Lm Strains Research Articles

Complete genome of the Listeria monocytogenes strain AUF, used as a live listeriosis veterinary vaccine

Listeria monocytogenes (Lm) is a highly pathogenic bacterium that can cause listeriosis, a relatively rare food-borne infectious disease that affects farm, domestic, wild animals and humans as well. The infected livestock is the frequent sources of Lm. Vaccination is one of the methods of controlling listeriosis in target farm animals to prevent Lm-associated food contamination. Here we report the complete sequence of the Lm strain AUF attenuated from a fully-virulent Lm strain by ultraviolet irradiation, successfully used since the 1960s as a live whole-cell veterinary vaccine. The de novo assembled genome consists of a circular chromosome of 2,942,932 bp length, including more than 2,800 CDSs, 17 pseudogenes, 5 antibiotic resistance genes, and 56/92 virulence genes. Two wild Lm strains, the EGD and the 10403S that is also used in cancer Immunotherapy, were the closest homologs for the Lm strain AUF. Although all three strains belonged to different sequence types (ST), namely ST12, ST85, and ST1538, they were placed in the same genetic lineage II, CC7.

Laser-induced 2D/0D graphene-nanoceria freestanding paper-based films for on-site hydrogen peroxide monitoring in no-touch disinfection treatments

A one-shot CO2 laser-based strategy to generate conductive reduced graphene oxide (rGO) decorated with nanoceria (nCe) is proposed. The 2D/0D rGO-nCe films, integrated as catalytic sensing layers in paper-based sensors, were employed for on-site monitoring of indoor fogging treatments against Listeria monocytogenes (Lm), a ubiquitous pathogenic bacterium. The rGO-nCe laser-assisted synthesis was optimized to preserve the rGO film morphological and electron-transfer features and simultaneously integrate catalytic nCe. The films were characterized by microscopical (SEM), spectroscopical (EDX, Raman, and FTIR), and electrochemical techniques. The most performing film was integrated into a nitrocellulose substrate, and the complete sensor was assembled via a combination of xurography and stencil printing. The rGO-nCe sensor's catalytic activity was proved toward the detection of H2O2, obtaining sensitive determination (LOD = 0.3 µM) and an extended linear range (0.5–1500 µM). Eventually, the rGO-nCe sensor was challenged for the real-time continuous monitoring of hydrogen peroxide aerosol during no-touch fogging treatment conducted following the EU’s recommendation for biocidal product use. Treatment effectiveness was proved toward three Lm strains characterized by different origins, i.e., type strain ATCC 7644, clinical strain 338, and food strain 641/6II. The sensor allows for discrimination and quantification treatments at different environmental biocidal amounts and fogging times, and correlates with the microbiological inhibition, promoting the proposed sensor as a useful tool to modulate and monitor no-touch treatments.Graphical

Molecular characterization of Listeria monocytogenes strains isolated from imported food in China from 14 countries/regions, 2003-2018.

Listeria monocytogenes (Lm) is associated with severe foodborne infections and ubiquitous in the nature. Identification of characteristics of Lm transmission through trading of food products is essential for rapidly tracking Lm sources and controlling dissemination of listeriosis. In this study, a total of 44 Lm strains were isolated from food products originating from 14 countries/regions during 2003-2018 at the Shanghai port. The genomes of these Lm strains were sequenced by high-throughput sequencing. Multilocus sequence typing (MLST) analysis showed that 43 isolates were divided into 17 sequence types (STs). The distribution of STs was decentralized, with the dominant ST2 accounting for only 18.18% of the strains. The LM63 strain did not match with any of the existing STs. Core-genome MLST (cgMLST) analysis based on 1748 core genes categorized the 44 strains into 30 cgMLST types (CTs), with CT10153 and CT7892 as the most predominant CTs. Notably, LM63 and LM67 shared the same CT in the cgMLST analysis. The phylogenetic analysis based on single-copy homologous genes revealed that the 44 Lm strains were primarily classified into two lineages. The SNP analysis also indicated that these strains were roughly divided into two clades, with strains in the first clade mainly collected earlier than those in the second clade, which were predominantly collected from 2010 onwards. The analysis using the virulence factor database (VFDB) indicated that the virulence gene inlJ was the most prevalent among these 44 strains. Notably, ddrA, msbA, and sugC were enriched in this dataset, requiring further clarification of their roles in Listeria through future studies. These results might provide a clue for understanding of the global epidemiology and surveillance of Lm and present insights for implementing effective measures to reduce or prevent Listeria contamination outbreaks in imported food products.

A Systematic Review of the Challenges of e-Learning Implementation in Sub-Saharan African Countries: 2016-2022

Even a cursory review of the research literature over the past two decades will reveal many e-learning implementation challenges facing sub-Saharan Africa Higher Education Institutions. The last available systematic review of these challenges was conducted in 2017. To address this gap and by following the PRISMA-ScR guidelines, our systematic review aimed to find answers to the following two research questions: (a) What e-learning challenges were identified in research conducted in Higher Education Institutions in sub-Saharan Africa from 2016, and (b) Which e-learning challenges identified before 2016 remain and which are new? Using search terms and synonyms associated with e-learning challenges, three reviewers performed a Boolean search in Google Scholar, Web of Science, and Scopus. After duplicates (113) were removed, a total of 353 potential articles were imported into a collaboration platform and filtered using the following inclusion criteria: (1) Higher Education Institutions in sub-Saharan Africa (2) English articles from 2016 to 2022 (3) user data on real-world e-learning implementations are available (4) use of a computer/mobile device excluding m-learning approaches and (5) blended or fully online courses. After a selection process where article sets were reviewed, rotated, and conflicts resolved, 25 articles that met the inclusion criteria formed the primary evidence base. After a data collection process and using selected Grounded Theory techniques, a total of 48 challenges, synthesised in 25 challenge categories, were identified across six themes: university-related (36 citations), instructor-related (20), access-related (14), LMS-related (10), student-related (9) and computer literacy-related (9). The main challenges categories were ICT infrastructure (14 citations), technical support (10) and student bandwidth (8), while the secondary challenges were mainly related to the instructor: commitment and capacity (7), capacity (6), LMS training (6) and instructional design and support (5). Suggestions are offered to address key challenges related to ICT infrastructure, technical support, and student bandwidth. Secondary challenges, which are mainly related to the instructor, are proposed to be resolved by a skilled and energetic instructional design and support team. Although many challenges identified before 2016 were reinforced in this investigation, no new challenges were identified. Overall, the number of citations for specific challenges shows that most challenges are context-specific, that is, limited to single HEIs. However, when combined in challenge categories and reflected as themes, the review reveals many challenges to remain persistent. To ensure that an institution, its instructors, and its students are geared and supported stepwise, a holistic top-down approach that considers all challenges is required. To achieve this, a recommendation for further research is the development of an updated SSA-specific checklist that can be used to rate e-learning readiness in terms of importance and context. The research aims to provide active researchers in the field with an update on the status of research on e-learning challenges in SSA, as well as to make decision-makers aware of potential challenges when implementing e-learning in the SSA context. This may result in better institutional strategies to support e-learning practitioners, as well as to ensure the effective development and implementation of e-learning management systems.

Multilab Validation Report for the Verification and Subtyping of Listeria monocytogenes Using qPCR

Listeria monocytogenes (Lm) is a Gram-positive bacterium that causes invasive listeriosis, an illness with high mortality and hospitalization rates. Due to the severity of illness associated with Lm, rapid identification and characterization of isolates from foods and the food-processing environment are critical to properly identify and track the pathogen and quickly remove adulterated foods from the market. Prior methods can rely on time-consuming biochemical or sera-agglutination assays to perform these tasks. Development of a high-throughput method that would rapidly perform these tasks is critical to improve response to contamination events. Previously, a single laboratory validation of a qPCR-based method was presented that could rapidly verify Lm isolates and characterize them into six molecular serogroups. In the current study, a multi-laboratory validation (MLV) was performed to evaluate the reliability of the qPCR method for identification and serogrouping of Lm isolates. Sixteen collaborating laboratories independently analyzed a panel of 43 blinded isolates plus three control strains using the qPCR method. This panel was comprised of representatives for non-Listeria (n = 7), Listeria sp. (n = 8), and Lm (n = 28) strains. The Lm isolates contained representatives of the six serogroups: 2A, 2B, 2C, 4B, NT, and 4bV/IVb-v1, with five strains for each serogroup except 4bV/IVb-v1 (n = 3). The results generated by 16 laboratories showed high sensitivity, specificity, and accuracy, generally ≥97%, for both the genus-species and serogrouping qPCRs. Results from one laboratory lowered the sensitivity of the non-Listeria group to 93%. These results indicated the method was highly reliable. However, only the previously evaluated serogroups were tested within the MLV panel, though there is the potential for other serogroup results. Sequence Read Archive (SRA) files for Lm isolates were evaluated to determine the frequency of other potential serogroup profiles. This effort identified a low percentage of isolates with atypical qPCR serogroups (0.30%) that are consistent with Lm and were generally associated with lineage II and the natural environment. In summary, the results indicate that the proposed qPCR method is reliable and has a high degree of sensitivity, accuracy, and specificity, while also decreasing hands-on analysis time and increasing throughput of the analysis.

Listeria monocytogenes Strains Persisting in a Meat Processing Plant in Central Italy: Use of Whole Genome Sequencing and In Vitro Adhesion and Invasion Assays to Decipher Their Virulence Potential.

In this study, we used both a WGS and an in vitro approach to study the virulence potential of nine Listeria monocytogenes (Lm) strains belonging to genetic clusters persisting in a meat processing plant in Central Italy. The studied clusters belonged to CC1-ST1, CC9-ST9, and CC218-ST2801. All the CC1 and CC218 strains presented the same accessory virulence genes (LIPI-3, gltA, gltB, and aut_IVb). CC1 and CC9 strains presented a gene profile similarity of 22.6% as well as CC9 and CC218 isolates. CC1 and CC218 showed a similarity of 45.2% of the same virulence profile. The hypervirulent strains of lineage I (CC1 and CC218) presented a greater ability to adhere and invade Caco-2 cells than hypovirulent ones (CC9). CC1 strains were significantly more adhesive and invasive compared with CC9 and CC218 strains, although these last CCs presented the same accessory virulence genes. No statistically significant difference was found comparing CC218 with CC9 strains. This study provided for the first time data on the in vitro adhesiveness and invasiveness of CC218-ST2801 and added more data on the virulence characteristics of CC1 and CC9. What we observed confirmed that the ability of Lm to adhere to and invade human cells in vitro is not always decipherable from its virulence gene profile.

Cell-surface anchoring of Listeria adhesion protein on L. monocytogenes is fastened by internalin B for pathogenesis

Listeria adhesion protein (LAP) is a secreted acetaldehyde alcohol dehydrogenase (AdhE) that anchors to an unknown molecule on the Listeria monocytogenes (Lm) surface, which is critical for its intestinal epithelium crossing. In the present work, immunoprecipitation and mass spectrometry identify internalin B (InlB) as the primary ligand of LAP (KD ∼ 42nM). InlB-deleted and naturally InlB-deficient Lm strains show reduced LAP-InlB interaction and LAP-mediated pathology in the murine intestine and brain invasion. InlB-overexpressing non-pathogenic Listeria innocua also displays LAP-InlB interplay. In silico predictions reveal that a pocket region in the C-terminal domain of tetrameric LAP is the binding site for InlB. LAP variants containing mutations in negatively charged (E523S, E621S) amino acids in the C terminus confirm altered binding conformations and weaker affinity for InlB. InlB transforms the housekeeping enzyme, AdhE (LAP), into a moonlighting pathogenic factor by fastening on the cell surface.

Change in antimicrobial susceptibility of Listeria spp. in response to stress conditions

Listeria species are exposed to various stressors throughout the food chain, which are crucial for microbe mitigation strategy in the food industry. However, the survival capabilities and development of antimicrobial resistance by Listeria spp. under different food processing environments (FPEs) stressors are not yet well understood. Hence, this study aims to determine the difference in survivability and antimicrobial susceptibility of L. monocytogenes (Lm) and other Listeria species (non-Lm) strains exposed to different FPEs stressors, including heat, acidic and alkaline pH, UV irradiation, and osmotic stress. For this, a collection of 11 Lm and 10 non-Lm strains were used to conduct experiments. This study showed that Lm strains were relatively more tolerant to environmental stresses than non-Lm strains (p > 0.05). Additionally, the evaluation of stress-induced resistance toward antimicrobials showed that anaerobic incubation, after exposition to environmental stresses, rendered Lm and non-Lm more resistant to antimicrobial agents than aerobic incubation. Furthermore, the study observed that different stressors induced an increase in minimum inhibitory concentrations (MICs) of certain antimicrobials. Specifically, heat stress persuaded an increase in MICs of tetracycline under aerobic incubation, and gentamicin and ciprofloxacin under anaerobic incubation. Acidic/alkaline pH induced an increase in MICs of gentamicin, ciprofloxacin, and trimethoprim-sulfamethoxazole, especially under anaerobic incubation. However, UV stress induced increase in MICs of tetracycline and trimethoprim-sulfamethoxazole under aerobic incubation and gentamicin, ciprofloxacin, and trimethoprim-sulfamethoxazole under anaerobic incubation. Additionally, osmotic stress induced an increase in MICs of tetracycline and ampicillin under aerobic incubation and gentamicin, tetracycline, and trimethoprim-sulfamethoxazole under anaerobic incubation. Collectively, this study highlights that stress tolerance may contribute to the predominance of Listeria species among FPEs and induce the development of antimicrobial resistance even without antibiotic selection pressure. The findings of this study may guide updated strategies to mitigate Listeria species in the food industry.

It takes a Village

Presentation: https://youtu.be/ff490WnkE2w
 Abstract:
 Many educational institutes have been challenged with the rapid response to developing appropriate “new norms” in the wake of the COVID-19 pandemic. This was complicated further at Auckland University of Technology (AUT) with the announced sundowning of the Learning Management System (LMS- Blackboard) in late 2020. With the forced transition to Canvas during a pandemic, AUT Learning Transformation LAB (altLAB) worked directly with academic and administrative staff to establish a collaborative approach to implementing Canvas tailored for each Faculty.
 The Faculty of Health and Environmental Sciences (FHES) established a Community of Practice (CoP) with staff from altLAB, “Canvas Champions”, Faculty Academic Advisors, and project Change Manager gaining direct relationships towards both the project management team and academics. CoP employs three key characteristics: (1) Joint Enterprise (shared domain of interest); (2) Mutual Engagement (engagement in joint activities and relationship building), and (3) Shared Repertoire (sharing of resources for practice) (Lave & Wenger, 1991; Nicolini et al., 2016; Wenger-Trayner et al., 2015). Joint Enterprise was readily established by inviting key members to a join a weekly “FHES Canvas Catch Up” with a shared aim of integrating well-designed curriculum and assessment into Canvas courses. Members of the group actively engaged in joint activities that both informed staff of the Canvas project, while opportunistically responding to the pandemic to “refresh” courses with pedagogically informed curriculum, assessment, and delivery of the Canvas builds. “How To” guides and Course Start Checklists were developed when gaps of learning were identified in the Canvas roll out, which were supported and reinforced by regular Q&A drop-in sessions and email-outs facilitated by the Canvas Champions.
 All 1837 AUT courses were transitioned for delivery in Canvas from Semester One, 2022. A standard template was developed and utilised to provide consistency across the university, with minor modifications to meet the needs of the individual faculties. As of October 2022, 97% of students (n=1898 responses) were accessing Canvas on a weekly basis with 66% daily; and 86% of students ‘satisfied’ (of which 36% were ‘extremely satisfied’). 83% of students perceived that they had a mostly consistent experience between courses.
 While student engagement and satisfaction were paramount and realised during the shift to Canvas (especially in disrupted times); the project was met with some challenges. For example, in response to COVID-19, the planned 18-month project was delayed resulting in a 15-month roll-out. Academics prioritise the transition to online delivery of teaching and learning over communications of the LMS implementation and training opportunities.
 This presentation will highlight some key learnings in the context of an LMS transition, with recommendations for future projects that may employ a Community of Practice for pedagogically informed curriculum and assessment design.
 References
 
 
 
 Lave, J., & Wenger, E. (1991). Situated Learning: Legitimate peripheral Participation. Cambridge University Press. https://doi.org/10.1017/CBO9780511815355
 
 
 
 
 
 Nicolini, D., Scarbrough, H., & Gracheva, J. (2016). Communities of Practice and Situated Learning in Health Care. Oxford University Press. https://doi.org/10.1093/oxfordhb/9780198705109.013.20
 
 
 
 
 
 Wenger-Trayner, E., Fenton-O'Creevy, M., Hutchinson, S., Kubiak, C., & Wenger-Trayner, B. (2015). Learning in landscapes of practice: boundaries, identity, and knowledgeability in practice-based learning. Routledge. https://doi.org/10.4324/9781315777122
 
 

Intra-Population Alteration on Voltinism of Asian Corn Borer in Response to Climate Warming.

The Asian corn borer (ACB) Ostrinia furnacalis (Guenée) can occur in one to seven generations annually from cool (48°00' N) to warm (18°10' N) region of corn cultivation in China. Although ACB is commonly known as a facultative larval diapause insect, the co-existence of various voltinism suggests that intra-population variation may have evolved for the nature of diapause, i.e., voltinism plasticity. Here, we conducted recurrent selection efforts to establish three strains of, respectively, univoltine (with obligate diapause), multivoltine (with facultative diapause), and non-diapausing ACB under various temperature and photoperiod environments. The univoltine (Lu) strain has evolved a stable univoltinism under a diapause suppressing condition (16 h daylength at 28 °C), with the diapause incidence constantly over 80% after three generations of selection. The multivoltine strain (Lm) under the high temperature (28 °C) was shown to have a typical facultative diapause induced by a range of short-day lengths (11-13.5 h). Diapause incidence was constantly <2.6% under the long day length (16 h) when the temperature was from 18 to 28 °C, i.e., low temperature could not enhance the diapause response in the Lm strain. However, the development was prolonged from 14.2 ± 0.3 d to 46.0 ± 0.8 d when the temperature was reduced from 28 °C to 18 °C. The majority (94.4%) of the developed Ln strain still maintained the non-diapausing nature under a diapause enhancing condition, i.e., a short (13 h) daylength at a low temperature (22 °C). Lm and Ln were able to complete their second generation in Heihe (50°14' N) if the first-generation moth oviposits before 18 June. The study suggests that ACB has evolutionary intra-population variation in voltinism. Under the climate change scenario warmer spring and summer might affect the proportion of sympatric voltine biotype populations that evolve toward being multivoltine.

Deciphering the global roles of Cold shock proteins in Listeria monocytogenes nutrient metabolism and stress tolerance.

Listeria monocytogenes (Lm) accounts for serious public health and food safety problems owing to its stress resilience and pathogenicity. Based on their regulatory involvement in global gene expression events, cold-shock domain family proteins (Csps) are crucial in expression of various stress fitness and virulence phenotypes in bacteria. Lm possesses three Csps (CspA, CspB, and CspD) whose regulatory roles in the context of the genetic diversity of this bacterium are not yet fully understood. We examined the impacts of Csps deficiency on Lm nutrient metabolism and stress tolerance using a set of csp deletion mutants generated in different genetic backgrounds. Phenotype microarrays (PM) analysis showed that the absence of Csps in ∆cspABD reduces carbon (C-) source utilization capacity and increases Lm sensitivity to osmotic, pH, various chemical, and antimicrobial stress conditions. Single and double csp deletion mutants in different Lm genetic backgrounds were used to further dissect the roles of individual Csps in these phenotypes. Selected PM-based observations were further corroborated through targeted phenotypic assays, confirming that Csps are crucial in Lm for optimal utilization of various C-sources including rhamnose and glucose as well as tolerance against NaCl, β-phenyethylamine (PEA), and food relevant detergent stress conditions. Strain and genetic lineage background-based differences, division of labour, epistasis, and functional redundancies among the Csps were uncovered with respect to their roles in various processes including C-source utilization, cold, and PEA stress resistance. Finally, targeted transcriptome analysis was performed, revealing the activation of csp gene expression under defined stress conditions and the impact of Csps on expression regulation of selected rhamnose utilization genes. Overall, our study shows that Csps play important roles in nutrient utilization and stress responses in Lm strains, contributing to traits that are central to the public health and food safety impacts of this pathogen.

Effects of experimental glaucoma in Lama1nmf223 mutant mice

To identify changes in response to experimental intraocular pressure (IOP) elevation associated with the laminin α1 nmf223 mutation in mice.Laminin mutant (LM) mice (Lama1nmf223) and C57BL/6J (B6) mice in two age groups each (4–5 months and >1 year) underwent intracameral microbead injections to produce unilaterally elevated IOP. We assessed axonal transport block of immunofluorescently labeled amyloid precursor protein (APP) after 3 days and retinal ganglion cell (RGC) axon loss after 6 weeks. Light, electron and fluorescent microscopy was used to study baseline anatomic differences and effects of 3-day IOP elevation in younger LM mice.In younger mice of both LM and B6 strains, elevated IOP led to increased APP block in the retina, prelaminar optic nerve head (preONH), unmyelinated optic nerve (UON), and myelinated optic nerve (MON). APP blockade not significantly different between younger B6 and LM mouse strains. Older LM mice had greater APP accumulation in both control and glaucoma eyes compared to older B6, however, accumulation was not significantly greater in LM glaucoma eyes compared to LM controls. Axon loss at 6 weeks was 12.2% in younger LM and 18.7% in younger B6 mice (difference between strains, p = 0.22, Mann Whitney test). Untreated LM optic nerve area was lower compared to B6 (nerve area, p < 0.0001, t-test). Aberrant axon bundles, as well as defects, thickening and reduplication of pia mater, were seen in the optic nerves of younger LM mice.Axonal transport blockade significantly differed between old B6 and old LM mice in control and glaucoma eyes, and younger LM mice had abnormal axon paths and lower optic nerve area.

Molecular typing and genome sequencing allow the identification of persistent Listeria monocytogenes strains and the tracking of the contamination source in food environments

The presence of Listeria monocytogenes (Lm) in the food processing environment (facilities and products) is a challenging problem in food safety management. Lm is one of the main causes of mortality in foodborne infections, and the trend is continuously increasing. In this study, a collection of 323 Lm strain isolates recovered from food matrices and food industry environments (surfaces and equipment) over four years from 80 food processing facilities was screened using a restriction site-associated tag sequencing (2b-RAD) typing approach developed for Lm. Thirty-six different restriction site-associated DNA (RAD) types (RTs) were identified, most of which correspond to lineage II. RT1, the most represented genotype in our collection and already reported as one of the most prevalent genotypes in the food environment, was significantly associated with meat processing facilities. The sequencing of the genomes of strains belonging to the same RT and isolated in the same facility in different years revealed several clusters of persistence. The definition of the persistent strains (PSs) allowed the identification of the potential source of contamination in the incoming raw meat that is introduced in the facility to be processed. The slaughterhouses, which, according to the European Union (EU) regulation, are not inspected for the presence of Lm could be hotspots for the persistence of Lm PSs.

An Implementation of Online Blended Learning Training and Learning Management System in The Covid-19 Pandemic

The existence of the COVID-19 pandemic outbreak has caused a high level of open unemployment in Indonesia. To overcome this problem, the Indonesian Ministry of Manpower held training to be given to people who do not have the skills and want to develop their skills. There are two kinds of training provided, namely blended learning training and online learning management system. This study aims to describe (1) the implementation of blended learning-based online training and LMS (2) Obstacles and solutions to the implementation of blended learning and LMS training. The research method used is a qualitative method. The research subjects included 2 training program managers, 2 training instructors, 2 training participants. The data collection techniques included in-depth interviews with 2 training program managers, 2 training instructors, 2 trainees, observation, and documentation related to the implementation of blended learning and LMS training. The validity of the data used triangulation of sources. The data analysis techniques consist of data collection, data reduction, data presentation and drawing conclusions. The results of the study explain that blended learning training was carried out with a portion of 30% online and 70% offline, while LMS was carried out online by combining an e-training platform as a learning support. The obstacle in implementing this training was the internet network connection. The solution provided was the training instructor reviewed the training material during offline training. In conclusion, BBPLK Semarang is one of the institutions that implement blended learning and LMS which has been held since the beginning of the pandemic by combining e-training. This implementation had several obstacles including internet connection and there were still some participants who did not have adequate gadgets. The implementation of this training was supported by the replacement of internet quota fees that could support blended learning and LMS training.

Pengembangan Learning Management System pada SMK PGRI 2 Badung

SMK PGRI 2 Badung is one of the schools that has been established in 1997. The current COVID-19 pandemic has resulted in the closure of schools as places to carry out teaching and learning activities. It forces the ministry of education to think of alternatives to continue the teaching and learning process. The utilization of e-learning can improve learning outcomes to the maximum. Likewise, SMK PGRI 2 Badung utilizes e-learning with an online learning platform, namely Google Classroom. Learning using Google Classroom encourages teachers to think creatively in delivering material and practicum. Online learning is done using Google Classroom with virtual face-to-face learning using Google Meet. However, for the collection of all school learning data, Google Classroom is lacking because the data storage of school learning activities is not centralized. For school archives, teachers still collect files to schools manually so that reporting is less structured and centralized. From the results of the analysis and interviews conducted, the author intends to do service with the title Training and mentoring for LMS development at SMK PGRI 2 Badung. The LMS that will be developed is an LMS with Moodle. Service activities are focused on developing the Moodle LMS, and training and mentoring the use of Moodle LMS. Activities are carried out using lecture, question and answer methods, and direct practice. From the LMS development and training activities, it was found that SMK PGRI 2 Badung has an independent LMS in the learning process, namely learning-smkpgri2badung. Sch. id and 80% of teachers can use Moodle LMS to help the learning process.

Retrospective Investigation of the Whole Genome of the Hypovirulent Listeria monocytogenes Strain of ST201, CC69, Lineage III, Isolated from a Piglet with Fatal Neurolisteriosis.

Listeria monocytogenes (Lm), the causative agent for both human and animal listeriosis, is considered to be a rare but potentially fatal foodborne pathogen. While Lm strains associated with current cases of human listeriosis are now being intensely investigated, our knowledge of this microorganism which has caused listerial infection in the past is still extremely limited. The objective of this study was a retrospective whole-genome sequence analysis of the Lm collection strain, 4/52-1953, isolated in the middle of the 20th century from a piglet with listerial neuroinfection. The multi-locus sequence typing (MLST) analysis based on seven housekeeping genes (abcZ, bglA, cat, dapE, dat, ldh, and lhkA) showed that the Lm strain 4/52-1953 was assigned to the sequence type 201 (ST201), clonal complex 69 (CC69), and phylogenetic lineage III. The strain 4/52-1953, similarly to other ST201 strains, probably originated from the ST9, CC69 via ST157. At least eight different STs, ST69, ST72, ST130, ST136, ST148, ST469, ST769, and ST202, were identified as the descendants of the first generation and a single one, ST2290, was proved to be the descendant of the second generation. Among them there were strains either associated with some sporadic cases of human and animal listerial infection in the course of more than 60 years worldwide or isolated from food samples, fish and dairy products, or migratory birds. Phylogenetic analysis based on whole genomes of all the Lm strains available in the NCBI GenBank (n = 256) demonstrated that the strain 4/52-1953 belonged to minor Cluster I, represented by lineage III only, while two other major Clusters, II and III, were formed by lineages I and II. In the genome of the strain 4/52-1953, 41 virulence-associated genes, including the Listeria pathogenicity island 1 (LIPI-1), and LIPI-2 represented by two internalin genes, the inlA and inlB genes, and five genes related to antibiotic resistance, were found. These findings can help to make the emergence of both hyper- and hypovirulent variants, including those bearing antibiotic resistance genes, more visible and aid the aims of molecular epidemiology as well.

Role of learning management system (canvas) training in the teacher’s performance and perception awareness - A case study of Bakhtar University, Kabul, Afghanistan

Inside the field of management, preparation and improvement of the institutional workplace is envisioned in improving the execution and awareness of people and groups within the organization. It is currently known by different names, counting personnel development, personnel training, and knowledge and education development. This review focused on researching the LMS (Canvas) preparation and improvement part in a case study employee practice at Bakhtar University in Kabul, Afghanistan. Universities commitments of other experts on the subject, preparation, preparation of strategies, benefits of preparing an LMS, personnel implementation, the relationship between preparation and improvement, and personnel application directed to interval examination were examined as a result of the study. The study took questions about the strategy of collecting information from as many as 125 people using tests from 96 staff at Bakhtar College in Afghanistan. Using published Tables to decide test measures based on distributed tables that give a set of test estimates for specific conditions. The test involved academic staff using the LMS (Canvas) to train college students. Using both formal and casual questions through 105 questionnaires, basic information was collected and disseminated to different staff across the faculties at the University. Because the researcher’s curiosity is to capture the thoughts, behaviors, and consequences of individuals to be considered, the researchers mainly uses clear research strategies and measures. The researchers relies on using basic and supporting information gathering strategies. It was well known that the preparation and development of the LMS (Canvas) had a positive role on both staff execution and awareness. This has contributed to tailored, convenient and high-quality LMS preparation. The researchers concluded that the preparation and progress of the LMS (Canvas) should be supported, considering the competitiveness, performance and perceptions of teachers, and student satisfaction. The study allows the assessment of training needs at Bakhtar University in Kabul, Afghanistan, on the basis of the goals of the organization as a whole. In addition, organizational policies should specify what LMS training programs should be planned for employees.

Atypical Serogroup IVb-v1 of Listeria monocytogenes Assigned to New ST2801, Widely Spread and Persistent in the Environment of a Pork-Meat Producing Plant of Central Italy.

In this study, we characterized 84 Listeria monocytogenes (Lm) strains having an atypical IVb-v1 profile and isolated in a meat producing plant of Central Italy. They were assigned to the new MLST type ST2801 (CC218). The new ST was widespread in the food-producing environment where it was able to persist for over a year even after cleaning and sanitation. Cluster analysis identified three main clusters genetically close to each other (0–22 allelic differences and 0–28 SNPs) from two different cgMLST types, suggesting a common source. The coexistence of closely related clusters over time could be the result of a different evolution path starting from a common ancestor first introduced in the plant and/or the consequence of the repetitive reintroduction of closely related clones probably by raw materials. All the strains presented several determinants for heavy metals resistance, stress response, biofilm production, and multidrug efflux pumps with no significant differences among the clusters. A total of 53 strains carried pLI100 and the j1776 plasmids, while in one strain, the pLM33 was found in addition to pLI100. Only the strains carrying plasmids presented cadA and cadC for cadmium resistance and the mco gene encoding a multicopper oxidase and gerN for an additional Na+/H+-K+ antiporter. All the strains presented a virulence profile including a full-length inlA gene and the additional LIPI-3. The isolation of a new ST with a large pattern of stress-adaptation genes and able to persist is an important contribution to deepening the current knowledge on the uncommon IVb-v1 and in general on the genomic diversity of Lm.

Listeria monocytogenes: Investigation of Fitness in Soil Does Not Support the Relevance of Ecotypes.

Listeria monocytogenes (Lm) is a ubiquitous bacterium that causes the serious foodborne illness listeriosis. Although soil is a primary reservoir and a central habitat for Lm, little information is available on the genetic features underlying the fitness of Lm strains in this complex habitat. The aim of this study was to identify (i) correlations between the strains fitness in soil, their origin and their phylogenetic position (ii) identify genetic markers allowing Lm to survive in the soil. To this end, we assembled a balanced panel of 216 Lm strains isolated from three major ecological compartments (outdoor environment, animal hosts, and food) and from 33 clonal complexes occurring worldwide. The ability of the 216 strains to survive in soil was tested phenotypically. Hierarchical clustering identified three phenotypic groups according to the survival rate (SR): phenotype 1 “poor survivors” (SR < 2%), phenotype 2 “moderate survivors” (2% < SR < 5%) and phenotype 3 “good survivors” (SR > 5%). Survival in soil depended neither on strains’ origin nor on their phylogenetic position. Genome-wide-association studies demonstrated that a greater number of genes specifically associated with a good survival in soil was found in lineage II strains (57 genes) than in lineage I strains (28 genes). Soil fitness was mainly associated with variations in genes (i) coding membrane proteins, transcription regulators, and stress resistance genes in both lineages (ii) coding proteins related to motility and (iii) of the category “phage-related genes.” The cumulative effect of these small genomic variations resulted in significant increase of soil fitness.

Scalable Manufacturing of Liquid Metal Circuits

AbstractLiquid‐metal (LM)‐based soft and stretchable electronics (SSEs) provide unique advantages for wearable computing, soft robotics, and implantable devices applications involving physical interaction with biological tissues and delicate objects. However, the lack of scalable manufacturing techniques prevents their mainstream adoption and commercialization. This paper introduces a scalable and reproducible manufacturing technique for wafer‐level fabrication of LM‐based SSEs, including LM‐only devices and hybrid circuits with LM and solid‐state microelectronics. The approach combines selective metal‐alloy wetting (SMAW) and controlled dip‐coating of eutectic gallium‐indium (EGaIn). The alloying of EGaIn and metal enables depositing EGaIn selectively onto the circuit layout defined by lithographically patterned copper traces on elastomer‐coated wafers. An automated, wafer‐level dip‐coating (DC) approach is developed to enable controllable and reproducible deposition of EGaIn. Wafer‐level simultaneous fabrication of many LM capacitors is demonstrated, and their geometric and electrical reproducibility is evaluated. The results indicate that the SMAW‐DC process can fabricate LM circuits reproducibly and rapidly. Hybrid SSEs are demonstrated by fabricating multiple wearable patches of ultra‐high frequency circuits with an LM dipole antenna, an LM strain gauge, and a solid‐state temperature sensor. Importantly, the presented technique can be integrated into the standard microfabrication flow for microelectronics, ensuring a high level of scalability.