Synergism between Basic Asp49 and Lys49 Phospholipase A2 Myotoxins of Viperid Snake Venom In Vitro and In Vivo

Two subtypes of phospholipases A₂ (PLA₂s) with the ability to induce myonecrosis, ‘Asp49’ and ‘Lys49’ myotoxins, often coexist in viperid snake venoms. Since the latter lack catalytic activity, two different mechanisms are involved in their myotoxicity. A synergism between Asp49 and Lys49 myotoxins from Bothrops asper was previously observed in vitro, enhancing Ca²⁺ entry and cell death when acting together upon C2C12 myotubes. These observations are extended for the first time in vivo, by demonstrating a clear enhancement of myonecrosis by the combined action of these two toxins in mice. In addition, novel aspects of their synergism were revealed using myotubes. Proportions of Asp49 myotoxin as low as 0.1% of the Lys49 myotoxin are sufficient to enhance cytotoxicity of the latter, but not the opposite. Sublytic amounts of Asp49 myotoxin also enhanced cytotoxicity of a synthetic peptide encompassing the toxic region of Lys49 myotoxin. Asp49 myotoxin rendered myotubes more susceptible to osmotic lysis, whereas Lys49 myotoxin did not. In contrast to myotoxic Asp49 PLA₂, an acidic non-toxic PLA₂ from the same venom did not markedly synergize with Lys49 myotoxin, revealing a functional difference between basic and acidic PLA₂ enzymes. It is suggested that Asp49 myotoxins synergize with Lys49 myotoxins by virtue of their PLA₂ activity. In addition to the membrane-destabilizing effect of this activity, Asp49 myotoxins may generate anionic patches of hydrolytic reaction products, facilitating electrostatic interactions with Lys49 myotoxins. These data provide new evidence for the evolutionary adaptive value of the two subtypes of PLA₂ myotoxins acting synergistically in viperid venoms.     

Differential predation on the two colour morphs of Nicaraguan Crater lake Midas cichlid fish: implications for the maintenance of its gold‐dark polymorphism

Predation can play an important role in the evolution and maintenance of prey colour polymorphisms. Several factors are known to affect predator choice, including the prey's relative abundance and conspicuousness. In polymorphic prey species, predators often target the most common or most visible morphs. To test if predator choice can explain why in Midas cichlid fish the more visible (gold) morph is also more rare than the inconspicuous dark morph, we conducted predation experiments using two differently coloured wax models in Nicaraguan crater lakes. Contrary to expectations, we observed an overall higher attack rate on the much more abundant, yet less conspicuous dark models, and propose frequency‐dependent predation as a potential explanation for this result. Interestingly, the attack rate differed between different types of predators. While avian predators were biased towards the abundant and less colourful dark morphs, fish predators did not show a strong bias. However, the relative attack rate of fish predators seemed to vary with the clarity of the water, as attack rates on gold models went up as water clarity decreased. The relative differential predation rates on different morphs might impact the relative abundance of both colour morphs and thus explain the maintenance of the colour polymorphism. © 2014 The Linnean Society of London, Biological Journal of the Linnean Society, 2014, 112 , 123–131.     

Age and size thresholds for pupation and developmental polymorphism in the browntail moth, Euproctis chrysorrhoea (Lepidoptera: Lymantriidae), under conditions that either emulate diapause or prevent it

Size and age thresholds for pupation are important life history traits of insects. They are the ultimate consequences of the underlying physiological mechanism that optimize resource allocation. Such thresholds may have a plastic response under time-varying environmental conditions, developmental polymorphism (i.e., plasticity in the number of instars before pupation) being a common strategy adopted by insects to overcome this challenging situation. In this study, we systematically explore the variables related with both age and size thresholds for pupation and developmental polymorphism in the browntail moth, Euproctis chrysorrhoea (Lepidoptera: Lymantriidae), by rearing a group of caterpillars under conditions that either emulate larval diapause or prevent it. As an innovative approach, we evaluated the importance of predictor variables by means of generalized linear modeling in a multi-model inference framework. Our results show that (i) rearing conditions affect fitness, (ii) rearing conditions, size of hatchlings, size and age at maturity and sex are related to the number of instars before pupation, and (iii) there are both age and size thresholds for pupation which differ between sexes and between larvae reared under different conditions. Results are discussed in the context of lepidopteran plasticity in life history traits and its relationship with optimal molting strategies.     

The role of the perch effect on the nucleation process in Mediterranean semi-arid oldfields

Oldfield succession in Mediterranean ecosystems has been studied extensively in mesic conditions. However, this phenomenon is still poorly understood in semi-arid Mediterranean areas, where reduced plant cover, the importance of facilitation processes and the role of abiotic factors make these environments distinct. We first test whether the carob tree (Ceratonia siliqua) generates nucleation patterns in semi-arid oldfields, and to what extent such patterns change with abandonment age. Then we test to what extent nucleation can be explained by the perch effect. And finally, we test whether the nucleated pattern around carob trees is a source of diversity in the oldfields studied. To answer these questions we located oldfields abandoned 25 and 50 years ago (20 in each case) in the Alacant Province (SE Spain, Iberian Peninsula) on the basis of aerial photographs and personal interviews with local landowners and managers. In each oldfield woody plant density and richness were sampled on two microsites: under the carob tree and in the open field. Analysis was performed on all woody plants and by separating the species in two functional groups: fleshy-fruited (with fleshy mesocarp) and non-fleshy-fruited species. The results suggest that woody vegetation colonising abandoned C. siliqua fields in SE Spain is not randomly distributed but follows a nucleation pattern with higher plant density under the trees. However, the nucleation pattern is only significant for fleshy-fruited species, suggesting that facilitative interactions alone cannot explain the nucleation pattern and that the perch effect plays an important role. The results also show that the nucleation pattern (total plant density and density of non-fleshy-fruited plants) did not increase with abandonment age, while the perch effect (density of fleshy-fruited plants) did increase significantly. Furthermore, the results also show that the nucleation pattern is not only a loci of high plant density but also a loci of high species richness. Thus we can conclude that the nucleation pattern found in oldfield succession is best explained by the perch effect, while facilitation has a secondary importance. This emphasises the key role that dispersal mode has on the dynamics of vegetation recovery in formerly cropped areas.     

Determinant role for Toll-like receptor signalling in acute mycobacterial infection in the respiratory tract

Toll-like receptors (TLRs) are a vital component of the innate branch of the immune system in its battle against mycobacterial infections. Extensive in vitro studies have demonstrated a role for both TLR2 and TLR4 in recognition of mycobacterial components, whereas the in vivo situation appears less clear, with results depending on the infection model. In the present work, the importance of TLR signalling in the course of mycobacterial infection was investigated in a human-like infection model using TLR-knockout mice. TLR2(-/-) and TLR4(-/-) mice infected with Mycobacterium tuberculosis by aerosol, or for the first time, intranasally with Mycobacterium bovis bacillus Calmette-Guérin (BCG), displayed increased susceptibility at an early stage of infection in the respiratory tract, while at a later stage of infection, the TLR deficiency appeared to be overcome. The higher susceptibility was correlated to impaired pro-inflammatory responses to BCG components, and reduced induction of anti-bacterial activity by infected macrophages from TLR2(-/-) mice, and to a lesser extent from TLR4(-/-) mice. These findings demonstrate a role for TLR signalling in protection against mycobacterial infection specifically in the respiratory tract at the acute phase, whereas the TLR deficiency can be compensated at a later stage of infection.     

Winter feeding leads to a shifted phenology in the browntail moth Euproctis chrysorrhoea on the evergreen strawberry tree Arbutus unedo

• 1 The browntail moth Euproctis chrysorrhoea is a highly polyphagous univoltine forest pest. Although its young larvae usually overwinter in diapause from early autumn to the beginning of spring, winter larval feeding has been reported when this species feeds on the evergreen woody shrub strawberry tree Arbutus unedo.
• 2 The present study investigated life‐history traits of four populations of E. chrysorrhoea feeding on A. unedo, including phenology of the different life stages, larval feeding activity and diapause incidence. By modelling the relationship between larval size and host plant leaf persistence, elevation and mean annual temperature, we also studied larval development in ten populations of this species sampled from a range of geographical locations in Spain, from both A. unedo and deciduous hosts.
• 3 The results obtained revealed that on A. unedo, E. chrysorrhoea phenology has shifted: from October to March, A. unedo larvae doubled their size, whereas, on deciduous Ulmus minor and Quercus faginea, larval size did not change. General linear models demonstrated that such differences were not related to environmental variables. We also found that on A. unedo larval feeding was arrested for 2 months, with this period representing a true diapause.
• 4 The results obtained in the present study suggest that E. chrysorrhoea populations are phenologically adapted to their local host plants, and that the presence of foliage is a key element in the phenological shift reported on A. unedo. These results may have implications with respect to the formation of E. chrysorrhoea host races.

     

Parent Kittiwakes experience a decrease in cell-mediated immunity as they breed

The maintenance of an effective immune system may entail physiological costs only affordable by individuals in good condition. Some species, like Black-legged Kittiwakes (Rissa tridactyla) in the high arctic, experience important physiological changes during breeding in order to fulfil their reproductive goals. We studied the effects of breeding effort on the cell-mediated immunity (CMI) of adult breeding Kittiwakes. Parent Kittiwakes showed a significant decrease in CMI from hatching to 15 days into the chick-rearing period, when both parents are known to experience a drastic increase in energy expenditure together with important physiological adjustments. Despite being a long-lived species with biparental care, parent Kittiwakes at the northern border of their breeding range, seem to experience important reproductive immunosupression, even during favourable breeding years.     

Mediterranean vegetation dynamics: modelling problems and functional types

Gap models have been applied to a wide range of ecosystems, mainly temperate and boreal forests, but rarely have such models been applied to Mediterranean ecosystems. In the present review we address some problems of gap models for predicting the long-term dynamics of Mediterranean ecosystems, and we suggest plant functional types suitable for modelling based on responses to disturbance. Most gap models do not take into account different life forms, interactions with fire (e.g., resprouting and stimulation of the germination), and underground structures. Long term human impact on Mediterranean ecosystems has made a significant impact on the current landscapes. That intense land use, involving long-lived slow-growing species, has had long-term consequences. It is not possible to understand Mediterranean vegetation and to validate any model without considering these factors. The lack of data for Mediterranean species may be overcome by taking into account correlations of traits and trade-offs between different functional types. A simple disturbance-based functional group system is discussed.     

Sexual dimorphisms in a neuromuscular system regulating courtship in the green anole lizard: Effects of season and androgen treatment

During the breeding season, male green anoles (Anolis carolinensis) court females by extending a red throat fan called a dewlap. Motoneurons controlling this sexually dimorphic behavior are located in two portions of the brain stem: (a) the vagal portion of nucleus ambiguus (AmbX), and (b) the region containing the glossopharyngeal portion of nucleus ambiguus and the ventral motor nucleus of the facial nerve (AmbIX/VIImv). These motoneurons project to the ceratohyoideus muscle via the ramus pharyngo‐laryngeus IX+X. To investigate the effects of season on and androgen regulation of neural and peripheral structures controlling dewlap extension, two experiments were conducted: (a) During the breeding and nonbreeding seasons, motoneuron number, soma size, and nucleus size were investigated in intact males and females and in castrated males treated with a testosterone propionate (TP) or a blank Silastic capsule. (b) Cross‐sectional area of the nerve and muscle fiber size, number, and density were investigated in the four treatment groups during the breeding season only. No significant differences were found in motoneuron number. In the breeding season, subtle male‐biased sex differences existed in both AmbX and AmbIX/VIImv soma size. Nerve cross‐sectional area and muscle fiber size and number were substantially larger in males than females. Muscle fiber density was higher in females. No consistent effects due to season or androgen treatment were detected, although characteristics of motoneurons were in some cases slightly larger in the nonbreeding season. These results suggest that, while parallels to behavior exist between the sexes, morphological changes in adulthood in the dewlap motoneurons and muscle do not normally regulate courtship behavior in the male green anole. © 1999 John Wiley & Sons, Inc. J Neurobiol 40: 202–213, 1999