Antimicrobial resistance (AMR) is a global health issue disproportionately affecting low- and middle-income countries. In Tanzania, multi-drug-resistant bacteria (MDR) are highly prevalent in clinical and community settings, inhibiting effective treatment and recovery from infection. The burden of AMR can be alleviated if antimicrobial stewardship (AMS) programs are coordinated and incorporate local knowledge and systemic factors. AMS includes the education of health providers to optimise antimicrobial use to improve patient outcomes while minimising AMR risks. For programmes to succeed, it is essential to understand not just the awareness of and receptiveness to AMR education, but also the opportunities and challenges facing health professionals. We conducted in-depth interviews (n = 44) with animal and human health providers in rural northern Tanzania in order to understand their experiences around AMR. In doing so, we aimed to assess the contextual factors surrounding their practices that might enable or impede the translation of knowledge into action. Specifically, we explored their motivations, training, understanding of infections and AMR, and constraints in daily practice. While providers were motivated in supporting their communities, clear issues emerged regarding training and understanding of AMR. Community health workers and retail drug dispensers exhibited the most variation in training. Inconsistencies in understandings of AMR and its drivers were apparent. Providers cited the actions of patients and other providers as contributing to AMR, perpetuating narratives of blame. Challenges related to AMR included infrastructural constraints, such as a lack of diagnostic testing. While health and AMR-specific training would be beneficial to address awareness, equally important, if not more critical, is tackling the challenges providers face in turning knowledge into action. Full article

Staphylococcus aureus was isolated from a total of 360 nasal and groin skin swabs from 180 systematic randomly-selected horses slaughtered for meat at Obollo-Afor, Enugu State, Southeast Nigeria and antimicrobial, methicillin and heavy metal resistance profile and virulence potentials of the isolates established. Baird-Parker agar with egg yolk tellurite was used for S. aureus isolation. S. aureus isolates were confirmed biochemically and serologically using a specific S. aureus Staphytect Plus™ latex agglutination test kit. The antimicrobial resistance profile, methicillin, vancomycin and inducible clindamycin resistance, and β-lactamase production of the isolates were determined with disc diffusion. Tolerance to Copper, Cadmium, Lead and Zinc was assessed using the agar dilution method and virulence potentials were determined using phenotypic methods. Forty-three (23.9%) of the 180 horses harbored S. aureus. Some 71 S. aureus were recovered from the 360 samples. Two (2.8%) of the 71 S. aureus were methicillin-resistant S. aureus (MRSA) and 69 (97.2%) were methicillin-susceptible. MRSA was recovered from 2 (1.1%) of the 180 horses. Some 9.4% of the isolates were multiple drug-resistant (MDR). The mean multiple antibiotic resistance indices (MARI) for the isolates was 0.24. Heavy metal resistance rate of the isolates ranged between 35.4–70.4%. The isolates, including the MRSA strains, displayed virulence potentials as clumping factor and catalase, gelatinase, caseinase, heamolysin, and biofilm was at the rate of 100%, 53.5%, 43.7%, 18.3% and 23.9%, respectively. This study showed that a considerable percentage of horses slaughtered in Obollo-Afor Southeastern Nigeria are potential reservoirs of virulent multiple drug- and heavy metal-resistant S. aureus, including MRSA, that could spread to humans and the environment. Full article

While there are many alternatives to antibiotics for the symptomatic treatment of urinary tract infections (UTIs), their application in practice is limited. Among other things, general practitioners (GPs) often feel pressure from patients to prescribe antibiotics. To gain a better understanding of why this happens and where this pressure originates from, we investigated experiences, expectations, motivations, and perspectives of patients with UTIs in general practice. During this qualitative study we performed 14 semi-structured online interviews among female UTI patients in general practice. Interviews were based on a topic list derived from sensitising concepts. All the interviews were recorded, transcribed, and analysed using a constant comparative technique. Three main categories emerged from the data; (1) experienced versus unexperienced patients with UTI, (2) patient’s lack of knowledge, and (3) patients feeling understood. Inexperienced patients consult a general practitioner for both diagnosis and symptom relief, while experienced patients seem to consult specifically to obtain antibiotics. In addition, patients have a lack of knowledge with regard to the diagnosis, treatment, self-care, and cause of UTIs. Finally, patients’ satisfaction is increased by involving them more in the process of decision making, so they feel understood and taken seriously. Patients’ expectations in UTI management in general practice often arise during their first experience(s) and play a major role in subsequent episodes. In conclusion, preventing misconceptions is especially important in the inexperienced patient group, as this may prevent future overtreatment of UTIs. In addition, involving patients in the decision making process will lead to greater understanding of the GP’s treatment choices. Full article

Electrolyte disorders (EDs) can disrupt normal bodily functions and lead to life-threatening complications. We evaluated whether piperacillin–tazobactam (TZP), a widely used antibiotic for moderate-to-severe infections, is associated with electrolyte imbalances via a disproportionality analysis of a self-reporting pharmacovigilance database. We searched The US Food and Drug Administration (FDA) Adverse Event Reporting System (FAERS) from 2004 to 2018 for EDs related to TZP and calculated three data-mining indices—the proportional reporting ratio (PRR), the reporting odds ratio (ROR), and the information component (IC)—compared to all other drugs. Signals were defined when one of the three criteria of the indices was met. For the signals detected in the initial analysis, further disproportionality analyses in relation to other penicillins were conducted using the same method. A total of 9829 reports related to TZP with 36,207 TZP–adverse event pairs were retrieved. Among 10 EDs, hypokalemia was detected as the only significant signal (PRR 2.61; ROR 2.61, 95% CI: 2.17–3.14; IC 95% lower CI: 1.11) compared to all other drugs. Compared with other penicillins, hypokalemia remained a significant signal for TZP using IC (95% lower CI: 0.26). In conclusion, TZP was significantly associated with hypokalemia. Full article

Salmonella enterica is an important foodborne pathogen that causes gastroenteritis and systemic infection in humans and livestock. Salmonella biofilms consist of two major components—amyloid curli and cellulose—which contribute to the prolonged persistence of Salmonella inside the host. Effective agents for inhibiting the formation of biofilms are urgently needed. We investigated the antibiofilm effect of Raspberry Ketone (RK) and its mechanism of action against Salmonella Typhimurium 14028 using the Congo red agar method, Calcofluor staining, crystal violet method, pellicle assay, and the TMT-labeled quantitative proteomic approach. RK suppressed the formation of different types of Salmonella biofilms, including pellicle formation, even at low concentrations (200 µg/mL). Furthermore, at higher concentrations (2 mg/mL), RK exhibited bacteriostatic effects. RK repressed cellulose deposition in Salmonella biofilm through an unknown mechanism. Swimming and swarming motility analyses demonstrated reduced motility in RK-treated S. typhimurium. Proteomics analysis revealed that pathways involved in amyloid curli production, bacterial invasion, flagellar motility, arginine biosynthesis, and carbohydrate metabolism, were targeted by RK to facilitate biofilm inhibition. Consistent with the proteomics data, the expressions of csgB and csgD genes were strongly down-regulated in RK-treated S. typhimurium. These findings clearly demonstrated the Salmonella biofilm inhibition capability of RK, justifying its further study for its efficacy assessment in clinical and industrial settings. Full article

Management of invasive fungal infections (IFI) and subsequent treatment choices remain challenging for physicians in the ICU. Documented evidence shows increased practice of the inappropriate use of antifungal agents in the ICU. Continuous education of healthcare providers (HCPs) represents the cornerstone requirement for starting an antifungal stewardship program (AFS). This study aimed at evaluating knowledge gaps in systemic antifungal prescribing among physicians and clinical pharmacists in a critical care setting. A cross-sectional, multi-center, survey-based study was conducted in five tertiary hospitals located in Al-Ahsaa, Saudi Arabia between January and May 2021. A self-administered questionnaire was distributed among the targeted clinicians. A total of 63 clinicians were involved (65.5% ICU physicians and 34.5% clinical pharmacists). It was noted that a minority of the participating HCPs (3.2%) had overall good knowledge about antifungal prescribing, but the majority had either moderate (46%) or poor (50.8%) knowledge. The difference in overall knowledge scores between the ICU physicians and the clinical pharmacists (p = 0.925) was not significant. However, pharmacists showed better scores for the pharmacokinetics of antifungal therapy (p = 0.05). This study has revealed a significant gap in the knowledge and practice of clinicians as regards prescribing antifungal therapy in our area. Although the results cannot be generalized, the outcome of this study has exposed the need for a tailored training program essential for carrying out an AFS program. Full article

Fosfomycin disodium is a potential therapeutic option to manage difficult-to-treat infections, especially when combined with other antimicrobials. In this study, we evaluated the activity of fosfomycin in combination with meropenem or polymyxin B against contemporaneous KPC-2-producing K. pneumoniae clinical isolates (KPC-KPN). Synergistic activity was assessed by checkerboard (CKA) and time–kill (TKA) assays. TKA was performed using serum peak and trough concentrations. The activity of these combinations was also assessed in the Galleria mellonella model. Biofilm disruption was assessed by the microtiter plate technique. CKA resulted in an 8- to 2048-fold decrease in meropenem MIC, restoring meropenem activity for 82.4% of the isolates when combined with fosfomycin. For the fosfomycin + polymyxin B combination, a 2- to 128-fold reduction in polymyxin B MIC was achieved, restoring polymyxin B activity for 47% of the isolates. TKA resulted in the synergism of fosfomycin + meropenem (3.0–6.7 log₁₀ CFU/mL decrease) and fosfomycin + polymyxin B (6.0–6.2 log₁₀ CFU/mL decrease) at peak concentrations. All larvae treated with fosfomycin + meropenem survived. Larvae survival rate was higher with fosfomycin monotherapy (95%) than that observed for fosfomycin + polymyxin B (75%) (p-value < 0.0001). Finally, a higher biofilm disruption was observed under exposure to fosfomycin + polymyxin B (2.4–3.4-fold reduction). In summary, we observed a synergistic effect of fosfomycin + meropenem and fosfomycin + polymyxin B combinations, in vitro and in vivo, against KPC-KPN, as well as biofilm disruption. Full article

The pathogenic microorganisms linked to fresh fruits and juices sold out in retail low-cost markets raise safety concerns as they may carry multidrug-resistant (MDR) genes. To evaluate the microbiological quality and safety of highly consumed fruits and derivatives in Imbabura Province, Ecuador, ready-to-eat strawberries (5 independent batches; n = 300 samples), and gooseberries (5 separate batches; n = 500 samples), purchased from a local fruit farm grower and low-cost retail market, along with 20 different natural fruit- and vegetables-based juices (3 independent batches; n = 60 samples) purchased from food courts located within the low-cost markets were analyzed. Bacteriological analysis showed that the microbial quality was lower as several indicators (n = 984) consisting of total coliforms (TCOL), total aerobes (AEROB), Enterobacter spp. (ENT), Shigella spp., (SHIGA), yeasts (YE), and molds (M) were detected. Staphylococcus spp. (STAPHY) was found in both fruits regardless of origin, while Escherichia coli (EC) isolates were found in strawberries but not gooseberries. Salmonella spp. (SALM) were detected in juices only. Antibiotic susceptibility testing showed multidrug resistance of several isolates. The hemolytic pattern revealed that 88.89% of EC and 61.11% of ENT isolates were beta-hemolytic. All STAPHY isolates were beta-hemolytic while SALM and SHIGA were alpha-hemolytic. Plasmid curing assay of MDR isolates (ENT, EC, SALM, and STAPHY) showed that the antibiotic resistance (AR) was highly indicative of being plasmid-borne. These results raise concerns about the consumption of MDR bacteria. However, good agricultural and industrial practices, behavioral change communication, and awareness-raising programs are necessary for all stakeholders along the food production and consumption supply chain. Full article

The aim of the study is to evaluate the effect of the addition of selected fruits and herbs belonging to the “superfoods” category for the bioactivity of a rapeseed honey matrix. Flavored creamed honeys with nine types of various additives (2 and 4% of content) were prepared and analyzed for the content of total phenols, flavonoids, antioxidant (FRAP, DPPH and ABTS) and antibacterial activity against four strains of bacteria. Additionally, the impact of three months of storage on the antioxidant properties of the products obtained was examined. The significant dose-dependent increase in the content of bioactive ingredients and antioxidant capacity in spiced honeys, as compared to control honey, was observed. The highest enrichment was obtained for the addition of powdered sea buckthorn leaves and black raspberry fruits. Honey with the addition of sea buckthorn leaves inhibited the growth of P. aeruginosa, S. aureus and K. pneumonia, whereas honeys with black raspberry and blackcurrant fruits showed activity only on the latter two strains. Furthermore, what is more interesting, honey supplemented with sea buckthorn leaf and black raspberry fruits inhibited S. aureus biofilm formation at the sub-minimum inhibitory concentrations (sub-MICs), showing a dose-dependent anti-biofilm effect. Full article

Klebsiella pneumoniae is a Gram-negative opportunistic pathogen responsible for a variety of community and hospital infections. Infections caused by carbapenem-resistant K. pneumoniae (CRKP) constitute a major threat for public health and are strongly associated with high rates of mortality, especially in immunocompromised and critically ill patients. Adhesive fimbriae, capsule, lipopolysaccharide (LPS), and siderophores or iron carriers constitute the main virulence factors which contribute to the pathogenicity of K. pneumoniae. Colistin and tigecycline constitute some of the last resorts for the treatment of CRKP infections. Carbapenemase production, especially K. pneumoniae carbapenemase (KPC) and metallo-β-lactamase (MBL), constitutes the basic molecular mechanism of CRKP emergence. Knowledge of the mechanism of CRKP appearance is crucial, as it can determine the selection of the most suitable antimicrobial agent among those most recently launched. Plazomicin, eravacycline, cefiderocol, temocillin, ceftolozane–tazobactam, imipenem–cilastatin/relebactam, meropenem–vaborbactam, ceftazidime–avibactam and aztreonam–avibactam constitute potent alternatives for treating CRKP infections. The aim of the current review is to highlight the virulence factors and molecular pathogenesis of CRKP and provide recent updates on the molecular epidemiology and antimicrobial treatment options. Full article

The emergence and global expansion of hyper-virulent and multidrug resistant (MDR) Klebsiella pneumoniae is an increasing healthcare threat worldwide. The epidemiology of MDR K. pneumoniae is under-characterized in many parts of the world, particularly Africa. In this study, K. pneumoniae isolates from hospitals in Khartoum, Sudan, have been whole-genome sequenced to investigate their molecular epidemiology, virulence, and resistome profiles. Eighty-six K. pneumoniae were recovered from patients in five hospitals in Khartoum between 2016 and 2020. Antimicrobial susceptibility was performed by disk-diffusion and broth microdilution. All isolates underwent whole genome sequencing using Illumina MiSeq; cgMLST was determined using Ridom SeqSphere+, and 7-loci MLST virulence genes and resistomes were identified. MDR was observed at 80%, with 35 isolates (41%) confirmed carbapenem-resistant. Thirty-seven sequence types were identified, and 14 transmission clusters (TC). Five of these TCs involved more than one hospital. Ybt9 was the most common virulence gene detected, in addition to some isolates harbouring iuc and rmp1. There is a diverse population of K. pneumoniae in Khartoum hospitals, harbouring multiple resistance genes, including genes coding for ESBLs, carbapenemases, and aminoglycoside-modifying enzymes, across multiple ST’s. The majority of isolates were singletons and transmissions were rare. Full article

Intestinal bacterial infections are a major threat to human and animal health. In this study, we found plant-derived antibacterial xanthones, particularly α-mangostin (AMG) from the mangosteen peel, exhibiting extraordinary activities against Clostridium perfringens. Structure–activity relationship analysis showed that prenylation modulated the activity of xanthones. The efficacy of AMG (4, 8, 20 mg/kg body weight) was also demonstrated in the broiler chicken necrotic enteritis model infected with Clostridium perfringens. In the models (n = 6 per group), feed supplementation of AMG maintained the homeostasis of the gut microbiome by reducing the colonization of clostridia and promoting the integrity of intestinal barriers via the upregulation of mucin expression. These results suggest that plant-derived xanthones may be a potential alternative to antibiotics for treating clostridial enteric infections in the clinic. Full article

Necrotic enteritis (NE), caused by Clostridium perfringens, is an emerging issue in poultry farming. New approaches, other than antibiotics, are necessary to prevent NE development and the emergence of multidrug-resistant bacteria. Enterococci are commensal microorganisms that can produce enterocins, antimicrobial peptides with activities against pathogens, and could be excellent candidates for protective cultures. This study aimed to screen and characterize Enterococcus strains of poultry origin for their inhibitory activity against C. perfringens. In total, 251 Enterococcus strains of poultry origin plus five bacteriocin-producing (BP+) E. durans strains of other origins were screened for antimicrobial activity against the indicator C. perfringens X2967 strain using the “spot on the lawn” method. We detected thirty-two BP+ strains (eleven Enterococcus faecium, nine E. gallinarum, eight E. faecalis, three E. durans, and one E. casseliflavus). We further studied the antimicrobial activity of the supernatants of these 32 BP+ strains using agar well diffusion and microtitration against a collection of 20 C. perfringens strains. Twelve BP+ enterococci that were found to exhibit antimicrobial activity against C. perfringens were characterized using whole genome sequencing. Among these, E. faecium X2893 and X2906 were the most promising candidates for further studies as protective cultures for poultry farming. Both strains belong to the sequence type ST722, harbor the genes encoding for enterocin A and enterocin B, do not possess acquired resistance genes, do not carry plasmids, and present the acm gene, which is implicated in host colonization. Further research is needed to determine the utility of these strains as protective cultures. Full article

Objectives: Fracture-related infection (FRI) is a major complication in orthopedic and trauma surgery. The management and choice of treatment can be difficult depending on multiple factors. Therefore, we implemented a weekly multidisciplinary team discussion to determine diagnostic and treatment strategies in FRI patients and aimed to analyze its effect on clinical outcomes. Methods: Clinical outcomes of FRI patients treated before and after implementation of a structured multidisciplinary treatment (MDT) approach with a weekly case discussion were compared at a follow-up of 12 months. Results: In total, n = 117 were eligible for enrolment, whereby n = 58 patients (72.4% male, mean age 56.7 ± 16.8 years) constituted the MDT group and n = 59 patients (72.9% male, mean age 55.0 ± 16.5 years) the control group. In the MDT group more cases were treated with local antibiotics (67.2% vs. 27.1%, p < 0.001) and significant less amputations (3.4% vs. 6.8%, p = 0.014), as well as less revision surgeries (1.5 ± 1.2 (0–5) vs. 2.2 ± 1.2 (0–7), p = 0.048) were performed. A trend towards less debridement, antibiotics and implant retention (DAIR) procedures, lower rates of recurrence of infection and less treatment failures in the MDT group was observable, even though not statistically significant. Conclusion: An MDT approach providing a patient tailored treatment concept in the treatment of FRI patients appears to be beneficial for the affected patients. Quality and efficacy of implemented MDT meetings should further be evaluated to provide sufficient evidence to further implement this valuable tool in clinical practice and decision making. Full article

Household cats have been identified as potential antimicrobial resistance (AMR) reservoirs, and the extended-spectrum β-lactamases (ESBL) producing E. coli circulating among cats has been more frequently reported globally, but the factors linked to its colonization remain poorly understood. Thus, the objectives of this study were to determine E. coli shedding and the occurrence of multidrug resistant (MDR)- and ESBL-producing E. coli, as well as to determine risk factors associated with colonization of MDR and ESBL-producing E. coli isolated from both healthy and diseased cats in the Eastern Province of Saudi Arabia. In a cross-sectional study, 2000 swabs were collected from five anatomical regions (anus, skin, ear canal, nares, and conjunctival sac) of 209 healthy and 191 diseased cats that were admitted to a veterinary clinic in the Eastern Province of Saudi Arabia. In addition, each cat owner filled out a questionnaire about their cat’s demographics, management, health status, and antimicrobial usage. E. coli was detected in 165 (41.3%) of all cats, including 59 (28.2%) healthy and 106 (55.5%) diseased cats. In total, 170 E. coli isolates were found in healthy (35.3%) and diseased (64.7%) cats. Susceptibility testing revealed that 123 (72.4%) of the E. coli isolates were resistant to at least one of the tested antimicrobials. Overall, 17.6% (30/170) of E. coli isolates were MDR, with 10 (5.9%) and 20 (11.8%) isolates found in healthy and diseased cats, respectively. However, only 12 (7.1%) E. coli isolates were resistant to cefotaxime and harbored the bla_CTX-M gene (ESBL-producer), with seven (4.1%) in healthy and five (2.9%) in diseased cats. Risk factor analysis showed that the odds of MDR and ESBL-producing E. coli were (20 and 17) and (six and eight) times higher when the family and cats were previously treated with antimicrobials, respectively. The presence of a child in the cat’s family was also linked to an increased risk of MDR E. coli colonization (OR = 3.4). In conclusion, a high frequency of MDR and ESBL-producing E. coli was detected among healthy and diseased cats in Saudi Arabia, raising concerns about transmission to humans and supporting the need of a “One Health” approach to address the potential threats of cats as AMR reservoirs. Full article