Detection of Penicillin-binding Proteins in the Endosymbiont of the Trypanosomatid Crithidia deanei

ABSTRACT. Growth by serial transfers of the trypanosomatid Crithidia deanei in culture medium containing 1 mg/ml of the β-lactam antibiotics ampicillin or cephalexin resulted in shape distortion of its endosymbiont. The endosymbiont first appeared as filamentous structures with restricted areas of membrane damage. An increase of electron lucid areas was also observed in the endosymbiont matrix. The continuous treatment with β-lactam antibiotics, resulted in endosymbiont membranes fragmentation; and later on the space previously occupied by the symbiont was identified as an electron lucid area in the host cytoplasm. The putative targets of β-lactam antibiotic were two membrane-bound penicillin-binding proteins (PBPs) detected in the Sarkosyl-soluble fraction of purified symbionts labeled with [³H]-benzylpenicillin. The apparent molecular weight of the proteins were 90 kDa (PBP1) and 45 kDa (PBP2). PBP2 represented 85% of the total PBP content in the membrane fraction of the endosymbionts. Competition experiments using the tested antibiotics and [³H]-benzylpenicillin showed that ampicillin and cephalexin have half saturating concentrations considerably higher than [³H]-benzylpenicillin and indicated that PBP1 is the probable lethal target of the antibiotics tested. These results suggest that a physiologically active PBP is present in the cell envelope of C. deanei endosymbionts and may play important roles in the control of processes such as cell division and shape determination.     

Undesirable side-effects of water hyacinth control in a shallow tropical reservoir

1. Based on a comprehensive data set collected monthly during 8 years (1997–2004), we evaluated the effects of mechanical removal of Eichhornia crassipes on the limnological characteristics and algal biomass of a polymictic shallow tropical reservoir. 2. Interrupted time series analyses indicated that the limnological responses to macrophyte removal can be classified as an ‘abrupt permanent impact’ implying that the overall mean of the time‐series shifted promptly after intervention. These analyses indicated a significant increase for pH, total phosphorus, total phytoplankton and cyanobacterial biomass, and a decrease in water transparency and CO₂ concentrations in the surface water; also, the increase in water stability, increase of bottom soluble reactive phosphorus (SRP) and decrease in bottom oxygen levels. 3. Cyclic anoxic periods previously observed during springs and summers were replaced by a persistent period of anoxic conditions in the sediment overlying water. Anoxic conditions were suitable for SRP release from sediments. Heavy cyanobacterial blooms became more persistent, maximum biomass (4229 mm³ L⁻¹) was 30 times larger, the blooms frequently reached 2 m and sometimes the bottom of the reservoir, contrasting to the preremoval period in which it reached at most 1 m deep. 4. The long‐term P dynamics in the system, initially driven by allochthonous nutrient loadings were replaced by internal ecological processes. Water hyacinth removal markedly accelerated the process of eutrophication due to internal feedback mechanisms, leading to a switch to a more turbid state. Biological feedback mechanisms were driven by cyanobacterial blooms by enhancing water stability, oxygen anoxia at the bottom and by increasing suitable conditions for P internal loading. These data support the hypothesis of the role of cyanobacterial blooms as an important factor impairing water quality and driving the ecosystem towards a stable degraded state. 5. These findings have important implications for the restoration of shallow stratifying eutrophic lakes, as the alternative degraded state is most likely to occur when compared with their non‐stratifying counterparts. Moreover, feedback mechanisms in tropical and subtropical shallow lakes seem to be stronger than in temperate ones, as stratification events are more likely to occur over the year, intensifying system resilience to restorative strategies.     

Ligands of boron in Pisum sativum nodules are involved in regulation of oxygen concentration and rhizobial infection

Boron (B) is an essential nutrient for N₂‐fixing legume–rhizobia symbioses, and the capacity of borate ions to bind and stabilize biomolecules is the basis of any B function. We used a borate‐binding‐specific resin and immunostaining techniques to identify B ligands important for the development of Pisum sativum–Rhizobium leguminosarum 3841 symbiotic nodules. arabinogalactan–extensin (AGPE), recognized by MAC 265 antibody, appeared heavily bound to the resin in extracts derived from B‐sufficient, but not from B‐deficient nodules. MAC 265 stained the infection threads and the extracellular matrix of cortical cells involved in the oxygen diffusion barrier. In B‐deprived nodules, immunolocalization of MAC 265 antigens was significantly reduced. Leghaemoglobin (Lb) concentration largely decreased in B‐deficient nodules. The absence of MAC 203 antigens in B‐deficient nodules suggests a high internal oxygen concentration, as this antibody detects an epitope on the lipopolysaccharide (LPS) of bacteroids typically expressed in micro‐aerobically grown R. leguminosarum 3841. However, B‐deprived nodules did not accumulate oxidized lipids and proteins, and revealed a decrease in the activity of the major antioxidant enzyme ascorbate peroxidase (APX). Therefore, B deficiency reduced the stability of nodule macromolecules important for rhizobial infection, and for regulation of oxygen concentration, resulting in non‐functional nodules, but did not appear to induce oxidative damage in low‐B nodules.     

Evaluation of Cage-Traps and Cable Restraint Devices to Capture Red Foxes in Spain

Abstract: A growing international concern for the welfare of animals, combined with the need to capture and handle specific species for conservation, management, or recreational purposes, is increasing the need for scientific evaluation of capture methods. We evaluated the efficiency, selectivity, and injury of cable restraint devices and cage-traps for capture of red foxes (Vulpes vulpes) in 4 sites of Castilla y León (Spain) during spring of 2006. All traps performed similarly at all sites, with no apparent site and trap interactions. Fox capture rates and mechanical efficiencies of the Belisle® (Edouard Belisle, Saint Veronique, PQ, Canada) and Collarum® (Wildlife Control Supplies, East Granby, CT; mention of product names does not infer endorsement) were similar, but both had higher capture rates than the cage-trap. Similar to previous studies, the Collarum was 100% selective for canids and had a selectivity of 94.4 overall, which was higher than that for the Belisle (63.4); both Collarum and Belisle were much more selective than the cage-trap (21.4). Fox injuries were statistically indistinguishable using injury scores, but the Collarum and the Belisle surpassed international standards for humane trapping; an insufficient number of animals were captured in cage-traps to allow evaluation. Both the Collarum and the Belisle may be useful for the capture of foxes in Spain, but training and experience with each may be necessary to ensure the highest efficiency while preventing injuries, especially to nontarget species. (JOURNAL OF WILDLIFE MANAGEMENT 72(3):830–836; 2008)     

Thermal tolerance and recovery behaviour of Thorectes lusitanicus (Coleoptera,Geotrupidae)

1. Population differences in physiological responses are examined in Thorectes lusitanicus, an endemic Iberian dung beetle species, by submitting individuals of different populations to the same experimental and acclimation conditions. 2. An infrared thermography protocol was used, consisting of three assays: start of activity, cold response, and heat response. Individuals of 12 populations were studied and the comparative explanatory capacities of several environmental factors in relation to the observed inter‐population differences were examined. 3. The heating rate from chill coma to the beginning of activity was the variable with the highest discrimination power among the studied populations, accounting for 94% of the observed variance. Regarding the heat response, only six of the 16 thermal variables reached significance (inter‐population differences accounted for 52–74% in these six thermal parameters). 4. From the three considered environmental factors (Mediterranean climate, land cover, and trophic characteristics) only land cover characteristics remain statistically significant, affecting the cold response of individuals. 5. Thorectes lusitanicus is a species characterised by a high diversity of thermotolerance and recovery traits across populations with a low degree of association with broad environmental factors. Finally, it is suggested that the apterous character of this species could be a determinant factor explaining the high diversity of ecophysiological traits related to thermal stress tolerance and the recovery time.     

Concordant phylogeography and cryptic speciation in two Western Palaearctic oak gall parasitoid species complexes

Little is known about the evolutionary history of most complex multi‐trophic insect communities. Widespread species from different trophic levels might evolve in parallel, showing similar spatial patterns and either congruent temporal patterns (Contemporary Host‐tracking) or later divergence in higher trophic levels (Delayed Host‐tracking). Alternatively, host shifts by natural enemies among communities centred on different host resources could disrupt any common community phylogeographic pattern. We examined these alternative models using two Megastigmus parasitoid morphospecies associated with oak cynipid galls sampled throughout their Western Palaearctic distributions. Based on existing host cynipid data, a parallel evolution model predicts that eastern regions of the Western Palaearctic should contain ancestral populations with range expansions across Europe about 1.6 million years ago and deeper species‐level divergence at both 8–9 and 4–5 million years ago. Sequence data from mitochondrial cytochrome b and multiple nuclear genes showed similar phylogenetic patterns and revealed cryptic genetic species within both morphospecies, indicating greater diversity in these communities than previously thought. Phylogeographic divergence was apparent in most cryptic species between relatively stable, diverse, putatively ancestral populations in Asia Minor and the Middle East, and genetically depauperate, rapidly expanding populations in Europe, paralleling patterns in host gallwasp species. Mitochondrial and nuclear data also suggested that Europe may have been colonized multiple times from eastern source populations since the late Miocene. Temporal patterns of lineage divergence were congruent within and across trophic levels, supporting the Contemporary Host‐tracking Hypothesis for community evolution.     

Sexual cannibalism in scorpions: fact or fiction?

Females of several scorpion species are currently deemed to be sexual cannibals, but this reputation is, however, largely built on anecdotal evidence from the older literature. Theoretical models predict that given the low rates of female-male encounter, males should allow themselves to be cannibalized after sperm transfer. The present paper examines the information available for a number of species to determine (a) whether sexual cannibalism is as widespread as previously suggested, and (b) whether it is likely to involve male self-sacrifice. Information on three scorpions currently regarded as sexual cannibals is presented: the bothriurids Bothriurus bonariensis (C.L. Koch) and Urophonius jheringii Pocock, and the buthid Leiurus quinquestriatus (Hemprich & Ehrenberg). No post-mating cannibalism was ever observed in these species, nor in another 19 representatives of four families. In L. quinquestriatus only two cases of cannibalism of the male by the female occurred, both in the beginning of courtship and after long periods of food deprivation. We suggest that if sexual cannibalism occurs in scorpions, it is most likely to be of an 'economic' type, where females perceive males as prey rather than mates, and not a 'self-sacrificial', post-mating cannibalism enhancing the fitness of the offspring of the male.