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Presence of extended-spectrum β-lactamases (ESBL)-producing Enterobacteriaceae in the dairy farm environment and food chain could be a possible interface for the exchange of antimicrobial resistance genes between humans and animals. A total of 600 samples comprised of raw bovine milk, faeces, feed, environmental swabs and water samples from 20 different bovine dairy farms in and around Hisar city, Haryana, India were analysed for presence of ESBL encoding genes. Out of 240 isolates of Escherichia coli obtained, 74 isolates were found to be ESBL producers. Maximum number of ESBL isolates were found from faeces (40.5%) followed by raw milk (37.8%) and environmental swabs (17.5%). Most of the ESBL E. coli isolates were sensitive to chloramphenicol (82.4%) and gentamicin (77.0%) antibiotics. The bla CTX-M gene was found to be most prevalent (52.0%) followed by bla TEM (9.45%) while bla SHV gene alone was not detected in any sample by simplex PCR. However, the co-expression of blaCTX-M + blaTEM (21.6%) and blaCTX-M + blaSHV (4.05%) genes were also observed. The housing system, milking method and the hygienic mangement practices followed at farm level are found to be significant risk factors of ESBL-producing E. coli in dairy farms of Haryana. [ABSTRACT FROM AUTHOR]

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ABSTRACT The spread of carbapenem-resistant Pseudomonas aeruginosa and carbapenemase-producing Enterobacterales (CPE) has dramatically impacted morbidity and mortality. COVID-19 pandemic has favored the selection of these microorganisms because of the excessive and prolonged use of broad-spectrum antibiotics and the outbreaks related to patient transfer between hospitals and inadequate personal protective equipment. Therefore, early CPE detection is considered essential for their control. We aimed to compare conventional phenotypic synergy tests and two lateral flow immunoassays for detecting carbapenemases in Enterobacterales and P. aeruginosa. We analyzed 100 carbapenem-resistant Gram-negative bacilli isolates, 80 Enterobacterales, and 20 P. aeruginosa (86 isolates producing KPC, NDM, OXA-48, IMP, and VIM carbapenemases and 14 non-carbapenemase-producing isolates). We performed a modified Hodge test, boronic acid and ethylenediaminetetraacetic acid (EDTA) synergy tests, and two lateral flow immunoassays: RESIST-4 O.K.N.V. (Coris Bioconcept) and NG Test Carba 5 (NG Biotech). In total, 76 KPC, seven VIM, one NDM, one OXA-48, and one isolate coproducing KPC + NDM enzymes were included. The concordance of different methods estimated by the Kappa index was 0.432 (standard error: 0.117), thus showing a high variability with the synergy tests with boronic acid and EDTA and reporting 16 false negatives that were detected by the two immunochromatographic methods. Co-production was only detected using immunoassays. Conventional phenotypic synergy tests with boronic acid and EDTA for detecting carbapenemases are suboptimal, and their routine use should be reconsidered. These tests depend on the degree of enzyme expression and the distance between disks. Lateral flow immunoassay tests are a rapid and cost-effective tool to detect and differentiate carbapenemases, improving clinical outcomes through targeted therapy and promoting infection prevention measures. IMPORTANCE Infections due to multidrug-resistant pathogens are a growing problem worldwide. The production of carbapenemases in Pseudomonas aeruginosa and Enterobacterales cause a high impact on the mortality of infected patients. Therefore, it is of great importance to have methods that allow the early detection of these multi-resistant microorganisms, achieving the confirmation of the type of carbapenemase present, with high sensitivity and specificity, with the aim of improving epidemiological control, dissemination, the clinical course to through targeted antibiotic therapy and promoting infection control in hospitals.

This study aimed to develop a relatively natural and safe botanical insecticide for controlling the storage pest Tribolium castaneum in the egg and pupal stages. It examined how Elsholtzia densa Benth. essential oil (EO) and its primary components, β -caryophyllene and limonene, affected T. castaneum eggs and pupae through contact and fumigation. Among th, the contact activities of β -caryophyllene against T. castaneum eggs and pupae are LD 50 (median lethal dose, 50%) = 0.156 mg/cm2 and ED 50 (median effective dose, 50%) = 16.35 mg/pupa respectively. The study also investigated the effect of β -caryophyllene and limonene on T. castaneum eggs and pupae through synergistic contact and fumigation. When the mixing ratio of β -caryophyllene and limonene was 7:1, the LD 50 value of contact activity against T. castaneum eggs was reduced to 0.100 mg/cm2, displaying an obvious synergistic effect. Experiments were conducted to investigate the antitoxic effect of β -caryophyllene on T. castaneum eggs and pupae, as well as its effects on the enzymatic activity of acetylcholinesterase, succinate dehydrogenase, glutathione S-transferase and carboxylesterase in T. castaneum pupae. Finally, the molecular docking techniques were employed to confirm the aforementioned effects on enzyme function. The findings of this study might help improve storage pest control with T. castaneum and create eco-friendly insecticides using E. densa EO, β -caryophyllene, and limonene. [Display omitted] • Elsholtzia densa Benth. EO and β -caryophyllene have excellent inhibitory activity against Tribolium castaneum eggs. • When β -caryophyllene and limonene are mixed in a ratio of 7:1, the contact activity on T. castaneum eggs is the best. • T. castaneum pupae showed certain antitoxic effects on β -caryophyllene. • β -Caryophyllene can significantly inhibit the enzyme activity of succinate dehydrogenase in T. castaneum pupae. [ABSTRACT FROM AUTHOR]

Background and Aim: Antibiotics’ resistance is the leading cause of complications in the treatment of urinary tract infections. This study aimed to screen the antimicrobial potential of 8 plants from Cameroon against multi-resistant uropathogenic (MRU) bacteria and to investigate their antibioresistance reversal properties. Method: Bioactive compounds were extracted from leaves of Leucanthemum vulgare, Cymbopogon citratus, Moringa oleifera and Vernonia amygdalina; barks of Cinchona officinalis and Enantia chlorantha barks and seeds of Garcinia lucida and leaves and seeds of Azadirachta indica using water and ethanol as solvents. The extracts were tested against Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 6538 and Candida albicans 10231 using the well diffusion and the broth microdilution methods. The antibiotic-resistance reversal activity was assessed against selected MRU bacteria. The phytochemical composition and the elemental composition of the most active extracts were assessed respectively using HPLC-MS/MS and X-ray fluorescence (XRF) spectrometry. Results: Among the most active plants, in decreasing order of antimicrobial activity we found ethanolic (EE) and aqueous extracts (AE) of E. chloranta bark (ECB), EE of L. vulgare leaves and G. lucida seeds. The best synergies between common antibiotics and extracts were found with EE-ECB which well-modulated kanamycin nitrofurantoin and ampicillin. All the compounds identified in EE-ECB were alkaloids and the major constituents were palmatine (51.63%), columbamine+7,8-dihydro-8-hydroxypalmatine (19.21%), jatrorrhizine (11.02%) and pseudocolumbamine (6.33%). Among the minerals found in EE-ECB (S, Si, Cl, K, Ca, Mn, Fe, Zn and Br), Br, Fe and Cl were the most abundant with mean fluorescence intensities of 4.6529, 3.4854 and 2.5942 cps/uA respectively. Conclusions: The ethanol extract of the bark of E. chlorantha has remarkable, broad-spectrum antimicrobial and contains several palmatine derivatives.

INTRODUCTION: The emergence of extended-spectrum beta-lactamase-producing Enterobacteriaceae (E-ESBLs) is currently a major public health problem in the world and, in particular, in developing countries. In Cameroon, data on E-ESBLs are rare, especially in Garoua and in the northern region of the country. The objective of this study is to document the epidemiology of E-ESBL infections in small children and to explore their associations with possible risk factors. METHODS: this was a cross-sectional, descriptive study conducted from June 14 to September 30, 2018, including small children with suspected urinary tract infections (UTI) attending the outpatient pediatric departments of two health facilities in the city of Garoua. Urine samples were analyzed at the Bacteriology Laboratory of the Pasteur Center of Cameroon, Annex Garoua. Bacterial culture was carried out on Bio-Rad UriSelect® chromogenic agar and the identification was confirmed by bioMérieux API 20E. The antibiotic susceptibility was determined using the bioMérieux ATB UR gallery and the ESBL phenotype was detected by the double disk synergy method according to the CA-SFM 2013 recommendations. The data was analyzed with the R Statistical Software version 2.15.2. RESULTS: a total of 57 urine samples were collected from children aged from one month to two years, 37 boys and 20 girls. Bacteria were detected by culture in 20 samples: Escherichia coli was the most frequently (75 %) isolated species followed by Klebsiella pneumoniae (25%). More than half of the infected samples (55%) contained E-ESBL. The presence of an ESBL was significantly associated with previous antibiotic intake up to 3 months prior current UTI (p=0.01664). The E-ESBL strains showed co-resistance to different antibiotics. CONCLUSION: this study reveals the important dissemination of E-ESBLs among small children in the community and a high rate of co-resistance to the different antibiotic families commonly used.

Objective: Combination therapy is needed to treat extensively drug-resistant (XDR) Acinetobacter baumannii infection. Colistin (Col) in combination with another drug is usually used for that purpose. The aim of this study was to determine the activity of antimicrobial combinations against XDR A. baumannii using both standard checkerboard (CB) method and E-test. E-test was also evaluated for application in a diagnostic bacteriology laboratory by comparing its efficacy with that of CB method. Methods: Eighty clinical isolates of XDR A. baumannii were used to determine the activity of the following antimicrobial combinations by CB method and E-test: Col+cefoperazone/sulbactam (Cps), Cps+moxifloxacin (Mox), and Col+Mox. Comparison of CB and E-test was also evaluated. Results: By CB method, Col+Cps yielded a synergistic effect rate (12.5%) higher than those of the other 2 combinations (CpS+Mox 5% and Col+Mox 0%). The majority of test results revealed additivity. Col+Cps, Cps+Mox, and Col+Mox exhibited additive effect against 78.75%, 85.0%, and 87.5% of isolates, respectively. Overall, E-test and CB yielded only 37.5% concordant rates. However, high concordant rates were specifically observed in additive effect of Col+Cps (73.8%) and Cps+Mox (80.4%). Conclusion: Col+Cps exhibited better activity than the other two combinations against XDR A. baumannii. E-test is the method that should be used, but its use is limited to the additive results of Col+Cps and Cps+Mox.

Introduction: the emergence of extended-spectrum beta-lactamase-producing Entero bacteriaceae (E-ESBLs) is currently a major public health problem in the world and, in particular, in developing countries. In Cameroon, data on E-ESBLs are rare, especially in Garoua and in the northern region of the country. The objective of this study is to document the epidemiology of E-ESBL infections in small children and to explore their associations with possible risk factors. Methods: this was a cross-sectional, descriptive study conducted from June 14 to September 30, 2018, including small children with suspected urinary tract infections (UTI) attending the outpatient pediatric departments of two health facilities in the city of Garoua. Urine samples were analyzed at the Bacteriology Laboratory of the Pasteur Center of Cameroon, Annex Garoua. Bacterial culture was carried out on Bio-Rad UriSelect® chromogenic agar and the identification was confirmed by bioMérieux API 20E. The antibiotic susceptibility was determined using the bioMérieux ATB UR gallery and the ESBL phenotype was detected by the double disk synergy method according to the CA-SFM 2013 recommendations. The data was analyzed with the R Statistical Software version 2.15.2. Results: a total of 57 urine samples were collected from children aged from one month to two years, 37 boys and 20 girls. Bacteria were detected by culture in 20 samples: Escherichia coliwas the most frequently (75%) isolated species followed by Klebsiella pneumoniae(25%). More than half of the infected samples (55%) contained E-ESBL. The presence of an ESBL was significantly associated with previous antibiotic intake up to 3 months prior current UTI (p=0.01664). The E-ESBL strains showed co-resistance to different antibiotics. Conclusion: this study reveals the important dissemination of EESBLs among small children in the community and a high rate of co-resistance to the different antibiotic families commonly used. [ABSTRACT FROM AUTHOR]

Infections due Scedosporium spp. in lung transplant recipients are associated with disseminated disease with high mortality rates. The adjunctive local antifungal therapy may be a useful option when systemic treatment is insufficient and/or surgery is not feasible. We present a case of mixed disseminated infection due Scedosporium apiospermum and S. prolificans in a lung transplant recipient. Combined local and systemic antifungal therapy provided an unusual long-term survival in the intensive care unit.

非顯而易知性的判斷標準，一直是美國專利法實務與學界想要嘗試解決的問題之一。本文認為，由於客觀證據與時間因素，可呈現出發明當時的狀態，因此，符合美國專利法第103 條的文義要求，而應被優先考量。因此，本文嘗試將最高法院於Graham 案中提出的「輔助性判斷因素」，加入美國學界所提出之「時間判斷法」判斷架構中，主張若有客觀證據可證明，同業與發明人面臨相同的發明條件，卻只有發明人成功解決他人無法克服的需求，則發明即可合理被認為非顯而易知。本研究之最大貢獻在於克服先前判斷標準之判斷困境，特別是在KSR 案中仍未解決的後見偏差問題。 The test of obviousness has long been a problem which the courts and scholars in U.S. are trying to resolve. This Article argues the objective indicia and the timing of invention should have priority when considering nonobviousness, because these factors can demonstrate the conditions at the time the invention was made and therefore fit the requirement of Section 103 of Patent Law. Hence, this Article tries to merge “Secondary Consideration” mentioned in Graham case by Supreme Court with “the Timing Approach” raised by scholar, and argues if there are objective evidences to prove that the person having ordinary skill in the art faces the same conditions of invention, only the inventor can resolve the problem successfully while others cannot, then the invention can be reasonably seen as nonobvious. The most contribution of this research is overcoming problems of previous tests of nonobviousness, especially the hindsight problem which was still not solved in KSR case. solved in KSR case.

Background/aim: Acinetobacter baumannii (A. baumannii) is one of the most common healthcare-associated infectious agents worldwide. The aim of this study was to investigate the in vitro synergistic activities of several antibiotic combinations against carbapenem-resistant (CR) A. baumannii isolates. Materials and methods: Eighteen CR A. baumannii strains were isolated from the patients who were hospitalized in the intensive care unit between June 2012 and August 2012. The in vitro effects of single and binary combinations of meropenem (MEM), colistin (CST), tigecycline (TGC), and sulbactam (SUL) on these isolates were determined using the Epsilometer test (E-test) method. Results: All 18 isolates were resistant to MEM and SUL and susceptible to CST. TGC was detected as susceptible in two of the isolates and intermediate susceptibility results were observed in the remaining isolates. With MEM-CST and MEM-TGC combinations, synergism was determined against all isolates. The synergistic and/or additive effect ratios were detected in MEM-SUL, CST-SUL, TGC-SUL, and CST-TGC combinations as 16.7%, 38.9%, 16.7%, and 5.6%, respectively. Conclusion: Among the tested antimicrobial combinations, the in vitro combination of MEM with TGC or CST was most effective against the CR A. baumannii strains. [ABSTRACT FROM AUTHOR]

Infections due Scedosporium spp. in lung transplant recipients are associated with disseminated disease with high mortality rates. The adjunctive local antifungal therapy may be a useful option when systemic treatment is insufficient and/or surgery is not feasible. We present a case of mixed disseminated infection due Scedosporium apiospermum and S. prolificans in a lung transplant recipient. Combined local and systemic antifungal therapy provided an unusual long-term survival in the intensive care unit. [ABSTRACT FROM AUTHOR]

Background and Aim: Antibiotics' resistance is the leading cause of complications in the treatment of urinary tract infections. This study aimed to screen the antimicrobial potential of 8 plants from Cameroon against multi-resistant uropathogenic (MRU) bacteria and to investigate their antibioresistance reversal properties., Method: Bioactive compounds were extracted from leaves of Leucanthemum vulgare , Cymbopogon citratus , Moringa oleifera and Vernonia amygdalina ; barks of Cinchona officinalis and Enantia chlorantha barks and seeds of Garcinia lucida and leaves and seeds of Azadirachta indica using water and ethanol as solvents. The extracts were tested against Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 6538 and Candida albicans 10231 using the well diffusion and the broth microdilution methods. The antibiotic-resistance reversal activity was assessed against selected MRU bacteria. The phytochemical composition and the elemental composition of the most active extracts were assessed respectively using HPLC-MS/MS and X-ray fluorescence (XRF) spectrometry., Results: Among the most active plants, in decreasing order of antimicrobial activity we found ethanolic (EE) and aqueous extracts (AE) of E. chloranta bark (ECB), EE of L. vulgare leaves and G. lucida seeds. The best synergies between common antibiotics and extracts were found with EE-ECB which well-modulated kanamycin nitrofurantoin and ampicillin. All the compounds identified in EE-ECB were alkaloids and the major constituents were palmatine (51.63%), columbamine+7,8-dihydro-8-hydroxypalmatine (19.21%), jatrorrhizine (11.02%) and pseudocolumbamine (6.33%). Among the minerals found in EE-ECB (S, Si, Cl, K, Ca, Mn, Fe, Zn and Br), Br, Fe and Cl were the most abundant with mean fluorescence intensities of 4.6529, 3.4854 and 2.5942 cps/uA respectively., Conclusions: The ethanol extract of the bark of E. chlorantha has remarkable, broad-spectrum antimicrobial and contains several palmatine derivatives., Competing Interests: The authors declare no conflict of interest., (© 2022 The Author(s). Published by IMR Press.)

We performed a retrospective extended-spectrum β-lactamase (ESBL) molecular characterization of Proteus mirabilis isolates recovered from urine of spinal cord injury patients. A incorrectly detected TEM-24-producing clone and a new weakly expressed TEM-derived ESBL were discovered. In such patients, ESBL detection in daily practice should be improved by systematic use of a synergy test in strains of P. mirabilis resistant to penicillins. [ABSTRACT FROM AUTHOR]

urate determination of resistance is important to ensure appropriate antimicrobial therapy in Stenotrophomonas maltophilia infections. This study was undertaken to evaluate the susceptibility results obtained by disc diffusion, E-test, Phoenix system, and reference agar dilution method and also to evaluate the in vitro activity of various antimicrobial combinations against multidrug-resistant S. maltophilia. Susceptibilities to several antimicrobial agents were determined by agar dilution, disc diffusion, and E-test according to the US Clinical Laboratory and Standards Institute (CLSI) guidelines. Results were also evaluated in the in Phoenix system for available agents. Twelve different antibiotic combinations were tested for synergy by the E-test method. Most synergic combinations were confirmed by microdilution checkerboard assay. Tigecycline, trimethoprim/sulfamethoxazole (TMP-SMX) and doxycycline were the most effective drugs against S. maltophilia. Poorest agreement was determined by disc diffusion and E-test against ticarcillin/clavulanate and ciprofloxacin ( κ < 0.4), by disc diffusion against colistin ( κ < 0.4), and by the Phoenix system against piperacillin/tazobactam ( κ < 0.4). Based on these data, disc diffusion seems to be unreliable for ticarcillin/clavulanate, ciprofloxacin, and colistin; E-test for ticarcillin/clavulanate and ciprofloxacin; and the Phoenix system for piperacillin/tazobactam for S. maltophilia susceptibility testing. Synergistic activity was detected predominantly with TMP-SMX + ticarcillin/clavulanate and TMP-SMX + ceftazidime. TMP-SMX + ceftazidime synergy was also supported by the checkerboard method. However, TMP-SMX + ticarcillin/clavulanate combination revealed indifferent effect by the checkerboard assay. As ticarcillin/clavulanate and ciprofloxacin E-test results were beyond the acceptable correlation limits, synergy testing performed with these agents was considered as unreliable. Further studies are required to standardize susceptibility testing, especially for colistin, ticarcillin/clavulanate, and ciprofloxacin for S. maltophilia. TMP-SMX-containing drug combinations seemed to be more synergistic on multidrug-resistant S. maltophilia; however, these results merit further evaluation. [ABSTRACT FROM AUTHOR]

The Vitek-2 extended-spectrum β-lactamase (ESBL) test was assessed using a collection of 94 ESBL-positive and 71 ESBL-negative non-duplicate isolates of Enterobacteriaceae. These isolates produced a wide diversity of well-characterised β-lactamases, including 61 different ESBLs, two class A carbapenemases and various species-specific β-lactamases. ESBL detection was performed using (i) the conventional synergy test as recommended by the Comité de l’Antibiogramme de la Société Française de Microbiologie, (ii) the CLSI phenotypic confirmatory test for ESBLs, and (iii) the Vitek-2 ESBL test. For Escherichia coli and klebsiellae, the sensitivity/specificity values were 97.3%/96.9% for the synergy test, 91.8%/100% for the CLSI phenotypic confirmatory test, and 91.8%/100% for the Vitek-2 ESBL test. For other organisms, the sensitivity/specificity values were 100%/97.4% for the synergy test, 90.5%/100% for the CLSI phenotypic confirmatory test, and 90.5%/100% for the Vitek-2 ESBL test. The Vitek-2 ESBL test seemed to be an efficient method for routine detection of ESBL-producing isolates of Enterobacteriaceae, including isolates producing AmpC-type enzymes. [ABSTRACT FROM AUTHOR]

The spread of carbapenem-resistant Pseudomonas aeruginosa and carbapenemase-producing Enterobacterales (CPE) has dramatically impacted morbidity and mortality. COVID-19 pandemic has favored the selection of these microorganisms because of the excessive and prolonged use of broad-spectrum antibiotics and the outbreaks related to patient transfer between hospitals and inadequate personal protective equipment. Therefore, early CPE detection is considered essential for their control. We aimed to compare conventional phenotypic synergy tests and two lateral flow immunoassays for detecting carbapenemases in Enterobacterales and P. aeruginosa. We analyzed 100 carbapenem-resistant Gram-negative bacilli isolates, 80 Enterobacterales, and 20 P. aeruginosa (86 isolates producing KPC, NDM, OXA-48, IMP, and VIM carbapenemases and 14 non-carbapenemase-producing isolates). We performed a modified Hodge test, boronic acid and ethylenediaminetetraacetic acid (EDTA) synergy tests, and two lateral flow immunoassays: RESIST-4 O.K.N.V. (Coris Bioconcept) and NG Test Carba 5 (NG Biotech). In total, 76 KPC, seven VIM, one NDM, one OXA-48, and one isolate coproducing KPC + NDM enzymes were included. The concordance of different methods estimated by the Kappa index was 0.432 (standard error: 0.117), thus showing a high variability with the synergy tests with boronic acid and EDTA and reporting 16 false negatives that were detected by the two immunochromatographic methods. Co-production was only detected using immunoassays. Conventional phenotypic synergy tests with boronic acid and EDTA for detecting carbapenemases are suboptimal, and their routine use should be reconsidered. These tests depend on the degree of enzyme expression and the distance between disks. Lateral flow immunoassay tests are a rapid and cost-effective tool to detect and differentiate carbapenemases, improving clinical outcomes through targeted therapy and promoting infection prevention measures. IMPORTANCE Infections due to multidrug-resistant pathogens are a growing problem worldwide. The production of carbapenemases in Pseudomonas aeruginosa and Enterobacterales cause a high impact on the mortality of infected patients. Therefore, it is of great importance to have methods that allow the early detection of these multi-resistant microorganisms, achieving the confirmation of the type of carbapenemase present, with high sensitivity and specificity, with the aim of improving epidemiological control, dissemination, the clinical course to through targeted antibiotic therapy and promoting infection control in hospitals.