Characterization of CRISPR-Cas Systems in Serratia marcescens Isolated from Rhynchophorus ferrugineus (Olivier, 1790) (Coleoptera: Curculionidae)

The CRISPR-Cas adaptive immune system has been attracting increasing scientific interest for biological functions and biotechnological applications. Data on the Serratia marcescens system are scarce. Here, we report a comprehensive characterisation of CRISPR-Cas systems identified in S. marcescens strains isolated as secondary symbionts of Rhynchophorus ferrugineus, also known as Red Palm Weevil (RPW), one of the most invasive pests of major cultivated palms. Whole genome sequencing was performed on four strains (S1, S5, S8, and S13), which were isolated from the reproductive apparatus of RPWs. Subtypes I-F and I-E were harboured by S5 and S8, respectively. No CRISPR-Cas system was detected in Si or S13. Two CRISPR arrays (4 and 51 spacers) were detected in S5 and three arrays (11, 31, and 30 spacers) were detected in S8. The CRISPR-Cas systems were located in the genomic region spanning from ybhR to phnP, as if this were the only region where CRISPR-Cas loci were acquired. This was confirmed by analyzing the S. marcescens complete genomes available in the NCBI database. This region defines a genomic hotspot for horizontally acquired genes and/or CRISPR-Cas systems. This study also supplies the first identification of subtype I-E in S. marcescens

Problem solving e negoziazione: un nuovo approccio alla gestione delle trattative sindacali. Il caso dell'Azienda Ospedaliero Universitaria Pisana.

Il presente elaborato ha come obiettivo quello di capire se le tecniche del Problem Solving possono essere adottate nell'ambito del processo di negoziazione e quanto esse potrebbero facilitare le trattative, raccogliendo anche il parere di soggetti che operano in questo settore. Il lavoro prende avvio definendo cosa debba intendersi per Problem Solving e spiegandone fasi e strumenti. Il focus dell'analisi è posto maggiormente sulle fasi in quanto in queste risiede il motivo del legame tra negoziazione e Problem Solving, infatti si metterà in evidenza quanto queste fasi tendano a coincidere in entrambi i processi. In seguito si introduce una nuova teoria della negoziazione chiamata “negoziazione di principi”, la quale permette proprio di mettere in evidenza il parallelismo tra Problem Solving e negoziazione e ci fa capire come attraverso l'utilizzo di strumenti tipici del Problem Solving la negoziazione possa essere condotta in maniera più efficace ed efficiente. Si passa quindi ad analizzare nel dettaglio la negoziazione, illustrando chi debba intendersi per “terza parte” e cosa ci si debba aspettare da questa, elencando i vari tipi di negoziazione, spiegandone, infine, le fasi che distinguono la negoziazione tradizionale dalla negoziazione di principi. La parte teorica dell'elaborato si conclude con l'analisi del contesto sindacale italiano. Essa parte dalla nascita del sindacato avente come obiettivo quello di facilitare la composizione degli interessi tra le parti nelle relazioni industriali, ne viene ripercorsa l'evoluzione e fornita una interpretazione di come questa altro non sia che un lungo viaggio verso il superamento delle originali contrapposizioni tra le parti alla ricerca di una comunanza di obiettivi. Al termine dell'elaborato sono illustrati i risultati di un'indagine condotta dalla scrivente presso l'Azienda Ospedaliero Universitaria Pisana, nella quale si è cercato di analizzare come davvero i problemi che si presentano all'interno dell'azienda vengano risolti e di capire se nel contesto delle trattative sindacali nelle amministrazioni pubbliche italiane sia possibile adottare un nuovo tipo di negoziazione basato sull'applicazione del Problem Solving in un'ottica di maggiore collaborazione

Identification of pigmented Serratia marcescens symbiotically associated with Rhynchophorus ferrugineus Olivier (Coleoptera: Curculionidae)

To characterize red pigment-producing bacteria (RPPB) regularly released during oviposition by red palm weevil (RPW), RPPB were recovered from eggs deposited in apples supplied as substrate for oviposition. The presence of RPPB was also detected from gut, the reproductive apparatus of dissected adult and virgin insects and from pupal cases collected within infested palms. RPPB were also identified all along the tissue of these palms. Analysis of the 16S rDNA, gyrB, rpoB, recA, and groEL sequences assigned RPPB to the species Serratia marcescens. RPPB exhibited an antimicrobial activity assessed by the agar well diffusion method against a number of gram-positive and gram-negative bacteria. In this study, we first report the identification of a red pigment-producing S. marcescens as extracellular symbiont of RPW. Route of transmission, detection within different organs, and a wide spread along the infested palm tissue, suggested S. marcescens is present as extracellular symbiont in different developmental stages of the RPW. Additionally, the antimicrobial activity exhibited versus Bacillus spp., Paenibacillus spp., and Lysinibacillus spp., reported as insect pathogens and potential candidates for biocontrol agents, could ascribe for S. marcescens a potential protective role

Antimicrobial resistance gene shuffling and a three-element mobilisation system in the monophasic Salmonella typhimurium strain ST1030

In this study we describe the genetic elements and the antimicrobial resistance units (RUs) harboured by the Salmonella Typhimurium monophasic variant 1,4,[5],12:i:- strain ST1030. Of the three identified RUs two were chromosomal, RU1 (IS26-blaTEM-1-IS26-strAB-sul2- IS26) and RU2 (IS26-tetR(B)-tetA(B)-ΔIS26), and one, RU3 (a sul3-associated class 1 integron with cassette array dfrA12-orfF-aadA2-cmlA1-aadA1), was embedded in a Tn21-derived element harboured by the conjugative I1 plasmid pST1030-1A. IS26 elements mediated the antimicrobial resistance gene (ARG) shuffling and this gave rise to pST1030-1A derivatives with different sets of ARGs. ST1030 also harboured two ColE1-like plasmids of which one, pST1030-2A, was mobilisable and the target of an intracellular translocation of the Tn21-derived element; the second (pST1030-3) was an orphan mob-associated oriT plasmid co-transferred with pST1030-1A and pST1030-2A. pST1030-2A and pST1030-3 also carried a parA gene and a type III restriction modification system, respectively. Overall analysis of our data reinforces the role played by IS26, Tn21-derived elements and non-conjugative plasmids in the spread of ARGs and supplies the first evidence, at least in Salmonella, for the identification of a natural isolate harbouring a three-element mobilisation system in the same cell

Filtering activity and nutrient release by the keratose sponge sarcotragus spinosulus schmidt, 1862 (Porifera, demospongiae) at the laboratory scale

Sponges are an important constituent of filter-feeder benthic communities, characterized by high ecological plasticity and abundance. Free bacteria constitute an important quota of their diet, making them excellent candidates in aquaculture microbial bioremediation, where bacteria can be a serious problem. Although there are studies on this topic, certain promising species are still under investigation. Here we report applied microbiological research on the filtering activity of Sarcotragus spinosulus on two different concentrations of the pathogenic bacterium Vibrio para-haemolyticus in a laboratory experiment. To evaluate the effects of the filtration on the surrounding nutrient load, the release of ammonium, nitrate, and phosphate was also measured. The results obtained showed the efficient filtration capability of S. spinosulus as able to reduce the Vibrio load with a maximum retention efficiency of 99.72% and 99.35% at higher and lower Vibrio concentrations, respec-tively, and remarkable values of clearance rates (average maximum value 45.0 ± 4.1 mL h−1 g DW−1 ) at the highest Vibrio concentration tested. The nutrient release measured showed low values for each considered nutrient category at less than 1 mg L−1 for ammonium and phosphate and less than 5 mg L−1 for nitrate. The filtering activity and nutrient release by S. spinosulus suggest that this species represents a promising candidate in microbial bioremediation, showing an efficient capability in removing V. parahaemolyticus from seawater with a contribution to the nutrient load

Bioinformatic survey of CRISPR loci across 15 Serratia species

The Clustered Regularly Interspaced Short Palindromic Repeats and CRISPR-associated proteins (CRISPR-Cas) system of prokaryotes is an adaptative immune defense mechanism to protect themselves from invading genetic elements (e.g., phages and plasmids). Studies that describe the genetic organization of these prokaryotic systems have mainly reported on the Enterobacteriaceae family (now reorganized within the order of Enterobacterales). For some genera, data on CRISPR-Cas systems remain poor, as in the case of Serratia (now part of the Yersiniaceae family) where data are limited to a few genomes of the species marcescens. This study describes the detection, in silico, of CRISPR loci in 146 Serratia complete genomes and 336 high-quality assemblies available for the species ficaria, fonticola, grimesii, inhibens, liquefaciens, marcescens, nematodiphila, odorifera, oryzae, plymuthica, proteomaculans, quinivorans, rubidaea, symbiotica, and ureilytica. Apart from subtypes I-E and I-F1 which had previously been identified in marcescens, we report that of I-C and the I-E unique locus 1, I-E*, and I-F1 unique locus 1. Analysis of the genomic contexts for CRISPR loci revealed mdtN-phnP as the region mostly shared (grimesii, inhibens, marcescens, nematodiphila, plymuthica, rubidaea, and Serratia sp.). Three new contexts detected in genomes of rubidaea and fonticola (puu genes-mnmA) and rubidaea (osmE-soxG and ampC-yebZ) were also found. The plasmid and/or phage origin of spacers was also established

ATP7B Variants as Modulators of Copper Dyshomeostasis in Alzheimer's Disease

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Molecular epidemiology and origin of cholera reemergence

Abstract In 1994 a cholera epidemic occurred in Italy and Albania after more than a decade of case absence. To investigate genotypic characteristics and the origin of the epidemic strains, 110 Vibrio cholerae O1 El Tor isolates from Italy and Albania were studied by randomly amplified polymorphic DNA analysis (RAPD), BglI ribotyping, and pulsed-field gel electrophoresis (PFGE) of genomic DNA. The Italian and Albanian strains were all ribotype 6 and their RAPD and PFGE patterns were identical as well. These findings indicated that the 1994 isolates belonged to the same clone and that the clone was part of the larger global spread of epidemic ribotype 6 strains, which started in southern Asia in 1990

Transcranial Magnetic Stimulation Studies in Alzheimer's Disease

Although motor deficits affect patients with Alzheimer's disease (AD) only at later stages, recent studies demonstrated that primary motor cortex is precociously affected by neuronal degeneration. It is conceivable that neuronal loss is compensated by reorganization of the neural circuitries, thereby maintaining motor performances in daily living. Effectively several transcranial magnetic stimulation (TMS) studies have demonstrated that cortical excitability is enhanced in AD and primary motor cortex presents functional reorganization. Although the best hypothesis for the pathogenesis of AD remains the degeneration of cholinergic neurons in specific regions of the basal forebrain, the application of specific TMS protocols pointed out a role of other neurotransmitters. The present paper provides a perspective of the TMS techniques used to study neurophysiological aspects of AD showing also that, based on different patterns of cortical excitability, TMS may be useful in discriminating between physiological and pathological brain aging at least at the group level. Moreover repetitive TMS might become useful in the rehabilitation of AD patients. Finally integrated approaches utilizing TMS together with others neuro-physiological techniques, such as high-density EEG, and structural and functional imaging as well as biological markers are proposed as promising tool for large-scale, low-cost, and noninvasive evaluation of at-risk populations