Triple lines gold nanoparticle-based lateral flow assay for enhanced and simultaneous detection of Leishmania DNA and endogenous control

A novel triple lines lateral-flow assay （LFA） with enhanced sensitivity for the detection of Leishmania infantum DNA in dog blood samples was designed and successfully applied. The enhanced LFA methodology takes advantage of the gold nanoparticle tags （AuNPs） conjugated to polyclonal secondary antibodies, which recognize anti-FITC antibodies. The polyclonal nature of the secondary antibodies allows for multiple binding to primary antibodies, leading to enhanced AuNP plasmonics signal. Furthermore, endogenous control consisting of the amplified dog 18S rRNA gene was introduced to avoid false negatives. Using this strategy, 0.038 spiked Leishmania parasites per DNA amplification reaction （1 parasite/100 μL of DNA sample） were detected. Detection limit of LFA was found to be lower than that of the conventional techniques. In summary, our novel LFA design is a universal and simple sensing altemative that can be extended to several other biosensing scenarios.

Identification of a suitable endogenous control gene in porcine blastocysts for use in quantitative PCR analysis of microRNAs

To obtain reliable results in quantitative PCR （qPCR） reactions, an endogenous control （EC） gene is needed to correct for systematic variations. In this study, a TaqMan low density array was used to quantify the expression levels of microRNA （rniRNA） genes in in vivo fertilized, in vitro fertilized, parthenogenetic and somatic cell nuclear transfer blastocysts. The aim was to identify suitable EC genes for the qPCR analysis of rniRNAs in porcine blastocysts. The results showed that thirty-six miRNAs were commonly expressed in the four kinds of embryos and the expression levels of eleven miRNAs were similar in the different embryo types （P-value〉0.05）. These 11 miRNAs were selected as candidate EC genes for further analysis and, of these, miR-16 was identified as the most stable EC gene by the GeNorm （a tool based on a pair-wise comparison model that calculates the internal control genes stability measure and determines the most reliable pair of EC genes） and NorrnFinder （an excel plug-in that uses an ANOVA-based model to estimate intra- and inter- group variation to indicate the single most stable EC gene） programs. In addition, a cell number normalization method validated miR-16 as a suitable EC gene for use in future qPCR analysis of miRNAs in porcine blastocysts.

Revealing the control of migratory fueling： An integrated approach combining laboratory and field studies in northern wheatears Oenanthe oenanthe

Migratory birds rely on fueling prior to migratory flights. Fueling in migrants is controlled by intrinsic as well as ex- trinsic factors. From captive studies we have started understanding the internal mechanisms controlling bird migration. Field studies have demonstrated the effects of external factors, such as food availability, weather, competitors, parasites or diseases, on the stopover behavior of migrants. However, an integrated approach is still missing to study coherently how the innate migration program interacts with the varying environmental cues and to estimate the contribution of the innate migration program and the environment to realized migration. The northern wheatear Oenanthe oenanthe offers a unique opportunity for integrated studies. It breeds across almost the whole Holarctic with just a ＂gap＂ between eastern Canada and Alaska. All breeding populations over- winter in sub-Saharan Africa which makes the northern wheatear one of the most long-distant migratory songbirds with extraor- dinary long non-stop flights across oceans. It is a nocturnal migrant which travels without parental or social aid/guidance. Thus, young birds rely entirely on endogenous mechanisms of timing, route selection and fueling on their first outbound migration. By establishing indoor housing under controlled conditions the endogenous control mechanisms of northern wheatear migration could be revealed. At the same time, environmental factors controlling fueling could be investigated in the field. On migration wheatears occur in a variety of habitats with sparse vegetation where their stopover behavior could be quantitatively studied in the light of ＂optimal migration＂ theory by the use of remote balances, radio-tagging and even experimentally manipulated food availability. The present paper summarizes our approach to understand the control of migration in northern wheatears by combin- ing field and laboratory studies at various spatial and temporal scales, and linking various sub-disciplines .

Individual repeatability in timing and spatial flexibility of migration routes of trans-Saharan migratory raptors

Satellite-tracking technology has allowed scientists to make a quantum leap in the field of migration ecology. Nowadays, the basic description of migratory routes of many species of birds has been reported. However, the investigation of bird migration at individual level （i.e. repeatability in migratory routes and timing） still remains seldom explored. Here, we investigated repeated migratory trips of a trans-Saharan endangered migratory raptor, the Egyptian Vulture Neophron percnopterus, tracked by GPS satellite telemetry. We compared between- and within-individual variation in migratory routes and timing in order to assess the degree of repeatability （or conversely, the flexibility） in migration. To this end, we analysed a dataset of 48 trips （23 springs and 25 autumns） recorded for six adult birds during 2007-2013. Our results showed consistent migration timing at the individual level, both in spring and autumn. Interestingly, there was a high degree of flexibility in the routes followed by the same individual in different years, probably due to variations in meteorological conditions. Contrary to expectations of a faster migration in spring than in autumn owing to a time-minimization strategy for breeding, birds spent less time in autumn migration （13 ± 2 days, range = 9-18 d） than in spring migration （19 ± 3 days, range = 13-26 d）, which can be explained by differences in environmental con- ditions en route. Egyptian vultures showed a consistent clockwise loop migration through western Africa, following more easterly routes in autumn than in spring. Finally, our results provide supporting evidence of low phenotypic plasticity in timing of migration （i.e. strong endogenous control of migration） and high flexibility in routes [Current Zoology 60 （5）： 642-652, 2014].