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312 result(s) for "bar coding"
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Impact of Anthropogenic Disturbance on Assembly Patterns of Termite Communities
Intensification of land‐use threatens biodiversity, especially in tropical ecosystems that harbor the planet's highest species richness. This negative impact of anthropogenic disturbance on species numbers is well established, but the mechanisms underlying the community assembly processes are less well understood. Termites are of fundamental importance in tropical ecosystems where they are critical for nutrient recycling and species diversity. We tested the impact of anthropogenic disturbance on termite species diversity and assembly processes in a West African savanna applying the newest techniques of phylogenetic community analyses. Species richness dropped in areas of intensive land‐use and compositional similarity between intensive land‐use areas was high. This contrasted with a protected National Park where communities were characterized by high species richness and intermediate species turnover between sites. Slightly disturbed areas in the buffer zone surrounding the park were intermediate, they still had high species richness but similarity between sites increased. Strikingly, the assembly pattern also changed with disturbance from more phylogenetic overdispersion to more clustering (coexisting species became phylogenetically more similar), but only when the fungus‐growing termite Macrotermes bellicosus was absent. Our data suggest that the major forces structuring termite communities depend: (1) on the presence of this dominant mound‐building termite; and (2) that they change to more environmental filtering with disturbance. Anthropogenic disturbance seems to function as a filter that allows only a specific subset of species to occur. Such an effect might be widespread in ecology but it is difficult to document quantitatively. Phylogenetic community analyses can help to contribute such evidence.
Alternaria and Curvularia leaf spot pathogens show high aggressivity on watermelon, and are emerging pathogens in cucurbit production
Fungal leaf spot pathogens of cucurbits cause significant yield losses. They cause extensive leaf necroses and defoliation, reducing host photosynthesis. They increase risks of fruit sunscald, and can cause substantial crop damage. Alternaria cucumerina has been recognized as the causal agent of leaf spot disease of cucurbits, and recent studies have identified other Alternaria species, and other emerging pathogens such as Curvularia. This study characterized 25 isolates obtained from infected watermelon and cucumber leaves from Hungary, Spain, and Kosovo. Morphological characterization and molecular analyses using TEF1-α, HIS3, and ITS gene regions identified Alternaria alternata and A. arborescens, and for the first time on this host, the genus Curvularia. Detached leaf assays of ten isolates on 73 watermelon accessions showed variation in isolate pathogenicity. The tested Curvularia isolate was the most aggressive, followed by the A. arborescens  and A. alternata isolates, although A. alternata was the most frequently identified  species. These results highlight the potential for emerging fungal pathogens causing cucurbit leaf spot, such as Curvularia sp., and show that these fungi can cause damage on economically important plants. This study also showed differing resistance within the watermelon collection, indicating potential for the plant introduction (PI) accessions as sources of resistance breeding.
Impact of Hospital Characteristics and Governance Structure on the Adoption of Tracking Technologies for Clinical and Supply Chain Use: Longitudinal Study of US Hospitals
Despite the increasing adoption rate of tracking technologies in hospitals in the United States, few empirical studies have examined the factors involved in such adoption within different use contexts (eg, clinical and supply chain use contexts). To date, no study has systematically examined how governance structures impact technology adoption in different use contexts in hospitals. Given that the hospital governance structure fundamentally governs health care workflows and operations, understanding its critical role provides a solid foundation from which to explore factors involved in the adoption of tracking technologies in hospitals. This study aims to compare critical factors associated with the adoption of tracking technologies for clinical and supply chain uses and examine how governance structure types affect the adoption of tracking technologies in hospitals. This study was conducted based on a comprehensive and longitudinal national census data set comprising 3623 unique hospitals across 50 states in the United States from 2012 to 2015. Using mixed effects population logistic regression models to account for the effects within and between hospitals, we captured and examined the effects of hospital characteristics, locations, and governance structure on adjustments to the innate development of tracking technology over time. From 2012 to 2015, we discovered that the proportion of hospitals in which tracking technologies were fully implemented for clinical use increased from 36.34% (782/2152) to 54.63% (1316/2409), and that for supply chain use increased from 28.58% (615/2152) to 41.3% (995/2409). We also discovered that adoption factors impact the clinical and supply chain use contexts differently. In the clinical use context, compared with hospitals located in urban areas, hospitals in rural areas (odds ratio [OR] 0.68, 95% CI 0.56-0.80) are less likely to fully adopt tracking technologies. In the context of supply chain use, the type of governance structure influences tracking technology adoption. Compared with hospitals not affiliated with a health system, implementation rates increased as hospitals affiliated with a more centralized health system-1.9-fold increase (OR 1.87, 95% CI 1.60-2.13) for decentralized or independent hospitals, 2.4-fold increase (OR 2.40, 95% CI 2.07-2.80) for moderately centralized health systems, and 3.1-fold increase for centralized health systems (OR 3.07, 95% CI 2.67-3.53). As the first of such type of studies, we provided a longitudinal overview of how hospital characteristics and governance structure jointly affect adoption rates of tracking technology in both clinical and supply chain use contexts, which is essential for developing intelligent infrastructure for smart hospital systems. This study informs researchers, health care providers, and policy makers that hospital characteristics, locations, and governance structures have different impacts on the adoption of tracking technologies for clinical and supply chain use and on health resource disparities among hospitals of different sizes, locations, and governance structures.
Exploring the veracity of food labels: a comparison of two countries with disparate levels of food control
Food fraud, including adulteration or mislabeling, is a significant global problem. The rate and type of food fraud may depend on the country. In the present study, we assessed the occurrence of two types of food fraud in Nigeria, a developing African country, and Finland, an EU country famous for its high food control. First, by LC‒MS/MS and ELISA methods, we determined melamine concentrations in infant formulas for sale in both countries as well as in whole pet foods available in Finland. Second, using bar-code end-point PCR and Sanger sequencing, we explored whether the species in minced beef products in Nigeria and Finland was correctly labeled. In all Nigerian and Finnish infant formulas and in cat and dog foods in Finland, melamine concentrations fell below the maximum acceptable limit. In contrast, all 10 Nigerian meat samples analyzed proved to be mislabeled, containing chicken, pork, or both, whereas all six Finnish meat products represented beef as labeled. These findings reveal that while melamine does not appear to be a risk in infant formulas today, even in Nigeria, mislabeling meat by substituting less expensive meat materials for beef seems to be there highly common. Therefore, broader nationwide product surveillance and stricter food control measures should be implemented in that country.
Barcoding quantitative PCR assay to distinguish between 'Aedes aegypti' and 'Aedes sierrensis'
The accurate identification of mosquito species is critical for effective mosquito surveillance and control, especially when presented with morphologically similar species like 'Aedes aegypti' and 'Aedes sierrensis'. Damaged specimens and morphologically similar life stages such as eggs and larvae make it difficult to distinguish 'Aedes aegypti' from 'Aedes sierrensis' using microscopy and taxonomic keys. To address this, the 'AegySierr'.ID-qPCR assay, a multiplex quantitative PCR assay that utilizes single-nucleotide polymorphisms within the 'mitochondrial cytochrome oxidase subunit I' gene, was developed to distinguish between these two species. The assay was tested on DNA extracted from the eggs, larvae, and adults of both species, as well as from environmental DNA (eDNA) collected from natural mosquito reproduction sites. It demonstrated a high diagnostic accuracy across multiple life stages, with a sensitivity exceeding 95% for most groups and specificity exceeding 90%, except for field-collected adult 'Ae. sierrensis' (75%). For eDNA samples, the assay achieved 100% sensitivity and 94% specificity for samples classified as 'Ae. sierrensis' and 91% sensitivity and 86% specificity for 'Ae. aegypti'. A two-graph receiver operating characteristic analysis was also used as an alternate method with which to establish Ct thresholds for interpreting results from unknown samples. The 'AegySierr'.ID-qPCR assay enables the rapid and sensitive identification of 'Ae. aegypti', and 'Ae. sierrensis' from specimens and eDNA, and may be of use in mosquito surveillance programs.
Wild and native plants and mushrooms sold in the open-air markets of south-eastern Poland
Background The study of plants and fungi sold in open-air markets is an important part of ethnobotanical enquiry. Only few such studies were carried out in Europe. Methods Four of the largest open-air markets of south-eastern Poland were visited regularly, and the plants sold in them were recorded between 2013 and 2015. The aim of the study was to record native and/or wild species sold in the markets. All the plants sold in the markets were photographed regularly. In each market, 25 sellers were interviewed. Voucher specimens were collected and fungi were identified using DNA barcoding. Results Altogether, 468 species of plants were recorded, 117 of them native to south-eastern Poland – 19 only collected from the wild and 11 both wild and cultivated. Seventeen of the species are under legal protection. Most protected plants were sold from cultivation, although proper authorization procedures had not been performed. Thirty-two species of fungi were sold (including two cultivated species), all of them for culinary purposes. Two species ( Lactarius quieticolor , Leccinum schistophilum ) are new to the mycobiota of Poland. Ornamental plants constituted a large section of the market, and they dominated the group of native species. Food plants dominated among wild-collected plants and were sold mainly as fruits for jams, juices and alcoholic drinks, or as culinary herbs. Very few medicinal or green vegetable plants were sold. An interesting feature of the markets was the sale of Ledum palustre as an insect repellent. Conclusions Finding two species of fungi which are new to Poland highlights the importance of DNA barcoding in ethnomycological studies. Most items in the markets are ornamental plants, or edible fruits and mushrooms. Very few medicinal plants and green vegetables are sold, which differentiates the markets from southern European ones. Such a pattern is probably the model for most central European markets.
Qualitative assessment of the flowering buds of Mesua ferrea Linn with special emphasize on HPTLC and universal DNA bar-coding technique and evaluation of its antimicrobial potential
Background The study has been scientifically exploring the powder sample and extracts of flowering buds of Mesua ferrea Linn (FBMF) based on pharmacognostic and phytochemical parameters. The medicinal plant Mesua ferrea Linn was identified by Botanical verification and the universal DNA bar-coding technique. The FBMF powder’s quality was performed by the micromeritics properties like bulk density, tapped density, angle of repose, Hausner ratio, Carr’s index, and optical microscopy method, and physicochemical evaluations were performed by the swelling index, foaming index, loss on drying, extractive values, and ash values. The extract’s preliminary phytochemical screening was accomplished by the alkaloids, flavonoids, tannins, steroids, carbohydrates, and glycoside tests. To ensure the presence of a bioactive compound of each FBMF extract by qualitative HPTLC study against the reference β-sitosterol Rf value of 0.83 was revealed at 254 nm with a developed solvent system toluene, ethyl acetate, and acetic acid (6:2:0.1) v/v. Many pharmacological benefits, including those related to wound healing, nonalcoholic fatty liver disease, analgesic, sedative effects, immunomodulatory, anticancer, antimicrobial, anti-inflammatory, anxiolytic, and atopic dermatitis, have been demonstrated by this phytosterol. An antimicrobial study was carried out by a well diffusion method and, lastly, measured minimum inhibitory concentration and compared to the marketed active pharmaceutical component. Numerous skin infections of more invasive, serious illnesses are caused by potential antimicrobial activity. Results The universal DNA bar-coding of plant Mesua ferrea Linn has shown a high percentage of identity. The micrometrics properties and physicochemical evaluation of the powder sample of FBMF were in an acceptable range. The preliminary phytochemical screening shows that all extracts contain steroids and carbohydrates. Moreover, flavonoids were found in ethyl acetate and ethanol, and a qualitative HPTLC study confirmed that every extract contains β-sitosterol. N-hexane extract of FBMF shows the potential antimicrobial activity with Staphylococcus aureus, and the MIC value was observed at 0.062 mg/ml. Conclusion Our research demonstrated that the plant Mesua ferrea Linn has been successfully authenticated by a DNA bar-coding technique, and all extracts of FBMF contain β-sitosterol. The n-hexane solvent extracts have shown the potential highest antimicrobial effects compared to other extracts. These results support using n-hexane extracts as a traditional medicine for treating several diseases. Graphical Abstract
Phylogeny of Canadian ergot fungi and a detection assay by real-time polymerase chain reaction
The ergot disease of cereals has become increasingly important in agricultural areas of Canada since 1999. Generally, this disease is considered to be caused by Claviceps purpurea, but the taxonomy of Claviceps from these areas has not been well studied. The objectives of this study were (i) to determine the phylogenetic lineages (phylogenetic species) present in agricultural areas of Canada and (ii) to develop a molecular assay that can separate the lineages on crops from other lineages. Genetic diversity of Claviceps collected from agriculture areas in Canada were investigated using multilocus sequence typing. The loci sequenced include nuc rDNA internal transcribed spacer (ITS1-5.8S-ITS2 = ITS), partial fragments of translation elongation factor 1-α (TEF1), RNA polymerase II second largest subunit (RPB2), β-tubulin (tubB), and two ergot alkaloid synthesis genes (easA, easE). Based on individual locus and concatenated alignments, phylogenetic analyses revealed seven lineages within the premolecular concept of C. purpurea, of which five corresponded with undescribed species (G2b and G4-7). Although lineages G2-7 had narrow host ranges, lineage G1 (= C. purpurea s.s.) had a broad host range that overlapped with other lineages. A molecular diagnostic quantitative polymerase chain reaction (qPCR) assay was developed and validated with 185 samples from a wide range of host plants and geographic origins, including 10 phylogenetic species in C. sect. Claviceps, 8 in C. sect. Pusillae, 1 in C. sect. Citrinae, and 1-2 species from Alternaria, Fusarium, and Penicillium. The assay can detect lineage G1 at a concentration of 7.5 pg/μL and distinguish it from other Claviceps species and lineages. This facilitates disease management by detecting the inocula from nonagriculture host plants.
Identification and Removal of Potential Contaminants in 16S rRNA Gene Sequence Data Sets from Low-Microbial-Biomass Samples: an Example from Mosquito Tissues
The study of tissue-associated microbiota from mosquitoes (primarily from the gut) has grown significantly in the last several years. Mosquito tissue samples represent a challenge for researchers given their low microbial biomass and similar taxonomic composition commonly found in the laboratory environment and in molecular reagents. The bacterial microbiota of the mosquito influences numerous physiological processes of the host. As low-microbial-biomass ecosystems, mosquito tissues are prone to contamination from the laboratory environment and from reagents commonly used to isolate DNA from tissue samples. In this report, we analyzed nine 16S rRNA data sets, including new data obtained by us, to gain insight into the impact of potential contaminating sequences on the composition, diversity, and structure of the mosquito tissue microbial community. Using a clustering-free approach based on the relative abundance of amplicon sequence variants (ASVs) in tissue samples and negative controls, we identified candidate contaminating sequences that sometimes differed from, but were consistent with, results found using established methodologies. Some putative contaminating sequences belong to bacterial taxa previously identified as contaminants that are commonly found in metagenomic studies but that have also been identified as part of the mosquito core microbiota, with putative physiological relevance for the host. Using different relative abundance cutoffs, we show that contaminating sequences have a significant impact on tissue microbiota diversity and structure analysis. IMPORTANCE The study of tissue-associated microbiota from mosquitoes (primarily from the gut) has grown significantly in the last several years. Mosquito tissue samples represent a challenge for researchers given their low microbial biomass and similar taxonomic composition commonly found in the laboratory environment and in molecular reagents. Using new and published data sets that identified mosquito tissue microbiota from gut and reproductive tract tissues (and their respective negative controls), we developed a simple method to identify contamination microbiota. This approach uses an initial taxonomic identification without operational taxonomic unit (OTU) clustering and evaluates the relative abundance of control sample sequences, allowing the identification and removal of purported contaminating sequences in data sets obtained from low-microbial-biomass samples. While it was exemplified with the analysis of tissue microbiota from mosquitos, it can be extended to other data sets dealing with similar technical artifacts.