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Inducible promoters are indispensable elements when considering the possibility to modulate gene expression on demand. Desirable traits of conditional expression systems include their capacity for tight downregulation, high overexpression, and in some instances for fine-tuning, to achieve a desired product's stoichiometry. Although the number of inducible systems is slowly increasing, suitable promoters comprising these features are rare. To date, the concomitant use of multiple regulatable promoter platforms for controlled multigene expression has been poorly explored. This work provides pioneer work in the human pathogenic fungus Aspergillus fumigatus, wherein we investigated different inducible systems, elucidated three candidate promoters, and proved for the first time that up to three systems can be used simultaneously without interfering with each other. Proof of concept was obtained by conditionally expressing three antifungal drug targets within the ergosterol biosynthetic pathway under the control of the xylose-inducible PxylP system, the tetracycline-dependent Tet-On system, and the thiamine-repressible PthiA system. IMPORTANCE In recent years, inducible promoters have gained increasing interest for industrial or laboratory use and have become key instruments for protein expression, synthetic biology, and metabolic engineering. Constitutive, high-expressing promoters can be used to achieve high expression yields; however, the continuous overexpression of specific proteins can lead to an unpredictable metabolic burden. To prevent undesirable effects on the expression host's metabolism, the utilization of tunable systems that allow expression of a gene product on demand is indispensable. Here, we elucidated several excellent tunable promoter systems and verified that each can be independently induced in a single strain to ultimately develop a unique conditional multigene expression system. This highly efficient, modular toolbox has the potential to significantly advance applications in fundamental as well as applied research in which regulatable expression of several genes is a key requirement.

Background and Objectives : To determine the antibiotic resistance rate of H. pylori among patients with peptic ulcer. Materials and Methods : A cross-sectional monocentric study was conducted from January to December 2021 among patients aged from 16 years with gastrointestinal symptoms and esophagogastroduodenoscopy. Gastric mucosa biopsies were collected at the edges of the ulcer or at lesion sites for H. pylori culture. Five antibiotics (amoxicillin (AMX), clarithromycin (CLR), metronidazole (MTZ), levofloxacin (LEV), and tetracycline (TET)) were selected for antibiotic susceptibility testing. Results : One hundred and twenty-five patients were included, and the sex ratio was 0.6. Their mean age was 47.3 ± 14.2 years. All of the participants had gastritis, and 24.0% had duodenitis. A total of 21.6% of patients had a duodenal ulcer, and 12.8% had an antral ulcer. A total of 40 specimens have grown in H. pylori culture. The proportion of resistance to AMX, CLR, MTZ, LEV, and TET was 27.5%, 50%, 67.5%, 35%, and 5%, respectively. The proportion of multidrug resistance was 22.5%. The proportion of double resistance to AMX + CLR was 20.0%, AMX + MTZ was 15.0%, AMX + LEV was 2.5%, CLR + MTZ was 32.5%, and TET + MTZ was 5.0%. Conclusions : Our research results show that the treatment with MTX-TET or LVX-AMOX has the highest sensitivity rate. Therefore, practitioners should refer to these regimes to eradicate H. pylori in patients with gastric and duodenal ulcers. The reports on H. pylori eradication from different geographic areas show heterogeneous results. Therefore, continuous monitoring of antibiotic resistance of H. pylori in each population is very important. Having evidence helps clinicians to treat patients most effectively, reduce treatment costs, and limit the rate of antibiotic resistance.

Bacterial pathogens are confronted with a range of challenges at the site of infection, including exposure to antibiotic treatment and harsh physiological conditions, that can alter the fitness benefits and costs of acquiring antibiotic resistance. Here, we develop an experimental system to recapitulate resistance gene acquisition by Staphylococcus aureus and test how the subsequent evolution of the resistant bacterium is modulated by antibiotic treatment and oxygen levels, both of which are known to vary extensively at sites of infection. We show that acquiring tetracycline resistance was costly, reducing competitive growth against the isogenic strain without the resistance gene in the absence of the antibiotic, for S. aureus under hypoxic but not normoxic conditions. Treatment with tetracycline or doxycycline drove the emergence of enhanced resistance through mutations in an RluD-like protein-encoding gene and duplications of tetL , encoding the acquired tetracycline-specific efflux pump. In contrast, evolutionary adaptation by S. aureus to hypoxic conditions, which evolved in the absence of antibiotics through mutations affecting gyrB , was impeded by antibiotic treatment. Together, these data suggest that the horizontal acquisition of a new resistance mechanism is merely a starting point for the emergence of high-level resistance under antibiotic selection but that antibiotic treatment constrains pathogen adaptation to other important environmental selective forces such as hypoxia, which in turn could limit the survival of these highly resistant but poorly adapted genotypes after antibiotic treatment is ended.

Background: Enterobacter bugandensis is an emerging human pathogen in which multidrug resistant strains have been continuously isolated from various environments. Thus, this organism possesses the potential to pose challenges in human healthcare. However, the mechanisms, especially the efflux pumps, responsible for the multidrug resistance in E. bugandensis remain to be well elucidated., Methods: The Enterobacter strain CMCC(B) 45301 was specifically identified using whole genome sequencing. The specific CMCC(B) 45301 homologues of the TolC dependent efflux-pump genes characterized in Escherichia coli were identified. The tolC deletion mutant in CMCC(B) 45301 was constructed and subjected to susceptibility tests using 26 different antimicrobial agents, along with the wild type strain. The synergistic effects combining the Bacillus crude extract (BCE) and several other TolC-affected compounds against CMCC(B) 45301 were assayed., Results: We reclassified the Enterobacter CMCC(B) 45301 strain from species cloacae to bugandensis, on the basis of its whole genome sequence. We found that the CMCC(B) 45301 TolC, AcrAB, AcrD, AcrEF, MdtABC, EmrAB, and MacAB exhibit high similarity with their respective homologues in E. coli and Enterobacter cloacae. Our results for the susceptibility tests revealed that lacking tolC causes 4- to 256-fold decrease in the minimal inhibitory concentrations of piperacillin, gentamicin, kanamycin, tetracycline, norfloxacin, ciprofloxacin, chloramphenicol, and erythromycin against CMCC(B) 45301. In addition, the inhibition zones formed by cefuroxime, cefoperazone, amikacin, streptomycin, minocycline, doxycycline, levofloxacin, florfenicol, trimethoprim-sulfamethoxazole, azithromycin, lincomycin, and clindamycin for the tolC mutant were larger or more obvious than that for the parent. Our data suggested the important role played by TolC in CMCC(B) 45301 susceptibility to common antibiotic families covering ß-lactam, aminoglycoside, tetracycline, fluoroquinolone, phenicol, folate pathway antagonist, macrolide, and lincosamide. Deletion for tolC also increased the susceptibility of CMCC(B) 45301 to berberine hydrochloride and BCE, two natural product-based agents. Finally, we found that erythromycin, norfloxacin, and ciprofloxacin can potentiate the antibacterial activity of BCE against CMCC(B) 45301., Discussion: The present study elaborated the comprehensive TolC effect on the antimicrobial susceptibility profile in E. bugandensis, which might contribute to the development of more therapeutic options against this nosocomial pathogen., Competing Interests: Author XL is employed by Panjin Guanghe Crab Industry Co., Ltd. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Li, Zhang and Li.)

To determine the prevalence of Escherichia coli and their drug resistance profiles in fresh pork sold at two retail outlets (open-air market and closed retail stores) in Alice, South Africa. Retail meat samples (n = 176) collected from four shops (two from open-air markets and two from closed stores) were analyzed by conventional biochemical and PCR-based molecular confirmatory tests. The confirmed isolates were profiled for antimicrobial susceptibility to a panel of 12 commercial antibiotics: tetracycline, ampicillin, sulphamethoxazole trimethoprim, erythromycin, gentamycin, colistin sulphate, cefotaxime, chloramphenicol, norfloxacin, ciprofloxacin, cefuroxime, and imipenem. Colistin, ampicillin, and erythromycin resistance genes were profiled with the gene-specific primers. Multidrug resistance (MDR) and the association of imipenem and colistin in the MDR profile were determined. A total of 68 (39.08%) E. coli isolates were confirmed by PCR analysis. Resistance was most common to erythromycin (100%), followed by cefotaxime (95.58%), ampicillin (88.23%), cefuroxime (88.23%), trimethoprim-sulphamethoxazole (88.23%), and tetracycline (60.29%). Overall, 27/68 (39.70%) were MDR (≥ 3antibiotics classes). MDR E. coli isolates associated with imipenem resistance (50.00%) and colistin resistance (33.82%) were detected. The resistance genes were detected among the isolates though not in all the phenotypically resistant isolates. The detection of colistin resistance among MDR E. coli isolates from retail meat is troubling as the drug is a last resort antibiotic. Overall, the epidemiological implications of the findings are of public health importance., (© 2022 Benson Chuks Iweriebor et al., published by Sciendo.)

Herein, we report a multifunctional hydrogel membrane with good mechanical properties, excellent antioxidant efficiency, and broad-spectrum antimicrobial activity. For this purpose, a series of chitosan-carboxymethyl cellulose-Pluronic P123 (CHT-CMC-P123) hydrogel membranes were prepared by blending various tetracycline hydrochloride (TCH) contents. The physicochemical and biological properties of CHT-CMC-P123 membranes were comprehensively investigated. With the increase of TCH content from 5 % to 20 %, hydrogel membranes presented a decreased water contact angle from 18.96° to 11.24°, and a decreased water vapor transmission rate from 171.8 to 156.1 g/m 2  h. Besides, with the increase of TCH content (5-20 %), the tensile strength (0.31-0.11 MPa) and elongation at break (10.57-4.82 %) of hydrogel membranes decreased while their thickness increased (113.5-324.3 μm). The data show that the release of TCH reached equilibrium after 26 days, with a cumulative percentage of approximately 28 %-87 %. Moreover, the hydrogel membranes exhibited a high antioxidant capacity of ~92 % for DPPH radical. Importantly, the incorporation of TCH significantly (~2.3 fold) enhanced the antimicrobial activity of the hydrogel membranes against Gram-positive, and Gram-negative bacteria and yeast. Based on our findings, these hydrogel membranes with superior properties may serve as effective food packaging and wound healing materials., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

With the development of medical drugs, the widely used tetracycline has brought many adverse effects on the ecosystem and human health. Tetracycline pollution of water environment is becoming more and more serious, and has become an emerging environmental problem. As single celled organisms, microalgae are not only model organisms for risk assessment of aquatic ecosystems, but also can efficiently purify sewage. Microalgae-mediated pollutant remediation has attracted more and more attention from researchers. In this paper, Chlamydomonas reinhardtii (C. reinhardtii) was used to remove tetracycline in aqueous solution, and the removal efficiency and mechanism of microalgae on tetracycline were studied. The results showed that the removal rates of tetracycline by active and inactivated microalgae at a density of 5 × 10 6  cells·mL -1 were 81.9% and 89.8%, respectively. C. reinhardtii removed tetracycline through biosorption and nonmetabolic processes. Microalgal cell supernatant and hydroxyl radicals could significantly promote the removal of tetracycline. The positively charged tetracycline was electrostatically adsorbed on the microalgae surface and extracellular polymeric substances. Microalgae biomass can promote the production of ROS and enhance the ability of microalgae to remove tetracycline., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Tetracycline (TC) antibiotic removal from water bodies is important to provide clean water and sanitation. Mesoporous graphitic carbon nitride (GCN) photocatalyst derived from three different types of precursors manages to remove TC effectively under visible light irradiation. Among urea, thiourea, and melamine precursors, melamine-prepared GCN (MGCN) via thermal polymerization has the highest efficiency to photodegrade tetracycline (TC) antibiotics up to 99.5% (0.0122 min -1 ) within 240 min. The COD for TC removal by using MGCN was up to 77.5% after 240 min of degradation. This is due to the slow charge recombination and rapid charge carrier migration. The MGCN encounters different properties such as high crystallinity, dense structure allowing fast charges migration, and nitrogen vacancies that create a defect state that suppresses charge recombination. It was found that the conduction band (CB) of MGCN was located at a more negative position (E CB  = -0.33 V) than (O 2 /O 2 •- ) and the valence band (VB) was placed at a more positive position (E VB  = 2.30 V) than (H 2 O/OH • ), which allows generation of both radicals for photodegradation. Based on the cell viability test, the photodegraded TC in the water how non-toxicity toward Balb/c 3T3 cells after being irradiated (λ > 420 nm) for 240 min under visible light. The MGCN prepared in this study demonstrated the highest effectiveness and recyclable photocatalyst for the removal of TC among all GCNs., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

The progressive increase in antibiotic resistance in recent decades calls for urgent development of new antibiotics and antibiotic stewardship programs to help select appropriate treatments with the goal of minimising further emergence of resistance and to optimise clinical outcomes. Three new tetracycline-class antibiotics, eravacycline, omadacycline, and tigecycline, have been approved within the past 15 years, and represent a new era in the use of tetracyclines. These drugs overcome the two main mechanisms of acquired tetracycline-class resistance and exhibit a broad spectrum of in vitro activity against gram-positive, gram-negative, anaerobic, and atypical pathogens, including many drug-resistant strains. We provide an overview of the three generations of tetracycline-class drugs, focussing on the efficacy, safety, and clinical utility of these three new third-generation tetracycline-class drugs. We also consider various scenarios of unmet clinical needs where patients might benefit from re-engagement with tetracycline-class antibiotics including outpatient treatment options, patients with known β-lactam antibiotic allergy, reducing the risk of Clostridioides difficile infection, and their potential as monotherapy in polymicrobial infections while minimising the risk of any potential drug-drug interaction. KEY MESSAGESThe long-standing safety profile and broad spectrum of activity of tetracycline-class antibiotics made them a popular choice for treatment of various bacterial infections; unfortunately, antimicrobial resistance has limited the utility of the early-generation tetracycline agents.The latest generation of tetracycline-class antibiotics, including eravacycline, tigecycline, and omadacycline, overcomes the most common acquired tetracycline resistance mechanisms.Based on in vitro characteristics and clinical data, these newer tetracycline agents provide an effective antibiotic option in the treatment of approved indications in patients with unmet clinical needs - including patients with severe penicillin allergy, with renal or hepatic insufficiency, recent Clostridioides difficile infection, or polymicrobial infections, and those at risk of drug-drug interactions.

The extensive use of tetracycline antibiotics has led to the widespread presence of tetracycline-resistance genes in Gram-negative bacteria and this poses serious threats to human and animal health. In our previous study, we reported a method for rapid detection of Tet(X)-producers using MALDI-TOF MS. However, there have been multiple machineries involved in tetracycline resistance including efflux pump, and ribosomal protection protein. Our previous demonstrated the limitation in probing the non-Tet(X)-producing tetracycline-resistant strains. In this regard, we further developed a MALDI-TOF MS method to detect and differentiate Tet(X)-producers and non-Tet(X)-producing tetracycline-resistant strains. Test strains were incubated with tigecycline and oxytetracycline in separate tubes for 3 h and then analyzed spectral peaks of tigecycline, oxytetracycline, and their metabolite. Strains were distinguished using MS ratio for [metabolite/(metabolite+ tigecycline or oxytetracycline)]. Four control strains and 319 test strains were analyzed and the sensitivity was 98.90% and specificity was 98.34%. This was consistent with the results obtained from LC-MS/MS analysis. Interestingly, we also found that the reactive oxygen species (ROS) produced by tetracycline-susceptible strains were able to promote the degradation of oxytetracycline. Overall, the MALDI Tet(X)-plus test represents a rapid and reliable method to detect Tet(X)-producers, non-Tet(X)-producing tetracycline-resistant strains, and tetracycline-susceptible strains.

Objectives: To analyse the genome sequences of four archival Acinetobacter nosocomialis clinical isolates (designated AC13, AC15, AC21 and AC25) obtained from Terengganu, Malaysia in 2011 to determine their genetic relatedness and basis of antimicrobial resistance., Methods: Antimicrobial susceptibility profiles of the A. nosocomialis isolates were determined by disk diffusion. Genome sequencing was performed using the Illumina NextSeq platform., Results: The four A. nosocomialis isolates were cefotaxime resistant whereas three isolates (namely, AC13, AC15 and AC25) were tetracycline resistant. The carriage of the bla ADC-255 -encoded cephalosporinase gene is likely responsible for cefotaxime resistance in all four isolates. Phylogenetic analysis indicated that the three tetracycline-resistant isolates were closely related, with an average nucleotide identity of 99.9%, suggestive of nosocomial spread, whereas AC21 had an average nucleotide identity of 97.9% when compared to these three isolates. The tetracycline-resistant isolates harboured two plasmids: a 13476 bp Rep3-family plasmid of the GR17 group designated pAC13-1, which encodes the tetA(39) tetracycline-resistance gene, and pAC13-2, a 4872 bp cryptic PriCT-1-family plasmid of a new Acinetobacter plasmid group, GR60. The tetA(39) gene was in a 2 001 bp fragment flanked by XerC/XerD recombination sites characteristic of a mobile pdif module. Both plasmids also harboured mobilisation/transfer-related genes., Conclusions: Genome sequencing of A. nosocomialis isolates led to the discovery of two novel plasmids, one of which encodes the tetA(39) tetracycline-resistant gene in a mobile pdif module. The high degree of genetic relatedness among the three tetracycline-resistant A. nosocomialis isolates is indicative of nosocomial transmission., (Copyright © 2022 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

The aim of this study was to evaluate 140 Salmonella Derby isolates collected over a 10-year period from porcine origins (environment, pig carcass, lymph nodes, intestinal content, and pork) for their phenotypic and genotypic antimicrobial resistance, their ability to produce biofilm, and their genetic relatedness. The minimum inhibitory concentration (MIC) was determined using microdilution broth method and antimicrobial resistance genes were investigated by PCR. The quantification of biofilm formation was performed in sterile polystyrene microtiter plates. Genetic relatedness was determined by Xba-I macrorestriction analysis. The highest frequencies of non-wildtype (nWT) populations were observed against tetracycline (75.7%), streptomycin (70%), and colistin (11.4%), whereas wildtype populations were observed against ciprofloxacin, ceftazidime, and gentamicin. The resistance genes found were bla TEM (ampicillin), aadA variant (streptomycin/spectinomycin), tetA (tetracycline), and floR (florfenicol). On 96-well polystyrene microtiter plate, 68.6% of the isolates proved to be biofilm producers. Among 36 S. Derby isolates selected to PFGE analysis, 22 were clustered with 83.6% of similarity. Additionally, 27 isolates were clustered in 11 pulsotypes, which presented more than one strain with 100% of similarity. Most of S. Derby isolates were able to form biofilm and were classified as nWT or resistant to tetracycline, streptomycin, and colistin. PFGE allowed the identification of closely related S. Derby isolates that circulated in pig slaughterhouses and pork derived products along a decade., (© 2022. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.)

Bacteriophage (phage) are bacterial predators that can also spread antimicrobial resistance (AMR) genes between bacteria by generalised transduction. Phage are often present alongside antibiotics in the environment, yet evidence of their joint killing effect on bacteria is conflicted, and the dynamics of transduction in such systems are unknown. Here, we combine in vitro data and mathematical modelling to identify conditions where phage and antibiotics act in synergy to remove bacteria or drive AMR evolution. We adapt a published model of phage-bacteria dynamics, including transduction, to add the pharmacodynamics of erythromycin and tetracycline, parameterised from new in vitro data. We simulate a system where two strains of Staphylococcus aureus are present at stationary phase, each carrying either an erythromycin or tetracycline resistance gene, and where multidrug-resistant bacteria can be generated by transduction only. We determine rates of bacterial clearance and multidrug-resistant bacteria appearance, when either or both antibiotics and phage are present at varying timings and concentrations. Although phage and antibiotics act in synergy to kill bacteria, by reducing bacterial growth antibiotics reduce phage production. A low concentration of phage introduced shortly after antibiotics fails to replicate and exert a strong killing pressure on bacteria, instead generating multidrug-resistant bacteria by transduction which are then selected for by the antibiotics. Multidrug-resistant bacteria numbers were highest when antibiotics and phage were introduced simultaneously. The interaction between phage and antibiotics leads to a trade-off between a slower clearing rate of bacteria (if antibiotics are added before phage), and a higher risk of multidrug-resistance evolution (if phage are added before antibiotics), exacerbated by low concentrations of phage or antibiotics. Our results form hypotheses to guide future experimental and clinical work on the impact of phage on AMR evolution, notably for studies of phage therapy which should investigate varying timings and concentrations of phage and antibiotics., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2022 Leclerc et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Campylobacter and non-typhoidal Salmonella are the major causes of bacterial gastrointestinal infections in humans. Although antimicrobial therapy is typically not recommended in many cases of these infections, it may be life-saving in patients with severe symptoms. Since chicken eggs and meat derived from layers are destined for human consumption, we investigated the prevalence and antimicrobial resistance of these two bacterial genera in 82 layer flocks at chicken processing plants in Honshu, Japan. Campylobacter was isolated from 77 flocks (93.9%). Resistance to ampicillin, tetracycline, and ciprofloxacin was documented in 42.3 (30/71), 16.9 (12/71), and 14.1% (10/71) of Campylobacter jejuni, respectively. Multilocus-sequence typing identified ST4389 and ST5262 as the most frequent C. jejuni sequence types. In C. coli, resistance to ampicillin, tetracycline, and ciprofloxacin was found in 20.0 (7/35), 20.0 (7/35), and 25.7% (9/35), respectively. The most frequent sequence type in C. coli was ST8292. Erythromycin resistance was not observed among Campylobacter species. Salmonella was isolated from 14 flocks (17.1%). The two most frequent serovars were Salmonella Corvallis and S. Braenderup. Neither S. Enteritidis nor S. Infantis were isolated. Streptomycin resistance was observed in six isolates (26.1%), and all isolates were susceptible to cefotaxime and ciprofloxacin. Thus, chicken eggs and meat derived from layers are possible sources of these bacterial infections in humans. The antimicrobial susceptibility of these isolates was maintained, reflecting restrictions on the use of antimicrobial agents on layers.

This study used surveillance data from a global program of clinical bacterial isolates to determine whether a tetracycline susceptible result can be used to predict an omadacycline susceptible result. Categorical agreement, very major error rates, and minor error rates were calculated for Staphylococcus aureus (MSSA and MRSA; n=38,364), S. lugdunensis (n=335), Streptococcus pneumoniae (n=11,725), S. pyogenes (n=3,390), S. anginosus group (n=622), Haemophilus spp. (n=6,419), Enterococcus faecalis (n=7,065), Klebsiella pneumoniae (n=10,313), and Enterobacter cloacae (n=4,418). Across the organisms, for which omadacycline has an FDA breakpoint established, a tetracycline susceptible result showed ≥96.3% categorical agreement in predicting an omadacycline susceptible result. The rates of very major errors were below the guideline-suggested level (<1.5%). Omadacycline retained activity against most (88.7-100% Gram-positive and 54-98.6% Gram-negative) tetracycline-resistant isolates. For laboratories that do not have the capability to perform susceptibility testing for omadacycline, one-sided surrogate testing for tetracycline can be a practical alternative., Competing Interests: Declaration of competing interests AWS, KW, and DA are employees and shareholders of Paratek Pharmaceuticals, Inc. TRK is a former consultant of Paratek Pharmaceuticals, Inc., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Re-osseointegration of an infected/contaminated dental implant poses major clinical challenges. We tested the hypothesis that the application of an antibiotic-releasing construct, combined with hard/soft tissue replacement, increases the efficacy of reconstructive therapy. We initially fabricated semi-flexible hybrid constructs of β-TCP/PHBHHx, with tetracycline (TC) (TC amounts: 5%, 10%, and 15%). Thereafter, using in vitro assays, TC release profile, attachment to rat bone marrow-derived stem cells (rBMSCs) and their viability as well as anti-bacterial activity were determined. Thereafter, regenerative efficacies of the three hybrid constructs were assessed in a rat model of peri-implantitis induced by Aggregatibacter actinomycetemcomitans biofilm; control animals received β-TCP/Bio-Gide and TC injection. Eight weeks later, maxillae were obtained for radiological, histological, and histomorphometric analyses of peri-implant tissues. Sulcus bleeding index was chronologically recorded. Serum cytokines levels of IL-6 and IL-1β were also evaluated by enzyme-linked immunosorbent assay. Substantial amounts of tetracycline, from hybrid constructs, were released for 2 weeks. The medium containing the released tetracycline did not affect the adhesion or viability of rBMSCs; however, it inhibited the proliferation of A. actinomycetemcomitans. Osteogenesis and osseointegration were more marked for the 15% hybrid construct group than the other two groups. The height of attachment and infiltration of inflammatory cells within fibrous tissue was significantly reduced in the experimental groups than the control group. Our protocol resulted in re-osseointegration on a biofilm-contaminated implant. Thus, an antibiotic releasing inorganic/organic construct may offer a therapeutic option to suppress infection and promote guided tissue regeneration thereby serving as an integrated multi-layer substitute for both hard/soft tissues., (© 2022 Wiley Periodicals LLC.)

ABSTRACT To address the inadequate removal of antibiotic resistance genes in wastewater treatment plants, this study investigated the impact of bioaugmentation with dried Eichhornia crassipes roots on removal of antibiotics sulfamethoxazole, tetracycline and erythromycin from pharmaceutical wastewater while optimizing potential for reclaiming value through biogas production, utilizing an anaerobic membrane bioreactor (AnMBR). Three sets of AnMBRs were set up for the experiment, C1 (inoculum), C2 (inoculum + antibiotics) and EC (inoculum + antibiotics +  E. crassipes ). The results showed that E. crassipes mitigated some of the toxic effects of antibiotics on the microbial community and prevented negative impact on the archaeal community, and significantly increased average biogas production (by 37% compared to control without antibiotics and 42% compared to control with antibiotics) as well as antibiotics removal. Furthermore, bioaugmented reactor showed significant reduction of erythromycin (97%) and tetracycline (83%) concentrations in effluent. Utilization of E. crassipes root offers a simple yet powerful tool for preventing the emergence of antimicrobial resistance and dissemination of such pollutants into the environment.

Abstract: Cow-calf production plays a significant role in the beef production chain. However, bacteria in these systems are not typically monitored for antimicrobial resistance (AMR). We determined the baseline level of AMR in fecal bacteria collected from preweaned calves prior to feedlot entry and evaluated the effects of type of graze and age on AMR occurrence. Two grazing experiments (16 cow-calf pairs each) were conducted on tall fescue or wheat. Fecal samples were cultured for the detection of tetracycline-resistant (TETr), third-generation cephalosporin-resistant (3GCr), and extended-spectrum β-lactamase (ESBL)-producing Escherichia coli. Isolates were characterized for resistance to other antibiotics and resistance mechanisms. Concentrations (P < 0.001) and prevalence (P = 0.007) of TETrE. coli isolates were significantly higher in the calves (5.1 log CFU/g and 93%, respectively) than in the cows (4.4 log CFU/g and 80%, respectively). Wheat grazing did not affect TETr isolates phenotypically; however, it significantly expanded (P = 0.005) the resistant population carrying tet(A) over that carrying tet(B). Fecal prevalence of 3GCr and ESBL-producing isolates was 31.3 and 3.4%, respectively, with no significant effects of age (P = 0.340) or wheat grazing (P = 0.597). All 3GCr and ESBL-producing isolates were multidrug resistant (resistant to at least three antimicrobial classes). 3GCr isolates were positive for blaCMY-2 (73%) or blaCTX-M (27%), and blaCTX-M-15 was the most prevalent gene (94%, n = 17) among the CTX-M-positive isolates. Wheat grazing significantly expanded (P < 0.001) the 3GCr population carrying blaCTX-M and reduced the population carrying blaCMY-2. Five of the seven ESBL-producing isolates were positive for blaCTX-M. Our study revealed age-dependent occurrence of TETrE. coli and that wheat grazing expanded the resistant population carrying certain resistance genes. Cow-calf production is a significant reservoir for antibiotic-resistant bacteria of significant public health importance such as 3GCr and CTX-M ESBL-producing E. coli., (Published 2022 by the International Association for Food Protection. Not subject to U.S. Copyright.)

In this work, NiFe 2 O 4 /g-C 3 N 4 heterostructure was prepared and used for the photocatalytic decomposition of tetracycline hydrochloride antibiotic and for inactivation of E. coli bacteria. The fabricated NiFe 2 O 4 /g-C 3 N 4 composite displayed enhanced ability for photodegradation of organic pollutants and disinfection activities compared to the bare samples, because of the enhancement of visible light absorbance, heterojunction formation and photo-Fenton process. The optimized sample 10%-NiFe 2 O 4 /g-C 3 N 4 has photodegraded 94.5% of tetracycline hydrochloride in 80 min. The active species trapping experiments revels that ·O 2 - , h + and •OH are key decomposing species participated in the antibiotic degradation. It is hoped that the present study will provide a better understanding to fabricate efficient photocatalysts for the decomposition of organic pollutants and disinfection of bacteria., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Black soldier fly larvae (larvae) can digest organic wastes and degrade contaminants such as oxytetracycline (OTC). However, compared to the kinetic processes and enhanced mechanisms used in the traditional microbial degradation of OTC, those employed by larvae are largely uncharacterized. To obtain further details, a combined analysis of larval development, larval nutritional values (crude protein, crude fat and the composition of fatty acids) and the expression of tetracycline resistance genes (TRGs) in the larval gut was performed for the degradation of OTC added to substrates and for oxytetracycline bacterial residue (OBR). When the larvae were exposed to the substrates, the degradation processes were enhanced significantly (P < 0.01), with a 4.74-7.86-fold decrease in the degradation half-life (day -1 ) and a 3.34-5.74-fold increase in the final degradation efficiencies. This result was attributed to the abundant TRGs (with a detection rate of 35.90%∼52.14%) in the larval gut. The TRGs presented the resistance mechanisms of cellular protection and efflux pumps, which ensured that the larvae could tolerate elevated OTC concentrations. Investigation of the TRGs indicated that enzymatic inactivation enhanced OTC degradation by larvae. These findings demonstrate that the larval degradation of antibiotic contaminants is an efficient method based on abundant TRGs in the larval gut, even though OTC degradation results in OBR. In addition, a more optimized system for higher reductions in antibiotic levels and the expansion of larval bioremediation to other fields is necessary., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Inc. All rights reserved.)

The extensive use of tetracyclines in human and veterinary medicine causes contamination in the environment that could contribute to the spread of antibiotic-resistant bacteria or competition between species of phytoplankton. In this study, Microcystis aeruginosa (a bloom-forming cyanobacterium) and Chlamydomonas microsphaera (common green alga) were selected to test the effects of different concentrations of tetracyclines (tetracycline and oxytetracycline) in monoculture and co-culture. The results showed that compared with monoculture, the cell growth of C. microsphaera decreased significantly in co-culture treated with different concentrations of tetracycline and oxytetracycline. The ratios of inhibition of M. aeruginosa exposed to 0.1, 2, and 10 mg L -1 of tetracycline varied between 17.7 and 31.37% in co-culture compared with monoculture, while the cell growth of M. aeruginosa was enhanced by treatment with 0.1, 2, and 7.25 mg L -1 of oxytetracycline in co-culture. However, the cell growth of C. microsphaera was significantly inhibited by all the treatments in co-culture. With the treatment of tetracycline, the specific growth rate of M. aeruginosa was 0.36 to 0.31 day -1 in monoculture and co-culture, while that of C. microsphaera ranged from 0.38 to 0.26 day -1 in monoculture, and it decreased from 0.25 day -1 (0 mg L -1 ) to 0.08 day -1 (10 mg L -1 ) in co-culture. With the treatment of oxytetracycline, the specific growth rate of M. aeruginosa was stimulated in co-culture, while that of C. microsphaera was significantly inhibited in co-culture compared with monoculture. Therefore, although M. aeruginosa significantly inhibited C. microsphaera in co-culture with the tetracycline-free treatment, the competitive advantage of M. aeruginosa expanded following the addition of low or high concentrations of tetracyclines., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The emergence of the plasmid-mediated high levels of the tigecycline resistance gene has drawn worldwide attention and has posed a major threat to public health. In this study, we investigated the prevalence of the tet (X4)-positive Enterobacterales isolates collected from a pig slaughterhouse and farms. A total of 101 tigecycline resistance strains were isolated from 353 samples via a medium with tigecycline, of which 33 carried tet (X4) (9.35%, 33/353) and 2 carried tet (X6) (0.57%, 2/353). These strains belong to seven different species, with Escherichia coli being the main host bacteria. Importantly, this report is the first one to demonstrate that tet (X4) was observed in Morganella morganii. Whole-genome sequencing results revealed that tet (X4)-positive bacteria can coexist with other resistance genes, such as bla NDM-1 and cfr . Additionally, we were the first to report that tet (X4) and bla NDM-1 coexist in a Klebsiella quasipneumoniae strain. The phylogenetic tree of 533 tet (X4)-positive E. coli strains was constructed using 509 strains from the NCBI genome assembly database and 24 strains from this study, which arose from 8 sources and belonged to 135 sequence types (STs) worldwide. We used Nanopore sequencing to interpret the selected 21 nonclonal and representative strains and observed that 19 tet (X4)-harboring plasmids were classified into 8 replicon types, and 2 tet (X6) genes were located on integrating conjugative elements. A total of 68.42% of plasmids carrying tet (X4) were transferred successfully with a conjugation frequency of 10 -2 to 10 -7 . These findings highlight that diverse plasmids drive the widespread dissemination of the tigecycline resistance gene tet (X4) in Enterobacterales of porcine origin. IMPORTANCE Tigecycline is considered to be the last resort of defense against diseases caused by broad-spectrum resistant Gram-negative bacteria. In this study, we systematically analyzed the prevalence and genetic environments of the resistance gene tet (X4) in a pig slaughterhouse and farms and the evolutionary relationship of 533 tet (X4)-positive Escherichia coli strains, including 509 tet (X4)-positive E. coli strains selected from the 27,802 assembled genomes of E. coli from the NCBI between 2002 and 2022. The drug resistance of tigecycline is widely prevalent in pig farms where tetracycline is used as a veterinary drug. This prevalence suggests that pigs are a large reservoir of tet (X4) and that tet (X4) can spread horizontally through the food chain via mobile genetic elements. Furthermore, tetracycline resistance may drive tigecycline resistance through some mechanisms. Therefore, it is important to monitor tigecycline resistance, develop effective control measures, and focus on tetracycline use in the pig farms.

Antibiotic tolerance not only enables bacteria to survive acute antibiotic exposures but also provides bacteria with a window of time in which to develop antibiotic resistance. The increasing prevalence of Campylobacter jejuni isolates resistant to clinically important antibiotics, particularly fluoroquinolones (FQs), is a global public health concern. Currently, little is known about antibiotic tolerance and its effects on resistance development in C. jejuni. Here, we show that exposure to ciprofloxacin or tetracycline at concentrations 10 and 100 times higher than the MIC induces antibiotic tolerance in C. jejuni, whereas gentamicin or erythromycin treatment causes cell death. Interestingly, FQ resistance rapidly develops in C. jejuni after tolerance induction by ciprofloxacin and tetracycline. Furthermore, after tolerance is induced, alkyl hydroperoxide reductase (AhpC) plays a critical role in reducing FQ resistance development by alleviating oxidative stress. Together, these results demonstrate that exposure of C. jejuni to antibiotics can induce antibiotic tolerance and that FQ-resistant (FQ R ) C. jejuni clones rapidly emerge after tolerance induction. This study elucidates the mechanisms underlying the high prevalence of FQ R C. jejuni and provides insights into the effects of antibiotic tolerance on resistance development. IMPORTANCE Antibiotic tolerance compromises the efficacy of antibiotic treatment by extending bacterial survival and facilitating the development of mutations associated with antibiotic resistance. Despite growing public health concerns about antibiotic resistance in C. jejuni, antibiotic tolerance has not yet been investigated in this important zoonotic pathogen. Here, our results show that exposure of C. jejuni to ciprofloxacin or tetracycline leads to antibiotic tolerance development, which subsequently facilitates the emergence of FQ R C. jejuni. Importantly, these antibiotics are commonly used in animal agriculture. Moreover, our study suggests that the use of non-FQ drugs in animal agriculture promotes FQ resistance development, which is crucial because antibiotic-resistant C. jejuni is primarily transmitted from animals to humans. Overall, these findings increase our understanding of the mechanisms of resistance development through the induction of antibiotic tolerance.

Microbial electrolysis cell (MEC) is a promising technology for effective energy conversion of wastewater organics to biogas. Yet, in swine wastewater treatment, the complex contaminants including antibiotics may affect MEC performance, while the high ammonia concentration might increase this risk by increasing cell membrane permeability. In this work, the responses of MECs on tetracycline (TC) with low and high ammonia loadings (80 and 1000 mg L -1 ) were fully investigated. The TC of 0 to 1 mg L -1 slightly improved MEC performance in current production and electrochemical characteristics with low ammonia loading, while TC ≥ 4 mg L -1 started to show negative effects. Generally, the high ammonia loading sensitized MECs to TC concentration, inducing the current and COD removal of MECs to sharply decline with TC ≥ 0.5 mg L -1 . The positive effect of high ammonia loading on MEC due to conductivity increase was counteracted with TC ≥ 1 mg L -1 . The co-contamination of TC and ammonia significantly decreased the bioactivity and biomass of anode biofilm. Although the high concentration of co-existing TC and ammonia inhibited MEC performance, the reactors still obtained positive energy feedback. The network analyses indicated that the effluent suspension contributed much to antibiotic resistance gene (ARG) transmission, while the microplastics (MPs) in wastewater greatly raised the risks of ARGs spreading. This work systematically examined the synergetic effects of TC and ammonia and the transmission of ARGs in MEC operation, which is conducive to expediting the application of MECs in swine wastewater treatment., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022. Published by Elsevier Ltd.)

The Gram-positive bacterium Staphylococcus aureus is responsible for a number of infections and has been described to exhibit resistance to antibacterial drugs through different resistance mechanisms. Among these, active efflux has been shown to be one of the main mechanisms of bacterial resistance associated with S. aureus. In this sense, the aim of the present study was to evaluate the ability of ellagic acid and gallic acid to reverse resistance by inhibiting the efflux pumps present in S. aureus strains IS-58 and K2068, which express the TetK and MepA flux pumps, respectively. In addition, the toxicity of both compounds was verified in Drosophila melanogaster. Broth microdilution assays were performed to obtain the minimum inhibitory concentration (MIC) values of ellagic acid and gallic acid, whereas efflux pump inhibition was tested using a subinhibitory concentration of standard efflux pump inhibitors, gallic acid and ellagic acid (MIC/8), where the ability of these compounds to decrease the MIC of ethidium bromide (EtBr) and antibiotics was verified. Toxicity was evaluated by mortality and negative geotaxis assays in D. melanogaster. Ellagic acid and gallic acid showed no direct antibacterial activity on S. aureus strains carrying the efflux pumps TetK and MepA. However, when we looked at the results for the TetK pump, we saw that when the two acids were associated with the antibiotic tetracycline, a potentiation of the antibacterial effect occurred; this behavior was also observed for the antibiotic ciprofloxacin in the MepA strain. For the efflux pump inhibition results, only the compound gallic acid showed potentiating effect on antibacterial activity when associated with the substrate EtBr for the IS-58 strain carrying the TetK efflux pump. Ellagic acid and gallic acid showed no toxicity on the model arthropod D. melanogaster. These results indicate the possible use of gallic acid as an adjuvant in antibiotic therapy against multidrug-resistant bacteria., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2022. Published by The Company of Biologists Ltd.)

Many contaminants were carried by dust, a common environment media that is easy to contact with human beings, and antibiotic resistance genes (ARGs) as an emergency pollutant also harbor in dust and pose serious threats to human health especially those carried by opportunistic pathogens because inactivation of antibiotics caused by ARGs may enhance pathogenicity. Considering there is a gap of investigation of dust ARGs, 16 S rRNA gene sequences and high-throughput quantitative PCR were employed to obtain information of microbial communities and accumulated ARGs in dust from different urban places, including the malls, hospitals, schools and parks, to investigate the distribution and influencing factors of ARGs and discover the potential hosts of ARGs in dust. Here, 9 types of ARGs such as sulfonamide, tetracycline, and beta-lactamase and 71 subtypes of ARGs like sul1, tetM-01, and drfA1 were detected in dust. ARGs had varying distribution in different public places and seasons in dust. The abundances of total ARGs, MLSB and tetracycline genes were higher in spring than summer. The diversity of ARGs was highest in malls, follow by hospitals, schools, and parks. Additionally, multi-drug resistance genes in dust were more abundant in hospitals than in schools and parks. The microbes were distinguished as the most important driving factors for ARGs in dust, followed by the mobile genetic elements (MGEs) and different places, while dust physicochemical parameters only exert a negligible impact. Notably, several opportunistic pathogens like the Streptococcus, Vibrio, and Pseudomonas were inferred as potential hosts of high-risk ARGs such as mecA, tetM-02, and tetO-01 in dust because of strongly positive co-occurrence. These results imply that dust is likely an important reservoir of ARGs. We should realize that ARGs may be harbored in some opportunistic pathogens occur in dust and endanger human health because of dust contacting to human easily., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Bacteria in the effluent of wastewater treatment plants (WWTPs) can transfer antibiotic resistance genes (ARGs) to the bacteria in receiving water through conjugation; however, there is a lack of quantitative assessment of this phenomenon in continuous cultures. Our objective was to determine the effects of background nutrient levels in river water column and growth rates of bacteria on the conjugation frequency of ARGs from effluent bacteria to river bacteria, as well as on the resulting resistance level (i.e., MICs) of the river bacteria. Chemostats were employed to simulate the discharge points of WWTPs into rivers, where effluent bacteria (donor cells) meet river bacteria (recipient cells). Both donor and recipient cells were Escherichia coli cells, and the donor cells were constructed by filter mating with bacteria in the effluent of a local WWTP. Results showed that higher bacterial growth rate (0.45 h -1 versus 0.15 h -1 ) led to higher conjugation frequencies (10 -4 versus 10 -6 transconjugant per recipient). The nutrient level also significantly affected the conjugation frequency, albeit to a lesser extent than the growth rate. The MIC against tetracycline increased from 2 mg/L in the recipient to 64 to 128 mg/L in transconjugants. In comparison, the MIC only increased to as high as 8 mg/L in mutants. Whole-genome sequencing showed that the tet -containing plasmid in both the donor and the transconjugant cells also occur in other fecal bacterial genera. The quantitative information obtained from this study can inform hazard identification related to the proliferation of wastewater-associated ARGs in surface water. IMPORTANCE WWTPs have been regarded as an important hot spot of ARGs. The discharge point of WWTP effluent, where ARGs may be horizontally transferred from bacteria of treated wastewater to bacteria of receiving water, is an important interface between the human-dominated ecosystem and the natural environment. The use of batch cultures in previous studies cannot adequately simulate the nutrient conditions and growth rates in receiving water. In this study, chemostats were employed to simulate the continuous growth of bacteria in receiving water. Furthermore, the experimental setup allowed for separate investigations on the effects of nutrient levels (i.e., simulating background nutrients in river water) and bacterial growth rates on conjugation frequencies and resulting resistance levels. The study generates statistically sound ecological data that can be used to estimate the risk of wastewater-originated ARGs as part of the One Health framework.

The tetracycline repressor (tetR)-regulated system is a widely used tool to specifically control gene expression in mammalian cells. Based on this system, we generated a human osteosarcoma cell line, which allows for the inducible expression of an EGFP fusion of the TAR DNA-binding protein 43 (TDP-43), which has been linked to neurodegenerative diseases. Consistent with previous findings, TDP-43 overexpression led to the accumulation of aggregates and limited the viability of U2OS. Using this inducible system, we conducted a chemical screen with a library that included FDA-approved drugs. While the primary screen identified several compounds that prevented TDP-43 toxicity, further experiments revealed that these chemicals abrogated the doxycycline-dependent TDP-43 expression. This antagonistic effect was observed with both doxycycline and tetracycline, and in several Tet-On cell lines expressing different genes, confirming the general effect of these compounds as inhibitors of the tetR system. Using the same cell line, a genome-wide CRISPR/Cas9 screen identified epigenetic regulators such as the G9a methyltransferase and TRIM28 as potential modifiers of TDP-43 toxicity. Yet again, further experiments revealed that G9a inhibition or TRIM28 loss prevented doxycycline-dependent expression of TDP-43. In summary, we have identified new chemical and genetic regulators of the tetR system, thereby raising awareness of the limitations of this approach to conduct chemical or genetic screening in mammalian cells., (© 2022 The Authors. FEBS Open Bio published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.)

Most bacteria in the natural environment have a biofilm mode of life, which is intrinsically tolerant to antibiotics. While until now, the knowledge of biofilm formation by Acinetobacter johnsonii is not well understood. In this study, the characteristics and the effect of a sub-inhibitory concentration of antibiotic on A. johnsonii biofilm and planktonic cells were determined. We discovered a positive relationship between biofilm formation and tetracycline resistance, and biofilms rapidly evolve resistance to tetracycline they are treated with. Persister cells commonly exist in both planktonic and biofilm cells, with a higher frequency in the latter. Further transcriptomic analysis speculates that the overexpression of multidrug resistance genes and stress genes were mainly answered to sub lethal concentration of tetracycline in planktonic cells, and the lower metabolic levels after biofilm formation result in high resistance level of biofilm cells to tetracycline. Altogether, these data suggest that A. johnsonii can adjust its phenotype when grown as biofilm and change its metabolism under antibiotic stress, and provide implications for subsequent biofilm control., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Antibacterial hydrogels have gradually become a powerful weapon to treat bacterially infected wounds and accelerate healing. In this paper, we designed a small-molecule self-healing antibacterial hydrogel containing 100% drug-loaded benzyl 3β-amino-11-oxo-olean-12-en-30-oate (GN-Bn), which was governed by π-π stacking, hydrogen bonding, and van der Waals forces. Due to the carrier-free design concept, the problems of interbatch variability during sample preparation and carrier-related toxicity can be effectively avoided. Moreover, the GN-Bn hydrogel exhibited promising antibacterial activities against multidrug-resistant Staphylococcus aureus (MRSA). The minimum inhibitory concentration (MIC) of the GN-Bn hydrogel was 1.5625 nmol/mL, which was lower than those against clinical agents such as norfloxacin, penicillin, and tetracycline. This is attributed to its unique antibacterial mechanism that aims at killing bacteria or preventing their growth by regulating arginine biosynthesis and metabolism through both transcriptomic (RNA-seq) analysis and quantitative polymerase chain reaction (qPCR) analysis. In addition, the GN-Bn hydrogel can also inhibit proinflammatory cytokines (TNF-α, IL-1β, and IL-6) to promote wound healing. Collectively, the GN-Bn hydrogel elicited dual therapeutic effects on an MRSA-infected full-thickness skin wound model through its antibacterial and anti-inflammatory activities, which is attributed to the fact that the GN-Bn hydrogel has multiple advantages including sufficient mechanical stability, biocompatibility, and unique antibacterial mechanisms, making it significantly accelerate MRSA-infected full-thickness skin wound healing as a wound dressing. In a word, the GN-Bn antibacterial hydrogel dressing with an anti-inflammatory and antibacterial bifunctional material holds great potential in clinical application.

Tetracycline-inducible systems are widely used control elements for mammalian gene expression. Despite multiple iterations to improve inducibility, their use is still compromised by basal promoter activity in the absence of tetracyclines. In a mammalian system, we previously showed that the introduction of the G72V mutation in the rtTA-M2 tetracycline activator lowers the basal level expression and increases the fold-induction of multiple genetic elements in a long chimeric antigen receptor construct. In this study, we confirmed that the G72V mutation was effective in minimising background expression in the absence of an inducer, resulting in an increase in fold-expression. Loss of responsiveness due to the G72V mutation was compensated through the incorporation of four sensitivity enhancing (SE) mutations, without compromising promoter tightness. However, SE mutations alone (without G72V) led to undesirable leakiness. Although cryptic splice site removal from rtTA did not alter the inducible control of the luciferase reporter gene in this simplified vector system, this is still recommended as a precaution in more complex multi-gene elements that contain rtTA. The optimized expression construct containing G72V and SE mutations currently provides the best improvement of fold-induction mediated by the rtTA-M2 activator in a mammalian system.

Bacterial infections and inflammation are two main factors for delayed wound healing. Coaxial electrospinning nanofibrous patches, by co-loading and sequential co-delivering of anti-bacterial and anti-inflammation agents, are promising wound dressing for accelerating wound healing. Herein, curcumin (Cur) was loaded into the polycaprolactone (PCL) core, and broad-spectrum antibacterial tetracycline hydrochloride (TH) was loaded into gelatin (GEL) shell to prepare PCL-Cur/GEL-TH core-shell nanofiber membranes. The fibers showed a clear co-axial structure and good water absorption capacity, hydrophilicity and mechanical properties. In vitro drug release results showed sequential release of Cur and TH, in which the coaxial mat showed good antioxidant activity by DPPH test and excellent antibacterial activity was demonstrated by a disk diffusion method. The coaxial mats showed superior biocompatibility toward human immortalized keratinocytes. This study indicates a coaxial nanofiber membrane combining anti-bacterial and anti-inflammation agents has great potential as a wound dressing for promoting wound repair.

Background and Objectives: In spite of the fact that antibiotics are considered to be the cornerstone of modern medicine, their use in the treatment of cancer remains controversial. In the present study, the main objective was to examine the effects of two antibiotics-tetracycline and ampicillin-on the viability, morphology, migration, and organization and structure of the nuclei and the actin fiber network of pharyngeal carcinoma cells-Detroit-562. Materials and Methods: In order to determine the viability of the cells, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method was applied after the cells were stimulated with five concentrations of tetracycline and ampicillin (10, 25, 50, 75, and 100 μM) for 72 h. A scratch assay was used to assess the migration ability of the cells. For the visualization of the nuclei and actin fibers, 4,6-diamidino-2-phenylindole (Dapi) and Rhodamine-Phalloidin were used. Results: There are different effects of tetracycline and ampicillin. Thus, tetracycline: (i) exhibited a concentration-dependent cytotoxic effect, decreasing cell viability to approximately 46%; (ii) inhibits cellular migration up to 16% compared to 60% for control cells; and (iii) induces changes in cell morphology as well as apoptotic changes in the nucleus and F-actin fibers. In contrast, in the case of ampicillin, an increase in viability up to 113% was observed at 10 μM, while a decrease in viability up to approximately 94% was observed at the highest concentration tested (100 μM). Conclusions: The results indicated a different effect regarding the impact on pharyngeal carcinoma cells. Thus, tetracycline has a concentration-dependent cytotoxic effect, while in the case of ampicillin a slight stimulation of cell viability was observed.

This study investigated the effects of microalgae growth on antibiotic removal and the attenuation of antibiotic resistance genes (ARGs)/ARGs host bacteria in algal-bacterial granular sludge (ABGS) system. In the presence of tetracycline (TC) and sulfadiazine (SDZ) mixture (2-4 mg/L), microalgae could grow on bacterial granular sludge (BGS) to form ABGS, with a chlorophyll-a content of 7.68-8.13 mg/g-VSS being achieved. The removal efficiencies of TC and SDZ by ABGS were as high as 79.0 % and 94.0 %, which were 4.3-5.0 % higher than those by BGS. Metagenomic analysis indicated that the relative abundances of TC/SDZ- related ARGs and mobile genetic elements (MGEs) in BGS were 56.1 % and 22.1 % higher than those in ABGS. A total of 26 ARGs were detected from the granules, and they were identified to associate with 46 host bacteria. 13 out of 26 ARGs and 13 out of 46 hosts were shared ARGs and hosts, respectively. The total relative abundance of host bacteria in BGS was 30.8 % higher than that in ABGS. Scenedesmus and Chlorella were the dominant microalgae that may reduce the diversity of ARGs hosts. Overall, ABGS is a promising biotechnology for antibiotic-containing wastewater treatment., (Copyright © 2022. Published by Elsevier B.V.)

Human non-typhoidal salmonellosis is among the leading cause of morbidity and mortality worldwide, resulting in huge economic losses and threatening the public health systems. To date, epidemiological characteristics of non-typhoidal Salmonella (NTS) implicated in human salmonellosis in China are still obscure. Herein, we investigate the antimicrobial resistance and genomic features of NTS isolated from outpatients in Shaoxing city in 2020. Eighty-seven Salmonella isolates were recovered and tested against 28 different antimicrobial agents, representing 12 categories. The results showed high resistance to cefazolin (86.21%), streptomycin (81.61%), ampicillin (77.01%), ampicillin-sulbactam (74.71%), doxycycline (72.41%), tetracycline (71.26%), and levofloxacin (70.11%). Moreover, 83.91% of isolates were resistant to ≥3 categories, which were considered multi-drug resistant (MDR). Whole-genome sequencing (WGS) combined with bioinformatic analysis was used to predict serovars, MLST types, plasmid replicons, antimicrobial resistance genes, and virulence genes, in addition to the construction of phylogenomic to determine the epidemiological relatedness between isolates. Fifteen serovars and 16 STs were identified, with the dominance of S . I 4, [5], 12:i:- ST34 (25.29%), S . Enteritidis ST11 (22.99%), and S . Typhimurium ST19. Additionally, 50 resistance genes representing ten categories were detected with a high prevalence of aac(6')-Iaa (100%), bla TEM-1B (65.52%), and tet(A) (52.87%), encoding resistance to aminoglycosides, β-lactams, and tetracyclines, respectively; in addition to chromosomic mutations affecting gyrA gene. Moreover, we showed the detection of 18 different plasmids with the dominance of IncFIB(S) and IncFII(S) (39.08%). Interestingly, all isolates harbor the typical virulence genes implicated in the virulence mechanisms of Salmonella , while one isolate of S . Jangwani contains the cdtB gene encoding typhoid toxin production. Furthermore, the phylogenomic analysis showed that all isolates of the same serovar are very close to each other and clustered together in the same clade. Together, we showed a high incidence of MDR among the studied isolates which is alarming for public health services and is a major threat to the currently available treatments to deal with human salmonellosis; hence, efforts should be gathered to further introduce WGS in routinely monitoring of AMR Salmonella in the medical field in order to enhance the effectiveness of surveillance systems and to limit the spread of MDR clones., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Chen, Ed-Dra, Zhou, Wu, Zhang and Yue.)

Tetracycline (TC) antibiotics can be detected worldwide in the aquatic environment due to their extensive use and low utilization efficiency, and they may affect the physiological processes of non-target organisms. In this study, the acute and sub-acute toxicities of TC on the freshwater microalga Scenedesmus obliquus were investigated with an emphasis on algal photosynthesis and transcription alterations during an 8 d TC exposure. The results showed that the IC 10 , IC 30 and IC 50 values were 1.8, 4.1 and 6.9 mg/L, respectively. During sub-acute exposure, the microalgae of the IC 10 treatment was able to recover comparable growth to that of the control by day 7, while significantly lower cell densities were observed in the IC 30 and IC 50 treatments at the end of the exposure. The photosynthetic efficiency F v / F M of S. obliquus first decreased as the TC concentration increased and then returned to a level close to that of the control on day 8, accompanied by an increase in photosynthetic activities, including light harvesting, electron transport and energy dissipation. Transcriptomic analysis of the IC 10 treatment (1.8 mg/L TC) revealed that 2157 differentially expressed genes were up-regulated and 1629 were down-regulated compared with the control. KEGG and GO enrichments demonstrated that 28 photosynthesis-related genes involving light-harvesting chlorophyll protein complex, photosystem I, photosystem II, photosynthetic electron transport and enzymes were up-regulated, which may be the factor responsible for the enhanced photosynthesis and recovery of the microalgae. Our work may be helpful not only for gaining a better understanding of the environmental risk of TC at concentrations close to the real levels in natural waters, but also for explaining photosynthesis and related gene transcription induced by antibiotics.

Lactobacilli are the most common probiotic bacteria found in the human gut microbiota, and the presence of acquired antibiotic resistance determinants carried on mobile genetic elements must be screened due to safety concerns. Unnecessary and inappropriate antibiotic therapy, as well as ingested antibiotic resistance bacteria (originating from food or food products), influence the abundance of antibiotic resistance genes in human guts, with serious clinical consequences. The current study looked into the antibiotic resistance of lactobacilli isolated from the guts of sepsis patients on long-term antibiotic therapy. The broth microdilution method was used to investigate the minimum inhibitory concentrations (MICs) of antibiotics such as imipenem, meropenem, erythromycin, tetracycline, cefepime, ciprofloxacin, and gentamycin, and the molecular genetic basis of resistance was studied based on the MIC values. The isolates were phenotypically resistant to tetracycline (20%), fluoroquinolone (20%), and macrolide (5%). Following that, resistance genes for tetracycline [tet(L), tet(O), tet(K), and tet(M)], macrolide [erm(B) and erm(C)], and beta-lactams [bla(CMY)] were investigated. Tetracycline or macrolide resistance genes were not found in the isolates, and only one isolate possessed the bla(CMY) resistance gene. The findings suggested that tetracycline and macrolide resistance may be linked to other resistance genes that were not investigated in this study. Because tetracyclines, fluoroquinolones, and macrolides are commonly used in clinics and animals, there has been concern about the spread of resistance in humans. If acquired antibiotic resistance is passed down through mobile genetic elements, it may serve as a reservoir of resistance for gut pathogens and other microbiome environments., (© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

The increasing antibiotic resistance of Helicobacter pylori infection is a globally urging problem. To investigate the H. pylori resistance situation in Nanjing, China, we enrolled patients in Nanjing First Hospital from January 2018 to May 2021. H. pylori strains were isolated from patients who had at least one positive 13C-urea breath or rapid urease result. Subsequently, we performed antibiotic susceptibility tests on the isolated strains to clarithromycin, metronidazole, levofloxacin, amoxicillin, furazolidone and tetracycline. ARMS-PCR was conducted to determine H. pylori clarithromycin resistance gene mutation. Our results demonstrated that the primary resistance rates of metronidazole, clarithromycin, levofloxacin, amoxicillin, furazolidone and tetracycline were 67.19% (1417/2109), 35.99% (759/2109), 24.23% (511/2109), 0.76% (16/2109), 0.28% (6/2109) and 0.09% (2/2109), respectively. The resistance rates of metronidazole, clarithromycin and levofloxacin elevated significantly after treatment and the three antibiotics composed the majority of multi-resistance patterns. However, the resistance rates of amoxicillin, furazolidone and tetracycline were still in low levels after treatment. ARMS-PCR showed a rather good consistency with antibiotic susceptibility test in detecting clarithromycin resistance, with a kappa value of 0.79. Overall, this study revealed the latest complex situation of antibiotic resistance of H. pylori infection in Nanjing and offered suggestions on clinical medication for curing H. pylori ., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Jiang, Qian, Wang, Dong, Pan, Zhang and Wang.)

The coexistence of multiple pollutants has become a distinctive feature of water pollution. However, there are a few strains that can remove nitrate and tetracycline (TC). Here, the efficiency of strain XS-18 in removing nitrate and TC was analyzed, and the mechanism of tolerance and removal of TC was investigated by infrared spectroscopy, three-dimensional fluorescence spectroscopy, and genome analysis. XS-18 could efficiently remove TC (0.40 mg·L -1 ·h -1 ) at pH 7.0-11.0 with auto-aggregation. TC was removed via extracellular polymeric substance (EPS) (55.90%) and cell surface (44.10%) adsorption. TC (10 mg/L) could stimulate XS-18 to secrete more polysaccharides and hydrophobic proteins to improve its auto-aggregation ability. The findings also confirmed that TC resistance genes were present. Furthermore, the bacterial flagellum, signal transduction of the chemotactic system and regulatory genes were shown to be related to the auto-aggregation of the strain. XS-18 has potential applications in the treatment of wastewater containing nitrate and TC., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Worldwide, the emergence of diarrhoea-causing multi-drug resistant (MDR) bacteria has become a crucial problem in everyday life. Tetracycline (TC) is a bacteriostatic agent that has a wide spectrum of antibacterial activity. One potential strategy to enhance the penetration and antibacterial activity of antibiotics is the use of nanotechnology. In this context, this study dealt with the synthesis of TC loading in biocompatible magnesium oxide nanoparticles (MgONPs), its characterization, and the potency of killing against diarrhoea-causing MDR bacteria E. coli and S. flexneri. TC loaded- MgONPs (MgONPs-TC) were characterized by DLS, SEM-EDS, UV-vis spectroscopy, and FTIR techniques with adequate physical properties. Antibacterial and antibiofilm studies indicate that this nanoparticle successfully eradicated both planktonic and sessile forms of those bacteria. It also significantly reduced the production of bacterial EPS, different levels of antioxidant enzymes, and induced reactive oxygen species (ROS) in the bacterial cell as a mode of antibacterial action. In particular, MgONPs-TC were efficient in reducing the colonization of MDR E. coli and S. flexneri in the C. elegans model. Therefore, all these data suggest that MgONPs-TC are a highly promising approach to combating diseases associated with diarrhoea-causing MDR bacteria in the medical field with limited health care budgets., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Bio-electrochemically assisted anaerobic methanogenic systems (An-BES) are highly effective in wastewater treatment for methane production and degradation of toxic compounds. However, information on the treatment of antibiotic-bearing wastewater in An-BES is still very limited. This study therefore investigated the effect of tetracycline (TC) on the performance, microbial community, as well as functional and antibiotic resistance genes of An-BES. TC at 1 and 5 mg/L inhibited methane production by less than 4.8% compared to the TC-free control. At 10 mg/L TC, application of 0.5 and 1.0 V decreased methane production by 14 and 9.6%, respectively. Under the effect of 1-10 mg/L TC, application of 1.0 V resulted in a decrease of current from 42.3 to 2.8 mA. TC was mainly removed by adsorption; its removal extent increased by 19.5 and 32.9% with application of 0.5 and 1.0 V, respectively. At 1.0 V, current output was not recovered with the addition of granular activated carbon, which completely removed TC by adsorption. Metagenomic analysis showed that propionate oxidizing bacteria and methanogens were more abundant in electrode biofilms than in suspended culture. Antibiotic resistance genes (ARGs) were less abundant in biofilms than in suspended culture, regardless of whether voltage was applied or not. Application of 1.0 V resulted in the enrichment of Geobacter in the anode and Methanobacterium in the cathode. TC inhibited exoelectrogens, propionate oxidizing bacteria, and the methylmalonyl CoA pathway, leading to a decrease of current output, COD consumption, and methane production. These findings deepen our understanding of the inhibitory effect of TC in An-BES towards efficient bioenergy recovery from antibiotic-bearing wastewater, as well as the response of functional microorganisms to TC in such systems., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Based on the prevalence of combined antibiotics and heavy metals contamination in the aquatic environment, this study utilized a microbial approach to achieve simultaneous removal of nitrate (NO 3 - -N), tetracycline (TTC), and Pb(II). Zoogloea sp. FY6 could achieve an optimal NO 3 - -N removal efficiency of 91.5% under C/N ratio of 2.0, at a pH of 6.3, and Fe(II) concentration of 20.23 mg L -1 based on response surface methodology. Additionally, strain FY6 was further found to achieve 89.9 and 81.7% removal of TTC and Pb(II) at 6 h under the optimal conditions. Finally, the results of Fluorescence excitation-emission matrix, X-ray diffraction, Fourier transform infrared spectrometer, and X-ray photoelectron spectroscopy further proved that the biologically formed nanoscale iron oxides and biological action jointly led to the removal of TTC and Pb(II). This study provided a theoretical basis for the application of microbially driven process to remove multi-pollutants in micro-polluted water bodies., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Several classes of antibiotics have long been known for protective properties that cannot be explained through their direct antimicrobial effects. However, the molecular bases of these beneficial roles have been elusive. In this issue of the JCI, Mottis et al. report that tetracyclines induced disease tolerance against influenza virus infection, expanding their protection potential beyond resistance and disease tolerance against bacterial infections. The authors dissociated tetracycline's disease-resistance properties from its disease-tolerance properties by identifying potent tetracycline derivatives with minimal antimicrobial activity but increased capacity to induce an adaptive mitochondrial stress response that initiated disease tolerance mechanisms. These findings have potential clinical applications in viral infections.

Background: Helicobacter pylori infection is responsible for considerable morbidity and mortality worldwide, and eradication rates are falling in many countries, primarily due to clarithromycin and metronidazole resistance., Aims: There is a paucity of contemporary Australian data, which we sought to address by evaluating local rates of resistance of H. pylori to amoxicillin, clarithromycin, metronidazole and tetracycline over the past 20 years., Methods: All gastric biopsy specimens collected at endoscopy to detect H. pylori infection at a single centre underwent routine culture and antibiotic susceptibility testing between 1998 and 2017. Specimens from 12 842 patients were cultured for H. pylori, of which 1473 positive cultures were tested for antibiotic susceptibility., Results: Antibiotic resistance to clarithromycin increased by 3.7% per year (incidence rate ratio [IRR] 1.037; P = 0.014) over 20 years, with a corresponding 5.0% annual increase in minimum inhibitory concentration (MIC) (odds ratio 1.050; P < 0.001). Since 2010, average clarithromycin resistance has exceeded 20%, with >25% of isolates resistant in the past 2 years of data capture. In contrast, rates of resistance to metronidazole (35.3%), amoxicillin (0.14%) and tetracycline (0.34%) and their MIC have remained stable. Review of a representative sample (n = 120; 8%) of these patients revealed that only 5% had documented prior H. pylori eradication therapy., Conclusions: Over the past 20 years there has been a substantial rise in clarithromycin resistance, with stable metronidazole resistance and low rates of resistance to amoxicillin and tetracycline. Current first-line H. pylori eradication therapy may fail to achieve adequate eradication rates, and optimal first-line therapy in Australia should be revisited., (© 2021 Royal Australasian College of Physicians.)

We investigated whether three extractable fractions of lemongrass (Cymbopogon citratus): aqueous and ethanol extracts and lemongrass essential oil exhibited any antimicrobial resistance modulatory effects if used in combination with selected antibiotics ampicillin, tetracycline, streptomycin, cefloxacin and amoxicillin on methicillin-resistant Staphylococcus aureus (MRSA). MRSA growth inhibition (zones of inhibition) was greatest for the lemongrass oil at concentrations of 1, 2, 5, 10 and 20 % (wt/vol). The MIC for lemongrass oil was 0.5 mg/mL, while it was 4 mg/mL for both the aqueous and ethanol extracts. Evaluation of extracts for antibacterial resistance modifying activities when used in combination with either of the five antibiotics at sub-inhibitory concentrations, showed that lemongrass oil highly potentiated the activities of three antibiotics; amoxicillin, streptomycin and tetracycline. The ethanol extract enhanced the activity of tetracycline and ampicillin, while the aqueous extract only increased the activity of tetracycline against MRSA. The activity of cefloxacin with the extracts was either indifferent. Analysis of the lemongrass oil by GC/MS showed the prominence of three compounds: the two isomers neral and geranial of citral and, the acetate geranyl acetate, which together made up 94 % of the composition. The compounds were also observed in the ethanol and water extracts but to a lesser extent when analyzed by HPLC-UV (λ 233 nm). Our study confirms the antibacterial properties of the extracts especially, lemongrass oil. It also demonstrates that lemongrass oil potentiates the activities of three antibiotics against the biofilm-forming MRSA. This biocidal, anti-biofilm disruption and antibiotic potentiating abilities are mainly attributable to citral and geranyl acetate, further evidence of lemongrass oil as a very useful source of phytochemicals, especially citral for the fight against antibiotic resistance., (© 2022 Wiley-VHCA AG, Zurich, Switzerland.)

Objectives: This study aimed to characterize livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) strains isolated from patients admitted to Policlinico San Matteo in Pavia, located in an Italian region with high livestock density., Methods: The starting dataset was composed by 353 MRSA strains isolated from blood cultures between 2011 and 2019 and in 954 MRSA isolated from nasal swabs, wound swabs, skin swabs, ulcer swabs, conjunctival swabs, urine and respiratory samples collected between 2018 and 2019. LA-MRSA was identified based on being MRSA resistant to tetracycline and negative for the PCR amplification of scn locus. Whole genome sequencing of the selected strains was performed, and virulence and resistance genes searched., Results: Five out of 353 MRSA isolates from blood cultures (1.4%) and nine out of 904 MRSA isolates obtained from other materials (1%) were resistant to tetracycline and negative for the scn locus. The 14 strains were also negative for the lukS-pv, tsst, eta and etb loci. Nine of the 14 strains belonged to ST398, the most common ST of LA-MRSA in Europe. ST398 isolates belonged to four spa-types, of which the prevalent was t899. Eight genomes had the cassette SCCmec type V, five genomes had SCCmec type IV and one genome lacked SCCmec, mecA and mecC., Conclusion: The frequency of LA-MRSA in the patients of this study (1.4% in blood cultures, 1% in other samples) is low but relatively constant over time prevalence and comparable to that found in the few studies performed on patients to date., Competing Interests: Declaration of Competing Interest No competing financial interests exist., (Copyright © 2022 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Soil salinity is a worldwide problem and is damaging soil functions. Meanwhile, increasing amounts of anthropogenic antibiotics are discharged to agricultural soils. Little is known about how soil salinity (e.g., NaCl) could influence the bioavailability of antibiotics to bacteria. In this study, a tetracycline-responsive Escherichia coli bioreporter grew on the surfaces of agar microcosms at the same tetracycline concentration (200 μg/L), but various NaCl concentrations (0.5-19.2 g/L) with estimated osmotic potential of -0.18 to -1.80 MPa, and agar content (0.3%-5%) with estimated intrinsic permeability of 38 to 32,928 nm 2 . These agar microcosms mimicked very fine textured soils with a range of NaCl salinity. Increasing agar content lowered the intrinsic permeability hence decreasing tetracycline bioavailability to E. coli, due likely to the reduced mass transfer of tetracycline via water flow. Intriguingly, tetracycline bioavailability increased with increasing NaCl concentration which caused the increase in osmotic stress. This is contradictory to the notion that osmotic stress reduces bacterial chemical uptake. Further analysis of E. coli membrane integrity demonstrated that the enhanced tetracycline bioavailability to bacteria could result from the compromised cell membranes and enhanced membrane permeability at higher NaCl salinity. Overall, this study suggests that high soil salinity (NaCl) may enhance the selection pressure exerted by antibiotics on bacteria., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Aim: The aim of this research was to assess the binding of fibrin clot to the teeth affected by periodontal disease following exposure to different root conditioning agents., Materials and Methods: A total of 60 human teeth with a solitary root that were subjected to extraction following severe periodontal disease were used as study samples in this research. Two analogous grooves were prepared on the proximal radicular surface of every sample employing a diamond-tapered fissure bur using an aerator handpiece beneath abundant irrigation. Every sample was assigned to one of the following groups: • Group I: Tetracycline hydrochloride solution • Group II: Ethylenediaminetetraacetic acid (EDTA) gel • Group III: Biopure MTAD™ Subsequent to conditioning, the samples were rinsed for 3 minutes with phosphate-buffered saline (PBS) and permitted to air-dry for 20 minutes. A drop of fresh human whole blood procured from a hale and hearty volunteer was coated onto the dentin blocks in all three groups. A scanning electron microscope under 5000× magnification at 15 kV was used to examine the samples. Kruskal-Wallis test and Mann-Whitney U test were performed to procure the inter- and intragroup assessments Results: The greatest fibrin clot union was noted in the EDTA gel group at 2.86 ± 0.14 in pursuit by Biopure MTAD™ group at 2.39 ± 0.08 as well as tetracycline hydrochloride solution group at 1.82 ± 0.10. A statistically significant difference was noted between the investigational groups ( p < 0.001)., Conclusion: This research arrived at a conclusion that the dentinal surfaces subjected to conditioning with EDTA gel group as well as coated with human whole blood resulted in appreciably superior fibrin clot bonding to dentin vs Biopure MTAD™ as well as the tetracycline hydrochloride solution group., Clinical Significance: Connective tissue attachment subsequent to surgical procedures causing the adhesion of a fibrin clot to the radicular surface as a result of initial wound healing processes is directly related to periodontal regeneration. It depends on biocompatibility for the fibrin clot and the periodontal pathosis-affected radicular surface to stick together, which can be procured with the aid of a variety of root conditioning measures in course of periodontal treatment.

The CuBi 2 O 4 /Bi 4 O 5 I 2 S-scheme heterojunction structure was constructed by a hydrothermal and subsequent calcination route. The combination of CuBi 2 O 4 and Bi 4 O 5 I 2 produced excellent photocatalytic performance under an LED light. A series of technical characterizations, including X-ray diffraction (XRD), scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS), were used to determine the successful construction of S-scheme CuBi 2 O 4 /Bi 4 O 5 I 2 composites. The improvement of photogenerated carrier separation efficiency helped to achieve the best photocatalytic performance of 37% CuBi 2 O 4 /Bi 4 O 5 I 2 , which can degrade tetracycline (TC) to 81.67% in 90 min, and completely inactivate Escherichia coli (E. coli) in 20 min and Staphylococcus aureus (S. aureus) in 40 min. The effects of some key parameters (such as the concentration of pollutants, the amount of catalyst, pH value of a solution, various inorganic anions and various water substrates) and the possible degradation path of tetracycline were systematically studied. Finally, the removal of pollutants and inactivation of bacterial mechanisms based on the S-scheme heterojunction (CuBi 2 O 4 /Bi 4 O 5 I 2 ) was proposed. This study provides insight into the synthesis of S-scheme heterojunction photocatalysts, which can efficiently degrade organic pollutants and inactivate bacteria under LED light irradiation., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Aliarcobacter butzleri is an emerging gastrointestinal pathogen found in many countries worldwide. In France, it has become the third most commonly isolated bacterial species from the stools of patients with intestinal infections. No interpretative criteria for antimicrobial susceptibility testing have been proposed for A. butzleri, and most strains are categorized using the recommendations of the Clinical and Laboratory Standards Institute or the European Committee on Antimicrobial Susceptibility Testing for Campylobacter or Enterobacterales . In the present study, the genomes of 30 resistant A. butzleri isolates were analyzed to propose specific epidemiological cut-off values for ampicillin, ciprofloxacin, erythromycin, and tetracycline. The identification of a β-lactamase and the T85I GyrA mutation associated with ampicillin and ciprofloxacin resistance, respectively, allowed us to adjust the disk diffusion (DD) and MIC cut-off values for these molecules. However, epidemiological cut-off values for erythromycin and tetracycline could not be estimated due to the absence of known resistance mechanisms. The present study paves the way for building a consensus for antimicrobial susceptibility testing for this concerning pathogen. IMPORTANCE Aliarcobacter butzleri is an emerging and concerning intestinal pathogen. Very few studies have focused on this particular species, and antimicrobial susceptibility testing (AST) is based on methods that have been mostly developed for Campylobacter spp. In fact, no disk diffusion and E-tests adapted cut-offs for A. butzleri are available which leads to misinterpretations. We have shown here that NGS approach to identify genes and mutations in close relation to phenotypic resistance levels is a robust way to solve that issue and precisely differentiate WT and NWT A. butzleri isolates for frequently used antimicrobials. MIC and DD cut-off values have been significantly adjusted and answer the need for a global consensus regarding AST for A. butzleri.