Isolation and characterization of microsatellite markers from the entomopathogenic fungus Metarhizium anisopliae

Fourteen polymorphic microsatellite markers were isolated from the entomopathogenic fungus, Metarhizium anisopliae, based on enriched genomic libraries. In order to assess allelic variability, the microsatellite loci were analysed in a collection of 34 isolates sampled from across Switzerland. The number of detected alleles in 14 loci ranged from two to eight and expected heterozygosity from 0.265 to 0.808. Because of the high expected heterozygosity, the 14 microsatellite loci are very useful for ecological studies and analysis of population diversity, and to identifying, monitoring, and tracking M. anisopliae strains applied as biological control agents.     

Sensitivity of respiratory metabolism and efficiency to foliar nitrogen during growth and maintenance

Scaling of respiration from the leaf to the canopy level currently depends on identification of physiological parameters that are tightly linked to respiration and that can readily be determined. Several recent studies have helped provide guides to predicting whole canopy respiration on the basis of foliar nitrogen (N). This approach is potentially powerful owing to the well‐described patterns of allocation of N that follow interception of radiation. In the present study, we investigated the sensitivity of the N–respiration correlation to environmental and developmental factors, in order to evaluate its usage for attempts to scale respiration to the organism and ecosystem level. We studied fully expanded, 1 and 2‐year‐old, and current‐year needles from canopies of Pinus radiata that had been treated (unthinned, thinned and thinned+fertilized treatments) in ways likely to induce a wide range of growth and respiratory responses. We examined respiration in detail during the growth period in spring and again at the end of summer, using calorespirometric methods (combined measurements of CO₂ and heat rates) to determine the respiration rates , instantaneous enthalpic growth rates (R_SGΔH_B, a measure of the conservation of electrons in anabolic products) and the enthalpy conversion efficiency (η_H) of needles differing in age. A general linear model revealed that was positively correlated with needle N, but this correlation was strongly dependent on the season and the needle age – indicating an important physiological difference between expanding young needles and fully expanded old needles. Furthermore, the strength of the correlation between needle N and respiration was comparatively weak for the current year, expanding foliage, indicating that factors other than foliage N significantly influenced the respiration of young needles. The analysis of instantaneous growth rates revealed two general processes. Older, nonexpanding foliage showed considerable rates of enthalpic growth (increases in enthalpy) that was mainly caused by the increment of lignin during secondary growth. Secondly, canopy development appeared dynamic and to be optimized according to environmental drivers and constraints – such as light and water availability. In late spring, needle extension slowed in the upper, but not the lower canopy, because the upper canopy appeared to be affected first by the onset of drought stress in late spring. Growth rates were reduced in the upper canopy despite greater rates of respiration, indicating higher demand of ATP for the maintenance of protein and for export of sugars. Consequently, the enthalpy conversion efficiency and enthalpic N productivity (enthalpic growth per unit N) were comparatively poor indicating advanced development of needles in the upper canopy. We suggest that the growth and maintenance paradigm of respiration is, at best, only moderately useful when applied to whole trees, and is not valid at the cellular level or that of the plant organ. A different concept, namely that of respiratory efficiency, seems a more suitable way to represent respiration in carbon (C) balance models and should help provide a better mechanistic understanding of how respiration affects the C conversion efficiency of plants, and ultimately the net primary productivity of ecosystems.     

SOS-red fluorescent protein (RFP) bioassay system for monitoring of antigenotoxic activity in plant extracts

An optical antigenotoxicity assay using genetically engineered red fluorescent bacteria is presented. Exposure of Escherichia coli RS4U to genotoxicants [mitomycin C (MMC), nalidixic acid (NA) and hydrogen peroxide (HP)] resulted in phenotypic red fluorescence proportional to the concentration of the inducer. Except for tannic acid (TA), co-treatment of the genotoxicant-activated bacteria with ascorbic acid (AA) and aqueous plant extracts (Mangifera indica, Psidium guajava and Syzygium cumini) afforded protection against all three genotoxicants. TA was effective in suppressing the genotoxic effect of MMC and HP. The antigenotoxic effect is seen as inhibition of the genotoxicant-triggered red fluorescence. The IC50 of the plant extracts and AA varied with the genotoxicant used. Rec assay verified the antigenotoxic activity of the plant extracts. Folin-Ciocalteu test, FeCl3 test and DPPH assay confirmed the presence of polyphenolic compounds and hydrolyzable tannins in the plant extracts and the antioxidant capacity of the plant samples.     

Selective lymphocyte depletion during the early stage of the immune response to foot-and-mouth disease virus infection in swine

Foot-and-mouth disease virus (FMDV) is the causative agent of a highly contagious vesicular disease of cloven-hoofed animals. In the present study we use FMDV serotype C infection of swine to determine, by analytical techniques, the direct ex vivo visualization of virus-infected immune cells during the first 17 days of infection. We report, for the first time, that FMDV C-S8c1 can infect T and B cells at short periods of time postinoculation, corresponding with the peak of the viremia. There is a significant lymphopenia that involves CD3(+) CD4(-) CD8(+/-), CD3(+) CD4(-) CD8(+)Tc, and CD3(+) CD4(+) CD8(+) memory Th but not CD3(+) CD4(+) CD8(-) na?ve Th lymphocytes. In addition, a profound depletion of the vast majority of peripheral T cells in lymph nodes and spleen is observed. This selective depletion of T cells is not due mainly to in situ death via apoptosis as visualized by the terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling (TUNEL) technique. Thus, early infection of T cells by FMDV may be the main cause of the observed T-cell depletion. Importantly, this lack of T cells is reflected in a reduced response to mitogen activation, which in many cases is totally eliminated. These data suggest a mechanism by which the virus causes a transient immunosuppression, subvert the immune systems, and spreads. These results have important implications for our understanding of early events in the development of a robust immune response against FMDV.     

The postcranial anatomy of Suminia getmanovi (Synapsida: Anomodontia), the earliest known arboreal tetrapod

The basal anomodont Suminia getmanovi Ivakhnenko, 1994 from the late Palaeozoic of Russia is highly specialized in its masticatory apparatus, and has been suggested to represent the earliest arboreal tetrapod in the fossil record. Its postcranial anatomy is described in detail for the first time, revealing a large number of autapomorphies for this small herbivore. These include a reduced number of presacral and therein dorsal vertebrae, an elongate neck, a long and possibly prehensile tail, a procoracoid with a notch at its ventromedial margin rather than a foramen, an iliac blade with a robust ridge at its anteromedial edge, a pubis with a puboischiadic fenestra and separate pubic foramen, and elongate limbs. Additional autapomorphic characters are displayed in the autopodium, which comprises about 40% of the entire limb length. These features include an enlarged, phalangiform distal carpal 1 and tarsal 1, a short and robust first metacarpal, a crescent‐shaped distal tarsal 4, and elongate penultimate phalangeal elements. The phylogenetic relationships of basal anomodonts are revisited using an expanded data set, with the addition of key taxa and several postcranial characters. Unlike dicynodonts, Suminia retained the plesiomorphic phalangeal formula for amniotes of 2‐3‐4‐5‐3 (manus) and 2‐3‐4‐5‐4 (pes). This pattern is achieved by the retention of disc‐like phalangeal elements between the proximal and penultimate phalanges in digits III, IV (manus and pes), and V (pes only). In light of the new material, Suminia can be recognized as the most complete basal anomodont, offering new insights into the early evolution of the group. © 2011 The Linnean Society of London, Zoological Journal of the Linnean Society, 2011, 162 , 661–698.     

Digestion versus abrasion features in rodent bones

The origin of most fossil small mammal assemblages is predation by avian or mammalian predators. Bone corrosion by gastric juices observed in these fossils is direct evidence of digestion, and traits of digestion indicate the type of predator involved. However, certain features observed in digested bones, such as rounding and polishing, are similar to the rounding and polishing produced by other processes, particularly abrasion, in which predation is not involved. Misidentification of digestion has major repercussions in palaeoenvironmental and palaeoclimatic interpretations as well as interpretations of biostratigraphy and potential reworking. Digestion is directional and progressive process, primarily affecting the most mineralized tissues (enamel) advancing from the tips to the centre of the anatomical element. In contrast, abrasion identically affects any type of osseous tissue homogeneously rounding the entire skeletal element. Microscopically, digested bones display a distinctive chemical corrosion (‘tornlike’ appearance), whilst abraded bones appear smooth with microstriations and pitting microwear. Here, we present the results of a series of experiments designed to establish new and clear criteria to distinguish bone rounding and polishing caused by digestion from that originating from abrasion.     

Physiology of pepper fruit and the metabolism of antioxidants: chloroplasts,mitochondria and peroxisomes

Background and Aims Pepper (Capsicum annuum) contains high levels of antioxidants, such as vitamins A and C and flavonoids. However, information on the role of these beneficial compounds in the physiology of pepper fruit remains scarce. Recent studies have shown that antioxidants in ripe pepper fruit play a key role in responses to temperature changes, and the redox state at the time of harvest affects the nutritional value for human consumption. In this paper, the role of antioxidant metabolism of pepper fruit during ripening and in the response to low temperature is addressed, paying particular attention to ascorbate, NADPH and the superoxide dismutase enzymatic system. The participation of chloroplasts, mitochondria and peroxisomes in the ripening process is also investigated.Scope and Results Important changes occur at a subcellular level during ripening of pepper fruit. Chloroplasts turn into chromoplasts, with drastic conversion of their metabolism, and the role of the ascorbate–glutathione cycle is essential. In mitochondria from red fruits, higher ascorbate peroxidase (APX) and Mn-SOD activities are involved in avoiding the accumulation of reactive oxygen species in these organelles during ripening. Peroxisomes, whose antioxidant capacity at fruit ripening is substantially affected, display an atypical metabolic pattern during this physiological stage. In spite of these differences observed in the antioxidative metabolism of mitochondria and peroxisomes, proteomic analysis of these organelles, carried out by 2-D electrophoresis and MALDI-TOF/TOF and provided here for the first time, reveals no changes between the antioxidant metabolism from immature (green) and ripe (red) fruits.Conclusions Taken together, the results show that investigation of molecular and enzymatic antioxidants from cell compartments, especially chloroplasts, mitochondria and peroxisomes, is a useful tool to study the physiology of pepper fruit, particularly in the context of expanding their shelf-life after harvest and in maintaining their nutritional value.     

Photobioreactors for the production of microalgae

Microalgae are produced today for human and animal markets, as food-feed and source of active compounds. Microalgae can be also used in wastewater treatment and they has been proposed as biofuels source to reduce global warming problem. Whatever the final application of microalgae its production is based on the same principles as light availability, enough mass and heat transfer and adequate control of culture parameters. In this paper these principals are revised. Moreover, the production must be carried out at adequate scale using photobioreactors. Design of photobioreactor is determined by the final use of biomass and quality required. Different designs today used are revised, including last designs proposed, identifying his characteristics parameters and applications. In addition, the obligation of adequate control strategies is discussed. Finally, the bottlenecks for the scale-up of the different technologies and thus of microalgae production are summarized.