Antimicrobial Resistance Status Research Articles

Association between respiratory disease pathogens in calves near feedlot arrival with treatment for bovine respiratory disease and subsequent antimicrobial resistance status.

This study assessed the risk of first treatment for bovine respiratory disease (BRD) given detection of nasopharyngeal bacteria (Mannheimia haemolytica, Pasteurella multocida, and Histophilus somni) and corresponding likelihood of antimicrobial susceptibility (C/S) at two time points during the early feeding period. Relationships between C/S results and later treatment for BRD were evaluated at both the calf-level and pen-level. The association between calf-level and pen-level C/S findings during the early feeding period and subsequent C/S results at BRD treatment were also reported. Auction-sourced, recently-weaned beef calves (n = 1,599 steers) were placed in adjacent feedlot pens (8 × 100 calves) in two subsequent years. Deep nasopharyngeal (DNP) swabs were collected from all calves at time of arrival processing (1DOF) and before metaphylaxis administration with either tulathromycin or oxytetracycline, 12 days later (13DOF), and at the time of first treatment for BRD. All samples were tested for C/S. Several pen-level and individual calf-level C/S measures of interest were associated with future treatment for BRD and C/S at the time of treatment. The median DOF for first BRD treatment was 24 days following tulathromycin metaphylaxis and 11 days following oxytetracycline. Overall, sampling at 13DOF resulted in the best fit for more models of subsequent treatment for BRD and C/S results at BRD treatment than for sampling at arrival. In individual calves, recovery of M. haemolytica, P. multocida, or H. somni at 13DOF was associated with subsequent treatment for BRD within 45DOF. Pen-level prevalence of Pasteurellacea bacteria with tetracycline or macrolide resistance at arrival and 13DOF were associated with detection of bacteria with antimicrobial resistance (AMR) at BRD treatment, as were individual calf results at 13DOF. These findings suggest that the bacteria and AMR outcomes recovered from cattle near two weeks on feed can inform the prediction of future BRD risk and concurrent antimicrobial susceptibility results at time of first BRD treatment. Notably, the associations between pen-level C/S results from previous testing and corresponding findings in calves with BRD from the same pen suggested potential testing strategies to inform antimicrobial use protocols for feedlot cattle.

Non-typhoidal Salmonella contributes to gastrointestinal infections in Morogoro: Evidence from patients attending Morogoro regional referral hospital in Tanzania.

Salmonella is one of the most common causes of food-borne outbreaks and infection worldwide. Non-typhoidal Salmonella (NTS) infections are common and remain a significant public health problem among important bacterial foodborne diseases. The current study aimed to establish the Non typhoidal Salmonella infection and antimicrobial resistance status among selected patients at Morogoro Regional Referral Hospital (MRRH), Morogoro Region, Tanzania, to inform clinical care management and public health interventions. A cross-sectional study was conducted using medical records and samples were collected from hospitalised and outpatients between October and December 2021. A total of 153 participants were enrolled in the study and 132 consented to being sampled. The collected samples were analysed using standard microbiological techniques. The isolates were subjected to molecular genotyping, where Polymerase Chain Reaction (PCR) was performed targeting the 16S rDNA gene. PCR products were then submitted for sequencing to establish phylogenetic relatedness. Antimicrobial susceptibility testing and resistance genes screening were also conducted. The phylogenetic analysis identified two Salmonella serovars; Salmonella Enteritidis and Salmonella Typhimurium. The isolates were from four adults and seven children patients. The isolates were tested against six antimicrobial agents: tetracycline, trimethoprim, gentamycin, ciprofloxacin, ampicillin and cefotaxime. Further antimicrobial assays were performed by screening 10 antimicrobial resistance genes using PCR. Overall, the highest resistance was observed in ampicillin (100%), whereas the lowest resistance was recorded for ciprofloxacin and gentamicin (9.1%). In addition, four (36.4%) of the isolates were resistant to cefotaxime and three (27.3%) to tetracycline and trimethoprim. The isolates also exhibit the presence of resistance genes for sulfamethoxazole 1&2, tetracycline (tet) A&B, Beta-lactamase CTXM, Beta-lactamase TEM, Beta-lactamase SHV, Gentamycine, Acra and acc3-1 in different occurrences. The overall prevalence of Salmonella species in Morogoro region was 8.3% (11/132) with Salmonella Enteritidis and Salmonella Typhimurium being the only serovars detected from adults and children stool samples. Our investigation showed that both children and adults had been exposed to Salmonella spp. However, the occurrence of NTS was higher in children (5.3% (7/132) compared to adults (3.0% (4/132). To stop zoonotic infections and the development of antimicrobial resistance in the community, this calls for Infection Prevention and Control (IPC) and stewardship programmes on rational use of antimicrobials in both health facilities and at the community level.

Antibiotic resistance profiles of sentinel bacteria isolated from aquaculture in Cambodia

ABSTRACT The misuse of antibiotics and the emergence of antimicrobial resistance (AMR) is a concern in the aquaculture industry because it contributes to global health risks and impacts the environment. This study analyzed the AMR of sentinel bacteria associated with striped catfish (Pangasisanodon hypophthalmus) and giant snakehead (Channa micropeltes), the two main fish species reared in the pond culture in Cambodia. Phenotypic and genotypic characterization of the recovered isolates from fish, water, and sediment samples revealed the presence of bacteria, such as 22 species belonging to families Aeromonadaceae, Enterobacteriaceae, and Pseudomonadaceae. Among 48 isolates, Aeromonas caviae (n = 2), Aeromonas hydrophila (n = 2), Aeromonas ichthiosmia (n = 1), Aeromonas salmonicida (n = 4) were detected. A. salmonicida and A. hydrophilla are known as fish pathogens that occur worldwide in both fresh and marine water aquaculture. Antibiotic susceptibility testing revealed antibiotic resistance patterns of 24 (50 %) isolates among 48 isolates with higher multiple antibiotic resistance index (> 0.2). All the isolates of Enterobacteriaceae were susceptible to ciprofloxacin. Ciprofloxacin is a frontline antibiotic that is not recommended to use in aquaculture. Therefore, its use has to be strictly controlled. This study expands our knowledge of the AMR status in aquaculture farms which is very limited in Cambodia.

Antimicrobials and antimicrobial resistance genes in the shadow of COVID-19 pandemic: A wastewater-based epidemiology perspective

Higher usage of antimicrobial agents in both healthcare facilities and the communities has resulted in an increased spread of resistant bacteria. However, the improved infection prevention and control practices may also contribute to decreasing antimicrobial resistance (AMR). In the present study, wastewater-based epidemiology (WBE) approach was applied to explore the link between COVID-19 and the community usage of antimicrobials, as well as the prevalence of resistance genes. Longitudinal study has been conducted to monitor the levels of 50 antimicrobial agents (AAs), 24 metabolites, 5 antibiotic resistance genes (ARGs) and class 1 integrons (intI 1) in wastewater influents in 4 towns/cities over two years (April 2020 - March 2022) in the South-West of England (a total of 1,180 samples collected with 87,320 individual AA measurements and 8,148 ARG measurements). Results suggested higher loads of AAs and ARGs in 2021–22 than 2020–21, with beta-lactams, quinolones, macrolides and most ARGs showing statistical differences. In particular, the intI 1 gene (a proxy of environmental ARG pollution) showed a significant increase after the ease of the third national lockdown in England. Positive correlations for all quantifiable parent AAs and metabolites were observed, and consumption vs direct disposal of unused AAs has been identified via WBE. This work can help establish baselines for AMR status in communities, providing community-wide surveillance and evidence for informing public health interventions. Overall, studies focused on AMR from the start of the pandemic to the present, especially in the context of environmental settings, are of great importance to further understand the long-term impact of the pandemic on AMR.

Clinical and economic burden of invasive pneumococcal disease and noninvasive all-cause pneumonia in hospitalized US adults: A multicenter analysis from 2015 to 2020

To evaluate the clinical and economic outcomes in adults hospitalized with invasive pneumococcal disease (IPD) and noninvasive all-cause pneumonia (ACP) overall and by antimicrobial resistance (AMR) status. Hospitalized adults from the BD Insights Research Database with an ICD10 code for IPD, noninvasive ACP or a positive Streptococcus pneumoniae culture/urine antigen test were included. Descriptive statistics and multivariable analyses were used to evaluate outcomes (in-hospital mortality, length of stay [LOS], cost per admission, and hospital margin [costs - payments]). The study included 88,182 adult patients at 90 US hospitals (October 2015-February 2020). Most (98.6%) had noninvasive ACP and 40.2% were <65 years old. Of 1450 culture-positive patients, 37.7% had an isolate resistant to ≥1 antibiotic class. Observed mortality, median LOS, cost per admission, and hospital margins were 8.3%, 6 days, $9791, and $11, respectively. Risk factors for mortality included ≥50 years of age, higher risk of pneumococcal disease (based on chronic or immunocompromising conditions), and intensive care unit admission. Patients with IPD had similar mortality rates and hospital margins compared with noninvasive ACP, but greater costs per admission and LOS. IPD and noninvasive ACP are associated with substantial clinical and economic burden across all adult age groups. Expanded pneumococcal vaccination programs may help reduce disease burden and decrease hospital costs.

Antibiotic Profiling of E. coli Borne UTI Infection in Tertiary Healthcare Settings.

Introduction In general, with frequent recurrence of urinary tract infections (UTIs), long-term antibiotic therapy is stipulated at a low dose. With this type of situation and with easy access to several classes of antibiotics in tertiary health care settings, the use of such drugs results in the development of resistant bacteria in patients.Escherichia coliis a frequent cause of UTI observed. Hence, it was proposed in the present study to assess the antimicrobial resistance status ofE. coliin UTI-infected patients. Methods This study was conducted among female patients diagnosed with UTI.About 80 urine samples were collected in an aseptic condition, Under the process of culture identification 44 samples were found to be positive for UTI infection. The positive samples were plated on blood agar. Out of 44 samples, 18 samples were found to be positive, and 26 samples were negative for E. coli infection. The 18 samples were screened on MALDI-TOF for identification. Further, the samples were assessed for susceptibility to antibiotic medication within the study area. Result The study identified different strains ofE. coli,and the CHB geneE. coli wasfound in eight samples. The sample showed pink oval-round spots in the culture medium and was resistant to nitrofurantoin, cephalosporin, and cephalexin antibiotics. Hence, antimicrobial susceptibility tests arenecessaryfor managing and treating bacterialE. coliinfections. Conclusion E. coliis a common bacterium found in the vaginal region of patients, suggesting a potential infection.E. colican be associated with UTIs in women. The results from this study conclude thatE. coliis rapidly becoming multidrug-resistant, as only higher antibiotics can inhibit its growth. To effectively manage infections caused byE. coliproper diagnosis, laboratory testing, and antibiotic treatment are required.

Genomic analysis of Salmonella enterica from Metropolitan Manila abattoirs and markets reveals insights into circulating virulence and antimicrobial resistance genotypes.

The integration of next-generation sequencing into the identification and characterization of resistant and virulent strains as well as the routine surveillance of foodborne pathogens such as Salmonella enterica have not yet been accomplished in the Philippines. This study investigated the antimicrobial profiles, virulence, and susceptibility of the 105 S. enterica isolates from swine and chicken samples obtained from slaughterhouses and public wet markets in Metropolitan Manila using whole-genome sequence analysis. Four predominant serovars were identified in genotypic serotyping, namely, Infantis (26.7%), Anatum (19.1%), Rissen (18.1%), and London (13.3%). Phenotypic antimicrobial resistance (AMR) profiling revealed that 65% of the isolates were resistant to at least one antibiotic, 37% were multidrug resistant (MDR), and 57% were extended-spectrum β-lactamase producers. Bioinformatic analysis revealed that isolates had resistance genes and plasmids belonging to the Col and Inc plasmid families that confer resistance against tetracycline (64%), sulfonamide (56%), and streptomycin (56%). Further analyses revealed the presence of 155 virulence genes, 42 of which were serovar-specific. The virulence genes primarily code for host immune system modulators, iron acquisition enzyme complexes, host cell invasion proteins, as well as proteins that allow intracellular and intramacrophage survival. This study showed that virulent MDR S. enterica and several phenotypic and genotypic AMR patterns were present in the food chain. It serves as a foundation to understand the current AMR status in the Philippines food chain and to prompt the creation of preventative measures and efficient treatments against foodborne pathogens.

Characterization of Virulence Genotypes, Antimicrobial Resistance Patterns, and Biofilm Synthesis in Salmonella spp Isolated from Foodborne Outbreaks.

Salmonella is the main bacterial pathogen that causes foodborne disease, particularly in developing countries. Nontyphoidal Salmonella (NTS) include Enteritidis and Typhimurium as the most prevalent strains which are one of the significant causes of acute gastroenteritis in children. Therefore, identifying the most predominant serovars, types of common contaminated food, and paying attention to their antibiotic resistance are the main factors in the prevention and control strategy of salmonellosis. This study was undertaken to evaluate the prevalence rate of serovars, the biofilm formation, antimicrobial resistance (AMR) status, and phenotypic virulence factors of Salmonella strains isolated from diarrhea samples in some cities of Iran. A total of 40 (10.41%) Salmonella isolates were recovered from 384 diarrhea samples processed and the most common serovar was Salmonella serovar Typhimurium (82.5). Also, all isolates belonging to serovar Typhimurium showed more virulence factors compared to other serovars. The isolates showed a high resistance rate to ampicillin (95%) and nalidixic acid (87.5%), while a low resistance rate was found for chloramphenicol (2.5%). Moreover, significant variances in the capacity of biofilm formation were found between different Salmonella serotypes. The resistance of NTS to extant choice drugs is a potential public health problem. Constant monitoring of AMR pattern and virulence profile of NTS serovars is suggested for the prevention of salmonellosis in humans.

Antimicrobial Resistance Profiles of E. coli Isolated From Pooled Samples of Sick, Farm, and Market Chickens in Nairobi County, Kenya.

Bacterial antimicrobial resistance (AMR) is a global threat to both human and animal health. This is mainly because the same antimicrobial molecules are used for the treatment and prophylaxis of bacterial diseases in both cases, and about 60% of human pathogens are shared with animals. For effective control of AMR in any country, the current situation has to be established; this is done through surveillance exercises. In Kenya, there is scanty data on the prevailing AMR situation, especially in animals. This paper reports on AMR profiles of 54 E. coli strains isolated from chickens in a cross-sectional study, out of which 36/54 (72%) were from clinically ill chickens, 11/54 (22%) were from farm chickens, and 7/54 (9.7%) were from slaughtered chicken, respectively. All 54 isolates exhibited varying antimicrobial resistance profiles with the majority showing resistance to Ampicillin (85.22%), Tetracycline (66.7%), Co-trimoxazole (57.4%), and Streptomycin (40.7%). Very few isolates were resistant to Amoxicillin and Gentamicin (each at 3.7%), Ampicillin (11.1%), and Nalidixic acid (24.1%). A total of 44/54 (81.5%) showed multiple resistance to up to 6 antimicrobial agents. This information will augment current data on the AMR status of bacteria harbored by chickens in Kenya. It will also inform policymakers in their fight against AMR.

Prevalence and molecular characterization of Multidrug-Resistant ESBL-Producing E. coli in commercial poultry

Background: Rural poultry farmers are gradually changing from the backyard poultry production to intensive production, which made poultry more vulnerable to diseases and lead to indiscriminate use of antimicrobials in poultry production. As per OIE, FAO and WHO, multiple spread antimicrobial resistant pathogenic bacteria are a serious global human and animal health problem. Hence the study has been carried out to know the antimicrobial resistance status of poultry in Tirunelveli, Tamil Nadu, India. Methods: The study was aimed to find the occurrence of extended spectrum beta lactamases (ESBLs) producing E. coli from poultry origin. Caecum samples from the broiler (n=180), desi chicken (n=180) and Japanese quail (n=180) were processed for the isolation and phenotypic identification of ESBL E. coli. The isolates were tested for antimicrobial susceptibility test and characterization of ESBL and virulence genes done by PCR. Result: On phenotypic confirmation of ESBL producing E. coli by combined disk diffusion test (CDDT), a total of 9.63% samples were positive for extended spectrum beta lactamase producer (ESBL). Multi drug resistance (MDR) was observed in 38.23% of the broiler isolates, 27.27% and 57.14% in desi chicken and Japanese quail isolates, respectively. All the phenotypically confirmed ESBL producing E. coli isolates (n=52) were positive for uspA gene by PCR confirming the isolates as E. coli. The overall presence of the blaTEM gene was 57.69%, with broiler isolates having the highest prevalence of 67.47%, followed by desi chicken 36.36% and Japanese quail 42.85%. In contrast, the blaSHV gene was found in only 17.64% of the broiler isolates, but none were positive for the blaCTXM and blaOXA-1 gene.

Extended-Spectrum Beta-Lactamases Genes in Klebsiella pneumoniae Isolates Obtained from Patients in Intensive Care Units

Background: Drug-resistant hospital-acquired infections (HAIs) are a growing concern in modern medicine throughout the world. Klebsiella pneumoniae is one of the most prominent causative agents of multidrug-resistant nosocomial infections. It is also widely recognized for having a high resistance level to many antibiotic classes, particularly beta-lactams. Carbapenemase-producing K. pneumoniae has been identified as a major global cause of HAIs with adverse clinical outcomes. Therefore, it is of the utmost importance to have an in-depth understanding of the antimicrobial resistance (AMR) genetic determinants of this bacterium to stop the spread of highly resistant K. pneumoniae in healthcare facilities and the resulting patient morbidity and mortality. Objectives: This study aimed to investigate the AMR pattern of K. pneumoniae isolates obtained from intensive care units (ICUs), with a focus on extended-spectrum beta-lactamases (ESBLs) genes blaCTX-M, blaGES, and blaIMP. Methods: A total of 105 K. pneumoniae isolates obtained from the sputum samples of ICU patients were identified and confirmed using standard microbiological tests and 16S rRNA polymerase chain reaction (PCR). The antibiotic susceptibility test was performed for all the isolates. The presence of ESBL genes was determined phenotypically and by PCR. Results: The highest level of resistance was observed against ceftazidime (100%), cefotaxime (99%), and imipenem (93.3%). Approximately 87.6% and 39% of the isolates were sensitive to colistin and gentamicin, respectively. Phenotypic ESBL production was observed in 16 isolates, and the prevalence of blaCTX-M was 86.7%. No blaGES and blaIMP genes were detected. Conclusions: Periodic investigation of AMR-mediating genes is essential due to the high prevalence of ESBL genes in HAIs. The presence of other ESBL genes needs to be investigated for a more accurate understanding of the AMR status of K. pneumoniae in healthcare settings.

Antimicrobial Resistance: Is There a 'Light' at the End of the Tunnel?

In recent years, with the increases in microorganisms that express a multitude of antimicrobial resistance (AMR) mechanisms, the threat of antimicrobial resistance in the global population has reached critical levels. The introduction of the COVID-19 pandemic has further contributed to the influx of infections caused by multidrug-resistant organisms (MDROs), which has placed significant pressure on healthcare systems. For over a century, the potential for light-based approaches targeted at combatting both cancer and infectious diseases has been proposed. They offer effective killing of microbial pathogens, regardless of AMR status, and have not typically been associated with high propensities of resistance development. To that end, the goal of this review is to describe the different mechanisms that drive AMR, including intrinsic, phenotypic, and acquired resistance mechanisms. Additionally, the different light-based approaches, including antimicrobial photodynamic therapy (aPDT), antimicrobial blue light (aBL), and ultraviolet (UV) light, will be discussed as potential alternatives or adjunct therapies with conventional antimicrobials. Lastly, we will evaluate the feasibility and requirements associated with integration of light-based approaches into the clinical pipeline.

Activity fingerprinting of AMR β-lactamase towards a fast and accurate diagnosis.

Antibiotic resistance has become a serious threat to global public health and economic development. Rapid and accurate identification of a patient status for antimicrobial resistance (AMR) are urgently needed in clinical diagnosis. Here we describe the development of an assay method for activity fingerprinting of AMR β-lactamases using panels of 7 β-lactam antibiotics in 35min. New Deli Metallo β-lactamase-1 (NDM-1) and penicillinase were demonstrated as two different classes of β-lactamases. The panel consisted of three classes of antibiotics, including: penicillins (penicillin G, piperacillin), cephalosporins (cefepime, ceftriaxone, cefazolin) and carbapenems (meropenem and imipenem). The assay employed a scheme combines the catalytic reaction of AMR β-lactamases on antibiotic substrates with a flow-injected thermometric biosensor that allows the direct detection of the heat generated from the enzymatic catalysis, and eliminates the need for custom substrates and multiple detection schemes. In order to differentiate classes of β-lactamases, characterization of the enzyme activity under different catalytic condition, such as, buffer composition, ion strength and pH were investigated. This assay could provide a tool for fast diagnosis of patient AMR status which makes possible for the future accurate treatment with selected antibiotics.

Prevalence, clonal diversity, and antimicrobial resistance of hypervirulent Klebsiella pneumoniae and Klebsiella variicola clinical isolates in northern Japan

Hypervirulent Klebsiella pneumoniae (hvKp) and Klebsiella variicola (hvKv) cause hospital/community-acquired infections, often associated with antimicrobial resistance (AMR). This study aimed to investigate the molecular epidemiology of hvKp and hvKv in northern Japan. A total of 500 K. pneumoniae and 421 K. variicola clinical isolates collected from August to December 2021 were studied. Prevalence of virulence factor-encoding genes, wzi sequence and associated K/KL type, sequence type (ST), and beta-lactamases and their types were characterized. Any virulence gene (rmpA, rmpA2, peg-344, iucA, iutA, and iroB) and/or magA was detected in 25% (n=125) of K. pneumoniae and 1% (n=5) of K. variicola. Among these hvKp/hvKv, 22 wzi types (18 and 4 types, respectively) and 24 STs (20 and 4 STs, respectively) were identified. Sequence types of hvKp were classified into some clonal groups (CGs), among which CG35, including six STs, was the most common (n=59; 47%), followed by CG23, and CG65. ST268 (CG35) associated with wzi95-K20 or wzi720 was the dominant lineage (n=43, 34%), while K1:ST23/ST249 and K2:ST65/ST86 accounted for 26% and 13% of hvKp, respectively. Extended-spectrum beta-lactamase (ESBL) genes (blaCTX-M-2, blaCTX-M-3, blaCTX-M-15, and blaCTX-M-27) were detected in only ST23 and CG35 (ST268 and ST412) hvKp. No isolate was resistant to carbapenems, without detection of the ESBL gene in K. variicola. Phylogenetically, wzi was differentiated into two main clusters of K. pneumoniae and K. variicola. A major clonal group CG347 was identified in K. variicola. Clonal structures were revealed for hvKp and hvKv clinical isolates with their AMR status in northern Japan.

Community-acquired antimicrobial resistance among Syrian refugees and the local population in Türkiye.

The long-standing antimicrobial resistance (AMR) pandemic has proven difficult to resolve and is becoming more complex, especially in the context of increasing forced migration, with little evidence around patterns of AMR in migrant communities. This study aimed to determine the frequency in the carriage of common types of antimicrobial-resistant bacteria between Syrian refugees and the local communities in Türkiye: extended-spectrum β-lactamase (ESBL), methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE). We collected nasal swabs and stool samples from the study participants, the local community members, and refugees, between September 2020 and March 2021. We screened clinical samples for the presence of ESBL, MRSA and VRE. Antimicrobial-resistant bacterial isolates were tested by phenotypic analysis to determine the AMR status. The study included a total of 3960 participants: 1453 individuals in the local community (36.2%) and 2525 Syrian refugees (63.8%). Overall, a significantly greater proportion of refugees (6.7%) carried MRSA compared to the local community (3.2%) (P < 0.001). The ESBL-positivity rate was 17.9% in Syrian refugees and 14.3% in the local community (P = 0.041). Carbapenemase activity was detected in three isolates from Syrian refugees. No VRE was detected in Syrian refugees or the local community. This large, community-based study on the frequency and the distribution of AMR among Syrian refugees and the local population is the first study in Türkiye.

Salmonella Enteritidis clades in South Africa: why we should be paying more attention

Salmonella Enteritidis is the most reported non-typhoidal Salmonella serovar and is implicated in both gastroenteritis and invasive non-typhoidal disease. Whole-genome sequence comparison of Salmonella Enteritidis isolates from across the world identified four definitive clades: Outlier, Global Epidemic, East African and West African. Three of these clades were implicated in epidemics: the Global Epidemic clade was linked to poultry-associated gastroenteritis, while the two African clades were related to invasive disease. Despite the recognition of different Salmonella Enteritidis clades, the distribution and epidemiology of these clades across Africa is poorly understood. In our proof-of-concept study, we classified 618 Salmonella Enteritidis isolates originating from four South African provinces over a period of two years (2012 – 2013) into clades using a clade-typing real-time PCR assay. Associations between clades and province of collection, antimicrobial resistance and HIV status were explored using statistical analysis. Majority of the South African isolates were classified within the Outlier clade (61.00%), with fewer classified within the Global Epidemic clade (38.83%) and only one isolate classified within the West African Clade (0.16%). Of note, were the isolates within the Global clade, which were strongly associated with ciprofloxacin resistance (15.42%, OR: 7.45, CI: 3.526 – 15.751) and invasive disease (58.33%, OR: 1.57, CI: 1.13 – 2.17) in humans. The increase in poultry consumption and importation in South Africa has placed the country at risk of a Salmonella Enteritidis epidemic. Thus, there is a necessity for routine monitoring of S. Enteritidis along the farm-to-fork continuum to implement preventative measures.

High prevalence of extensively drug resistant and extended spectrum beta lactamases (ESBLs) producing uropathogenic Escherichia coli isolated from Faisalabad, Pakistan.

Urinary tract infections (UTIs) are predominantly caused by uropathogenic Escherichia coli (E. coli). There is rapid increase in antimicrobial resistance in UTIs, also declared as a serious health threat by World Health Organization (WHO). Present study was designed to investigate the antimicrobial resistance status with specific focus on ESBLs and carbapenemases in local uropathogenic E. coli (UPEC) isolates. E. coli isolates were characterized from patients of all ages visiting diagnostic laboratories for urine examination. Demographic data was also recorded for each patient. Antibiograms were developed to observe antibiotic resistance in UPEC using Kirby Bauer disc diffusion technique. Double Disc Synergy test (DDST) was used for phenotypic ESBL test. ESBLs and carbapenemases genes were detected in UPEC using PCR. The PCR results were confirmed by sequencing. The UPEC isolates under study exhibited 78%, 77%, 74%, 72% and 55% resistance against cefotaxime, amoxicillin, erythromycin, ceftriaxone and cefixime, respectively. Resistance against colistin and meropenem was observed in 64% and 34% isolates, respectively. Phenotypic DDST identified 48% isolates as ESBLs producers. Genotypic characterization identified 70%, 74.4% and 49% prevalence of CTXM-1, TEM-1 and CTXM-15 genes respectively. One isolate was observed exhibiting co-existence of all ESBL genes. TEM-1 + CTXM-1 and TEM-1 + CTXM-1 + CTXM-15 + OXA-1 gene patterns were dominant among ESBLs. For carbapenem-resistance, 14% isolates indicated the presence of KPC whereas GES and VIM was detected in 7% and 3.4% isolates, respectively. In conclusion, our results present a high prevalence of extensively drug resistant UPEC isolates with a considerable percentage of ESBL producers. These findings propose the need of continuous surveillance for antimicrobial resistance and targeted antimicrobial therapy.

Comparison of antimicrobial resistance among Salmonella enterica serovars isolated from Canadian turkey flocks, 2013 to 2021

The emergence of antimicrobial resistance (AMR) in Salmonella from turkeys has raised a food safety concern in Canada as certain serovars have been implicated in human salmonellosis outbreaks in recent years. While several studies evaluated AMR in broiler chickens in Canada, there are limited studies that assess AMR in turkey flocks. This study analyzed data collected between 2013 and 2021 by the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS) farm turkey surveillance program to determine the prevalence of AMR and differences in resistance patterns among Salmonella serovars recovered from turkey flocks. Salmonella isolates were tested for susceptibility to 14 antimicrobials using a microbroth dilution method. Hierarchical clustering dendrograms were constructed to compare the individual AMR status of Salmonella serovars. Differences in the probability of resistance between Salmonella serovars were determined using generalized estimating equation logistic regression models to account for farm-level clustering. Of the 1,367 Salmonella isolates detected, 55.3% were resistant to at least one antimicrobial and 25.3% were multidrug resistant (MDR) (resistant to ≥3 antimicrobial classes). The Salmonella isolates exhibited high resistance to tetracycline (43.3%), streptomycin (47.2%), and sulfisoxazole (29.1%). The 3 most frequently occurring serovars were S. Uganda (22.9%), S. Hadar (13.5%), and S. Reading (12.0%). Streptomycin-sulfisoxazole-tetracycline (n=204) was the most frequent MDR pattern identified. Heatmaps showed that S. Reading exhibited coresistance to the quinolone class antimicrobials, ciprofloxacin, and nalidixic acid; S. Heidelberg to gentamicin and sulfisoxazole; and S. Agona to ampicillin and ceftriaxone. Salmonella Hadar isolates had higher odds of resistance to tetracycline (OR: 152.1, 95% CI: 70.6-327.4) while the probability of being resistant to gentamicin and ampicillin was significantly higher in S. Senftenberg than in all the other serovars. Moreover, S. Uganda had the highest odds of being MDR (OR: 4.7, 95% CI: 3.7-6.1). The high resistance observed warrants a reassessment of the drivers for AMR, including AMU strategies and other production factors. Differences in AMR patterns highlight the need to implement serovar-specific mitigation strategies.

Prevalence of antimicrobial resistance in fecal Escherichia coli and Enterococcus spp. isolates from beef cow-calf operations in northern California and associations with farm practices.

Antimicrobials are necessary for the treatment of bacterial infections in animals, but increased antimicrobial resistance (AMR) is becoming a concern for veterinarians and livestock producers. This cross-sectional study was conducted on cow-calf operations in northern California to assess prevalence of AMR in Escherichia coli and Enterococcus spp. shed in feces of beef cattle of different life stages, breeds, and past antimicrobial exposures and to evaluate if any significant factors could be identified that are associated with AMR status of the isolates. A total of 244 E. coli and 238 Enterococcus isolates were obtained from cow and calf fecal samples, tested for susceptibility to 19 antimicrobials, and classified as resistant or non-susceptible to the antimicrobials for which breakpoints were available. For E. coli, percent of resistant isolates by antimicrobial were as follows: ampicillin 100% (244/244), sulfadimethoxine 25.4% (62/244), trimethoprim-sulfamethoxazole 4.9% (12/244), and ceftiofur 0.4% (1/244) while percent of non-susceptible isolates by antimicrobial were: tetracycline 13.1% (32/244), and florfenicol 19.3% (47/244). For Enterococcus spp., percent of resistant isolates by antimicrobial were as follows: ampicillin 0.4% (1/238) while percent of non-susceptible isolates by antimicrobial were tetracycline 12.6% (30/238) and penicillin 1.7% (4/238). No animal level or farm level management practices, including antimicrobial exposures, were significantly associated with differences in isolate resistant or non-susceptible status for either E. coli or Enterococcus isolates. This is contrary to the suggestion that administration of antibiotics is solely responsible for development of AMR in exposed bacteria and demonstrates that there are other factors involved, either not captured in this study or not currently well understood. In addition, the overall use of antimicrobials in this cow-calf study was lower than other sectors of the livestock industry. Limited information is available on cow-calf AMR from fecal bacteria, and the results of this study serve as a reference for future studies to support a better understanding and estimation of drivers and trends for AMR in cow-calf operations.

Genomic Diversity, Antimicrobial Susceptibility, and Biofilm Formation of Clinical Acinetobacter baumannii Isolates from Horses.

Acinetobacter (A.) baumannii is an opportunistic pathogen that causes severe infections in humans and animals, including horses. The occurrence of dominant international clones (ICs), frequent multidrug resistance, and the capability to form biofilms are considered major factors in the successful spread of A. baumannii in human and veterinary clinical environments. Since little is known about A. baumannii isolates from horses, we studied 78 equine A. baumannii isolates obtained from clinical samples between 2008 and 2020 for their antimicrobial resistance (AMR), clonal distribution, biofilm-associated genes (BAGs), and biofilm-forming capability. Based on whole-genome sequence analyses, ICs, multilocus (ML) and core-genome ML sequence types (STs), and AMR genes were determined. Antimicrobial susceptibility testing was performed by microbroth dilution. A crystal violet assay was used for biofilm quantification. Almost 37.2% of the isolates were assigned to IC1 (10.3%), IC2 (20.5%), and IC3 (6.4%). Overall, the isolates revealed high genomic diversity. We identified 51 different STs, including 22 novel STs (ST1723-ST1744), and 34 variants of the intrinsic oxacillinase (OXA), including 8 novel variants (OXA-970 to OXA-977). All isolates were resistant to ampicillin, amoxicillin/clavulanic acid, cephalexin, cefpodoxime, and nitrofurantoin. IC1-IC3 isolates were also resistant to gentamicin, enrofloxacin, marbofloxacin, tetracycline, and trimethoprim/sulfamethoxazole. All isolates were susceptible to imipenem. Thirty-one multidrug-resistant (MDR) isolates mainly accumulated in the IC1-IC3 groups. In general, these isolates showed less biofilm formation (IC1 = 25.0%, IC2 = 18.4%, IC3 = 15.0%) than the group of non-IC1-IC3 isolates (58.4%). Isolates belonging to the same ICs/STs revealed identical BAG patterns. BAG blp1 was absent in all isolates, whereas bfmR and pgaA were present in all isolates. At the level of the IC groups, the AMR status was negatively correlated with the isolates' ability to form a biofilm. A considerable portion of equine A. baumannii isolates revealed ICs/STs that are globally present in humans. Both an MDR phenotype and the capability to form biofilms might lead to therapeutic failures in equine medicine, particularly due to the limited availability of licensed drugs.