Foliar nematode,Litylenchus crenatae ssp. mccannii,population dynamics in leaves and buds of beech leaf disease‐affected trees in Canada and the US

A foliar nematode, Litylenchus crenatae ssp. mccannii, is associated with beech leaf disease (BLD) symptoms. Information about the types of tissues parasitized and how nematode populations fluctuate in these tissues over time is needed to improve surveys as well as understand the nematodes role in BLD. During this study, the nematode was detected throughout the known range of BLD by researchers at both Canadian and US institutions using a modified pan method to extract nematodes. Monthly collections of symptomatic and asymptomatic leaves during the growing season (May–October), and leaves and buds between growing seasons (November–March), revealed that nematodes were present in all tissue types. Progressively larger numbers of nematodes were detected in symptomatic leaves from Ohio and Ontario, with the greatest detections at the end of the growing season. Smaller numbers of nematodes were detected in asymptomatic leaves from BLD‐infected trees, typically at the end of the growing season. The nematode was detected overwintering in buds and detached leaves. The discovery of small numbers of nematodes in detached leaves at one location before BLD was detected indicates that nematodes may have been present before disease symptoms were expressed. Other nematodes, Plectus and Aphelenchoides spp., were infrequently detected in small numbers. Our findings support the involvement of the nematode in BLD and indicate that symptoms develop only when certain requirements, such as infection of buds, are met. We also found that the nematode can be reliably detected in buds and leaves using the modified pan extraction method.     

Responses of Australian sea lions,Neophoca cinerea,to anthropogenic activities in the Perth metropolitan area,Western Australia

1. Tourist‐based activities, partly due to their rapid increase, have raised concerns regarding the impacts of anthropogenic activity on marine fauna. Documented effects on pinnipeds in proximity to humans include changes in behaviour, site use and potentially higher aggression levels towards people. Effects vary considerably between populations and sites, thus requiring separate assessment of human impacts on activity and energy budgets.
2. Responses of the endangered Australian sea lion, Neophoca cinerea, to human visitation were recorded from November 2013 through April 2014. Exposure levels and response types to anthropogenic activities were assessed at two easily accessible locations with different management schemes, Seal (landing prohibited) and Carnac (landing permitted) islands, Western Australia. Exposure levels were measured as both stimulus type (i.e. ‘People’, ‘Paddlers’, ‘Small’, ‘Medium’, and ‘Large vessels’, ‘Tour vessels’, and ‘Jet skies’), and people (‘Direct’, ‘Attract’, ‘Interact’, ‘View’, ‘Incidental’, ‘Water’, ‘Low‐level’), and vessel activities (‘Interact’, ‘Approach/Follow’, ‘Anchor noise’, ‘Engine noise’, ‘Close to beach’, ‘Moderate/Fast travel’, ‘Slow travel’, ‘Transit’, ‘Drift/At anchor’, ‘Aircraft noise’).
3. Exposure levels varied significantly between the islands in numbers, stimuli type, duration and minimum approach distances. The instantaneous behaviours of ‘Lift head’, ‘Interact’ and ‘Sit’ were the most frequent responses. ‘Aggressive’ and ‘Retreat’ responses, the highest disturbance levels measured, occurred on Carnac approximately once per day, but rarely on Seal Island. ‘Aggressive’ behaviour towards ‘People’ was observed only on Carnac Island and elicited only by ‘People’. ‘People’, ‘Tour vessels’, and scenic ‘Aircrafts’ on both islands as well as ‘Jet skis’ on Carnac Island had the highest probability of triggering responses. Owing to their relatively high visitation at Seal Island, ‘Paddle powered vessels’, followed by ‘Tour vessels’ elicited the highest number of responses, compared with ‘People’, ‘Small’, and ‘Medium vessels’ at Carnac Island. The majority of responses occurred when any stimulus type was at short‐range (≤10 m), and ‘People’ ‘Viewing’ N. cinerea elicited most. Vessels triggered more responses at larger ranges than ‘People’.
4. To limit close‐range access to N. cinerea, one possibility is to close the beach at Carnac Island to human visitation and increase the minimum approach distance by vessels and ‘People’ by installing marker buoys at least 15 m from the shore.

© 2016 The Authors. Aquatic Conservation: Marine and Freshwater Ecosystems published by John Wiley & Sons, Ltd.     

Toward metrological traceability for DNA fragment ratios in GM quantification. 2. Systematic study of parameters influencing the quantitative determination of MON 810 corn by real-time PCR

This paper is part of a set of three papers investigating metrological traceability of the quantification of DNA fragments as, for instance, used for quantification of genetic modifications. This paper evaluates the possible impact of several factors on results of real-time Polymerase Chain Reaction (PCR) measurements. It was found that the particle size of the powder samples does not have an influence, whereas the nature of the calibrant (plasmidic or genomic DNA) has a significant effect. Moreover, two real-time PCR detection methods (construct-specific and event-specific) for MON 810 corn were compared. The results obtained in a specifically designed interlaboratory study revealed a significant influence of the DNA extraction method on measurement results when the MON 810 construct-specific real-time PCR detection method was applied. Statistical analyses confirmed the importance of validating DNA extraction methods in conjunction with real-time PCR methods.     

Extraction, processing, and storage effects on curcuminoids and oleoresin yields from Curcuma longa L. grown in Jamaica

Aromatic diarylheptanoid compounds from Curcuma longa Linn grown in Jamaica were quantified by UV-vis spectrophotometry and high-performance liquid chromatographic (HPLC) analyses. The oleoresin yields from ethanolic extracts were quantified and evaluated with regard to the effects of the type of postharvesting process and the type of extraction method conducted on the plant material. Fresh samples that were hot solvent extracted provided the highest oleoresin yields of 15.7% +/- 0.4 ( n = 3), and the lowest oleoresin yields of 7.8% +/- 0.2 ( n = 3) were from the dried milled samples that were cold solvent extracted. Data from the ASTA spectrophotometer assay confirmed that dried samples contained the highest curcuminoid content of 55.5% +/- 2.2 ( n = 6) at the fifth month of storage, and the fresh samples showed a curcuminoid content of 47.1% +/- 6.4 ( n = 6) at the third month of storage. A modified HPLC analysis was used to quantify curcumin content. Data from the HPLC analysis confirmed that the dried treated, hot extracted, room temperature stored samples had the highest curcumin content of 24.3%. A novel high-performance thin layer chromatography (HPTLC) method provided a chemical fingerprint of the C. longa with the use of a commercial curcumin standard.     

Quantitative extraction of macro-invertebrates from temperate and tropical leaf litter and soil: efficiency and time-dependent taxonomic biases of the Winkler extraction

Winkler extractors, a simple device presumed to extract macro-invertebrates efficiently from soil and litter samples, is being used increasingly in ecological surveys and functional studies of soil macro-invertebrate communities. In this study the extraction efficiency and taxonomic bias of the Winkler extraction are evaluated for extraction periods of 3 h up to 7 weeks, calibrated by hand-sorting after 7 weeks. The method extracts most macro-invertebrates completely or to a proportion of over 90% except Isopoda, Diplopoda and Mollusca. However, for an exhaustive result, a long extraction period of several weeks is necessary. For the most speciose group (adult beetles) and for the commonly most abundant group (ants), a short extraction of 3 days was sufficient to get 70% of the individuals and nearly all species. Three days was also sufficient to recover the rank abundance order of beetle families, while for ‘higher taxa’ and for Chilopoda species, 4 and 3 weeks were necessary, respectively. Optimum extraction times for the abundant macro-invertebrate groups and possible adjustment factors for the soil macro-invertebrates of temperate woodlands are proposed to compensate the taxonomic bias caused by short extraction periods. However, for recording an accurate snapshot of the soil and litter fauna at a particular time, shorter extraction periods are advisable because of the short life cycle of many soil invertebrates causing emergence of later stages or a second generation during longer extraction periods. The problem of contamination of samples is also discussed.     

Repellent effects of five monoterpenoid odours against Tribolium castaneum (Herbst) and Rhyzopertha dominica (F.) in Calabar, Nigeria

Secondary plant compounds are recognised as important components of plant defence system against herbivores and pathogens. Five monoterpenoids, (R)-linalool, 1,8-cineole, (S)-2-heptyl acetate, (S)-2-heptanol and citral, which are natural components of the essential oils of Aframomum melegueta (K. Schum) and Zingiber officinale (Roscoe), were tested at the ratios in which they occur naturally for repellent activity against Tribolium castaneum (Herbst.) and Rhyzopertha dominica (F.) in a 4-way olfactometer. The results showed the repellent properties of the compounds as both beetles spent less time in the olfactometer arm containing the test stimuli. (R)-linalool and (S)-2-heptanol were stronger repellent compounds than the others. Linalool showed good repellent activity against T. castaneum (P = 0.001) as the insect spent 1.22 min in the test arm compared to the control arms (2.78 min), and R. dominica (P = 0.001) with 0.89 min in the test arm compared to 2.87 min in the control arms. With (S)-2-heptanol, T. castaneum spent 1.23 min in the test arm compared to 2.83 min in the control arms. R. dominica spent 1.61 min in the test arm and 2.69 min in the control arms. For the number of entries or visits made, while both insects were significantly repelled (P < 0.05) by the linalool-treated arm than the control, only R. dominica was repelled by the (S)-2-heptanol-treated arm (P = 0.038) compared to the control arms. The results indicate that A. melegueta and Z. officinale essential oils and their components could be suitable as safer repellents or fumigants against T. castaneum and R. dominica.     

Dereplication and Chemotaxonomical Studies of Marine Algae of the Ochrophyta and Rhodophyta Phyla

Dereplication and chemotaxonomic studies of six marine algae of the Ochrophyta and one of the Rhodophyta phyla resulted in the detection of 22 separate compounds. All 16 secondary metabolites, including four new compounds (16–19), could be rapidly dereplicated using HPLC-NMR and HPLC-MS methodologies in conjunction with the MarinLit database. This study highlights the advantages of using NMR data (acquired via HPLC-NMR) for database searching and for the overall dereplication of natural products.