Abstract Background Shotgun sequencing-based metagenomics is a useful approach to profiling microbiomes in environmental and patient samples. In a clinical setting, metagenomic techniques have the advantage of identifying organisms, which cannot be readily cultured or confirmed by other techniques. We have developed a clinical-grade, streamlined metagenomics-based pipeline that includes regulatory compliant method considerations, such as an internal control followed by a machine-based learning (ML) process to identify pathogens in urine samples. Methods We built an optimized novel end-to-end NGS assay pipeline that harnesses pathogen-specific genome data to detect bacterial species. We processed de-identified clinical urine specimens, collected from patients symptomatic for urinary tract infection (UTI). This workflow includes an IPC, QIACube-MDx extraction, library preparation and Illumina NextSeq 550 sequencing and a novel interpretable ML based analytic approach, Biotia-DX. Clinical culture results and qPCR were used as a baseline for the assay to train the ML model and to establish accuracy relative to the clinical standard of care. Results We clinically validated over 40 key uropathogens and conducted clinical studies of specificity, intra/inter reproducibility, accuracy in urine specimens (n=300), and limit of detection in E. coli, K. pneumoniae, P. mirabilis, S. aureus, E. faecalis and Candida. Additionally, the implementation of an internal control coupled with our Biotia-DX software provides an accurate (F1 score 94.3%) and highly sensitive clinical grade diagnostic tool. Conclusion Urine has historically presented a challenge for diagnostics via culturing, with a high rate of culture-negative results (∼30% on average). We improved the clinical utility of an NGS urine assay by leveraging an IPC and ML software. This decreased the rate of false positive species called in a sample relative to other NGS techniques and allows for greater sensitivity and taxonomic specificity. This assay may be especially useful for low colony-count or negative-culture samples to diagnose and guide patient treatment. Disclosures Mara Couto-Rodriguez, MS, Biotia Inc.: Employee of Biotia Inc. a for-profit biotechnology company David C. Danko, PhD, Biotia Inc.: Employee of Biotia Inc. a for-profit biotechnology company Xavier O. Jirau Serrano, MS, Biotia Inc.: Employee of Biotia Inc. a for-profit biotechnology company Taylor Paisie, MS, Biotia Inc.: Employee of Biotia Inc. a for-profit biotechnology company John C. Papciak, BS, Biotia Inc.: Employee of Biotia Inc. a for-profit biotechnology company Eszter Szollosi, BS, Biotia Inc.: Employee of Biotia Inc. a for-profit biotechnology company Christopher E. Mason, PhD, Biotia Inc.: Advisor/Consultant|Biotia Inc.: Board Member|Biotia Inc.: Ownership Interest Caitlin Otto, PhD, D(ABMM), Biotia Inc.: Advisor/Consultant Niamh B. O'Hara, PhD, Biotia Inc.: Board Member|Biotia Inc.: Ownership Interest Dorottya Nagy-Szakal, MD PhD, Biotia Inc.: Employee of Biotia Inc. a for-profit biotechnology company|Biotia Inc.: Stocks/Bonds.
Although avian malarial parasites are globally distributed, the factors that affect the geographical distribution and local prevalence of different parasite lineages across host populations or species are still poorly understood. Based on the intense screening of avian malarial parasites in nine European blue tit populations, we studied whether distribution ranges as well as local adaptation, host specialization and phylogenetic relationships can determine the observed prevalences within populations. We found that prevalence differed consistently between parasite lineages and host populations, indicating that the transmission success of parasites is lineage specific but is partly shaped by locality-specific effects. We also found that the lineage-specific estimate of prevalence was related to the distribution range of parasites: lineages found in more host populations were generally more prevalent within these populations. Additionally, parasites with high prevalence that were also widely distributed among blue tit populations were also found to infect more host species. These findings suggest that parasites reaching high local prevalence can also realize wide distribution at a global scale that can have further consequences for host specialization. Although phylogenetic relationships among parasites did not predict prevalence, we detected a close match between a tree based on the geographic distance of the host populations and the parasite phylogenetic tree, implying that neighbouring host populations shared a related parasite fauna.
Az oroklott tulajdonsagok mellett az utodok ratermettseget a korai kornyezeti (anyai) hatasok jelentősen modosithatjak. Madaraknal vizsgaltuk, hogy a tojo altal a tojasba juttatott anyagok (ivari hormonok, vitaminok, karotinoidok, immunanyagok) hogyan segitik a fiokak fejlődeset ill. az anyagok allokacios mintazatat milyen (proximalis es ultimalis) valtozok magyarazhatjak. Megallapitottuk, hogy az orvos legykaponal a tojasmeretet a kornyezeti tenyezők es a tojo minősege, a tojas IgG-szintjet a tojo minősege prediktalta. A karotinoidok, az E- es A-vitamin, valamint a biliverdin (tojashej pigment) deponalasa a tojo parjanak koratol fuggott. Kompenzacios mechanizmusra utal, hogy a tapasztalatlan fiatal himek feszekaljaiban a tojok noveltek a tesztoszteront es az antioxidansokat. Kiserletesen bizonyitottuk, hogy a tesztoszteron depoziciojara hat, a karotinoidok es az IgG depoziciojara viszont nem hat a szocialis kornyezet valtozasa. Az orvos legykapo tojasok biopszias vizsgalataibol es japanfurj tojasok hormonkoncentraciojanak kiserletes megemeleseből arra kovetkeztethetunk, hogy a tojasokban levő androgenek (T, A4, DHT) rovid tavon elterően hatnak az utodok ratermettsegere. A kek cinegeknel pozitiv kapcsolatot talaltunk a himek egy ivari szignalja (fejtető UV visszaverese) es a tojasok becsult karotinoidmennyisege kozott. Vegul a szencinege tobbszoros ivari szignalizaciojat elemezve ramutattunk a szignalizacio es az anyai hatasok kornyezetfuggősegere. | In addition to genetic effects, the fitness of offspring is considerably modified by early environmental (maternal) effects. We examined how egg constituents deposited by the female (sexual hormones, vitamins, carotenoids, immune mediators) help offspring development and what proximate and ultimate factors may explain the allocation pattern of these substances. In the collared flycatcher we found that egg size was predicted by environmental factors and female quality, while IgG level by the quality of the female. The deposition of carotenoids, biliverdin, vitamins A and E depended on the characteristics of the female's mate. Consistent with a compensation mechanism, females increased the concentration of egg testosterone and antioxidants in clutches laid for inexperienced young males. We experimentally demonstrated that changing social environment modifies the deposition of testosterone but not that of carotene metabolites and IgG. By conducting biopsy studies of collared flycatcher eggs and experimentally elevating sex steroid levels in Japanese quail eggs, we found that different androgens in the egg (T, A4, DHT) have different short term effects on offspring fitness. In blue tits, we found a positive relationship between a sexual signal of males (crown UV reflectance) and the estimated carotenoid levels of eggs. Finally, analyses of multiple sexual signals in great tits indicated the environment-dependence of signalization and maternal effects.
Several hypotheses predict that the major histocompatibility complex (MHC) drives mating preference in females. Olfactory, colour or morphological traits are often found as reliable signals of the MHC profile, but the role of avian song mediating MHC-based female choice remains largely unexplored. We investigated the relationship between several MHC and acoustic features in the collared flycatcher (Ficedula albicollis), a European passerine with complex songs. We screened a fragment of the class IIB second exon of the MHC molecule, of which individuals harbour 4-15 alleles, while considerable sequence diversity is maintained at the population level. To make statistical inferences from a large number of comparisons, we adopted both null-hypothesis testing and effect size framework in combination with randomization procedures. After controlling for potential confounding factors, neither MHC allelic diversity nor the presence of particular alleles was associated remarkably with the investigated qualitative and quantitative song traits. Furthermore, genetic similarity among males based on MHC sequences was not reflected by the similarity in their song based on syllable content. Overall, these results suggest that the relationship between features of song and the allelic composition and diversity of MHC is not strong in the studied species. However, a biologically motivated analysis revealed that individuals that harbour an MHC allele that impairs survival perform songs with broader frequency range. This finding suggests that certain aspects of the song may bear reliable information concerning the MHC profile of the individuals, which can be used by females to optimize mate choice.
In a polygynous mating system, females settling with already mated males often experience low mating success due to the reduced parental contribution of the male. However, there are numerous factors that may still make it advantageous for some females to choose this mating status. Facultative polygyny is believed to be dominated by male advertisement and female choice. Although quality differences and competition among females are increasingly recognized as important determinants of polygynous settlement patterns, the importance of signals of female quality in this mating system is largely unknown. Here we examined the relationship of the white wing patch size (WPS) of female collared flycatchers, a phenotypically plastic and age-dependent ornament, with social mating status, while controlling for settlement date and age. At the population level, monogamous, primary, and secondary females did not differ in WPS. However, the primary female of individual males was more ornamented than the secondary female, and this difference declined with increasing distance between primary and secondary nests. Secondary female ornamentation increased, whereas that of the primary female did not change with nest distance. These results suggest a subtle role for female ornamentation at polygynous mating. Future studies should therefore take into account mating status when assessing the costs and benefits of female signals. Moreover, patterns in quality-indicating female traits may contribute to the explanation of differences in reproductive success among females of different mating status.
Many hypotheses have been proposed to explain why female birds start to incubate before clutch completion (IBCC). Some of those suggest that the resulting hatching asynchrony (HA) is adaptive because it increases the size hierarchy among offspring and in turn reduces nestling competition and energy demands during the peak feeding period. Others argue that IBCC is a good strategy in unpredictable environments. When food conditions deteriorate, the large size hierarchy quickly results in the death of the last hatched nestlings, allowing the remaining ones to survive and fledge in better condition. In comparison, under favorable conditions, all nestlings can fledge independent of hatching order. To test these hypotheses, we performed a brood size manipulation experiment (as a simulation of good and bad years) in collared flycatchers Ficedula albicollis and examined the effect of size hierarchy on offspring and brood performance. We found that chicks with an initial size disadvantage experienced reduced body mass growth and had shorter feathers at fledging in both reduced and enlarged broods. In enlarged broods, they also fledged with a smaller skeletal size. Although broods on average or parents could possibly still benefit from HA when food is scarce, this was not seen in the current study. Parental survival was not related to the size hierarchy in the broods, and the average body mass growth of the nestlings was slower in broods with a high initial size variance. We therefore conclude that HA and the resulting size hierarchy are probably detrimental for the growth of nestlings in both good and bad years, at least in species where nestling mortality does not occur early in life.
Abstract Many vocalizing animals produce the discrete elements of their acoustic signals in a specific sequential order, but we know little about the biological relevance of this ordering. For that, we must characterize the degree by which individuals differ in how they organize their signals sequentially and relate these differences to variation in quality and fitness. In this study, we fulfilled these tasks in male collared flycatchers (Ficedula albicollis). We characterized the sequential order of syllables with a network analysis approach and studied the consistency of network variables on distinct time scales (within day, between days, and between years), and assessed their relationship with such quality indicators like age, body condition, arrival date, and fitness related proxies like survival to the next year and pairing success. We found that the syllables were associated nonrandomly with one another and both the frequency differences of consecutive syllables and the number of motif types were higher in the original than in randomized syllable sequences. Average degree and small-worldness showed considerable among-individual differences and decreasing repeatability with increasing time scale. Furthermore, we found relationships between male age and average degree among and within individuals. Accordingly, older males produce syllable sequences by using common syllables less often than younger individuals. However, the network variables showed no relationship with fitness-related variables. In conclusion, the sequential organization of birdsong has the potential to encode individual-specific characteristics, which thus could be used as signal in social interactions and thus potentially could be subject to sexual selection.