Adapting the IUCN Red List criteria for invertebrates

The IUCN Red List is the most useful list of species that are at risk for extinction worldwide, as it relies on a number of objective criteria. Nevertheless, there is a taxonomic bias that excludes species with small body sizes, narrow distribution ranges and low dispersal abilities, which constitute the vast majority of the planet’s biota, particularly local endemics.By evaluating each IUCN criterion separately, we (i) identify the shortcomings for invertebrate applications, (ii) explain how risk categories may be wrongly applied due to inapplicable and/or misleading thresholds, (iii) suggest alternative ways of applying the existing criteria in a more realistic way and (iv) suggest possible new criteria that were not considered in the current evaluation framework but that could allow a more comprehensive and effective assessment of invertebrates.By adapting the criteria to rely more explicitly on the Area of Occupancy and the Extent of Occurrence, their respective trends and by using ecological modelling methods, the criteria’s applicability would be increased. The change in some thresholds or, eventually, the creation of sub-categories would further increase their adequacy. Additionally, co-extinction could be introduced as an explicit part of the classification process.As a case study, we evaluated 48 species of Azorean arthropods and Iberian spiders according to the current criteria. More than one-quarter (27%) of all evaluated species were classified as Critically Endangered, 19% as Endangered, 6% as Vulnerable and 8% as Least Concern. The remaining 40% did not have enough data to reach a classification.     

Biomass allocation and foliage heteroblasty in hard pine species respond differentially to reduction in rooting volume

Rooting space is considered as a resource in plants, but comparative studies on the biomass allocation plasticity in response to rooting volume (RV) are rare. We compared responses in growth, biomass allocation and ontogenetic heteroblasty in nine hard pine species of contrasted ecology. Seedlings were cultivated in containers of 0.2, 0.5, 1.0, 2.8 and 7 L for two growing seasons (425 days). Reduction in RV caused a reduction in plant absolute and relative growth rate and biomass allocated to stems but it increased biomass allocated to roots. RV affected to a lesser extent and in a less consistent direction allocation to leaves. Species that grew faster (higher relative growth rate) had a steeper decrease in growth with the reduction in RV. Ontogenetic heteroblasty, evaluated as the proportion of secondary needles in the needle biomass, showed highly different plasticity patterns in response to RV. Decrease in RV caused negligible or no change either in the most ontogenetically delayed Mediterranean pines or in the most ontogenetically advanced pines, the mesic Pinus sylvestris and P. uncinata. By contrast, ontogenetically intermediate species showed steep reaction norms in response to reduction in RV. While P. pinaster and P. brutia showed marked rejuvenation, P. nigra accelerated the development of adult foliage.     

A dog model to study ovary, ovarian ligament and visceral pain

Objective A dog model was developed to study visceral pain by stimulating the ovarian ligament. Study design Prospective experimental trial. Animals Twelve 1‐year old female hound dogs weighing 25.7 ± 3.6 kg. Methods Dogs were anesthetized with sevoflurane. The right ovary was accessed via laparoscopy. A suture was placed around the ovarian ligament and exteriorized through the abdominal wall for stimulation. The noxious stimulus consisted of pulling the ovary and ovarian ligament with a force transducer. The response to noxious stimulation was determined using the anesthetic minimum alveolar concentration requirement (MAC) for sevoflurane. The ovarian MAC was compared to the standardized somatic noxious stimulation tail clamp MAC. The results are depicted as mean ± SD and corrected to sea‐level. Results The stimulus–response curve during ovarian stimulation in three dogs was hyperbolic and best represented by a three‐parameter logistic growth curve model. The curve plateaued at 7.12 ± 4.19 N. From the stimulus‐response curve, we chose 6.61 N to test the consistency and repeatability of the model in nine dogs. The ovarian stimulation MAC for sevoflurane in these dogs was 2.16 ± 0.46%. The ovarian stimulation confidence interval and limits are comparable to the results from tail stimulation MAC. The tail stimulation MACs before and after laparoscopy surgery were not different (1.86 ± 0.28% and 1.77 ± 0.38% respectively; p > 0.05) but lower when compared to the ovarian MAC (p < 0.01). The dogs recovered from anesthesia without complications. Conclusions and clinical relevance The ovarian stimulation model is an adequate and repeatable means of producing visceral stimulation to determine MAC. The model may provide a humane mechanism to study the effectiveness of analgesics for acute ovarian pain.     

Expression of 14-3-3 σ protein in normal and neoplastic canine mammary gland

14-3-3 σ protein is a negative cell cycle regulator, with both reduced and elevated levels associated with cancer in humans. This study assessed the expression of this protein in canine mammary tissues using immunohistochemistry and Western blotting. 14-3-3 σ was detected in 97% of the mammary tissue samples examined and was found in both myoepithelial (MECs) and epithelial (ECs) cells. Expression levels were elevated and reduced in neoplastic ECs and MECs, respectively (P < 0.001). Intense expression of 14-3-3 σ was detected in neoplastic ECs infiltrating blood vessels and lymph nodes and suggests a possible role for this protein in the malignant transformation of mammary neoplasms. Moreover, double immunostaining for 14-3-3 σ and the MEC – specific marker p63, confirmed that 14-3-3 σ is a highly sensitive marker of MECs since all p63 – positive cells were also positive for 14-3-3 σ. However, this protein is not exclusive to MECs as ECs also labelled positively.     

Field trial measuring the compatibility of methoxyfenozide and flonicamid with Orius laevigatus Fieber (Hemiptera: Anthocoridae) and Amblyseius swirskii (Athias-Henriot) (Acari: Phytoseiidae) in a commercial pepper greenhouse

BACKGROUND: Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae) and Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) are among the most serious pests of sweet peppers in greenhouses. Chemical control is difficult because of their high reproductive rates and insecticide resistance, and seasonal inoculative releases of Orius laevigatus (Fieber) and Amblyseius swirskii (Athias‐Henriot) are commonly used to reduce their populations. As chemical treatments are often needed in the crop against other pests, the side effects of methoxyfenozide (an insect growth regulator against lepidopteran pests) and flonicamid (a selective feeding inhibitor against sucking insects) were studied in both beneficial organisms in a commercial greenhouse. RESULTS: Orius laevigatus and A. swirskii were released at commercial rates (4–5 and 100 m^?2), and a strong establishment and a very homogeneous distribution were reached. One pesticide treatment with the maximum field recommended concentration of methoxyfenozide and flonicamid (96 and 100 mg AI L^?1) was done when they were well established, and their population levels were not affected either immediately or up to 30 days after treatment. CONCLUSION: The results are indicative of no impact of methoxyfenozide and flonicamid on the two natural enemies in the field, and both can be considered as potential alternatives to be included in IPM programmes in sweet pepper. Copyright © 2011 Society of Chemical Industry     

Hydroxytyrosol 4-beta-D-glucoside,an important phenolic compound in olive fruits and derived products

There is increasing interest in olive polyphenols because of their biological properties as well as their contribution to the color, taste, and shelf life of olive products. However, some of these compounds remain unidentified. It has been shown that hydroxytyrosol 4-beta-D-glucoside (4-beta-D-glucosyl-3-hydroxyphenylethanol) coeluted with hydroxytyrosol [(3,4-dihydroxyphenyl)ethanol] under reversed phase conditions in the phenolic chromatograms of olive pulp, vegetation water, and pomace of olive oil processing. A method to separate this compound from hydroxytyrosol by HPLC has been developed. The concentration of this glucoside increased in olive pulp with maturation and could be the main phenolic compound in mature olives. In contrast, the presence of this compound was not detected in olive oil by using HPLC-MS. The compound must be considered both in table olives and olive oil processing because of its glucose and hydroxytyrosol contribution to these products.     

Polyphenol changes during fermentation of naturally black olives

The individual evolution of phenolic compounds has been studied during the natural fermentation of black olives for the first time. Cyanidin 3-rutinoside and cyanidin 3-glucoside were the main anthocyanins identified in fresh olives, and they were not detected after 1 month of storage either in brine or in olive. The fruit colors were different when aerobic or anaerobic conditions were used and as a consequence of the different anthocyanin polymerizations that took place. At time zero, the polyphenols observed in the olive juice were hydroxytyrosol-4-beta-glucoside, oleuropein, hydroxytyrosol, tyrosol, salidroside, and verbascoside and, after 12 months, the main phenol was hydroxytyrosol. The polyphenol content in the oil phase of olives was also analyzed. The dialdehydic form of elenolic acid linked to hydroxytyrosol and tyrosol, oleuropein aglycon, and ligstroside aglycon were the main compounds found at the beginning of fermentation but were not detected after 3 months. In contrast, hydroxytyrosol, hydroxytyrosol acetate, tyrosol, and tyrosol acetate were the main polyphenols detected in the oil phase of the final product. The acid hydrolysis of the initial glucosides (in olive juice) and the aglycons (in oil phase) was, therefore, the main reaction that took place during fermentation.     

Antibacterial and antioxidant activities of essential oils isolated from Thymbra capitata L. (Cav.) andOriganum vulgare L

Antilisterial activities of Thymbra capitata and Origanum vulgare essential oils were tested against 41 strains of Listeria monocytogenes. The oil of T. capitata was mainly constituted by one component, carvacrol (79%), whereas for O. vulgare three components constituted 70% of the oil, namely, thymol (33%), gamma-terpinene (26%), and p-cymene (11%). T. capitata essential oil had a significantly higher antilisterial activity in comparison to O. vulgare oil and chloramphenicol. No significant differences in L. monocytogenes susceptibilities to the essential oils tested were registered. The minimum inhibitory concentration values of T. capitata essential oil and of carvacrol were quite similar, ranging between 0.05 and 0.2 microL/mL. Antioxidant activity was also tested, the essential oil of T. capitata showing significantly higher antioxidant activity than that of O. vulgare. Use of T. capitata and O. vulgare essential oils can constitute a powerful tool in the control of L. monocytogenes in food and other industries.     

Usefulness of amino acid composition to discriminate between honeydew and floral honeys. Application to honeys from a small geographic area

With the aim of finding methods that could constitute a solid alternative to melissopalynological and physicochemical analyses to determine the botanical origin (floral or honeydew) of honeys, the free amino acid content of 46 honey samples has been determined. The honeys were collected in a small geographic area of approximately 2000 km(2) in central Spain. Twenty-seven honey samples were classified as floral and 19 as honeydew according to their palynological and physicochemical analyses. The resulting data have been subjected to different multivariant analysis techniques. One hundred percent of honey samples have been correctly classified into either the floral or the honeydew groups, according to their content in glutamic acid and tryptophan. It is concluded that free amino acids are good indicators of the botanical origin of honeys, saving time compared with more tedious analyses.