Feeding Ecology of Juvenile Spiny Lobster, Panulirus interruptus, on the Pacific Coast of Baja California Sur, Mexico

Many aspects of the early life history of the red lobster Panulirus interruptus are little known, including the relationship between habitat structure, food resource availability, and nutrition of juveniles. We investigated the spatial and temporal differences in food intake, diet composition, and nutritional condition of juveniles at two sites along the Pacific coast of the Baja California Peninsula (Mexico) with contrasting oceanographic and biological conditions. One site (Arvin) is located inside a protected bay, Bahia Tortugas, where the waters are cooler and temperate seagrasses and macroalgae are the dominant benthic flora. The second site (Queen) in Bahia Sebastian Vizcaino was located along a more open coastline where seawater temperatures were higher and the benthic flora more indicative of warmer seas. At both sites, we randomly sampled epifauna within vegetated habitats to estimate the seasonal availability of food resources for juvenile lobsters from autumn 2001 until summer 2002. Concurrently, we used traps to sample P. interruptus juveniles for stomach content analysis. At both sites, Amphipoda, Gastropoda, and Polychaeta dominated the epifauna assemblages, as determined by an Index of Importance. Juvenile P. interruptus primarily consumed crustaceans (mostly amphipods and isopods) and vegetal material (surf-grass Phyllospadix spp. and calcareous algae), but their food spectrum was wide. Manly\u27s Index of Resource Selection indicated that lobsters preferred some prey (e.g., Brachyurans) over others despite their low Index of Importance. Despite marked differences in the types of food and their availability between sites, there were no significant differences in the nutritional condition (e.g., relative weight of the digestive gland) of lobsters at the two sites. However, the nutritional condition of lobsters was effected during some seasons. In particular, their condition deteriorated during the spring (April 2002) at Arvin, as did the proportion of individuals with empty stomachs. This study shows the feeding adaptation capacity of the juvenile California spiny lobster P. interruptus to different environmental conditions prevalent in Centre Baja California Peninsula

Performance of ELISA and Western blot to detect antibodies against HSV-2 using dried blood spots

Background and aims: Herpes simplex virus type 2 (HSV-2) is a sexually transmitted agent and is detected worldwide. HSV-2 is the main cause of genital ulcers and is diagnosed mainly with serological tests. The objective of current study was to evaluate the use of DBS samples to detect HSV-2 antibodies using commercial ELISA and Western blot tests. Materials and methods: IgG-G2 ELISA (Human® Diagnostics, Germany) and Western blot IgG/IgM (EUROLINE-WB, Euroimmun® Germany) tests were modified to use DBS samples. Samples were processed by both methods to determine ELISA cutoff points using ROC curves. ELISA was performed with 100 μl and the Western blot with 200 μl of eluted DBS. A 1:5 dilution was used and the incubation times were increased for the Western blot. Results: 908 DBS samples were processed and the following cutoff points were determined: negative (0–3.79), undetermined (3.8–4.6) and positive (≥4.61), with sensitivity and specificity close to 95%. Conclusion: Modifications of the cutoff points of the ELISA test were obtained with technical adjustments done to detect HSV-2 antibodies by ELISA and Western blot using DBS samples. Keywords: Herpes simplex virus type 2, Dried blood spots, Western blot, ELIS

Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field