A Multiplex Real-Time PCR with High Resolution Melting Analysis for the Characterization of Antimicrobial Resistance in Neisseria gonorrhoeae.

Resistance to antibiotics used against Neisseria gonorrhoeae infections is a major public health concern. Antimicrobial resistance (AMR) testing relies on time-consuming culture-based methods. Development of rapid molecular tests for detecting AMR determinants could provide valuable tools for surveillance, epidemiological studies and to inform individual case management. We developed a fast (<1.5 hrs) SYBR-green based real-time PCR method with high resolution melting (HRM) analysis. One triplex and three duplex reactions included two sequences for N. gonorrhoeae identification and seven determinants of resistance to extended-spectrum cephalosporins (ESCs), azithromycin, ciprofloxacin, and spectinomycin. The method was validated by testing 39 previously fully-characterized N. gonorrhoeae strains, 19 commensal Neisseria spp., and an additional panel of 193 gonococcal isolates. Results were compared with culture-based AMR determination. The assay correctly identified N. gonorrhoeae and the presence or absence of the seven AMR determinants. There was some cross-reactivity with non-gonococcal Neisseria species and the detection limit was 10(3)-10(4) gDNA copies/reaction. Overall, the platform accurately detected resistance to ciprofloxacin (sensitivity and specificity, 100%), ceftriaxone (sensitivity 100%, specificity 90%), cefixime (sensitivity 92%, specificity 94%), azithromycin and spectinomycin (both sensitivity and specificity, 100%). In conclusion, our methodology accurately detects mutations generating resistance to antibiotics used to treat gonorrhea. Low assay sensitivity prevents direct diagnostic testing of clinical specimens but this method can be used to screen collections of gonococcal isolates for AMR more quickly than with current culture-based AMR testing.

High-resolution pollen record of ODP Hole 175-1078C

To address the connection between tropical African vegetation development and high-latitude climate change we present a high-resolution pollen record from ODP Site 1078 (off Angola) covering the period 50-10 ka BP. Although several tropical African vegetation and climate reconstructions indicate an impact of Heinrich Stadials (HSs) in Southern Hemisphere Africa, our vegetation record shows no response. Model simulations conducted with an Earth System Model of Intermediate Complexity including a dynamical vegetation component provide one possible explanation. Because both precipitation and evaporation increased during HSs and their effects nearly cancelled each other, there was a negligible change in moisture supply. Consequently, the resulting climatic response to HSs might have been too weak to noticeably affect the vegetation composition in the study area. Our results also show that the response to HSs in southern tropical Africa neither equals nor mirrors the response to abrupt climate change in northern Africa.

A high-resolution Lateglacial and Holocene palaeoceanographic record from the Greenland Sea

We present an unprecedented multicentennial sediment record from the foot of Vesterisbanken Seamount, central Greenland Sea, covering the past 22.3 thousand years (ka). Based on planktic foraminiferal total abundances, species assemblages, and stable oxygen and carbon isotopes, the palaeoenvironments in this region of modern deepwater renewal were reconstructed. Results show that during the Last Glacial Maximum the area was affected by harsh polar conditions with only episodic improvements during warm summer seasons. Since 18?ka extreme freshwater discharges from nearby sources occurred, influencing the surface water environment. The last major freshwater event took place during the Younger Dryas. The onset of the Holocene was characterized by an improvement of environmental conditions suggesting warming and increasing ventilation of the upper water layers. The early Holocene saw a stronger Atlantic waters advection to the area, which began around 10.5 and ended quite rapidly at 5.5?ka, followed by the onset of Neoglacial cooling. Surface water ventilation reached a maximum in the middle Holocene. Around 3?ka the surface water stratification increased leading to subsequent amplification of the warming induced the North Atlantic Oscillation at 2?ka.