Clinical Utility of Universal Broad-Range Polymerase Chain Reaction Amplicon Sequencing for Pathogen Identification: A Retrospective Cohort Study

We assessed the real-world utility of universal broad-range polymerase chain reaction sequencing for pathogen detection. Among 1062 clinical samples, 107/1062 (10.1%) had a clinically significant, positive result, with substantial variation by specimen type. Clinical management was changed in 44/1062 (4.1%). These data can help maximize utility of this emerging diagnostic.

Should activated sludge models consider influent seeding of nitrifiers? Field characterization of nitrifying bacteria

This study revealed the presence of nitrifying bacteria in influent municipal wastewaters reaching full-scale biological wastewater treatment plants. Respirometric assays showed that the influent nitrifiers were active following a 5- to 8-hour period of metabolic induction. Diversity analyses by pyrosequencing of functional gene PCR (polymerase chain reaction) amplicon suggested that the nitrifiers in the influent stream likely seeded activated sludge bioreactors since the most abundant operational taxonomic units in the influent and mixed liquor were the same. Based on the estimated seeding intensity of 0.3 g of nitrifiers per day per gram of nitrifiers already present, the absolute minimum solids retention time (SRT) was reduced by approximately 56% at 5 °C as compared to non-seeding conditions. This can have important repercussions on the design and sizing of bioreactors operating in cold climates and calls for a need to fine-tune process modelling by considering the contribution of autotrophic nitrifying biomass from municipal influent streams.

BseGI Restriction of the Polymerase Chain Reaction Amplicon Th444 Is Required to Distinguish Biotypes of Trichoderma aggressivum Causing Serious Losses in Mushroom (Agaricus bisporus) Production

Green mold is a serious disease of the cultivated mushroom causing losses in production of economical importance. In the present study, digestion of a Th444 amplicon with endonuclease BseGI was useful to discriminate Trichoderma aggressivum f. aggressivum (T.a.f.a) from the T. aggressivum f. europeanum (T.a.f.e.). The informative restriction fragments of 260 and 300 bp were revealed in the corresponding reference strains T.a.f.a. and T.a.f.e. The 300-bp marker was found in all 28 Polish mushroom isolates tested.