Heterologous expression of bacteriocin E-760 in Chlamydomonas reinhardtii and functional analysis

The use of antimicrobial peptides (AMPs) synthesized by bacteria (bacteriocins) is an alternative for combating multidrug resistant bacterial strains and their production by recombinant route is a viable option for their mass production. The bacteriocin E-760 isolated from the genus Enterococcus sp. has been shown to possess inhibitory activity against Gram-negative and Gram-positive bacteria. In this study, the expression of a chimeric protein coding for E-760 in the nucleus of C. reinhardtii was evaluated, as well as, its antibacterial activity. The synthetic gene E-760S was inserted into the genome of C. reinhardtii using Agrobacterium tumefaciens. A transgenic line was identified in TAP medium with hygromycin and also by PCR. The increment in the culture medium temperature of the transgenic strain at 35 °C for 10 minutes, increased the production level of the recombinant protein from 0.14 (Noninduced culture, NIC) to 0.36% (Induced culture, IC) of total soluble proteins (TSP); this was quantified by an ELISA assay. Recombinant E-760 possesses activity against Staphylococcus aureus in 0.34 U log, Streptococcus agalactiae in 0.48 U log, Enterococcus faecium in 0.36 U log, Pseudomonas aeruginosa in 2 U log and for Klebsiella pneumoniae, the activity was 0.07 U log. These results demonstrate that the nucleus transformation of C. reinhardtii can function as a stable expression platform for the production of the synthetic gene E-760 and it can potentially be used as an antibacterial agent.     

Bird's nest ferns promote resource sharing by centipedes

Bird's nest ferns (Asplenium spp.) support large numbers of invertebrates, including centipedes. As top invertebrate predators, centipedes drive ecosystem function, for example, by regulating decomposer populations, but we know little of their ecology in forest canopies. We provide the first detailed observations of the diversity and structure of the centipede communities of bird's nest ferns, revealing the importance of these epiphytes as nurseries for centipedes. We collected 305 centipedes equating to ˜11,300 mg of centipede biomass from 44 bird's nest ferns (22 of which were from the high canopy and 22 from the low canopy) in primary tropical rainforest in Sabah, Malaysian Borneo. Most abundant were the Scolopendromorpha (n = 227 individuals), followed by the Geophilomorpha (n = 59), Lithobiomorpha (n = 14), and Scutigeromorpha (n = 5). Although we observed very little overlap in species between the forest strata, scolopendromorph centipedes dominated throughout the canopy. Null model analysis revealed no significant competitive interactions; on the contrary, we observed centipedes sharing nest sites within the ferns on three of the ten occasions that we found nests. All nests belonged to centipedes of the family Scolopendridae, which are typically aggressive, and usually show negative spatial association. This study reveals a diverse community of canopy centipedes, providing further evidence of the importance of bird's nest ferns to a wide range of animals, many of which use the ferns at critical life stages. Future conservation strategies should regard these ubiquitous epiphytes as umbrella species and protect them accordingly in landscape management decisions.     

Mechanical performance of rat, mouse and mole spring traps, and possible implications for welfare performance

Lethal spring traps are widely used for killing small mammals in the UK. Many require government approval, based primarily on humaneness. However, mole traps and break-back traps for rats and mice are exempt; those available vary widely in price and apparent quality. The EU is considering implementing a Trapping Directive that would alter UK legislation, and a recent report advised the EU that trapping legislation should cover all trapped species and encourage improvement of traps. Mechanical trap performance is often used as an indicator of welfare impact. We examined the mechanical evidence for scope to improve the welfare standards of rat, mouse and mole spring traps. We measured mechanical performance among a range of rat, mouse and mole traps. Impact momentum values varied 6-8 fold, and clamping force values 4-5.5 fold, among traps for killing each species. There was considerable overlap in the performance of rat and mouse traps. Trap-opening angle and spring type were related to impact momentum and clamping force in traps for both species. There was no relationship between price and mechanical performance in traps for any species, except talpa mole traps. We are unable to judge the direct welfare impact of the traps tested, but rather the potential welfare threat associated with their exemption from approval. The wide variation in mechanical performance in traps for each species, overlap in performance between rat and mouse traps and increasing availability of weaker plastic rodent traps indicate considerable scope for improving the humaneness of spring traps for rats, mice and moles. We conclude that all such traps should be subject to the UK approval process. New welfare categories might improve trap standards further. Our results could also help improve rodent trap design and assist consumers in selecting more powerful traps. Many thousands of rats, mice and moles might benefit.     

Integration of EST-SSR markers of Medicago truncatula into intraspecific linkage map of lentil and identification of QTL conferring resistance to ascochyta blight at seedling and pod stages

Microsatellite markers have been extensively utilised in the leguminosae for genome mapping and identifying major loci governing traits of interest for eventual marker-assisted selection (MAS). The lack of available lentil-specific microsatellite sequences and gene-based markers instigated the mining and transfer of expressed sequence tag simple sequence repeat (EST-SSR)/SSR sequences from the model genome Medicago truncatula, to enrich an existing intraspecific lentil genetic map. A total of 196 markers, including new 15 M. truncatula EST-SSR/SSR, were mapped using a population of 94 F₅ recombinant inbred lines produced from a cross between cv. Northfield (ILL5588)?×?cv. Digger (ILL5722) and clustered into 11 linkage groups (LG) covering 1156.4?cM. Subsequently, the size and effects of quantitative trait loci (QTL) conditioning Ascochyta lentis resistance at seedling and pod/maturity stages were characterised and compared. Three QTL were detected for seedling resistance on LG1 and LG9 and a further three were detected for pod/maturity resistance on LG1, LG4 and LG5. Together, these accounted for 34 and 61% of the total estimated phenotypic variation, respectively, and demonstrated that resistance at the different growth stages is potentially conditioned by different genomic regions. The flanking markers identified may be useful for MAS and for the future pyramiding of potentially different resistance genes into elite backgrounds that are resistant throughout the cropping season.     

Contributions of the hippocampus and medial prefrontal cortex to energy and body weight regulation

The effects of selective ibotenate lesions of the complete hippocampus (CHip), the hippocampal ventral pole (VP), or the medial prefrontal cortex (mPFC) in male rats were assessed on several measures related to energy regulation (i.e., body weight gain, food intake, body adiposity, metabolic activity, general behavioral activity, conditioned appetitive responding). The testing conditions were designed to minimize the nonspecific debilitating effects of these surgeries on intake and body weight. Rats with CHip and VP lesions exhibited significantly greater weight gain and food intake compared with controls. Furthermore, CHip-lesioned rats, but not rats with VP lesions, showed elevated metabolic activity, general activity in the dark phase of the light-dark cycle, and greater conditioned appetitive behavior, compared with control rats without these brain lesions. In contrast, rats with mPFC lesions were not different from controls on any of these measures. These results indicate that hippocampal damage interferes with energy and body weight regulation, perhaps by disrupting higher-order learning and memory processes that contribute to the control of appetitive and consummatory behavior.     

Deletion of a ribosomal ribonucleic acid operon in Escherichia coli.

One (rrnE) of the seven operons which codes for ribosomal ribonucleic acid in Escherichia coli was deleted. No significant change in phenotype was observed even under maximum laboratory growth conditions.