often show differences in foraging due to experience, skills or life history strategy, but such differences in hoarding under spatio-temporally varying environmental conditions have rarely been studied in the wild. We studied hoarding behaviour of Eurasian pygmy owls (Glaucidium passerinum) during 2003–2016 in western Finland, where the abundance of their main prey (voles) fluctuates in three-year population cycles. In 14 years, 1056 food stores were found during the hoarding season (Oct–Dec) and 330 pygmy owls were trapped at these stores. The number of stores per individual did not vary in relation to age, sex or vole abundance. Adults (+ 1-year old) had their stores farther apart than yearlings. Both the number of stores per year and the biomass of stored prey items increased with vole abundance. Females and yearlings had larger and heavier stores than males and adults, respectively. The same individuals stored more food as yearlings than as adults. These sex- and age-differences in hoarding indicate that it is not constrained by experience or skills. It rather seems that less-experienced yearlings rely more on stored food than adults. Females may need more food due to their larger size and need to accumulate energy reserves before reproduction. A detailed knowledge of age- and sex-related differences in hoarding behaviour under fluctuating abundances of main foods is fundamental to better understand a population response to climate change and forest management. Significance statement: The hoarding behaviour of animals has evolved to cope with the problem of food limitation. On the basis of 14-year data from pygmy owls, we show that the number of stores per year and the biomass of prey items per store increased with vole abundance in the environment. Adults had stores farther apart than yearlings, and females and yearlings stored more prey items and biomass compared to males and adults, respectively. These results indicate that hoarding behaviour responds to the available main prey abundance and varies with traits such as age and sex. Because different age and sex classes might respond differently to variation in food abundance, due to habitat alterations or climate change, a detailed knowledge of hoarding behaviour can be of particular importance to understand changes in body condition, reproductive success and survival of pygmy owls under changing climate and management of boreal forest

Habitat choice of a secondary cavity user indicates higher avoidance of disturbed habitat during breeding than during food-hoarding

Tree cavities are an essential requirement for reproduction and survival of secondary cavity nesters. Little is known, however, about how cavity distribution in space and time may limit their use during and outside of the breeding season. Using long-term (17 yr) data on the use of nest-boxes resembling tree cavities by individually marked Eurasian pygmy owls Glaucidium passerinum, we compared the habitat and the distances among the nest-boxes used during two different seasons. We found that pygmy owls are more demanding in their requirements for breeding than for food-hoarding habitat, as the habitat surrounding the nesting sites had a higher proportion and biomass of coniferous forests, especially spruce, and nesting sites were farther from houses than food-hoarding sites. Pygmy owls mainly used different nest-boxes for nesting and food-hoarding and they also used different nest-boxes in different years. The distance between the nest site and food-hoarding sites was longer when the main food availability (voles) was low than when it was high. The distance was also longer in females than in males, probably indicating that females need to move further to find suitable hunting areas. In conclusion, these results highlight the importance of spruce forests as a nesting habitat of pygmy owls, and the need of several tree cavities within their home range. Forest management should secure a sufficient supply of these resources at the scale of pygmy owl home ranges

Maternal transfer of androgens in eggs is affected by food supplementation but not by predation risk

Mothers may affect the future success of their offspring by varying allocation to eggs and embryos. Allocation may be adaptive based on the environmental conditions perceived during early breeding. We investigated the effects of food supplementation and predation risk on yolk hormone transfer in the pied flycatcher Ficedula hypoleuca. In a food supplementation experiment, females were food-supplemented prior to and during egg-laying and androgen concentrations were measured throughout the laying order. Predation risk was investigated in three different studies combining both correlative data, where flycatchers bred in close proximity to two different predator species that prey upon adult flycatchers (either Tengmalm's owl Aegolius funereus or pygmy owl Glaucidium passerinum), and an experimental manipulation, where flycatchers were exposed to cues of a nest predator (least weasel Mustela nivalis). Females receiving food supplementation laid eggs with lower concentrations of androstenedione (A4) than females not receiving food supplements. Yolk testosterone (T) concentration showed the same pattern but the difference was not statically significant. Testosterone (but not A4) concentration increased within clutches, from the first to the last egg, independently of the food supplementation. Females breeding under high predation risk did not differ from control females in their yolk androgen levels (A4, T or progesterone). However, concentrations of A4 tended to be lower in the proximity of pygmy owls, which could indirectly increase offspring survival after fledging. Food supplementation during egg-laying seems to have a stronger impact on maternal transfer of androgens than predation risk. Food availability and predation risk could differentially affect the trade-offs of androgen allocation for the offspring when raised in good vs. dangerous environments