Academic literature on the topic 'Wing genotyping'

OBJECTIVETo determine the source of aLegionella pneumophilaserogroup 5 nosocomial outbreak and the role of the heat exchanger installed on the hot water system within the previous year.SETTINGA 400-bed tertiary care university hospital in Sherbrooke, Canada.METHODSHot water samples were collected and cultured forL. pneumophilafrom 25 taps (baths and sinks) within wing A and 9 taps in wing B. Biofilm (5) and 2 L water samples (3) were collected within the heat exchangers forL. pneumophilaculture and detection of protists. Sequence-based typing was performed on strain DNA extracts and pulsed-field gel electrophoresis patterns were analyzed.RESULTSFollowing 2 cases of hospital-acquired legionellosis, the hot water system investigation revealed a large proportion ofL. pneumophilaserogroup 5 positive taps (22/25 in wing A and 5/9 in wing B). High positivity was also detected in the heat exchanger of wing A in water samples (3/3) and swabs from the heat exchanger (4/5). The outbreak genotyping investigation identified the hot water system as the source of infections. Genotyping results revealed that all isolated environmental strains harbored the same related pulsed-field gel electrophoresis pattern and sequence-based type.CONCLUSIONSTwo cases of hospital-acquired legionellosis occurred in the year following the installation of a heat exchanger to preheat hospital hot water. No cases were reported previously, although the sameL. pneumophilastrain was isolated from the hot water system in 1995. The heat exchanger promotedL. pneumophilagrowth and may have contributed to confirmed clinical cases.Infect. Control Hosp. Epidemiol.2016;1475–1480

The honey bee Apis mellifera L. colony is headed by a single and indispensable queen, whose duty it is to ensure brood production and provide pheromonal stability within the colony. This study presents a non-invasive method that allows the identification of the queen maternal lineage and subspecies using the remaining tissue of her clipped wing. The DraI mtDNA COI-COII (DmCC) test was applied to various sizes of queen and worker wings and the results were compared with data obtained from other bee tissues. Furthermore, we propose a new method allowing in silico transition of the DmCC test and haplotype identification based on extended sequencing of the tRNAleu and COII genes. Our results show that DNA extracted by Chelex 10% from one-third of a queen’s wing is deemed adequate for a successful identification of her maternal evolutionary lineage, haplotype and subspecies. The in silico method proposed in this study fully adheres to the established guidelines of the DmCC, provides a universal standard for haplotype identification, and offers faster and more precise results by reconciling both cleaved amplified polymorphic sequences (CAPS) and Sanger sequencing approaches.

Abstract We address the problem of microsatellite genotyping errors associated with polymerase chain reaction (PCR) amplification from degraded and dilute template DNA and provide suggestions for improving the accuracy of genotype data in studies using older museum specimens as a source of DNA. In the course of a population genetics study of African indigobirds (Vidua spp.), we used replicate PCR to evaluate genotyping reliability for nine microsatellite loci in relation to PCR fragment length and DNA template concentration (DNA extracted from the calamus of one vs. two wing feathers). Complete amplification failure and the dropout of one allele from heterozygous genotypes were the predominant problems encountered. For samples with heterozygous genotypes, allele dropout occurred in 19.2 and 12.1% of PCR using extracts derived from one and two feathers, respectively. The amplification of artifact bands was less frequent (affecting 4.9 and 1% of positive PCR reactions with one- and two-feather extracts, respectively). Those results indicate that multiple replicates per sample and locus are required to obtain accurate genotype data from museum feather samples. Although higher DNA concentration improved success, PCR fragment size had a much stronger influence on the success and repeatability of microsatellite amplification, which suggests that the accuracy and efficiency of genotyping can be improved most easily by designing primers that amplify smaller DNA fragments.

Abstract Blue-black Grassquits (Volatinia jacarina) are small, granivorous, Neotropical birds that are abundant in central Brazil. During the reproductive season, the socially monogamous males acquire a blue-black plumage and defend very small, clustered territories that resemble leks. They execute a conspicuous courtship display that consists of a leap, revealing white under-wing patches, synchronized with a vocalization. We collected data on the morphology and behavior of banded males and characteristics of their territories to determine how these factors may influence acquisition of mates and nesting. For a second group of birds in the area, we used microsatellite genotyping to test the breeding synchrony hypothesis, which predicts that tropical species that breed synchronously should exhibit high rates of extra-pair fertilization (EPF). We found that males that successfully formed a pair bond differed from unsuccessful males in their behavior, but not in morphological attributes or territory features. Successful males spent more time in their territories, executed displays for longer periods and at greater rates, and their display leaps were higher. These results point to the greater importance of behavior relative to other factors in the successful reproduction of Blue-black Grassquit males. In the second group of birds, EPFs occurred in 63% of 11 nests and involved 50% of the 20 chicks sampled. This exceptionally high incidence of EPF in the small sample analyzed occurred in diverse contexts, including intraspecific parasitism and quasi-parasitism, a rare type of maternity loss where the resident female is parasitized by other females that were fertilized by the resident male. A short and highly synchronized breeding season, clustered breeding in small territories, and granivorous habits may contribute to the high rates of EPF in this tropical species.

AbstractAraucaria angustifolia is a dioecious and wind pollinated conifer that typically occurs in higher attitudes of Southern Brazil. After a significant reduction of its population during the twentieth century, public policies have enabled natural populations to recover. As new studies focus on the genetics of the species it is important to investigate Mendelian inheritance, genetic linkage, and genotypic disequilibrium for the microsatellite loci developed for the species. Here we analyze ten microsatellite loci developed for A. angustifolia by genotyping 295 adult trees and 13 open pollinated progenies from a forest fragment in Santa Catarina, Brazil. The likelihood G-test shows a perfect 1:1 Mendelian segregation for all ten loci, indicating that these molecular markers are genetic markers. Significant genetic linkage between pairwise loci was detected in only 3% of the tests, suggesting that these loci are not located in the same linkage groups within the chromosomes. However, genotypic disequilibrium was detected in 51% of pairwise loci for adult trees, probably due to the strong spatial genetic structure of the population. Our results indicate that the ten loci analyzed can be used in studies on genetic diversity and structure, mating system, and gene flow of the species.

The hypothesis of this study was that different plant hosts of the medfly Ceratitis capitata may cause variability as a prerequisite for its invasiveness. The main objective was to determine population variability based on medfly wing shape in three favorable medfly host plants (peach, fig and mandarin) from different agroecological growing areas with different pest management practices, and to evaluate phenotypic plasticity as a basis for future expansion into new areas and new hosts. Using geometric morphometric methods across 14 specific landmarks on the medfly wings, 10 populations were tested from infested peach, fig and mandarin fruits, as well as laboratory-grown sterile populations. The studies led to the following main findings: (1) all of the medfly populations that were studied exhibited sexual dimorphism in wing shape; (2) the hosts in which the medfly develops influence wing shape and condition its variability; (3) there is significant variability between laboratory mass-reared sterile and wild individuals in male and female populations; (4) a high phenotypic plasticity of medfly populations was observed along the study sites. Even the low but clearly detected variability between different agroecological conditions and localized variability indicate genotypic stability and high phenotypic plasticity, which can be considered as a prerequisite for medfly invasiveness and dispersal to new areas.

In two related experiments, total RNA was extracted from wounded and unwounded bark of young aspen ramets for Northern and dot blot analyses. Wound-inducible genes isolated from other plant species were hybridized to blots, and mRNA levels were estimated. Analyses of variance (ANOVA) were used to examine the significance of experimental factors (wounding, time after wounding, and genotype) affecting variation in mRNA levels. The first experiment examined the timing (0.5-96 h after wounding) of expression of wound-inducible genes in bark tissue of a single Populus tremuloides Michx. genotype. Wounding and variation among RNA samples significantly (p < 0.05) affected mRNA levels of two chitinases (win6, win8) and phenylalanine ammonia-lyase (PAL). The second experiment examined interclonal variation of wound-induced win6 and PAL expression in aspen bark. Ramets of four P. tremuloides and one Populus grandidentata Michx. genotypes were wounded and bark was collected 4, 8, or 12 h later. Genotype, wounding, and time after wounding all significantly affected win6 and PAL mRNA levels, with levels increasing as a result of wounding.

Wine quality is determined by the particular yeast strains prevailing at various stages of fermentation. Therefore, the ability to make an easy, fast, and unambiguous discrimination of yeasts at the strain level is of great importance. Here, the tandem repeat-tRNA (TRtRNA) method with the 5GAC or ISSR-MB primer sets and random amplified polymorphic DNA (RAPD) analysis with (GTG)3, R5, and RF2 oligonucleotides were tested on various non-Saccharomyces wine yeast species. The TRtRNA-PCR employing ISSR-MB showed the highest capacity in discriminating Lachancea thermotolerans and Metschnikowia pulcherrima isolates. RAPD with RF2 was the most efficient method in resolving Starmerella bacillaris isolates, although it produced few polymorphic bands. RAPD with R5 showed the highest capacity to discriminate among the Issatchenkia orientalis, Hanseniaspora guilliermondii, and Pichia anomala isolates. RAPD with either R5 or RF2 exhibited the highest ability to discriminate among the Torulaspora delbrueckii isolates. RAPD with (GTG)3 was the most discriminating method for the H. uvarum isolates. Here we concluded that both TRtRNA-PCR and RAPD-PCR offer rapid means for typing non-Saccharomyces species. However, each method performs better for a given species when paired with a particular primer set. The present results can be useful in wine research for the fast fingerprinting of non-Saccharomyces yeasts.

AbstractFunctional wing length (wing length/head-capsule width) of female Sweltsa revelstoka (Jewett) from streams was measured for 19 sites that have been free of Wisconsin glacial ice for about 15 000 years and possibly longer, and from 23 sites that have been ice free for about 10 000 years. At the former sites brachypterous populations were common and there was a significant negative relationship between functional wing length and elevation, and a positive relationship between functional wing length and stream size. In the area that deglaciated more recently, populations were not or only slightly brachypterous and there was no significant relationship between wing length and elevation or between wing length and stream size. Functional wing length was not related to body size. These analyses indicate that the brachypterous condition is probably genotypic in origin. I suggest that streams were colonized by macropterous forms shortly after deglaciation, and that brachyptery takes several millennia to develop at small, high-elevation streams.

Honey bees play an important role in modern agriculture as farm animals and crop pollinators and represent an animal model in scientific research. Since few decades, managed honey bees are facing large scale losses worldwide due to the interaction of several biotic and abiotic factors such as the spread of pathogens and parasites, the habitat loss, the use of pesticides, and the occurrence of climate changes. For years, beekeepers have controlled deadly pathogens such as Varroa destructor with acaricides (pyrethroid), but widespread chemical resistance is manifesting. Alternative management strategies could be developed to characterize and select bees with heritable traits allowing them to resist mites and diseases. Hygienic behaviour (HB) is a heritable phenotype that confers colony level resistance against brood disease, with a potential effect on the damaging parasitic mite V. destructor. Unfortunately, breeding such bees is complicated as the assays involved to characterize such phenotype are time-consuming and expensive if performed on a consistent number of colonies. Additionally, the mechanisms behind social immunity are not yet fully understood. These issues have motivated the scientific community to develop research tools that can offer insight into the causes of declining bee health as well as identify biomarkers to guide breeding programs. The studies presented in this thesis are the results of the collaboration between the Animal Genetic Group of the University of Milan and an Italian beekeeping company, which made available its animals for the majority of the studies presented in this work. The first two chapters of this thesis focus on three important aspects regarding hygienic behaviour: in-field phenotypic implementation of a method to measure HB on large testing populations, its genetic parameters estimation and developing molecular tools for the identification of potential HB genetic markers from breeding queens. Queens are the only fertile females of the colony, responsible for laying eggs and maintaining the cohesion of the colony. The quality of the queen may have an important effect on a colony’s development, productivity and survival and queen failure or loss is considered a cause of the decline of colonies worldwide. The queen quality, resulting from her genetic background, developmental condition, mating success and environment, can be assessed by some morphological measures. We investigated the genetic parameters of some traits that describe the quality of bee queens in the third chapter. Beside queen morphology, in the fourth chapter, we investigated also the morphology of the worker bees with Computed Tomography (CT). In this study, a non-invasive CT technique and image analysis approach coupled with brood manual inspection was used to clarify the relationship between honey bee pupa length and its varroa mite infestation status, developmental status and spatial position within the brood comb. The results of this chapter suggest that the CT-scan may represent a suitable non-invasive tool to investigate the morphology and developing status of honey bee brood. Finally, since honey yield is one of the major breeding goal for a beekeeping company, we investigated the genetic parameters for honey yield in a small testing population of honey bees in Northern Italy both considering the total honey yield and the single harvests of a colony within a year on data collected over a period of three years.