Aeromonas caviae, Pseudomonas monteilii, Shewanella algae, Vibrio alginolyticus, V. brasiliensis, V. natriegens, V. parahaemolyticus, V. shilonii, V. xuii, Zobellella denitrificans which are Gram-negative, and Bacillus licheniformis and B. pumilus which are Gram-positive. Notably, six strains identified as Acinetobacter venetianus might be a concern of risk for shrimp industry. The antibiotic resistance pattern reveals that the strain YWO8-97 (identified as P. monteilii) was resistant to all twelve antibiotics. Ceftazidime was the most powerful antibiotic since most of the studied strains were sensitive against it. The six strains of A. venetianus showed multiple antibiotic resistance patterns. MAR index were ranged from 0.08 to 1.0, and values of 26 isolates were more than 0.2 which means prior high exposure to the antibiotics. From the present study, it can be concluded that shrimp PL nurseries in southern part of Bangladesh are getting contaminated with antibiotic resistant pathogenic bacteria.Shrimp aquaculture has been accomplished with breeding and nursing of shrimp in an artificial environment to fulfill the increasing demand of shrimp consumption worldwide. However, the microbial diseases appear as a serious problem in this industry. The study was designed to identify the diverse bacteria from shrimp PL (post-larvae) nurseries and to profile antibiotic resistance patterns. The rearing water  and shrimp PL were collected from eight nurseries of south-west Bangladesh. Using selective agar plates, thirty representative isolates were selected for 16S rRNA gene sequencing, antibiotic susceptibility test and MAR index calculation. Representative isolates were identified as  Penaeus monodon contributed 67% to the total earning from exported shrimp and prawn . With the intensification of shrimp farming, the demand of shrimp PL has been increased . In earlviz. Photobacterium phosphorum, P. leiognathi, Vibrio fischeri, V. harveyi, V. splendidus and V. vulnificus affect the hatcheries.Moreover, Monodon baculovirus (MBV), external fouling organisms, V. harveyi, V. anguillarum, V. vulnificus are found in shrimp eggs, PL, rearing tank water (treated water), marine water source (raw water) and feed (Artemia nauplii and microcapsulated feed)  were the main Gram-positive bacteria  . Some ofd shrimp . The bacbacteria . Eleven  in Asia . A sever harveyi . P. indi culture . The pre monodon . Early mientalis  and P. m monodon . However shrimps . Acinetoaculture . Still, Thus, proper biosecurity measures should be applied to prevent the diseases in shrimp aquaculture caused by several microorganisms. The implication of biosecurity in shrimp industry defines the practice of specific pathogen exclusion from broodstock, hatcheries, grow out farms, and from the entire area for disease anticipation . AlthougIn addition, antimicrobial resistance (AMR) has become a potential danger to public health, and the indiscriminate use of antibiotics in animals has been recognized as a major issue nowadays. Different types of antibiotics and other antimicrobial agents are used in shrimp production, predominantly in shrimp hatcheries of Bangladesh and elsewhere .Thus, the objective of this study was to identify diverse bacterial strains from the water and shrimp PL samples from different nurseries of shrimp farming regions of Bangladesh. Moreover, the antibiotic susceptibility and multiple antibiotic resistance profiles were also investigated for the representative bacterial isolates.viz. raw seawater, treated (after filtration and disinfection) water and outlet water were collected from eight nurseries of south-west region of Bangladesh , EMB (eosin methylene blue), SS , and MSA  agar media. For the isolation of Vibrio spp. the processed samples were inoculated in alkaline peptone water (APW) and incubated for 6–18 h at 37 °C for enrichment.PL samples were homogenized with physiological saline using sterile mortar-pestle at 1:9 ratios and taken separately into sterile zipper bags. Then, 100 μL of each sample solution was taken with 900 μL sterile physiological saline separately and diluted up to 10−1 and 10−3 dilution were inoculated on sterile TCBS , SS , MSA , and EMB  agar plates. Only from treated water, 100 μL of processed undiluted water sample and 10−2 dilution sample was inoculated on different agar plates. Then, the samples were spread thoroughly by using a sterile glass spreader. The plates were transferred into an incubator at 37 °C for 18–24 h. After incubation, the well discrete colonies from different plates were selected for subculture on TCBS, SS, MSA and EMB agar plates. Overall, 98 colonies were selected primarily and sub-cultured on respective selective agar plates for isolated single colonies. The selected colonies from respective agar plates were taken and streaked on the fresh respective plates to get isolated single colony for pure culture. Based on the colony morphology on selective agar plates, thirty representative isolates were cultured by streaking plate technique on TSA plates for further works.From diluted PL samples, raw water and outlet water samples, 100 μL of 10The heat extraction method was performed for the DNA extraction from 30 representative bacterial isolates . The tarA neighbor-joining phylogenetic tree was constructed using MEGA X software  for the The Kirby-Bauer disc diffusion technique  was perfThirty representative isolates were inoculated in Mueller-Hinton Broth (MHB)  and incubated for 24 h and then the broth culture was spread on the surface of the Mueller-Hinton Agar (MHA). The antibiotic discs  were applied on the surface of the agar plates and incubated for 24 h at 37 °C. Finally, the zone of inhibition was measured to detect susceptibility of the bacteria .The multidrug resistance and MAR index of the representative isolates were calculated against twelve antibiotics. The bacterial strains resistant against three or more antibiotics were considered as multidrug resistant strains . The MARA total of thirty representative isolates were selected from four selective agar plates for further study on the basis of different colors and sizes. The colony morphologies of representative isolates are presented in Vibrio spp. deposited in the NCBI database. YPL1-2 showed 100% identity with V. xuii strain DSM 17185. The isolates YPL1-4, YWO1-14, YPL2-19 and YPL4-52 revealed their similarity with V. shilonii strain having 100% query coverage and identity. YPL2-16 showed 100% identity and query cover with V. natriegens. The 16S rRNA gene sequence of YPL-18 was identified as V. parahaemolyticus. After searching the sequences in the GenBank, six isolates  redirect their 100% identity with V. alginolyticus. YWR2-22 and YWT5-55 showed identity more than 99% with V. brasiliensis. On the other hand, 15 remaining isolates showed their identity with eight different species. YWO1-12 and YWR2-23 were identified as Bacillus licheniformis and B. pumilus, respectively. These two isolates are Gram-positive.Based on the colony morphology, 30 representative bacterial isolates were selected for 16S rRNA gene sequencing in the present study. After amplification, all the representative isolates showed positive band estimated at 1,500 bp in gel electrophoresis . After sAcinetobacter venetianus. It is noteworthy that these isolates were screened mainly on EMB agar plates showing pinkish colored small colonies except YPL6-75, which was isolated from SS agar plate showing cream colored small size colony. The isolates YWO5-61 and YPL5-62 were identified as Zobellella denitrificans and Aeromonas caviae, respectively. The sequences of YPL5-64, YPL7-89 and YWT8-94 showed 100% query cover and identity with Shewanella algae. The sequence of YWO8-97 reflects 100% identity and query cover with Pseudomonas monteilii, and YPL8-98  showed similarity with Pseudomonas spp. in the NCBI database.Besides, the isolates YPL3-35, YWT4-39, YPL6-75, YWR7-79, YWO7-86 and YWR8-91 showed their 100% similarity with MT368010 to MT368038.The reported sequences in this study have been deposited in the NCBI GenBank database under accession numbers from The phylogenetic tree was constructed based on the neighbor-joining method using the 16S rRNA gene sequences of 29 representative isolates (YPL8-98 was excluded because of relatively short sequence), and other reference sequences downloaded from the GenBank . The treThe identified bacterial strains have been summarized in the P. monteilii) from outlet water of one nursery of Satkhira district that was resistant to all 12 antibiotics. On the other hand, V. shilonii, V. brasiliensis, B. licheniformis, V. natriegens, V. parahaemolyticus, B. pumilus, Z. denitrificans were quite sensitive to the tested antibiotics. In this study, among six isolates identified as V. alginolyticus, five isolates were resistant to ampicillin and gentamycin. All isolates of V. alginolyticus were resistant to penicillin. V. xuii  had the highest MAR index since the strain was resistant to all tested antibiotics  of culture ponds in Taiwan  in cultured shrimp  and Artemia nauplii disclosed that V. brasiliensis can cause mortality up to 100%  PL nurseries of Khulna and Satkhira districts of Bangladesh. A. venetianus is a novel species which was isolated from the sea in Japan, seawater in Israel, oil in Italy and aquaculture ponds in Denmark  cultured in freshwater  is characterized with a four-gene-cluster for which the bacterium can effectively synthesize biodegradable polyhydroxybutyrate  in salty environment  and nitrite-nitrogen (NO2-N), therefore no requirement to change the culture water  and the tetracycline class (38.7%); same study revealed that multidrug resistance was associated with 29% of Vibrio isolates , cefotaxmine (95%), ciprofloxacin (94%), piperacillin and tazobactam (98%)  . HoweverA. venetianus in the shrimp PL nurseries of southwest regions of Bangladesh. In China whiteleg shrimp, P. vannamei has been reported to be affected recently by A. venetianus causing red leg disease and the strain showed resistances to different antibiotics with multiple resistances against chloramphenicol, quinolones and tetracyclines . Therefore, the antimicrobial resistance emerges as a global threat, and to fight this battle the World Health Organization (WHO) launched a Global Action Plan (GAP) based on a ‘One Health’ approach which highlights the interdependence of human health, animal health and the environment (MAR index values ≥0.2 indicates that the bacterial strains were previously exposed to heavy antibacterial contamination, and therefore, are at high risk of multi-drug resistance . The preanalysis . Those sanalysis . Our stusistance . Howeversistance . Antimicironment .A. venetianus. The antibiotic susceptibility test proved that there were antibiotic resistant bacteria present in the study sites and most of the isolates showed multidrug resistance. From this study, it may be concluded that shrimp PL nurseries in southern part of Bangladesh are getting contaminated with the potentially pathogenic bacteria either through sea water they are getting from the open sea for rearing the PL or from vertical transmission of broodstock to PL, the feed they used for PL rearing and poor hygiene maintenance. Therefore, it is very important to pay attention to good rearing practice, hygiene, controlled use of antibiotic supplemented feed and overall bio-security measures in the shrimp PL nurseries and rearing farms.This study concludes the presence of both nonpathogenic and potentially pathogenic bacteria in shrimp PL and water samples collected from the nurseries of south-west region of Bangladesh. Specifically, there were some bacteria which have been proved in other studies as human pathogen. So, it can be said that there remains risk of bacterial disease while handling them. There were also some bacterial strains in the present study which were proved as pathogenic for shrimp from previous reports, especially


BackgroundThe aim of this study was to assess the predictive value of the Vandenberg and Kuse Mental Rotation Test (MRT) [1] on performance of novice medical students for manipulation of a nasal endoscope on a cadaveric model . Regarding spatial abilities, MRT is one of the most widely used [1]. The aim is to identify correct reproductions of a 3-dimensional reference figure.Materials and methodsWe randomly selected 39 medical students who had never handled a nasal endoscope and subjected them to the MRT. General information including experience in manual, technical, or surgical activities and testing of anatomical knowledge were collected [4]. They were then asked to perform series of cadaveric model exercises using a nasal endoscope. Their cadaver performance was evaluated by 2 blinded observers. We used an adapted version of the 7-point scale established by Lindquist et al . This is a standardized Global Rating Scale using a 5-point Likert-type scale with different dimensions . The overall score is calculated as the sum of the scores from the 7 subdimensions.ResultsWe found that medical students with higher MRT score had significantly increased endoscopic sinus performance . Higher anatomy score was also associated with better endoscopic sinus performance (P=.0141). Other parameters had no impact on endoscopic sinus performance .ConclusionsThe score obtained on the MRT was correlated with the practical performance of manipulating the nasal endoscope in cadaver. It could therefore be a useful spatial ability tool for directing targeted training in rhinology.AcknowledgementsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this abstractReferences1. Caissie AF, Vigneau F, Bors DA. What does the mental rotation test measure? An analysis of item difficulty and item characteristics. Open Psychol J. 2009, 2, 94-102.2. Reznick RK. Teaching and testing technical skills. Am J Surg. 1993;165:358-361.3. Jelovsek JE, Kow N, Diwadkar GB. Tools for the direct observation and assessment of psychomotor skills in medical trainees: a systematic review. Med Educ. 2013;47(7):650-673.4. Yamauchi Y, Yamashita J, Morikawa O, et al. Surgical skill evaluation by force data for endoscopic sinus surgery training system. MICCAI 2002, LNCS 2488, 44-51;2002.5. Lindquist NR, Leach M, Simpson MC, Antisdel JL. Evaluating simulator-based teaching methods for endoscopic sinus surgery. Ear Nose Throat J. 2019;98(8):490-495.6. Laeeq K, Waseem R, Weatherly RA, et al. In-training assessment and predictors of competency in endoscopic sinus surgery. Laryngoscope. 2010;120(12):2540-2545.BackgroundCrisis and disastrous events might have various impact in terms of human, material and financial costs [1]. Disaster medicine aims at optimizing resources management which the number of victims is high and resources limited, notably by using triage systems [2]. However, many difficulties persist when it comes to practice triage in real life [3], re-emphasizing the importance to train efficiently the healthcare professionals. In this context, simulation appears to be offer real interests. Within the framework of the INTERREG project "International Knowledge and Information Centre" (IKIC), a virtual reality environment simulating a train accident has been created and funded. We tested the impact of this training on participants’ satisfaction and learning.Materials and methodsThis quantitative study included 17 nurses specialised in emergency medical aid. These students participate in our training course which proceeded through several steps: pre-course e-learning, pre-briefing, briefing, virtual reality scenario and debriefing. Questionnaires were distributed before and after the simulation to assess satisfaction, self-efficacy and key learnings. The focus of the training was on the pre-triage of victims.ResultsOur training increased participants' sense of self-efficacy . We also noticed high levels of satisfaction and confidence in acquired learning and a high sense of presence. The majority (70.59%) of students felt that the environment had a positive impact on their performance. However, knowledge did not change after the training .ConclusionsIn this experience, using a simulation training with a virtual reality environment had a positive impact on participants satisfaction and perceived self-efficacy. The fidelity of the environment was underlined by the participants. In future research, it would be interesting to compare such virtual reality training with conventional training.References1. Merciefca JM, Ammirati CH, Amsallem C. La formation des infirmiers en médecine de catastrophe. Société française de médecine d’urgence. 2010 ; 1173- 1183.2. Jouffroy R, Nahon M, Delpech P, Puidupin A, Tourtier J-P, Carli P, Vivien B. Comment organiser les soins pré- et intrahospitaliers en cas d’affluence massive de patients ?. Réanimation. 2015 July; 24: 557-572.3. Cicero M, Whitfill T, Overly F, Baird J, Walsh B, Yarzebski J, Riera A, Adelgais K, Meckler G, Baum C, Cone D, Auerbach M. Pediatric Disaster Triage: Multiple Simulation Curriculum Improves Prehospital Care Providers' Assessment Skills’. Prehosp Emerg Care. 2017 Mar-Apr;21(2):201-208.BackgroundThe present study aims to demonstrate an improvement of intravenous cannulation skills after skill lab implementation for final year veterinary students.Materials and methodsPrevious experience, confidence to place an intravenous (iv) cannula and the success rate of iv cannula placement were evaluated in final-year veterinary students by using a questionnaire. First, the students enrolled in the study filled a questionnaire containing questions to determine previous experience and their personal opinion on the usefulness of mannequin experience, theoretical courses and their confidence to perform the cannulation successfully. Next, all students practiced iv cannulation during clinical rotation and their performance was recorded by auto-evaluation. Results were analysed by Chi-square tests where appropriate.ResultsSixty-one students enrolled in the “non-experienced” group (NonExp), 38 students enrolled in the “clinical experience” (ClinExp) group, 35 students enrolled in the “skill lab group” (SkillLab), and 12 students in the group with experience in both, clinical and skill lab mannequin (ClinSkil). Previous experience was significantly different among groups with 77 % of the NonExp students have placed less than 5 catheters in dogs and 74% of the ClinExp group have placed more than 5 catheters before starting their clinical rotation. Students of all groups felt equally confident to place an iv cannula successfully. In the SkillLab group, 43 % of the students felt that mannequin training helped them placing an iv cannula, while 34% thought that mannequin experience did not help them and 23% had no opinion on this question. Success rate of iv cannula placement during clinical rotation was not significantly different among groups but tended to be higher in students that felt more confident.ConclusionThe preliminary data presented here fails to demonstrate a difference between skill lab and clinical experience in iv cannula placement in final year veterinary students.BackgroundTechnical skills (TS) and non-technical skills (NTS) play a major role in ensuring patient safety , this study examines the impact of NTS on the management of the third stage of scorpion envenomation in a high-fidelity health simulation session.Materials and methodsThis is a descriptive study that targeted 24 students  in a master's degree in advanced health care at the Higher Institute of Health Sciences in Settat (Morocco). The main objective of the simulation session was to evaluate student’s TS and NTS that are crucially important in daily confronted situations by nurses (e.g the scorpion envenomation cases). TS were evaluated based on the national scorpion envenomation management algorithm [3]. The Anaesthetists Non-technical skills scale (ANTS) was used to assess NTS [4].ResultsIt was found that the high performing groups mastered all modalities of soft skills, TS and NTS were significantly correlated to each other . Also, learners expressed a positive attitude towards simulation-based learning.ConclusionsThis study showed that NTS are associated with TS. Mastering NTS is a key component to ensure effective decision-making, and particularly, in the third stage of scorpion envenomation case.References1. Riem N, Boet S, Bould MD, Tavares W, Naik VN. Do technical skills correlate with non-technical skills in crisis resource management: A simulation study. Br J Anaesth. 2012;109(5):723–8.2. Peltonen V, Peltonen LM, Salanterä S, Hoppu S, Elomaa J, Pappila T, et al. An observational study of technical and non-technical skills in advanced life support in the clinical setting. Resuscitation [Internet]. 2020;153:162–8. Available from: https://doi.org/10.1016/j.resuscitation.2020.06.0103. Bencheikh RS, Khattabi A, Faraj Z, Semlali I. Conduite à tenir devant une piqûre de scorpion au Maroc. Ann Fr Anesth Reanim. 2008;27(4):317–22.4. Fletcher G, Flin R, McGeorge M, Glavin R, Maran N, Patey R. Anaesthetists’ non-technical skills (ANTS): Evaluation of a behavioural marker system. Br J Anaesth. 2003;90(5):580–8.BackgroundEclampsia is a life-threatening emergency that causes approximately 60,000 maternal deaths per year worldwide [1]. Although global health training is developing with the increasing needs of the population [2]. Simulation has proven its importance as an innovative teaching strategy in the education of health science students [3]. And to asses these students, the Objective Structured Clinical Examination (OSCE) has been known as one of the most credible methods [4].Materials and methodsThis study aims to describe the impact of using high-fidelity simulation in the management of eclampsia amongst midwifery students, and to evaluate this impact through OSCE pre and post simulation. To do that, 31 second year midwifery students, were given a diagnostic assessment through the pre-simulation OSCE, followed by a high-fidelity simulation session, and then, the students were benefited from a post-simulation OSCE session for a summative evaluation. The general objective of all OSCE and simulation sessions, was the management of eclampsia crisis. Adapted assessment grids have been developed and validated according to the specific objectives of each OSCE station, and then a well-adapted evaluation grid that includes items describing technical skills, and non-technical skills was mobilized during the simulation session. The total score of all the grids was 20.ResultsThe average OSCE grid score for all three stations increased from 9.16/20 to 16.2/20 after the high-fidelity simulation session (p<0.001). The results of the simulation session grid show that the mean total score was 14.54±1.12 . There was a statistically significant relationship between the technical and non-technical skill scores (P<0.001).ConclusionThe present study joins simulation and OSCE in a common experience presenting the management of eclampsia. The simulation and OSCE must to be strongly integrated in the learning curriculum of midwifery students given their added value in the learning and assessment of midwifery students.References1. Raney JH, Morgan MC, Christmas A, Sterling M, Spindler H, Ghosh R, et al. Simulation-enhanced nurse mentoring to improve preeclampsia and eclampsia care: an education intervention study in Bihar, India. BMC Pregnancy Childbirth. dec 2019;19(1):41.2. Kynes JM, Kauffmann R, Walters CB, Sizemore C, Banerjee A. The Preparing Residents for International Medical Experiences (PRIME) Simulation Workshop: Equipping Surgery and Anesthesia Trainees for International Rotations. MedEdPORTAL [Internet]. 2021 ;17. Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7880254/3. Akalin A, Sahin S. The impact of high‐fidelity simulation on knowledge, critical thinking, and clinical decision‐making for the management of pre‐eclampsia. Int J Gynecol Obstet. sept 2020;150(3):354‑60.4. Kolivand M, Esfandyari M, Heydarpour S. Examining validity and reliability of objective structured clinical examination for evaluation of clinical skills of midwifery undergraduate students: a descriptive study. BMC Med Educ. déc 2020;20(1):96.BackgroundCOVID-19 pandemic has disrupted the real "face to face" clinical simulation which has been exchanged for the virtual "face to face"[1]. Virtual simulation as Screen based simulator (SBS) became the new paradigm for nursing students education to ensure pedagogical continuity. What are the benefits of SBS on the motivation, satisfaction, as an additional tool in training, preventing stress or perceived capacity to reinvest theoretical knowledges in clinical practice (CP)?Materials and methodsAn online questionnaire was sent to 323 French nursing schools (NS) who had free access to the 9 SBS from Simforhealth society. Students who did not have the benefit of these SBSs were able to view a video presentation so that they could answer the questions based on their perception as compared to students who had a real experience on SBS.ResultsAmong 323 NS, 220 transferred the survey to their students (68%) and 1600 students participated to the study. Some  were excluded for completeness. 870 students (63.8%) did not use SBS: 31(6.6%) for lack of interest, time  or information but wanted to use it . Both groups using or not SBS were totally agreed to consider SBS as an additional tool , no difference was found for motivation and satisfaction to use it F (1.1339) = 0.42, p = 0.52; F (1.1339) = 2.06, p = 0.15 reciprocally. Students who used SBS  were estimated their ability to reinvest their knowledges in CP with a high mean score . SBS could reduce stress before CP .ConclusionsStudents who did not have the opportunity to use the SBS during lockdown reported being as motivated and satisfied to use them in initial training as others to reduce their stress prior to CP.References1. Luctkar-Flude ML-F, Tyerman J. The Rise of Virtual Simulation: Pandemic Response or Enduring Pedagogy? 2021;57:1–2. doi:10.1016/j.ecns.2021.06.008.2. Foronda CL. What Is Virtual Simulation? Clinical Simulation In Nursing. 2021;52:8. doi:10.1016/j.ecns.2020.12.004.3. Lioce L, Lopreiato J, Downing D, Chang T, Robertson J, Anderson M, et al. Healthcare simulation dictionary . Agency for Healthcare Research and Quality. 2020. doi:DOI: 10.23970/simulationv2.BackgroundThe patient partnership model of Montreal has revealed new practices towards the acknowledgement of the patient's experiential knowledge. This model can be applied in healthcare, education and research settings [1]. Currently, the term "patient-partner" is not included in the SSH dictionary [2] or among the standards of best practices in simulation [3]. Regarding the simulation, the terms that can be found are the standardized patient, whose behaviour is standardized for summative evaluations, and simulated patient, whose behaviour is more authentic and more flexible to the needs of the learners. These two terms can be used to describe both the patients, but also the actors. For this reason, the naming can therefore lead to confusion when it comes to simulation : should we use patient-partner or actor as a “simulated patient”?Case reportOn the basis of the observations of simulation experiments (+/- 50 sessions in 5 years) carried out as part of an initial training programme in medical communication, we have defined the benefits and limitations of using patient-partner or simulated patients in the context of communication simulation. Indeed, in order to use patient-partners in simulation, several criteria should be fulfilled: the training of the patient but also an adequacy between the objective of the simulation and the patient profile. Also, the scenario should not include too many aspects of emotional patient management when involving the patient-partner instead of the simulated patient.In complex communication situations (bad news announcement), the patient partner can complete the role-play talking about his own experience. His involvement as a simulated patient must be thought through with all partners : teacher, old student, patient partner, simulated patient-actor and simulation trainer.ConclusionsThe applications of human simulation are as broad as the objectives they serve, and therefore require better definition of the contours of a partnership with patients.ReferencesMontreal model » : enjeux du partenariat relationnel entre patients et professionnels de la santé. Santé Publique 2015/HS (S1), pages 41 à 501. Pomey MP, Flora L. et al. Le « 2. Lioce L., Lopreiato J. et al. Healthcare Simulation Dictionary. Second Edition 2020. The Society for Simulation in Healthcare: {http://www.ssih.org/Dictionary}3. INACSL Standards Committee. INACSL Standards of Best Practice: Simulation (SM) - Simulation Glossary. Clinical Simulation Nursing. 2016; 12, S39-S47.BackgroundHealth simulation has become essential in many medical and paramedical disciplines. Our experience describes a discovery scenario used to introduce a one-day simulation training which was organized ten times. The goals are to reduce the participants’ stress and to create an atmosphere that encourages discussion and learning during the debriefing [1].Materials and methodsParticipants: two actors, the first is playing a physiotherapist supervisor and the second is playing a geriatric patient, they are also the simulation instructors, and four learners taking on their role of interns [2]. Discovery scenario: This scenario takes place in a private hospital room, in a geriatrics or pneumology department. The physiotherapist supervisor takes his trainees to put a patient back to bed and to administer an aerosol in a context of swallowing disorders and dementia. The supervisor is not focused on the situation; the patient is disoriented and uncompliant with the instructions. The supervisor lets the learners deal with the patient and doesn’t give them any support [3]. During the debriefing, the following topics are discussed: hierarchical relationship and its impact on stress and reflexivity. Finally, the impact on the stress level is assessed by the feeling expressed by the participants before and after the simulation.ResultsThe adjectives named by participants show that the stress has been reduced (before vs after): stress vs reassured, nervous vs comprehension of what is expected, worry vs impatient to discover the next simulation.ConclusionsWe observed that our discovery scenario has allowed the students to reduce their stress, to become involved in the simulation, and it has promoted communication and group cohesion. In order to further assess the stress level of this scenario on the learners, it would be interesting to conduct a study on this subject.References1. Gonzalez-Caminal, G., Camps, A., Lorza, G., & Jubany, J. (2018). Simulation as an experience to prepare physiotherapy students for clinical practice - Preliminar perspectives at faculty of Health Sciences at Manresa. 12th Internaional Technology, Education and development Conference. Valencia. doi:10.21125/inted.2018.16142. Kameg, K. M., Szpak, J. L., Cline, T. W., & Mcdermott, D. S. . Utilization of Standardized Patients to Decrease Nursing Student Anxiety. Clinical Simulation in Nursing, 10(11), pp. 567 - 573. doi:http://dx.doi.org/10.1016/j.ecns.2014.09.006.3. Nielsen, B., & Harder, N. . Causes of Student Anxiety during Simulation: What the Literature Says. Clinical Simulation in Learning, 9(11), pp. e507 - e512. doi:http://dx.doi.org/10.1016/j.ecns.2013.03.003BackgroundMany studies have addressed the consequences of public health crises . However, very few authors have explored the impact of these crises on hospitals organization. The aim of this study was to identify changes in the organizational and managerial context in the hospital environment following the Covid crisis.Materials and methodsA mixed methods approach with a convergent design was adopted. For the quantitative phase (n=98), the validated COMEt questionnaire was administered to a convenience sample of physicians, nurses and caregivers to measure the managerial and organizational context before and after the crisis. For the qualitative phase (n=102), a convenience sample of the same healthcare workers and managers were recruited to attend interviews. Questions were asked about the hospital crisis management using the plus/delta method. Data were then integrated through a data transformation model.ResultsResults showed multiple tendencies following the first wave of the Covid crisis. These changes may have impacted Covid and Non-Covid units differently. Interprofessional relationships tended to improve , as well as relationships with managers . Interviewees stated a fracture between lower and top-level management. Staff satisfaction  and perceived performance of the unit  also seemed to decrease while burnout was felt higher after the crisis . Although there is no consensus, the results highlighted a better crisis management and resilience among Covid units compared to Non-Covid .ConclusionsThe Covid crisis seemed to have changed the hospital organizational and managerial context. This experience should encourage improved communication within hospitals management levels and better team spirit. The multidisciplinary approach should also be given more consideration to improve work environment to speak as teams rather than functions.References1. Tisdell CA. Economic, social and political issues raised by the COVID-19 pandemic. Econ Anal Policy 2020;68:17–28. doi:10.1016/j.eap.2020.08.002.2. Pak A, Adegboye OA, Adekunle AI, Rahman KM, McBryde ES, Eisen DP. Economic Consequences of the COVID-19 Outbreak: the Need for Epidemic Preparedness. Front Public Heal 2020;8:19. doi:10.3389/fpubh.2020.00241.


Escherichia coli and Klebsiella pneumoniae are seldom studied in Vietnam. In this study, we investigated ESBL-producing Enterobacterales in the birth canal of pregnant women. Between 2016 and 2020, vaginal swabs were collected from 3104 pregnant women  and inoculated onto MacConkey agar plates. Colonies were subjected to direct identification and antimicrobial susceptibility testing using the VITEK®-2 automated compact system and disk diffusion. ESBL production was determined phenotypically. E. coli, Klebsiella species were identified in 30% (918/3104) of the vaginal swabs, with E. coli being the most common . ESBL-production was detected in 47% (432/918) of Enterobacterales, with frequent multidrug-resistant phenotype. The overall prevalence of carbapenem resistance was low (8%). Over 20% of Klebsiella spp. were carbapenem-resistant. Pregnant women had a high prevalence of colonization and may transmit ESBL-E to neonates at birth, an important risk factor to be considered. The high rate of ESBL-producers and carbapenem resistance in Enterobacterales in Vietnam emphasizes the need for consequent surveillance and access to molecular typing.Extended-spectrum β-lactamase-producing Enterobacterales (ESBL-E) resistance to commonly prescribed drugs is increasing in Vietnam. During pregnancy, ESBL-E may predispose women to reproductive tract infections and increases the risk for neonatal morbidity. Vaginal colonization and infections by  Antimicrobial resistance (AMR) is increasingly recognized as a global public health issue in many countries. With a population of more than 96 million and a high burden of infectious diseases, Vietnam faces a significant increase in AMR in the last years.Escherichia coli and Klebsiella pneumoniae [E.coli and K. pneumoniae being the predominant contributors of ESBL-E in Vietnam [In low and middle-income countries, antibiotic resistance is increasing with high morbidity and mortality, but resistance data has been poorly documented. The WHO recently declared that the group of bacteria most commonly causing hospital-acquired infections (HAIs) in Asia as priority 1 includes  Vietnam .E. coli infections of 59% and 81%, respectively [Reports from 2002 until 2011 suggest an exponential increase in resistance due to ESBLs ESBL from 39 to 55% in Southeast Asia [ectively . A pointectively .E. coli and K. pneumoniae as vaginal colonizers is alarming and of clinical relevance due to limited antibiotic options to treat pregnant women and neonatal early onset sepsis [Maternal colonization with group B streptococci (GBS) and Enterobacterales is considered a significant risk factor for the acquisition of neonatal sepsis . While At sepsis .E. coli and Klebsiella spp. isolates in vaginal swabs of pregnant women have not been studied. The knowledge of the burden of colonization in pregnant women and the extent of maternal-neonatal transmission is important for AMR surveillance and subsequent prevention strategies [In Vietnam, the incidence of ESBL-producing strains among clinical rategies . In thisA total of 3863 pregnant women with a mean age of 30 years (17–46) and a mean gestational age of 31 weeks (range 16–41 weeks) were enrolled in the study. Vaginal swab specimens were collected from 3104 pregnant women and were subsequently investigated .E. coli and Klebsiella spp. Among them, E. coli was predominant , followed by K. pneumoniae , and K. oxytoca . The rates of colonization by ESBL-positive and ESBL-negative Enterobacterales in all swab samples were 14% (n = 432) and 16% (n = 486), respectively. ESBL production was detected in 47% (432/918). Among the 432 ESBL-positive isolates, E. coli was predominant , followed by K. pneumoniae , and K. oxytoca . There were no significant differences in the distribution of E. coli and Klebsiella spp. between the ESBL-positive and negative isolates  isolates were positive for isolates .E. coli and Klebsiella spp., 97% (n = 891/918) showed antibiotic resistance to at least one antimicrobial agent. Due to nonuniformity in the availability of susceptibility data, only resistance data with complete antimicrobial susceptibility for ampicillin, ceftriaxone, ertapenem, gentamicin, ciprofloxacin and trimethoprim/sulfamethoxazole (n = 904/918) were considered for better comparability  were ESBL producers. MDR was frequent in ESBL-positive E. coli than ESBL-negative E. coli . Co-resistances to gentamicin, ciprofloxacin and trimethoprim/sulfamethoxazole were frequent. Only 3% (22/657) of E. coli tested were resistant to carbapenem  and of 8 K. oxytoca, 50% (4/8) were ESBL-producers. Like E. coli, MDR was more frequent in ESBL-producers . In ESBL-producers, the prevalence of carbapenem resistance was low (3% towards ertapenem and 9% towards imipenem). Surprisingly, a significantly higher percentage of ESBL-negative Klebsiella spp. were carbapenem-resistant   and ArgeE. coli and K. pneumoniae are the most commonly isolated gram-negative Enterobacterales and were known to be the major causative agents of neonatal ESBL-E infections [E. coli and K. pneumoniae as the two most common resistant priority pathogens [E. coli colonization rate of 21% observed in this study is a risk factor for preterm birth [E. coli colonization is associated with obstetric infections and neonatal infections. The colonization rate was higher than other studies, e.g., 20% in Lithuania [K. pneumoniae isolates observed in this study is lower than those reported from Bangladesh (9%) [E. coli and K. pneumoniae vary depending on the regions [E. coli in this study was 11%, which is significantly higher [In this study, fections ,21. Furtfections  and are fections ,24. The athogens . The vagrm birth . Vaginalithuania , 14% in ithuania . The 8% esh (9%)  and in Nesh (9%) . The rat regions  and ESBLy higher .E. coli. A recent review showed a high prevalence of antibiotic resistance to commonly used antibiotics in commensal E. coli isolates from humans in low-middle-income countries (LMICs), including Vietnam; ampicillin (72%), oxytetracycline (78%), tetracycline (67%), streptomycin (58%) and trimethoprim (67%), chloramphenicol (45%), ciprofloxacin (17%), co-trimoxazole (63), nalidixic acid (30%) and third-generation cephalosporin–cefotaxime (27%) [E. coli resistance to ampicillin was 92% and 56% to third-generation cephalosporin, higher than another study [Klebsiella sp. are resistant to third-generation cephalosporin, and over 36% were ESBL-producers, which is in line with previously reported epidemiological data from this region [K. pneumoniae and K. oxytoca were carbapenem-resistant, which was much higher than anticipated. The acquisition of carbapenem-resistant Enterobacterales in Vietnam through contact with hospital settings has been described previously [Klebsiella spp. is alarming and warrants further investigations.In the presented study, the Enterobacterales isolates were observed to be more resistant to commonly prescribed drugs, such as ciprofloxacin (41%), trimethoprim/sulfamethoxazole (69%), and third-generation cephalosporins  . In thiser study . Furthers region . Unexpeceviously . The higK. pneumoniae infections has led to the rapid emergence of carbapenem resistance [Most of the commonly prescribed drugs revealed high resistance; drugs such as broad-range penicillin, cephalosporins and carbapenems, may be considered as an alternative treatment option. Carbapenems are considered drugs of choice for infections caused by ESBL-producing enterobacteria ,37. Howesistance . TherefoThere are a few study limitations. A major limitation of this study is that AMR data were generated in the routine microbiological diagnostic and molecular characterization could not be performed routinely due to limited resources. Since bacterial isolates were not cryopreserved, retrospective genotypic analysis for ESBL-producers could not be performed. Nonetheless, the presented findings highlight the ongoing AMR problem in Vietnam and raise important issues to access molecular typing methods/facilities in low and middle-income countries. Genotypic analysis of the observed drug-resistant isolates would have been valuable to verify the findings. Other limitations include that the study did not examine when pregnant women became colonized with ESBL-E. Second, the study did not examine potential risk factors that promote ESBL-E colonization/infection, which would have allowed for any intervention for clinical management. Finally, the study does not document antibiotics taken as self-medication or prescribed during pregnancy.n = 3104) were collected from hospitalized pregnant women and screened for Gram-negative bacteria. All clinical and demographic parameters are summarized in Informed written consent was obtained from all study participants. The study was approved by the Institutional Review Board of Vietnam Military Medical University, Hanoi, Vietnam. The study was conducted between January 2016 and November 2020 at the National Hospital of Obstetrics and Gynecology (NHOG) in Hanoi, Vietnam, the largest healthcare center and a referral hospital for northern Vietnam, serving a population of ~55 million people with 450,000 deliveries annually. Vaginal swab samples  with VITEK® 2 GN ID card  for species-level identification of clinically important Gram-negative bacilli, including a broad range of Gram-negative Enterobacterales, non-Enterobacterales and other highly pathogenic organisms.Vaginal samples were processed for detection of Enterobacterales according to routine laboratory protocols. In brief, vaginal swab samples were inoculated onto MacConkey agar plates  and incubated at 37 °C for 24 h to grow Gram-negative enteric bacteria. The identified colonies were subjected to direct identification and antimicrobial susceptibility testing using the VITEKE. coli ATCC 25922 and K. pneumoniae ATCC 700603 (for β-lactam/β-lactamase inhibitor combination).ESBL phenotypic confirmation was performed using a combined disc diffusion method recommended by Clinical and Laboratory Standards Institute-2018  . Briefly® 2. In case of unavailability of VITEK® 2 test kits, susceptibility testing was performed by disk diffusion.A large proportion of the bacterial isolates were tested for susceptibility to different classes of antibiotics, including β-lactams , cephalosporins/cephamycins , carbapenems , aminoglycosides , tetracycline, quinolones , sulfonamides (trimethoprim/sulfamethoxazole), fosfomycin and nitrofurans (nitrofurantoin) using VITEKp-value of 0.05 was considered statistically significant. Comparisons of associations between Enterobacterales carriage, antibiotic susceptibility were computed using a chi-squared test.Statistical analyses were performed using Stata 13.1  and R 4.0.2 . All tesKlebsiella spp. in Vietnam is alarming and highlights the necessity for implementing AMR surveillance in Vietnamese hospitals. Most importantly, access to molecular typing methods in low- and middle-income countries is desperately needed to better understand spreading AMR in the global context.Pregnant women had a high prevalence of colonization and may transmit ESBL-E to neonates at birth, an important risk factor for early-onset neonatal sepsis. However, further studies are needed to validate our findings and to assess the necessity of ESBL screening in pregnant women. The high prevalence of ESBL-producing Enterobacterales and carbapenem resistance in


Fusobacterium nucleatum is among the most studied bacteria in digestive tract cancers and has been described as a poor prognostic factor in several digestive cancers. We retrospectively analyzed surgical samples from a homogeneous multicenter cohort of 166 patients with ASCC who underwent abdominoperineal resection. This study showed that F. nucleatum was an independent predictor of favorable overall survival and disease-free survival. This allowed the identification of a patient subgroup with a good prognosis (upper tercile). Our current work strengthens the new insight into the prognostic role of intratumoral F. nucleatum in cancer patients. Validation of these findings would allow to guide therapeutic strategies in dedicated trials by proposing intensification or de-escalation of systemic treatments and follow-up according to F. nucleatum loads.The main prognostic factors of localized/locally advanced anal squamous cell carcinoma (ASCC) are insufficient to predict 10–20% of metastatic relapses. Fusobacterium nucleatum has been associated with poor prognosis in several digestive cancers. In this study, we assessed the association between intratumoral F. nucleatum load and clinico-pathological features, relapse, and survival in patients with ASCC who underwent abdominoperineal resection (APR) after RT/CRT. We retrospectively analyzed surgical samples from a cohort of 166 patients with ASCC who underwent APR. F. nucleatum 16S rRNA gene sequences were quantified using real-time quantitative PCR. We associated F. nucleatum load with classical clinicopathological features, overall survival (OS), disease-free survival (DFS), and metastasis-free survival (MFS) using Cox regression univariate and multivariate analyses. Tumors harboring high loads of F. nucleatum (highest tercile) showed longer OS and DFS . High F. nucleatum load was a predictor of longer OS  and DFS  in multivariate analysis. High F. nucleatum load is an independent favorable prognostic factor in patients with ASCC who underwent APR.Main prognostic factors of anal squamous cell carcinoma (ASCC) are tumor size, differentiation, lymph node involvement, and male gender. However, they are insufficient to predict relapses after exclusive radiotherapy (RT) or chemoradiotherapy (CRT).  Anal canal cancer (squamous cell carcinoma in 95% of cases) is a rare disease accounting for 2.5% of digestive cancers . Human pMost patients 95%) present with local/locoregional disease at diagnosis. The aim of the treatment of localized/locally advanced ASCC is to cure the patient and achieve the best local control while maintaining a functional anal sphincter. The treatment is based on radiotherapy (RT), usually combined with 5-fluorouracil-based chemotherapy , and achieves approximately 80% of complete pathological response with a recurrence-free survival at 3 years of approximately 70% . Surgery% presentFusobacterium nucleatum is among the most studied bacteria in digestive tract cancers and has been described as a poor prognostic factor in esophageal [F. nucleatum load was associated with longer survival in oral squamous cell carcinoma (OSCC) and was associated with a favorable immune microenvironnement [The main prognostic factors of localized/locally advanced ASCC are tumor size, differentiation, lymph node involvement, HPV status, and male gender ,8. Howevophageal , gastricophageal , pancreaophageal , and colophageal ,16,17 caonnement .F. nucleatum load and clinicopathological features, relapse, and survival in a homogeneous multicenter cohort of patients with ASCC who underwent APR after the failure of RT or CRT.In this study, we assessed the association between intratumoral This retrospective multicenter study involved nine French centers and included all consecutive ASCC patients who underwent APR for tumor persistence or local relapse after RT or CRT from January 1996 and February 2016. We selected all patients with complete clinical and histological data and a follow-up of at least 2 years. The diagnosis of ASCC was confirmed by histology in all cases. Demographic, clinical data and tumor features, details on initial treatment by RT or CRT, indication for APR , and histological parameters from the APR were collected. After completion of RT or CRT, a persistent ulceration or a re-emergence of the anal lesion within 6 months of completion of RT was classified as persistent disease, while lesions appearing after 6 months post-RT were classified as a relapse.Relapse was defined by the first occurrence of one of the two following events after APR: local for pelvic relapse, and metastatic for distant relapse. The study was conducted in accordance with the ethics principles of the Declaration of Helsinki and the General Data Protection Regulation (GDPR). According to French regulations, this study did not need informed consent. Patients were informed of the study by each investigator and did not express opposition.F. nucleatum analysis, we included negative controls (buffers/reagents without tumor samples) and the samples were manipulated under a hood with masks and gloves.For each patient, six tissue sections of 6 μm thickness were obtained from FFPE samples and a seventh tissue section was stained with HE. The tumor-rich areas were macrodissected using a single-use blade and the samples underwent proteinase K digestion in a rotating incubator at 56 °C for 3 days. DNA was extracted with the NucleoSpin kit  according to supplier recommendations. DNAs were quantified using Nanodrop spectrophotometer ND-1000 . In order to rule out external contaminations for F. nucleatum was quantified using a real-time quantitative PCR according to the same protocol as our previous study [F. nucleatum levels on the basis of JUN contents [us study . Brieflycontents .HPV detection and genotyping were performed using Real-time PCR and specific primers for HPV16, and PCR to detect HPV L1 DNA and Sanger sequencing for HPV16-negative samples as previously described .n > 60) and Fisher’s exact test for small samples (n < 60). Statistical significance was set at p < 0.05.Associations among binary variables were assessed by the Chi-squared test for large samples (F. nucleatum are very heterogeneous among the population and one-third of the population has extremely low quantities < 0.001 (n = 52)  . Cuttingp < 0.05) were entered into the final multivariable Cox regression model, after considering redundancy between variables. Gender and initial stage were included in the model as it is a known prognostic factor.Survival endpoints were defined according to the DATECAN consensus . OverallUnivariate and multivariate Cox regression analyses and Kaplan–Meier curves were computed using the survival R package. Forest plots used for multivariate analysis were drawn through the forest model R package.F. nucleatum status. Survival analysis was restricted to 154 patients evaluable for OS and 153 patients for DFS , with moderate/high differentiation (78%), vascular (61%) and lymphatic (66%) invasion, and R0 resection margins (79%). Here, 16% of tumors were associated with HIV infection, 80% with HPV16 infection and 11% were HPV-negative.The median OS was 39.4 months from APR (64.3 months from diagnosis) and the median DFS from APR was 20.7 months.F. nucleatum loads were not statistically different according to individual centers (p = 0.30) (p = 0.49) ( = 0.30)  or to th = 0.49) .F. nucleatum were enriched in initial stage II ASCC (p = 0.02) and not significantly associated with other clinicopathological factors (High loads (upper tercile) of  factors .F. nucleatum load was significantly associated with better OS compared to lower terciles   .F. nucleatum load was associated with better DFS compared to low/intermediate terciles   .F. nucleatum load were associated with better MFS compared to the lowest tercile  (F. nucleatum loads for OS (p = 0.032), DFS (p = 0.009), and MFS (p = 0.02) (The highest and intermediate terciles of  0.0054) . We also = 0.02) .n = 102) showed a significant prognostic value for OS (p = 0.03) and DFS (p = 0.015) but not MFS (p = 0.27).After excluding patients treated with RT alone, analyses performed in the subgroup of patients treated with CRT (p < 0.001), lymph node invasion , perineural invasion , lymphatic invasion , and tumor invasion depth  were significantly associated with shorter OS in the univariate analysis. Age ≤ 65 years, tumor invasion depth, vascular emboli, perineural invasion, positive resection margins and lymph node invasion were also significantly associated with poor DFS  in the univariate analysis. Age ≤ 65 years, perineural invasion, and lymph node invasion were associated with poor MFS  in the univariate analysis. The highest tercile of F. nucleatum load was significantly associated with longer OS , DFS  but not MFS .Positive resection margins   .p < 0.05). Age ≤ 65 years was significantly associated with shorter DFS and MFS (p < 0.05).Among the clinicopathological parameters tested, positive resection margins and perineural invasion remained associated with poor OS, DFS, and MFS . Other independent prognostic indicators included lymph node invasion, positive resection margins, and tumor invasion depth, as previously reported [In this work, we assessed the association between intratumoral reported .F. nucleatum load and improved survival was unexpected as this bacteria is usually associated with poor prognosis in digestive cancers [F. nucleatum was associated with better survival in OSCC [F. nucleatum showed negative prognostic effect  are adenocarcinoma and not exposed to RT. In our study, survival was analyzed by taking the date of surgery as the starting point. However, the univariate analysis goes in the same direction when we take the date of diagnosis as the starting point (The association between  cancers ,13,14,15 cancers ,17. Howe in OSCC ,21. OSCCng point .F. nucleatum on the immune microenvironment are conflicting. Most studies showed the pro-inflammatory and immunosuppressive properties of F. nucleatum through expansion of myeloid-derived immune cells, Tregs, and M2 macrophages, and inhibition of cytotoxic T-cells [F. nucleatum loads were associated with a specific immune microenvironment poor in M2 macrophages, CD4 lymphocytes, fibroblasts, TLR4, OX40 ligand, and TNFRSF9, but high in TNFSF9 and IL-1ß allowing M1 polarization [F. nucleatum may be associated with a tumor microenvironment insensitive to pro-inflammatory signals resulting in favorable clinical outcomes. Another recent study also reported a positive prognostic value of intratumoral F. nucleatum in head and neck cancers [This positive survival effect may be mediated by modulation of intratumoral immunity , previou T-cells ,25,26,27rization . This su cancers .F. nucleatum in cancer patients. The underlying mechanisms warrant further investigation. Yet our study has some limitations. First, we had only one cohort of patients with ASCC without a validation cohort. However, our cohort is multicentric and unique: it is the largest cohort of ASCC treated by APR and the sample size was significant given the rarity of the disease. In addition, we selected patients who required surgical intervention after the failure of RT/CRT, bringing great homogeneity to our cohort but possibly introducing a selection bias. Nevertheless, the prognostic value of a new parameter is of particular interest in a population at high risk of relapse. Finally, immune microenvironment analysis was not available in our study; it could be of interest to assess the correlation between intratumoral F. nucleatum expression and immune components in further studies. Besides this, F. nucleatum may be a predictive marker for immunotherapy response and need to be assessed in ancillary studies of immunotherapy trials. Immunotherapy is indeed being developed in ASCC but seems to be active only in a small subset of patients, and predictive biomarkers to identify patients who may benefit the most from this approach are needed.Our current work strengthens the new insight into the prognostic role of intratumoral F. nucleatum and ASCC patient survival warranting further validation in larger prospective cohorts. Validation of these findings would allow to guide therapeutic strategies in dedicated trials by proposing intensification or de-escalation of systemic treatments and follow-up according to F. nucleatum loads. This can also give a rationale for further exploration of the role of F. nucleatum in ASCC carcinogenesis and response to treatment, particularly immunotherapy.In conclusion, we highlight a unique association between


Achromobacter xylosoxidans, leading to an infectious endocarditis. Achromobacter xylosoxidans (A. xylosoxidans) is an aerobic Gram-negative rod-shaped bacterium, which has been associated with multiple nosocomial opportunistic diseases in human medicine. One such manifestation of A. xylosoxidans infection is endocarditis. A. xylosoxidans infections are challenging to treat due to the reduced effectiveness of a wide range of antimicrobial agents. To date, only a few case reports of infections with A. xylosoxidans in animals have been described. This is the first case report of A. xylosoxidans endocarditis in a dog. Whole-genome sequencing was performed to determine the sequencing type and to gain more information about this bacterium regarding its intrinsic resistance genes. With this case report, we seek to increase awareness of A. xylosoxidans as an opportunistic nosocomial pathogen in dogs and to provide a short summary regarding the current state of general knowledge and known resistance patterns.Infectious endocarditis (IE) in dogs is often associated with a high mortality rate as diagnostic work-up as well as antibiotic treatment might be challenging. The present case describes bacteremia in a dog caused by  Achromobacter xylosoxidans (A. xylosoxidans) is an aerobic Gram-negative rod-shaped bacterium, first isolated in 1971 from the ear discharge of a human patient [A. xylosoxidans has been associated with multiple opportunistic diseases in human medicine [A. xylosoxidans infection is endocarditis [medicine ,3,4,5. Ocarditis .A. xylosoxidans in animals have been described [A. xylosoxidans endocarditis in a dog, to the best of our knowledge. This manuscript introduces A. xylosoxidans as a possible opportunistic nosocomial pathogen in dogs suffering from infectious endocarditis in veterinary medicine. Therefore, we give a short overview of A. xylosoxidans in general and its known resistance pattern. Furthermore, this manuscript discusses the important questions of possible infection routes and the risk of zoo–anthropogenic transmission.To date, only a few case reports of infections with escribed ,7,8,9. TA large, spayed, 12-year-old, mixed-breed female dog was referred to the university clinic due to intermittent lameness, fever, and a newly detected heart murmur. According to past medical history, the dog had been repeatedly infested with ticks within the previous year. In addition, the dog had lost weight over the last 3 months after suffering an episode of diarrhea, which was treated with an intravenous infusion at a private veterinary practice.During the initial physical examination, the dog showed tachycardia (180 bpm), a bounding pulse, and a body temperature of 39.9 °C. A systolic and diastolic heart murmur grade 4/6 audible on the left chest side was identified on auscultation. The orthopedic examination revealed an alternating lameness between all four extremities and a painful palpation of the right hip.9/L; 6–15 × 109/L) with neutrophilia without left shift ; monocytosis ; moderate hypoalbuminemia ; severely increased C-reactive protein ; mildly increased troponin I ; as well as severe hypocobalaminemia . Due to the aforementioned tick exposure, tests for the detection of antibodies against Anaplasma spp., Ehrlichia spp., Borrelia burgdorferi, the antigen of Dirofilaria immitis , as well as PCR for Bartonella spp. were performed. All of these proved negative. Urine analysis, including dipstick, sediment, bacterial culture, and urine protein/creatinine ratio, was unremarkable.Hematology and blood biochemistry revealed mild, non-regenerative, normocytic, and normochromic anemia ; mild leukocytosis  and a normal systolic function (FS 36.6–38%) were detected. Left atrial size was normal (La/Ao 1.3).Based on the clinical signs and the results of the echocardiography, endocarditis with secondary polyarthritis was suspected. Subsequent diagnostics included blood culture, arthrocentesis of the carpal and tarsal joint followed by cytological and microbiological examinations, and a cytological examination of the spleen. Ten milliliters of blood was collected from the left jugular vein following shaving and disinfection with chlorhexidine and alcohol, which was then transferred aseptically into a blood culture bottle . Afterward, the bottle was gently mixed and incubated at 37 °C.An ultrasound-guided fine-needle aspiration of the spleen revealed splenitis with the predominance of small lymphocytes and neutrophils as the main cell type. Cytology of the synovial fluid revealed a pyogenic sterile arthritis. The microbiological examination, including mycoplasmas, was negative. Based on these results, secondary immune-mediated polyarthritis was suspected.Due to the poor clinical condition of the dog, antibiotic treatment with amoxicillin-clavulanic acid (22 mg/kg TID IV) and marbofloxacin (4 mg/kg SID IV) was initiated as reported elsewhere for suspected IE with negative blood culture results . A multiA. xylosoxidans, identified at the species level by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry . According to the literature [A. xylosoxidans. We recommended continuing the therapy with meropenem, however, the owner decided to discontinue antibiotic treatment due to financial and personal issues. The dog was euthanized a few days later as a result of clinical deterioration.The dog showed a transient improvement in the clinical signs, but the fever continued despite antibiotic therapy. After seven days, the initial blood cultures tested positive for terature ,12 and dA. xylosoxidans strain. De novo assembly of raw reads and WGS data analysis were performed as previously described [Achromobacter MLST Databases hosted at PubMLST  [A. xylosoxidans strain belonged to the sequence type (ST) 201.Whole-genome sequencing (WGS) was performed by isolating and sequencing bacterial DNA as previously described  to gain escribed ,15. Specescribed . Classicer 2020) . Based ohttps://card.mcmaster.ca/home, accessed on 26 May 2020) [A. xylosoxidans strain harbored genes for a narrow-spectrum class D-lactamase blaOXA114a and the efflux pumps AxyXY-OprZ. The minimum inhibitory concentration (MIC) was determined by Etest  for imipenem, meropenem, and ceftazidime, according to the manufacturer’s recommendation, resulting in an MIC of 1 µg/mL for imipenem and meropenem, and 2 mg/L for ceftazidime.The identification of acquired resistance genes and chromosomal mutations was performed using the Comprehensive Antibiotic Resistance Database  . The A. A. xylosoxidans.This is the first case describing IE in a dog with bacteremia due to Staphylococcus spp. (24%) and Streptococcus spp. (37%)), Gram-negative rods (22%), facultative Gram-positive rods (5%), Actinomyces spp. (2%), Mycobacterium spp. (2%), and Bartonella spp. (20%) [A. xylosoxidans in dogs exist, even though it belongs to the Gram-negative class of bacteria.IE is an infrequently documented disease in dogs ) , and it p. (20%) . IE assop. (20%) , in compA. xylosoxidans in dogs. A. xylosoxidans has been isolated from a wide range of clinical material [A. xylosoxidans is unknown, the mode of transmission in hospitals is considered to be through contaminated fluids used for diagnostics or therapy, or indwelling devices [A. xylosoxidans endocarditis was mostly associated with catheter-related bacteremia in a human case series [A. xylosoxidans in animals have been mostly nosocomial infections following invasive interventions: infection of a total hip prosthesis in a dog [A. xylosoxidans was detected in the intestine of ticks (Ixodes spp.) as part of the unspecific bacterial flora [According to the literature, this is the first case report of bacterial endocarditis due to material ,20 assocmaterial ,4 in hummaterial ,20,21. A devices ,23. In pin a dog , hemolytin a dog , cathetein a dog , and lowin a dog . Other iin a dog . Furtheral flora . WhetherAchromobacter, 485 different sequence types (ST) have been reported according to public records . The detected ST201 in our case was first reported in a human patient suffering from cystic fibrosis (CF) [Achromobacter spp. have emerged in CF patients, with A. xylosoxidans showing the highest prevalence (7.4–10.4%) [A. xylosoxidans was also detected in the domestic and outdoor environment of patients with CF [Achromobacter spp. between cystic fibrosis patients was observed [Regarding the genetic diversity of the genus 4–10.4%)  and main4–10.4%) . A. xylo with CF . To the observed , it requA. xylosoxidans infections are challenging to treat due to the reduced effectiveness of a wide range of antimicrobial agents [A. xylosoxidans, as a non-fermentative Gram-negative bacterium, is intrinsically resistant to benzylpenicillin, first- and second-generation cephalosporins, glycopeptides, lipoglycopeptides, fusidic acid, macrolides, lincosamides, streptogramins, rifampicin, and oxazolidinones [A. xylosoxidans [A. xylosoxidans [A. xylosoxidans is commonly associated with two efflux pumps, AxyABM and AxyXY-OprZ, and a narrow-spectrum class D β-lactamase OXA-114 [A. xylosoxidans strain of the present case harbored genes for the narrow-spectrum class D β-lactamase OXA-114 and the efflux pump AxyXY-OprZ. The efflux pump AxyXY-OprZ was associated with higher minimum inhibitory concentration (MIC) values of aminoglycosides, cefepime, carbapenems, some fluoroquinolones, tetracyclines, and erythromycin, while the class D β-lactamase OXA-114 showed efficient hydrolysis of piperacillin and, to a lesser extent, ticarcillin [A. xylosoxidans infections. However, newly developing resistance genes, as aforementioned [l agents ,2,20,29.idinones . Based osoxidans . In totaarcillin . Due to arcillin , carbapeentioned , may redThe choice of an appropriate antibiotic should be based on microbiological criteria, such as the minimum inhibitory concentration (MIC), and pharmacological criteria, including the achievable concentration of the antibiotic at the clinical site as well as clinical efficacy. All these parameters take clinical breakpoints into account: clinical breakpoints are the concentrations of antibiotics used to define whether infection by a particular bacterial strain/isolate is likely to be treatable in a patient .A. xylosoxidans, a semi-quantitative susceptibility test such as the agar disk diffusion test could not be performed in the present case. To obtain MICs, an Epsilometer test (Etest) was performed with three antibiotics  based on the aforementioned intrinsic resistance pattern for A. xylosoxidans. Due to its known effectiveness against A. xylosoxidans infections in humans and its superior bactericidal activity compared to cephalosporins [A. xylosoxidans in dogs, it is unknown whether a serum concentration of meropenem was achieved to inhibit A. xylosoxidans in our case. However, one pharmacokinetic study reported that bacteria having an MIC of 0.12 µg/mL could be treated with dosages as low as 8 mg/kg, SC every 12 h in infections of the soft tissues or the urinary tract [A. xylosoxidans, especially for endocarditis, need to be developed, as all effective antibiotics such as meropenem belong to the critically important antibiotic substance in human medicine [A. xylosoxidans.Because of the missing clinical breakpoints for osporins ,33, meroosporins . Due to ry tract . Neverthry tract . Based ory tract ; so, tremedicine . CarbapeA. xylosoxidans as a potential pathogen causing infectious endocarditis in dogs. Further investigations are needed to confirm possible transmission routes in animals and to determine the risk of zoo–anthropogenic transmission. Known intrinsic resistance and current in vitro susceptibility tests must be considered for adequate antibiotic treatment, especially if they are refractory to standard antibiotic treatment regimens. In vivo MICs and clinical breakpoints should be established for bacteria such as A. xylosoxidans in order to select adequate antibiotics with appropriate effects in target tissues.In conclusion, our case report highlights the ST 201 of


Arachis pintoi Krapov. & W.C.Greg. is an important leguminous forage grass species that have extremely wide ranges of distribution in the tropical and sub-tropical regions. It has high feeding value and horticultural value. In this study, we sequenced and assembled the complete chloroplast genome of A. pintoi. The chloroplast genome is 156,185 bp in length, containing a pair of inverted repeated (IR) regions of 25,820 bp that are separated by a large single copy (LSC) region of 85,637 bp, and a small single copy (SSC) region of 18,908 bp. The complete chloroplast genome contains 112 unique genes, including 80 protein-coding genes, 28 transfer RNA genes (tRNAs), and four ribosomal RNA genes (rRNAs). The overall GC content was 36.4%. The phylogenetic analysis demonstrated that A. pintoi formed a single branch among genus Arachis. The whole chloroplast genome of A. pintoi will be a useful resource for future studies on phylogeny and conservation in Arachis. Arachis pintoi Krapov. & W.C.Greg. is a perennial herb of the genus Arachis in Leguminosae  for DNA extraction. Voucher specimen was deposited at Zhejiang Academy of Agricultural Sciences with the specimen voucher number of FP200817. Total genomic DNA was extracted from sampled leaves using the Plant Genomic DNA Kit  following the manufacturer’s instructions and sequenced based on the Illumina pair-end technology. Then, a total of 49,464,196 raw reads were obtained by next-generation sequencing, conducting on the Illumina Hiseq 4000 Platform . The complete chloroplast genome was assembled via NOVOPlasty  used as a reference sequence. The mean sequencing depth was 1030 × for the chloroplast genome of A. pintoi. The annotation was performed using an online GeSeq (https://chlorobox.mpimp-golm.mpg.de/geseq.html/), and the gene map of A. pintoi chloroplast genome was obtained by using OGDRAW (https://chlorobox.mpimp-golm.mpg.de/OGDraw.html). Finally, a complete chloroplast genome of A. pintoi and annotation information had been submitted to GenBank with accession number MW561116.Fresh and young leaves of A. pintoi is 156,185 bp in length, consists of a large single-copy region , a small single-copy region , and a pair of inverted repeat regions . The overall GC content was 36.4%. The complete chloroplast genome contains 112 unique genes, including 80 protein-coding genes, 28 transfer RNA (tRNA) genes, and four ribosomal RNA (rRNA) genes. Among these genes, fifteen of them contain one intron and three of them contains two introns.The complete chloroplast genome of A. pintoi, a phylogenetic analysis was performed based on the complete chloroplast genome sequences of 18 Papilionoideae species. All of the chloroplast genome sequences were aligned by using MAFFT (Kazutaka and Standley Stylosanthes viscosa was used as the outgroup. As shown in the ML phylogenetic tree (A. pintoi formed a single branch among genus Arachis with 100% bootstrap support value. The results provide valuable information for future studies on phylogenetic and evolution adaptation studies on A. pintoi and its related species.To examine the phylogenetic position of tic tree , the restic tree . The res


Pinus species has focused on specialized metabolites and axial resin ducts, but the role of lignin in defense within diverse systems is unclear. We investigated lignin levels in the outer bark and phloem of P. longaeva, P. balfouriana, and P. flexilis; tree species growing at high elevations in the western United States known to differ in susceptibility to mountain pine beetle . Pinus longaeva and P. balfouriana are attacked by MPB less frequently than P. flexilis, and MPB brood production in P. longaeva is limited. Because greater lignification of feeding tissues has been shown to provide defense against bark beetles in related genera, such as Picea, we hypothesized that P. longaeva and P. balfouriana would have greater lignin concentrations than P. flexilis. Contrary to expectations, we found that the more MPB-susceptible P. flexilis had greater phloem lignin levels than the less susceptible P. longaeva and P. balfouriana. No differences in outer bark lignin levels among the species were found. We conclude that lignification in Pinus phloem and outer bark is likely not adaptive as a physical defense against MPB.A key component in understanding plant-insect interactions is the nature of host defenses. Research on defense traits among  Bark beetles  are forest disturbance agents globally and include many tree-killing species . OvercomDendroctonus ponderosae Hopkins, Coleoptera: Curculionidae, Scolytinae) is an ecologically and economically significant bark beetle with an extensive distribution across western North America [Pinus species are considered MPB hosts [P. longaeva (Great Basin bristlecone pine) and P. balfouriana (foxtail pine) are rare [P. flexilis (limber pine) [P. longaeva in both field [P. longaeva relative to P. flexilis [P. longaeva and P. balfouriana also have dense sapwood and heartwood and possess high concentrations of constitutive specialized metabolite defense compounds relative to co-occurring P. flexilis [The mountain pine beetle (MPB)  –13. In ath field  and laboth field , and extflexilis . P. longflexilis .Specialized metabolites as well as anatomical structures are fundamental in conifer defense. They can be expressed constitutively or upregulated upon attack as needed to maximize the economy of available resources –20. VariLignin, a fundamental plant structural element, is the second most naturally abundant biopolymer in plant cell walls, after cellulose ,35. LignPinaceae, sclerenchyma cells of the phloem occur as large stone cells that are primarily comprised of lignin [Picea sitchensis Bongard) were associated with decreased spruce weevil (Pissodes strobi Peck) growth rate, survival, and fecundity, and disruption of larval establishment [Dendroctonus micans Kugelann) larvae were also associated with increased lignin concentrations [Hylobius abietis (L.) [P. strobi [In the family f lignin ,52,53. Ilishment . Decreastrations ,57 and ntis (L.) . Moreove. strobi ,53,59 it. strobi ,60,61 by. strobi .Pinus, specifically, is known to have evolved various defensive strategies against phloem-feeding insects, such as specialized metabolites [Pinus species [P. longaeva, P. balfouriana and P. flexilis from multiple sites and compared lignin concentrations within and among species and between the two tissue types. We hypothesized that the more MPB-resistant P. longaeva and P. balfouriana would have greater lignin concentrations than co-occurring P. flexilis.The genus abolites –64 and rabolites ,65,66, b species ,68. LittP. longaeva and P. balfouriana, four of them in stands with co-occurring P. flexilis  ranged from 30–46 cm. Study sites without signs of MPB or pathogen activity were chosen to avoid an influence of induced defenses. Permission for sampling was obtained through the Inyo, Klamath, Sierra Nevada and Humboldt-Toiyabe National Forests.Between June and September 2016, trees were sampled at five sites across the ranges of flexilis ; Table 1TM). Four samples were taken on the north, south, west, and east aspects of the tree trunk and pooled to account for potential within-tree variation. Upon tissue removal, phloem thickness (mm) was measured from the north and south aspect samples. Outer bark and phloem tissues were then separated and placed immediately in a sealed vial in a cooler with dry ice for transport to the Rocky Mountain Research Station  for cold storage (-40°C).To assess lignin levels (mg/g fresh weight) in outer bark and phloem, trees were sampled by boring into the tree at breast height with a 1” diameter circular hole saw  , as descDifferences among tree species in phloem and outer bark lignin concentrations, phloem thickness, and DBH were assessed with a hierarchical mixed effect analysis of variance (ANOVA), that accounts for variation among sites, using the package “lme4”  in R verP. longaeva and P. balfouriana, but, contrary to our hypotheses, P. flexilis had significantly higher (~2-fold) phloem lignin concentrations than the other two species , although P. flexilis outer bark lignin concentration differed at two sites  with varying resistance to the phloephagous emerald ash borer (Agrilus planipennis Fair.) [F. americana and F. pennsylvanica phloem/outer bark was associated with resistance to the emerald ash borer [Pinus species we sampled has not been investigated and should be part of future studies.Contrary to our expectations, s Fair.) ,76. Althsh borer . The potPinus flexilis has consistently been found to have less constitutive and induced LMW specialized metabolites  than other species, including P. longaeva and P. balfouriana at the sites sampled for this study [P. contorta and P. ponderosa [P. aristata . Although interspecific differences in selective pressure may have led to differences in investment in phloem specialized metabolite defenses [P. flexilis had thinner phloem, but greater lignin concentrations and absolute abundance than P. longaeva and P. balfouriana, the latter two having thicker phloem. Moreover, P. flexilis with the thickest phloem had the lowest lignin concentrations, further suggesting a negative relationship between phloem thickness and lignification. That outer bark lignin concentrations did not differ among the tree species but phloem concentrations did, suggest that lignification within the phloem may be under different selective pressures relative to outer bark. Trait associations and underlying mechanisms facilitating phloem lignification may be unique to the functions of nutrient transport or defense against invading bacteria or pathogens [is study , P. contonderosa , and thedefenses ,79,80, odefenses ,31. In oathogens .Pinus, higher lignin levels would be expected within both outer bark and phloem tissues of species considered less susceptible to MPB. This expectation is supported by prior research in Picea that was focused on species that are generally not considered primary mortality agents of mature trees, including Pissodes larva that feed in terminal buds [H. abietis that girdle seedlings [D. micans [P. flexilis had greater phloem lignin concentrations relative to the less MPB-susceptible P. longaeva and P. balfouriana suggest that the defensive function of lignin may be dependent on the plant tissue consumed and aggressiveness of the insect. We also found that the species with the greatest constitutive phloem lignin concentrations, P. flexilis, was previously found to produce lower levels of constitutive LMW specialized metabolites than the other two species. While increased tissue lignification may have an additive effect with specialized metabolites on host defenses against MPB, there may be metabolic tradeoffs that are not accounted for between LMW specialized metabolites and lignin. Therefore, greater lignification within feeding tissues does not appear to be generally adaptive as a defense against MPB. Moreover, interspecific differences in phloem but not outer bark lignin concentrations highlight that the benefits and costs of lignification in Pinus are likely specific to phloem tissue. High elevation Pinus species are increasingly threatened by MPB as a result of warming temperatures. Our results enhance the important knowledge base of defense strategies employed by MPB-susceptible high elevation Pinus.In summary, if defense against bark beetle attack were a strong selective driver for higher lignification in nal buds –56,69, Heedlings , and the. micans ,57. MPB S1 FigAll phloem samples were clear and colorless and therefore assumed pure . Outer bark samples were assumed to be pure when clear and colorless to light pink , but incompletely digested and/or contaminated when dark red .(JPG)Click here for additional data file.S1 TableEffect size (Est.) and 95% confidence interval (95% CI) estimates between comparison samples are shown. P-values (p) presented describe the likelihood of statistical difference with values < 0.05 presented in bold.(DOCX)Click here for additional data file.