Antimicrobial Susceptibility Assays Research Articles

Pomegranate Rhizosphere Microbial Diversity Revealed by Metagenomics: Toward Organic Farming, Plant Growth Promotion and Biocontrol?

Food production must undergo systems change to meet the sustainable development goals (SDGs). For example, organic farming can be empowered by soil microorganisms with plant growth promotion (PGP) and biocontrol features. In this context, there have been limited studies on pomegranate. We investigated microbial diversity in rhizosphere of the pomegranate "Bhagwa" variety and its potential role in PGP and biocontrol. Both bulk and rhizosphere soil samples were analyzed for their physicochemical properties. Whole metagenome sequencing was conducted using the Illumina NovaSeq6000 platform. Surprisingly, we found that bulk and rhizosphere soil samples had comparable microbial diversity. Metagenome sequencing revealed the abundance of Streptomyces indicus, Bradyrhizobium kalamazoonesis, and Pseudomonas cellulosum in the rhizosphere that are reported here for the first time in agricultural literature. Pathway prediction analysis using KEGG (Kyoto Encyclopedia for Genes and Genomes) and COG (clusters of orthologous genes) databases identified metabolic pathways associated with biocontrol properties against pathogens. We confirmed the metagenome data in vitro, which demonstrated their PGP potential and antimicrobial properties. For instance, S. indicus produced high concentration of indole-3-acetic acid, a PGP phytohormone, that can stimulate plant growth. In addition, an antimicrobial susceptibility assay suggested that bacterial extracts displayed activity against Xanthomonas, a primary pathogen causing the pomegranate wilt disease. In conclusion, this study suggests that S. indicus, B. kalamazoonesis, and P. cellulosum can potentially be PGP and biocontrol agents that may contribute to increased crop productivity in pomegranate cultivation. These agents and their combinations warrant future research with an eye on SDGs and so as to enable and innovate organic farming and pomegranate agricultural practices.

Phenotypic and molecular characterization of clinically isolated antibiotics-resistant S. aureus (MRSA), E. coli (ESBL) and Acinetobacter 1379 bacterial strains.

Antibiotic-resistant bacteria causing nosocomial infections pose a significant global health concern. This study focused on examining the lipid profiles of both non-resistant and clinically resistant strains of Staphylococcus aureus (MRSA 1418), E. coli (ESBL 1384), and Acinetobacter 1379. The main aim was to investigate the relationship between lipid profiles, hydrophobicity, and antibiotic resistance so as to identify the pathogenic potential and resistance factors of strains isolated from patients with sepsis and urinary tract infections (UTIs). The research included various tests, such as antimicrobial susceptibility assays following CLSI guidelines, biochemical tests, biofilm assays, and hydrophobicity assays. Additionally, gas chromatography mass spectrometry (GC-MS) and GC-Flame Ionization Detector (GC-FID) analysis were used for lipid profiling and composition. The clinically isolated resistant strains (MRSA-1418, ESBL-1384, and Acinetobacter 1379) demonstrated resistance phenotypes of 81.80%, 27.6%, and 63.6%, respectively, with a multiple antibiotic resistance index of 0.81, 0.27, and 0.63. Notably, the MRSA-1418 strain, which exhibited resistance, showed significantly higher levels of hemolysin, cell surface hydrophobicity, biofilm index, and a self-aggregative phenotype compared to the non-resistant strains. Gene expression analysis using quantitative real-time PCR (qPCR). Indicated elevated expression levels of intercellular adhesion biofilm-related genes (icaA, icaC, and icaD) in MRSA-1418 (pgaA, pgaC, and pgaB) and Acinetobacter 1379 after 24h compared to non-resistant strains. Scanning electron microscopy (SEM) was employed for structural investigation. These findings provide valuable insights into the role of biofilms in antibiotic resistance and suggest potential target pathways for combating antibiotic-resistant bacteria.

Smelly shark, smelly ray: what is infecting you?

Although elasmobranchs are consumed worldwide, bacteriological assessments for this group are still sorely lacking. In this context, this study assessed bacteria of sharks and rays from one of the most important landing ports along the Rio de Janeiro coast. Bacteria were isolated from the cloacal swabs of the sampled elasmobranchs. They were cultured, and Vibrio, Aeromonas, and Enterobacterales were isolated and identified. The isolated bacteria were then biochemically identified and antimicrobial susceptibility assays were performed. Antigenic characterizations were performed for Salmonella spp. and Polymerase Chain Reaction (PCR) assays were performed to identify Escherichia coli pathotypes. Several bacteria of interest in the One Health context were detected. The most prevalent Enterobacterales were Morganella morganii and Citrobacter freundii, while Vibrio harveyi and Vibrio fluvialis were the most prevalent among Vibrio spp. and Aeromonas allosacharophila and Aeromonas veronii bv. veronii were the most frequent among Aeromonas spp. Several bacteria also displayed antimicrobial resistance, indicative of Public Health concerns. A total of 10% of Vibrio strains were resistant to trimethoprim-sulfamethoxazole and 40% displayed intermediate resistance to cefoxitin. Salmonella enterica strains displayed intermediate resistance to ciprofloxacin, nalidixic acid and streptomycin. All V. cholerae strains were identified as non-O1/non-O139. The detected E. coli strains did not exhibit pathogenicity genes. This is the first study to perform serology assessments for S. enterica subsp. enterica isolated from elasmobranchs, identifying the zoonotic Typhimurium serovar. Salmonella serology evaluations are, therefore, paramount to identify the importance of elasmobranchs in the epidemiological salmonellosis chain. The detection of several pathogenic and antibiotic-resistant bacteria may pose significant Public Health risks in Brazil, due to high elasmobranch consumption rates, indicating the urgent need for further bacteriological assessments in this group.

Prevalence, Virulence Profiles, and Genetic Relatedness of Escherichia coli O45 from Raw meats, Southern Thailand

Non-O157 Enterohaemorrhagic Escherichia coli (EHEC) has become a major public health concern all around the globe. This study investigated the E. coli serogroup O45 from raw meats in southern Thailand using the immunomagnetic separation approach (IMS). Forty-nine E. coli O45 strains were obtained from 13 positive meat samples (13/105), showing the prevalence of O45 contamination as 12.4%. They were not members of EHEC. Exploration of other virulence genes exhibited that fimH, responsible for bacterial adherence, was found in all strains, while astA encoding for EAST-1 toxin was 63.2% of the strains. Moreover, lpf encoding long polar fimbriae was found as 30.6%. Phylogenetic group analysis revealed that the majority of E. coli O45 belonged to group D (88%), followed by group A (8%) and group B1 (4%). However, none belonged to group B2. The intactness of stx2-phage integration sites showed that sbcB was occupied by prophages at the highest rate, followed by the Z2577 site. Antimicrobial susceptibility assay demonstrated relatively high bacterial resistance to cephalothin (78%), streptomycin (51%), cotrimoxazole (39%), tetracycline (31%), and chloramphenicol (23%). Furthermore, multi-drug resistant ability was uncovered in O45 strains at 49%. DNA profiling of E. coli O45 by BOX-PCR analyzed at 80% genetic similarity revealed 5 distinguishable clusters. More importantly, the strains from different samples and time intervals showed identical DNA fingerprints, suggesting that they may have originated from the same bacterial clone.

Aurones and derivatives as promising New Delhi metallo-β-lactamase (NDM-1) inhibitors

Bacterial resistance is undoubtedly one of the main public health concerns especially with the emergence of metallo-β-lactamases (MBLs) able to hydrolytically inactivate β-lactam antibiotics. Currently, there are no inhibitors of MBLs in clinical use to rescue antibiotic action and the New Delhi metallo-β-lactamase-1 (NDM-1) is still considered as one of the most relevant targets for inhibitor development. Following a fragment-based strategy to find new NDM-1 inhibitors, we identified aurone as a promising scaffold. A series of 60 derivatives were then evaluated and two of them were identified as promising inhibitors with Ki values as low as 1.7 and 2.5 µM. Moreover, these two most active compounds were able to potentiate meropenem in in vitro antimicrobial susceptibility assays. The molecular modelling provided insights about their likely interactions with the active site of NDM-1, thus enabling further improvement in the structure of this new inhibitor family.

Enteropathogenic Escherichia coli is a predominant pathotype in healthy pigs in Hubei Province of China.

This study aims to investigate the prevalence of intestinal pathogenic Escherichia coli (InPEC) in healthy pig-related samples and evaluate the potential virulence of the InPEC strains. A multiplex PCR method was established to identify different pathotypes of InPEC. A total of 800 rectal swab samples and 296 pork samples were collected from pig farms and slaughterhouses in Hubei province, China. From these samples, a total of 21 InPEC strains were isolated, including 19 enteropathogenic E. coli (EPEC) and 2 shiga toxin-producing E. coli (STEC) strains. By whole-genome sequencing and in silico typing, it was shown that the sequence types and serotypes were diverse among the strains. Antimicrobial susceptibility assays showed that 90.48% of the strains were multi-drug resistant. The virulence of the strains was first evaluated using the Galleria mellonella larvae model, which showed that most of the strains possessed medium to high pathogenicity. A moderately virulent EPEC isolate was further selected to characterize its pathogenicity using a mouse model, which suggested that it could cause significant diarrhea. Bioluminescence imaging (BLI) was then used to investigate the colonization dynamics of this EPEC isolate, which showed that the EPEC strain could colonize the mouse cecum for up to 5 days.

Effect of Ginkgo biloba and Anacyclus pyrethrum against Streptococcus species isolated from Dental caries and Periodontitis

Streptococcusis the main etiological agents of dental caries and periodontitis. This study aims to evaluate the antimicrobial activity of Anacyclus pyrethrum and Ginkgo biloba extracts on the isolated Streptococcus speciesfrom dental caries and periodontitis samples. Antimicrobial susceptibility assays such as the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the extract against Streptococcus species were performed. S. mutans resistance was also determinedby plasmid curing. The extract of A. pyrethrum and G. biloba exhibited 82.55% and 81.73% of inhibition against Streptococcus mutans at MIC 8 mg/ml and 9mg/ml respectively. Other Streptococcus species are resistant to A. pyrethrum (>77%) and G. biloba (>74%). MBC of A. pyrethrum extract was 20mg/ml for S. mutans and 22mg/ml for other Streptococcus species. Meanwhile, MBC was 24mg/ml and 26mg/ml for G. bilobaagainst S. mutansand other Streptococcus species respectively. The plasmid curing by faint plasmid band analysis indicated that these herbal extracts partially reversed the resistance against S. mutans. Our observations indicated that the A. pyrethrum and G.biloba extracts combat resistance against oral streptococcus. Further study needs to be conducted to check the potential active ingredients of these herbal extracts and to be formulated in oral care applications.

Antimicrobial susceptibility assays for Neisseria gonorrhoeae: a proof-of-principle population-based retrospective analysis

Neisseria gonorrhoeae treatment guided by molecular antimicrobial susceptibility assays could improve treatment options and antimicrobial stewardship; however, few commercial assays are available. We aimed to investigate antimicrobial susceptibility of N gonorrhoeae isolates in New South Wales, Australia, and estimate the potential usefulness of hypothetical combinations of rapid molecular antimicrobial susceptibility assays. In this proof-of-principle, population-based, retrospective analysis, we assessed N gonorrhoeae susceptibility data for ceftriaxone, azithromycin, ciprofloxacin, and penicillin. Isolates were previously collected as part of the Australian Gonococcal Surveillance Programme between Jan 1, 2008, and Dec 31, 2019. All cultured N gonorrhoeae isolates with susceptibility data to all four antimicrobials were included. However, only one isolate was included if several isolates originated from the same individual within 13 days of the previous isolate originating from that individual, and there were less than two standard double-dilution minimum inhibitory concentrations between the isolates. We assessed the use of different combinations of hypothetical antimicrobial susceptibility assays and treatment combinations in terms of their ability to minimise overall ceftriaxone use, and use specifically in isolates with decreased susceptibility to ceftriaxone, compared with standard non-assay-guided empirical ceftriaxone treatment. We included 23 089 N gonorrhoeae isolates. The prevalence of antimicrobial sensitivity fluctuated significantly during the study. Isolates with decreased susceptibility to ceftriaxone were more likely to be resistant to one or more antimicrobials than isolates without decreased susceptibility (782 [98·6%] of 793 vs 10 661 [47·8%] of 22 296), particularly ciprofloxacin (p<0·0001) and penicillin (p<0·0001). Compared with empirical ceftriaxone treatment, we estimated that strategies based on the use of hypothetical antimicrobial susceptibility would reduce ceftriaxone use (p<0·0001). However, because of co-resistance, most assay-directed treatment strategies, including those involving use of assays for two antibiotics, would result in only moderate reductions in ceftriaxone use among isolates with decreased susceptibility to ceftriaxone. Individualised treatment guided by molecular antimicrobial susceptibility diagnostics could help to reduce overall ceftriaxone use in gonorrhoea. However, the use of these assays needs to be informed by the non-random nature of co-resistance among circulating N gonorrhoeae strains. Australian Government and Queensland Government.

Analysis of an IncR plasmid carried by carbapenem-resistant Klebsiella pneumoniae: A survey of swine Klebsiella pneumoniae in Jilin Province

This study was conducted in Jilin Province to investigate the mechanism involved in the antibiotic resistance and pathogenicity of Klebsiella pneumoniae. Lung samples were collected from large-scale pig farms in Jilin Province. Antimicrobial susceptibility and mouse lethality assays were carried out. K. pneumoniae isolate JP20, with high virulence and antibiotic resistance, was chosen for whole-genome sequencing. The complete sequence of its genome was annotated, and the virulence and antibiotic resistance mechanism were analysed. A total of 32 K. pneumoniae strains were isolated and tested for antibiotic resistance and pathogenicity. Among them, the JP20 strain showed high levels of resistance to all tested antimicrobial agents and strong pathogenicity in mice (lethal dose of 1.35×1011 CFU/mL). Sequencing of the multidrug-resistant and highly virulent K. pneumoniae JP20 strain revealed that the antibiotic resistance genes were mainly carried by an IncR plasmid. We speculate that extended-spectrum β-lactamases and loss of outer membrane porin OmpK36 play an important role in carbapenem antibiotic resistance. This plasmid contains a mosaic structure consisting of a large number of mobile elements. Through genome-wide analysis, we found that an lncR plasmid carried by the JP20 strain may have evolved in pig farms, possibly leading to multidrug resistance in the JP20 strain. It is speculated that the antibiotic resistance of K. pneumoniae in pig farms is mainly mediated by mobile elements (insertion sequences, transposons, and plasmids). These data provide a basis for monitoring the antibiotic resistance of K. pneumoniae and lay a foundation for an improved understanding of the genomic characteristics and antibiotic resistance mechanism of K. pneumoniae.

Phenotypic and Genetic Analysis of KPC-49, a KPC-2 Variant Conferring Resistance to Ceftazidime-Avibactam and Maintaining Resistance to Imipenem and Meropenem.

Klebsiella pneumoniae, a gram-negative bacterium, poses a severe hazard to public health, with many bacterial hosts having developed resistance to most antibiotics in clinical use. The goal of this study was to look into the development of resistance to both ceftazidime-avibactam and carbapenems, including imipenem and meropenem, in a K. pneumonia strain expressing a novel K. pneumoniae carbapenemase-2 (KPC-2) variant, referred to as KPC-49. After 1 day of incubation of K1 on agar containing ceftazidime-avibactam (MIC = 16/4 mg/L), a second KPC-producing K. pneumoniae strain (K2) was recovered. Antimicrobial susceptibility assays, cloning assays, and whole genome sequencing were performed to analyse and evaluate antibiotic resistance phenotypes and genotypes. K. pneumoniae strain (K1), that produced KPC-2, was susceptible to ceftazidime-avibactam but resistant to carbapenems. The K2 isolate harboured a novel bla KPC-49 variant, which differs from bla KPC-2 by a single nucleotide (C487A), and results in an arginine-serine substitution at amino acid position 163 (R163S). The mutant K2 strain was resistant to both ceftazidime-avibactam and carbapenems. We demonstrated the ability of KPC-49 to hydrolyse carbapenems, which may be attributed to high KPC-49 expression or presence of an efflux pump and/or absence of membrane pore proteins in K2. Furthermore, blaKPC-like was carried on an IncFII (pHN7A8)/IncR-type plasmid within a TnAs1-orf-orf-orf-orf-orf-orf-ISKpn6-bla KPC-ISKpn27 structure. The bla KPC-like gene was flanked by various insertion sequences and transposon elements, including the Tn3 family transposon, such as TnAs1, TnAs3, IS26, and IS481-ISKpn27. New KPC variants are emerging owing to sustained exposure to antimicrobials and modifications in their amino acid sequences. We demonstrated the drug resistance mechanisms of the new mutant strains through experimental whole genome sequencing combined with bioinformatics analysis. Enhanced understanding of laboratory and clinical features of infections due to K. pneumoniae of the new KPC subtype is key to early and accurate anti-infective therapy.

Antibiogram Screening of Bacterial Pathogens Obtained From Respiratory ‎Manifested Patients in Great Cairo ‎

Respiratory tract infections are the most prevalent infections and implemented in hundred thousand deaths among humans globally, especially in the last years. The goal of the current investigation was to determine the prevalence and antibiotic sensitivity pattern of microorganisms isolated from respiratory samples obtained from laboratories in Great Cairo between January to June 2022. A total of 107 samples distributed as fifteen naso-oral, forty three throat and forty nine sputum were collected. Among them, 91 samples (85.04%) exhibited significant microbial growth on culture media and near equal rate in patients with male and female gender (46 and 45 respectively). The most susceptible ages were both under 12 and over 50 years. The microorganism identification and antibiotic susceptibility testing were conducted. Out of 91isolates species, 56 (61.54%) were Gram-negative bacteria, 35 (38.46%) were Gram-positive bacteria. Prime microorganisms isolated were Klebsiella pneumonia 28 (30.77%), Streptococcous pneumonia 26 (28.57%), Staphylococcous aureus 9 (9.9%), Pseudomonas aeruginosa 8 (8.79%), Escherichia coli (E. coli) 8 (8.79%), Citrobacter spp. 7 (7.69%), Proteus vulgaris 4 (4.4%) and one sample positive for non-lactose fermenting (NLF) E. coli (1.1%). In antimicrobial susceptibility assay, the isolates were mostly resistant to many antibiotics; particularly amoxicillin-clavulanic acid, ampicillin, ceftazidime, clindamycin, oxicillin and trimethoprim-sulfamethoxazole. On the other hand the isolates exposed maximum susceptibility to imipenem and piperacillin-tazobactam (100%), as well as cefoperazone-sulbactam (96.66%) and amikacin (85%).In conclusion, the precise identification of causative microorganisms and their susceptibility profiles to antibiotics is pivotal for selecting targeted and effective antibiotic therapy in respiratory tract infections, and to hinder the emergence of multidrug-resistant bacteria. Gram-negative bacteria exhibited the species most commonly obtained. A high rate of drug resistance was observed in this investigation which may be of public health significance.

Physicochemical Properties Predict Retention of Antibiotics in Water-in-Oil Droplets.

Water-in-oil droplet microfluidics promises capacity for high-throughput single-cell antimicrobial susceptibility assays and investigation of drug resistance mechanisms. Every droplet must serve as an isolated environment with a controlled antibiotic concentration in such assays. While technologies for generation, incubation, screening, and sorting droplets mature, predictable retention of active molecules inside droplets remains a major outstanding challenge. Here, we analyzed 36 descriptors of the antibiotic molecules against experimental results on the cross-talk of antibiotics in droplets. We show that partition coefficient and fractional polar surface area are the key physicochemical properties that predict antibiotic retention. We verified the prediction by monitoring growth inhibition by antibiotic-loaded neighboring droplets. Our experiments also demonstrate that transfer of antibiotics between droplets is concentration- and distance-dependent. Our findings immediately apply to designing droplet antibiotic assays and give deeper insight into the retention of small molecules in water-in-oil emulsions.

Peptide Stability Is Important but Not a General Requirement for Antimicrobial and Antibiofilm Activity In Vitro and In Vivo.

Peptide stability to proteases has been a major requirement for developing peptide therapeutics. This study investigates the effects of peptide stability on antimicrobial and antibiofilm activity under various conditions. For this purpose, two human cathelicidin-derived peptides differing in stability to proteases were utilized. While GF-17, a peptide derived from the major antimicrobial region of human LL-37, can be rapidly cleaved by proteases, the engineered peptide 17BIPHE2 is resistant to multiple proteases. In the standard antimicrobial susceptibility, killing kinetics, and membrane permeabilization assays conducted in vitro using planktonic bacteria, these two peptides displayed similar potency. The two peptides were also similarly active against methicillin-resistant Staphylococcus aureus (MRSA) USA300 prior to biofilm formation. However, 17BIPHE2 was superior to GF-17 in disrupting preformed biofilms probably due to both enhanced stability and slightly higher DNA binding capacity. In a wax moth model, 17BIPHE2 better protected insects from MRSA infection-caused death than GF-17, consistent with the slower degradation of 17BIPHE2 than GF-17. Here, peptide antimicrobial activity was found to be critical for in vivo efficacy. When incorporated in the nanofiber/microneedle delivery device, GF-17 and 17BIPHE2 displayed a similar effect in eliminating MRSA in murine chronic wounds, underscoring the advantage of nanofibers in protecting the peptide from degradation. Since nanoformulation can ease the requirement of peptide stability, it opens the door to a direct use of natural peptides or their cocktails for antimicrobial treatment, accelerating the search of effective antibiofilm peptides to treat chronic wounds.

Identification, 16S rRNA-based characterization, and antimicrobial profile of Gallibacterium isolates from broiler and layer chickens.

Gallibacterium spp., particularly G. anatis, have received much attention as poultry pathogens in recent years. We report here the presence and antimicrobial resistance profile of 69 Gallibacterium isolates obtained from 2,204 diagnostic submissions of broiler and layer chickens in 2019-2021. Gallibacterium-positive chickens had lesions primarily in the respiratory tract, reproductive tract, and related serosal surfaces. Gallibacterium spp. were initially identified based on their typical cultural characteristics on blood agar. The isolates were confirmed by a genus-specific PCR spanning 16S-23S rRNA and MALDI-TOF mass spectrometry. Phylogenetic analysis based on 16S rRNA gene sequence revealed distinct clades. Of the 69 isolates, 68 clustered with the reference strains of G. anatis and 1 with Gallibacterium genomospecies 1 and 2. Antimicrobial susceptibility testing of 58 of the 69 isolates by a MIC method showed variable responses to antimicrobials. The isolates were all susceptible to enrofloxacin, ceftiofur, florfenicol, and gentamicin. There was a high level of susceptibility to trimethoprim-sulfamethoxazole (98.0%), streptomycin (98.0%), amoxicillin (84.0%), sulfadimethoxine (71.0%), and neomycin (71.0%). All of the isolates were resistant to tylosin. There was resistance to penicillin (98.0%), erythromycin (95.0%), clindamycin (94.0%), novobiocin (90.0%), tetracycline (88.0%), oxytetracycline (76.0%), and sulfathiazole (53.0%). A high rate of intermediate susceptibility was observed for spectinomycin (67.0%) and sulfathiazole (40.0%). Our findings indicate a potential role of G. anatis as an important poultry pathogen and cause of subsequent disease, alone or in combination with other pathogens. Continuous monitoring and an antimicrobial susceptibility assay are recommended for effective treatment and disease control.

IG1, a Mansonone F Analog, Exhibits Antibacterial Activity against Staphylococcus aureus by Potentially Impairing Cell Wall Synthesis and DNA Replication.

Infection caused by Staphylococcus aureus, especially methicillin-resistant S. aureus (MRSA), is very common in communities and hospitals, which poses a great challenge to human health. Therefore, increasing attention has been paid to finding effective antimicrobial agents. Mansonone F is a natural compound which has an oxaphenalene skeleton and anti-S. aureus activity, but its sources are limited and its synthesis is difficult. Thus, IG1, a C9-substituent mansonone F analog, was assessed for its activity against Staphylococcus aureus and its mechanism of action was investigated. Antimicrobial susceptibility assays showed that IG1 has strong antibacterial activity against S. aureus, including MRSA, with minimum inhibitory concentrations (MICs) ranging from 0.5 to 2 μg/mL, which were very close to those of vancomycin, and these changed little, even with an increase in the amount of the inoculum. To further explore the antibacterial properties of IG1, time-kill experiments were conducted. Compared with vancomycin and moxifloxacin, treatment with different concentrations of IG1 reduced the viability of organisms in a very similar manner and the reduction was not significant, which indicated that IG1 is a potentially strong anti-S. aureus agent. Finally, the antibacterial mechanism was analyzed, with flow cytometric analysis revealing that IG1 treatment resulted in a time-dependent decrease in the DNA content of S. aureus. Transmission electron microscopy (TEM) analysis showed that very few dividing cells could be found and the cell wall was damaged in the field of IG1-treated cells. These results indicate that IG1 is a potential new antibacterial agent against S. aureus, including MRSA.

Indigenous Plant-Based Alternatives for Treating Cattle Skin Ailments: A Case Study from the Mnisi Tribal Authority of Mpumalanga Province, South Africa

The study was carried out in the Mnisi Tribal Authority, Mpumalanga Province, South Africa, between August and October 2021. Ethno-veterinary data were collected using pre-structured questionnaires and field observations with cattle owners. The data were analyzed and summarized using descriptive statistics. Frequently mentioned plants belonged to the families Vitaceae (Cissus quandrangularis), Euphorbiaceae (Synadenium cupulare), Fabaceae (Dichrostachys cinerea), Solanaceae (Nicotiana batacum), Euphorbiacae (Jatropha zayheri), and Apocynaceae (Sarcostemma viminate). Antimicrobial susceptibility assays with ampicillin as positive control were carried out on extracts of Synadenium cupulare (aerial parts and leaves), Dichrostachys cinerea (seeds/ fruits), and Cissus quadrangularis (aerial parts and leaves). Aqueous extracts of S.cupulare (L) were the most effective against E. faecalis with the lowest MIC of 0.73 mg/ml, followed by aqueous extracts of Dichrostachys cinerea with an MIC of 1.36. Open wounds were the most common ailment (35%), followed by abscesses (23%) and abrasions (17%). Documenting Indigenous knowledge can be a basis for the development of sustainably used medicinal plants in the area and the validation of these plant preparations for veterinary use since the low cost of these preparations makes them useful for local communities.

Cosmeceutical Potential of Extracts Derived from Fishery Industry Residues: Sardine Wastes and Codfish Frames.

The fishery industry generates large amounts of waste (20–75% (w/w) of the total caught fish weight). The recovery of bioactive compounds from residues and their incorporation in cosmetics represents a promising market opportunity and may contribute to a sustainable valorisation of the sector. In this work, protein-rich extracts obtained by high-pressure technologies (supercritical CO2 and subcritical water) from sardine (Sardina pilchardus) waste and codfish (Gadus morhua) frames were characterized regarding their cosmeceutical potential. Antioxidant, anti-inflammatory and antibacterial activities were evaluated through chemical (ORAC assay), enzymatic (inhibition of elastase and tyrosinase), antimicrobial susceptibility (Klebsiella pneumoniae, Staphylococcus aureus and Cutibacterium acnes) and cell-based (in keratinocytes-HaCaT) assays. Sardine extracts presented the highest antibacterial activity, and the extract obtained using higher extraction temperatures (250 °C) and without the defatting step demonstrated the lowest minimum inhibitory concentration (MIC) values (1.17; 4.6; 0.59 mg/mL for K. pneumoniae, S. aureus and C. acnes, respectively). Codfish samples extracted at lower temperatures (90 °C) were the most effective anti-inflammatory agents (a concentration of 0.75 mg/mL reduced IL-8 and IL-6 levels by 58% and 47%, respectively, relative to the positive control). Threonine, valine, leucine, arginine and total protein content in the extracts were highlighted to present a high correlation with the reported bioactivities (R2 ≥ 0.7). These results support the potential application of extracts obtained from fishery industry wastes in cosmeceutical products with bioactive activities.

Antibiotics Resistance Pattern of Food-Borne Bacteria Isolated from Ice Cream in Bangladesh: A Multidisciplinary Study

Ice cream is one of the most popular food items consumed during the summer season in Bangladesh by all ages but mostly school-going students. Due to the ingredients and handling process of ice cream, it acts as a good shelter for pathogenic and nonpathogenic microorganisms. Therefore, we aimed to investigate the microbial count, prevalence, isolate and characterize multidrug-resistant bacteria in ice cream samples collected from nearby shops of schools in Tangail district, Bangladesh. Ice-cream consumer and nonconsumer students were selected by providing questionnaires. Total viable count (TVC) and total coliform count (TCC) were determined by pour plate methods, where conventional methods were performed for bacterial identification. The Kirby-Bauer disk diffusion method was used to determine the antimicrobial susceptibility of bacterial isolates. Kado and Liu method, with some modifications, was used to extract plasmid from the isolated bacteria and visualized through gel electrophoresis. The demographic characteristics showed that the degree of symptoms regarding microorganisms mediated disorders and rate of antibiotics intake in ice cream consumers were significantly higher than the nonconsumers. The range of TVC and TCC in the ice cream samples was found 0–9.9 × 109 CFU/ml and 0–900 CFU/ml, respectively. Interestingly, 93.75% of the total ice cream samples also showed fungal positive. A total of 12 different bacterial species were identified, including Proteus spp, E. coli, V.cholera, Pseudomonas spp, Shigella spp, Klebsiella spp, Aeromonas spp, V. Parahemolyticus. Salmonella paratyphi, Citrobacterspp, Plesiomonasspp, and Staphylococcus aureus. The antimicrobial susceptibility assay showed the multiple resistance frequency of these isolates to different antimicrobial drugs. All individual isolates were screened for plasmid DNA, and we found that seven strains harbored a single or more than two plasmids sized approximately between 1.9 and 140 MDa, indicating a possible connection between resistance phenotype pattern and genotype.

Prevalence and antimicrobial susceptibility profiles of ESBL-producing Klebsiella Pneumoniae from broiler chicken farms in Shandong Province, China

Klebsiella pneumoniae (K. pneumoniae) is a conditionally pathogenic bacterium present in the intestinal or the respiratory tract of animals, and it is a common factor in acquired infections and a major threat to public health. Increased production of extended-spectrum β-lactamases (ESBLs) has become a serious issue in the treatment of K. pneumoniae infections. In this study, we examined the serotypes and antibiotic resistance profiles of K. pneumoniae isolated from broiler chickens on farms in Shandong Province, China. The K. pneumoniae isolation rate was 4.67% (33/707), and the serotype Capsular K54 (42.42%, 14/33) was the most prevalent serotype in broilers in Shandong. The antimicrobial susceptibility assay revealed that the 33 isolates were resistant to 28 antimicrobial drugs to varying degrees; among these, the highest resistance rate was observed for tetracyclines (90.91%), and the lowest rate of resistance was observed for moxifloxacin and fosfomycin (0%). The multidrug resistance (MDR) rate was 87.88% (29/33). The carrying rate of β-lactam-resistance genes was as high as 100%, with blaSHV having the highest rate (93.94%). It is worth noting that one carbapenem-resistant K. pneumoniae (CRKP) isolate carrying blaNDM-1 and one colistin-resistant K. pneumoniae (COLR-KP) isolate carrying mcr-3 were found in broiler chickens. This study indicates that ESBL-producing CRKP isolates and COLR-KP isolates have emerged on poultry farms in Shandong and could be a potential threat to food safety and public health.

Dissemination and Genetic Relatedness of Multidrug-Resistant and Extensively Drug-Resistant Acinetobacter baumannii Isolates from a Burn Hospital in Iraq.

Acinetobacter baumannii is an aggressive opportunistic bacterial pathogen that causes severe nosocomial infections, especially among burn patients. An increasing number of hospitals-acquired infections have been reported all over the world. However, little attention has been paid to the relatedness between A. baumannii isolates from different hospital environments and patients. In this study, 27 isolates were collected from the Burn and Plastic Surgery Hospital of Al Sulaymaniyah City, Iraq, from January through December 2019 (11 from patients and 16 from the wards environment), identified to species level as A. baumannii using Vitek 2 system and molecular detection of 16S rRNA gene, and then confirmed by targeting the blaOXA-51 gene. Moreover, the isolates were characterized by means of automated antimicrobial susceptibility assay, antimicrobial-resistant patterns, a phenotypic method using a combined disk test, and molecular methods for the detection of class A and C β-lactamase genes, and finally, the genetic relatedness was classified. Antimicrobial susceptibility testing showed that 63% (17/27) of the retrieved A. baumannii isolates were extensively drug-resistant to 8/9 antimicrobial classes. Furthermore, 37% (10/27) of the isolates were classified as multidrug-resistant; 8 isolates exhibited similar resistant patterns and the other two isolates showed 2 different patterns, while resistance was greater in isolates from patients than from the ward environment. Combined disk test showed that two isolates contained extended-spectrum β-lactamase. All isolates carried blaTEM-1, and two copies of the blaCTX-1 gene were indicated in one isolate, while blaSHV was absent in all isolates. Twenty-four isolates carried the blaAmpC gene; among them, 3 isolates harbored the insertion sequence ISAba-1 upstream to the gene. Using Enterobacterial Repetitive Intergenic Consensus PCR, the isolates were clustered into 6 distinct types; among them, two clusters, each of four strains, were classified to contain isolates from both patients and environments. The clusters of similar genotypes were found in inpatients as well as the environments of different wards during time periods, suggesting transmission within the hospital. Identification of possible infection sources and controlling the transmission of these aggressive resistance strains should be strictly conducted.