Antibacterial Resistance Profile Research Articles

Mechanistic insight into the synergistic antimicrobial potential of Fagonia indica Burm.f. extracts with cefixime

Fagonia indica Burm.f. is known for its anti-infective character and has been studied in the present work as a synergistic remedy against resistant bacterial strains. Initially, phytochemicals were quantified in n-Hexane (n-Hex), ethyl acetate (E.A), methanol (MeOH), and aqueous (Aq.) extracts by Total Phenolic Content (TPC), Total Flavonoid Content (TFC) and Reverse Phase High Performance Liquid Chromatography (RP-HPLC) analysis. Later, after establishing an antibacterial resistance profile for extracts and antibiotics against gram-positive and gram-negative strains, synergism was evaluated in combination with cefixime through time-kill kinetics and bacterial protein estimation studies. Topographic images depicting synergism were obtained by scanning electron microscopy for Methicilin-resistant Staphylococcus aureus (MRSA) and Resistant Escherichia coli (R.E. coli). Results showed the presence of maximum phenolic (28.4 ± 0.67 μg GAE/mg extract) and flavonoid (11 ± 0.42 μg QE/mg extract) contents in MeOH extract. RP-HPLC results also displayed maximum polyphenols in MeOH extract followed by E.A extract. Clinical strains were resistant to cefixime whereas these were moderately inhibited by all extracts (MIC 150–300 µg/ml) except Aq. extract. E.A and n-Hex extracts demonstrated maximum synergism (Fractional inhibitory concentration index (FICI) 0.31) against R.E. coli. The n-Hex extract displayed total synergism against R.P. a with a 4-fold reduction in cefixime dose. Time-kill kinetics showed maximum inhibition of gram-negative bacterial growth from 3 to 12 h when treated at FICI and 2FICI values with > 10-fold reduction of the extracts’ dose. All combinations demonstrate > 70 % protein content inhibition with bacterial cell wall disruption in SEM images. Fortunately, FICI concentrations have low hemolytic potential (<5%). Conclusively, F. indica extracts can mitigate antimicrobial resistance against cefixime and can be investigated in detail by in vivo and mechanistic studies.

Genetic diversity of Staphylococcus aureus isolated from ear infections in Iran: Emergence of CC8/ST239-SCCmec III as major genotype.

Increase in antibiotic resistance in Staphylococcus aureus isolated from ear infection is a serious public health problem. The objective of this investigation was to determine the antibacterial resistance profile and genetic variability of the S. aureus isolated from adult patients with otitis externa (OE) and otitis media (OM) infections, Tehran- Iran. The disk diffusion was employed to detect the susceptibility of 45 S. aureus strains. Biofilm production was evaluated by microtiter plate assay. Genetic diversity of the isolates was determined by staphylococcal cassette SCCmec, spa, and MLST techniques. Resistance to mupirocin and vancomycin were identified in 40 and 2.2% of isolates. Out of the 45 S. aureus isolates, 41 (91.2%) strains were considered as positive biofilm strains at different levels. According to our results, S. aureus isolated from OM (44.4%, 20/45) were including CC8/ST239-SCCmecIII corresponded to spa types t860, t030, t037, t234, t421 (70%, 14/20) and CC/ST30-SCCmecIV corresponded to spa types t605 and t019 (30%, 6/20) while S. aureus isolated from OE (55.6%, 25/45) were including CC/ST30-SCCmecIV corresponded to spa types t605, t345 and t1130 (52%, 13/25), CC/ST22-SCCmecIV corresponded to spa type t790 (20%, 5/25), CC8/ST8-SCCmecIV corresponded to spa type t008 (16%,4/25), and CC/ST45-SCCmecIV corresponded to spa types t004 and t038 (12%, 3/25). This study highlighted genetic variability and strong biofilm formation ability among our isolates revealing its crucial role in enhancing the resistance of this bacteria to drugs. Thus, it is necessary to continue the epidemiological analysis to improve the control of ear infections related to S. aureus.

Curcusinol from the fruit of Carex baccans with antibacterial activity against multidrug-resistant strains

Ethnopharmacological relevanceCarex baccans, known as Shan-Bai-Zi or Ye-Gao-Liang in China, is a traditional medicinal herb used by several ethnic groups in Yunnan Province. It is utilized for the treatment of wound infections, ulcers, and dysentery. However, there is currently a dearth of research reports on its antimicrobial potential. Aim of the studyThe substance basis of the antimicrobial activity of C. baccans will be unveiled, and the in vitro and in vivo antibacterial activities against multidrug-resistant bacteria of its major active compounds, as well as their preliminary mechanisms of action, will be investigated. Materials and methodsAn antibacterial bioactivity-guided isolation method was used to isolate and identify the active compound curcusinol from C. baccans. UPLC-DAD-MS was employed for the quantitative analysis of curcusinol. The antibacterial activity, resistance profile, synergistic effects, anti-biofilm activity, and potential mechanisms of action of curcusinol against methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Enterococcus (VRE), and other multidrug-resistant bacteria (Escherichia coli, Pseudomonas aeruginosa, Acinetobacter baumannii) were investigated using various methods, including the broth microdilution method, scanning electron microscopy, time kill assay, multi-generational resistance induction assay, checkerboard synergy assay, anti-biofilm assay, and metabolomics. Furthermore, the therapeutic efficacy of curcusinol was assessed in vivo by establishing an animal skin wound infection model of MRSA. ResultsCurcusinol was isolated from the fruit of C. baccans, which accounts for 3.1% of the dry weight of the fruit. Curcusinol exhibited significant bactericidal and anti-biofilm activities against antibiotic-resistant Gram-positive bacteria in vitro. Furthermore, curcusinol acted as an antibiotic adjuvant to enhance the activity of various commonly used antibiotics against both Gram-positive and Gram-negative antibiotic-resistant bacteria without cytotoxicity to mammalian cells (A549 and RAW264.7) at 64 μM. Moreover, curcusinol affected arginine biosynthesis, cysteine and methionine metabolism, and alanine, aspartate, and glutamate metabolism significantly in MRSA cells under stress. Additionally, curcusinol effectively treated MRSA-infected mouse skin wounds and accelerated wound healing in vivo. ConclusionsThe results of this study not only support the traditional uses of C. baccans but also demonstrate that its major active compound, curcusinol, is an effective plant-derived bactericidal agent and antibacterial adjuvant with potential applications in the treatment of skin infections.

Evidence of Antibiotic Resistance and Virulence Factors in Environmental Isolates of Vibrio Species.

The outbreak of waterborne diseases such as cholera and non-cholera (vibriosis) is continuously increasing in the environment due to fecal and sewage discharge in water sources. Cholera and vibriosis are caused by different species of Vibrio genus which are responsible for acute diarrheal disease and soft tissue damage. Although incidences of cholera and vibriosis have been reported from the Vaishali district of Bihar, India, clinical or environmental strains have not been characterized in this region. Out of fifty environmental water samples, twelve different biochemical test results confirmed the presence of twenty Vibrio isolates. The isolates were found to belong to five different Vibrio species, namely V. proteolyticus, V. campbellii, V. nereis, V. cincinnatiensis, and V. harveyi. From the identified isolates, 65% and 45% isolates were found to be resistant to ampicillin and cephalexin, respectively. Additionally, two isolates were found to be resistant against six and four separately selected antibiotics. Furthermore, virulent hlyA and ompW genes were detected by PCR in two different isolates. Additionally, phage induction was also noticed in two different isolates which carry lysogenic phage in their genome. Overall, the results reported the identification of five different Vibrio species in environmental water samples. The isolates showed multiple antibacterial resistance, phage induction, and virulence gene profile in their genome.

Resistance Profile of Bovine Mastitis Isolates, Presence of the mecA Gene and Identification of ESBL Producing Strains from Small Rural Dairy Properties

Bovine mastitis is an inflammation of the mammary gland in response to invasion by opportunistic agents. Due to the high economic importance of dairy production and the complexity related to animal health, the objective of this work was to identify and evaluate the antibacterial resistance profile of samples of mastitis milk, milking hand and milking equipment from small rural dairy farms belonging to the northwest region of the state of Paraná, Brazil. Five small, non-technical dairy farms in the municipalities of Boa Esperança, Juranda and Tapejara, all belonging to the northwest region of the state of Paraná, Brazil, were selected. The properties had Holstein and/or crossbred herds, carried out a bucket-by-foot milking system and all had the presence of animals with subclinical mastitis confirmed by the California Mastitis Test. Samples of sterile swabs from the milking insufflators, the milking hand and milk samples were collected-and later, isolation tests and phenotypic characterization of the samples, sensitivity tests to antimicrobials and phenotypic tests for the detection of beta-producing strains were performed with extended-spectrum beta-lactamase (ESBL), molecular identification of Staphylococcus aureus isolates and mecA gene research. Of the 199 samples collected from the 15 selected properties in the municipalities of Boa Esperança, Tapejara and Juranda, 72 (36.20%) were classified as multiresistant. Isolated from milkers' hands and milking machines, which phenotypically produce extended-spectrum beta-lactamase (ESBL), the presence of the mecA gene was also observed in 11 isolates of Staphylococcus spp. of milk samples, machines and milking hands. Mastitis can be spread to the herd through the milking process by the milkers' instruments and hands, and adequate management measures can prevent its transmission and the conscious use of antibiotics decreases the prevalence of multidrug-resistant pathogens. In this work, different pathogenic bacteria were detected in mastitic milk, milking equipment and milking hand with a high percentage (36.20%) of isolates classified as multidrug resistant. In addition, the presence phenotypically (ESBL) and molecularly (mecA gene) of isolates carrying resistance genes was also verified. These results directly reflect on the health of the animals, the health of the workers and the health of the respective environment, which can enable the continuity of the propagation of the etiological agents involved in the mastitis infection. The awareness of producers and workers on these properties about the disease, transmission, sanitary aspects and adequate management and treatment are essential for improving milk production and production efficiency.

Investigation of Salmonella types and its antibiotic resistance profile and determination of parasite carrier in free-living hedgehogs in Istanbul, Türkiye

Hedgehogs, which are kept as pets worldwide, are prohibited from being sold or kept as pets in Türkiye. The aim of the study was to investigate Salmonella and endo-parasite carriage in hedgehogs and to determine the antibacterial resistance profiles of isolated Salmonella agents. Free-range hedgehogs living in parks and gardens of Istanbul that were brought to veterinary clinics by animal lovers for check-up and treatment were used for sampling. Thirty faecal samples were taken from the animals. For bacteriological examinations, samples were seeded on selective media, and then the isolates were identified by conventional methods as Salmonella spp. Identification and antibiotic susceptibilities of the isolates were determined by the BD Phoenix Automated Microbiology System. All faecal samples were also investigated by PCR for Salmonella spp. specific gene regions. The aminoglycoside and quinolone resistance profiles of the strains were analysed by PCR. On parasitological examination, samples were analysed for parasites’ eggs, oocysts, and larvae. Two Salmonella isolates were determined as Salmonella enterica serovar Typhimurium (S. Typhimurium), whose antigenic formula was 4,5,12:i:1,2. The isolates were phenotypically resistant to ciprofloxacin but did not harbour any of the genes examined. Out of 30 faecal samples, 19 (63.3%) were positive for one or more genera of parasites. The hedgehogs were infected with the species of Capillaria spp. (60%), Crenosoma spp. (13.3%), Eimeria spp. (10%) and Acanthocephala spp. (6.6%). This study is the first report of Salmonella carrier and internal parasite fauna of hedgehogs in Istanbul, Türkiye.

Antibiotic resistance and virulence factor gene profile of A. hydrophila isolated from carp (Cyprinidae) suspected with hemorrhagic septicemia in Gilan, Iran.

The present study was conducted to determine the antibacterial resistance profile of Aeromonas hydrophila (n = 42) isolated from the 100 hemorrhagic septicemia-suspected carp in Gilan, Iran. The prevalence of class 1 and 2 integrons, antibiotic resistance genes (ARG) and virulence factor genes (VFG) among these isolates was investigated using PCR. Also, the possible association between the presence of VFGs and the antibiotic resistance profile of isolates was assessed. The majority of A. hydrophila isolates (83·33%) exhibited multi-drug resistance (MDR) profile, and all isolates were resistant to clindamycin, while all isolates were susceptible to amikacin. intI1 and intI2 gene was found in 26·2 and 4·8% isolates, respectively. This is the first report of the presence of the intI2 gene in A. hydrophila isolates in Iran. The blaTEM (40·5%) and tetA (33·3%) genes were found as the predominant ARGs. The most frequently detected VFGs were lip and ahh1(90·5%), while the examined isolates carrying at least three VFGs and the most prevalent VFGs profile was ast+, act+, alt+, ahhl+, aerA+, ahyB+ and lip+. The results of this study indicate a positive association between the presence of VFGs and antibiotic resistance, and most MDR A. hydrophila isolates showed high frequencies of VFGs.

Incidence of Vancomycin-Resistant Staphylococcus aureus Strains among Patients with Urinary Tract Infections.

There has been a substantial rise in the number of vancomycin-resistant Staphylococcus aureus (VRSA) strains during the last several years. The proportion of vancomycin-resistant strains among isolated S. aureus has risen steadily in recent years, with the first spike occurring in critical care units and thereafter in general hospital wards. S. aureus isolates from urinary tract infection patients were studied for their prevalence and antibiotic resistance. From 292 urine samples, 103 bacterial strains (35.3%) were identified as S. aureus. Various antibiotics were used to test the isolates’ antibacterial resistance profiles. Antibiotic resistance to erythromycin was found in most bacterial isolates, whereas tobramycin antibiotic sensitivity was found in most of them. Vancomycin resistance was found in 23 of all S. aureus isolates in this study. Analysis for β-lactamase found that 71% of S. aureus isolates were positive in all isolates. There was a single plasmid with a molecular weight of 39.306 Kbp in five selected VRSA isolates that was subjected to plasmid analysis. There was evidence of vancomycin resistance among the S. aureus isolates collected from UTI patients in this investigation. This vancomycin resistance pretenses a challenge in the treatment of S. aureus infections and the need to precisely recognize persons who require last-resort medication such as tobramycin.

STAPHYLOCOCCUS AUREUS RESISTANCE AGAINST CEPHALOSPORIN ANTIBIOTIC IN ADEN-YEMEN

The study was carried out to determine antibacterial resistance profiles of methicillin-resistant Staphylococcus aureus (MRSA), isolated from clinical samples (n: 352) of patients during the period, from January 2019 to July 2020 in five governmental and private medical laboratories of Aden governorate, Yemen. The results showed the percentage for resistance ratio is differentiated between samples and the category of cephalosporin antibiotic groups. The highest percentage of resistance was in the wound sample for Cefadroxil, Cefuroxime, and Ceftriaxone at (100%), in addition to 100% in the pus, CSF, and sputum samples for Ceftriaxone antibiotic. Moreover, Cefadroxil has 100 % of resistance in semen, ear, and sputum samples. However, the Staphylococcus aureus isolated from the pharyngeal sample not showed any resistance to all cephalosporin antibiotic groups.

Mastitisli Sığırlardan İzole Edilen Stafilokoklarda Antibiyotik Direnci ve Virülens Gen Profilleri

Staphylococcus spp. are the important bacterial agents of subclinical and clinical mastitis cases. This study was aimed to determine the vancomycin resistances, antibacterial resistance profiles, some virulence genes, and pheno- and genotyping of staphylococci from mastitis. For this aim, 121 staphylococcal isolates were analyzed. The identifications of isolates were confirmed with PCR for being Staphylococcus spp. and Staphylococcus aureus. The antibiotic resistance patterns were determined by Kirby bauer Disc Diffusion Tests and according to the resistance profiles, the isolates were antibiotyped. The vancomycin resistance genes were determined by PCR for investigating the vanA, vanH, vanR, vanS, vanZ, vanY and vanX genes. The vancomycin resistant isolates were genotyped with RAPD-PCR. The nuc gene was detected in 86 of 121 staphylococcal isolates examined and named as Staphylococcus aureus. The remaining 35 isolates were defined as Staphylococcus spp. S. aureus isolates were found to be resistant to penicillin G, amoxicillin/clavulanic acid, oxacillin, tetracycline, cefaperazone, teicoplanin, vancomycin and trimethoprim-sulfomethaxazole at 50% (43/86), 40.7% (35/86), 34.9% (30/86), 23.3% (20/86), 22.1% (19/86), 18.6% (16/86) 10.5% (9/86) and 8.1% (7/86) respectively. On the other hand, 4 (11.4%) of 35 Staphylococcus spp. isolates were resistant to penicillin G, 3 (8.6%) to amoxicillin/clavulanic acid, 2 (5.7%) to trimethoprim-sulfometaxazole, 1 (2.9%) to oxacillin, cefaperazone, teicoplanin, while all isolates were sensitive to vancomycin and tetracycline. Of the 9 S. aureus isolates that were phenotypically resistant to vancomycin, vanA gene was detected in 2 and vanR gene in 3 isolates. Multiple resistances to three or more antibiotics were determined in 42 of 86 S. aureus isolates. In addition, coa gene was detected in 61 (70.9%) of 86 S. aureus isolates. 10 different gene polymorphisms were detected in coa gene positive isolates. While the spa gene was determined in all S. aureus isolates, it was revealed that they had 4 spa gene polymorphisms. Nine different genotypes with a similarity between 51-75% were detected in the genotyping of vancomycin resistant 9 isolates. In conclusion, multiple antibiotic resistance rates in S. aureus isolates investigated were found to be important for mastitis treatment. The results obtained from this study show that milk and dairy products containing these factors pose a public health risk due to the determination of vancomycin resistance in mastitis-derived Staphylococcus strains.

S. aureus and K. pneumoniae on the Surface and within Core of Tonsils in Adults with Recurrent Tonsillitis.

Background and Objectives: Recurrent tonsillitis is an infection of the palatine tonsils. Samples for microbiological testing are usually obtained from the inflamed surface of the tonsils. Colonizing the surface bacteria does not always correlate with pathogens causing recurrent tonsillitis and there is no consensus or this in research studies. The aim of the study was to compare whether Staphylococcus aureus (S. aureus) and Klebsiella pneumoniae (K. pneumoniae) differ when isolated from the tonsillar surface or tonsillar crypts in patients with recurrent tonsillitis. Materials and Methods: a case series study was conducted at a tertiary referral center among 25 patients diagnosed with recurrent tonsillitis. An evaluation of S. aureus and K. pneumoniae incidence, biofilm formation and antibacterial susceptibility was performed. Results: There was a statistically significant association between surface and punch biopsy samples for S. aureus (Fisher’s Exact test p = 0.004) and K. pneumoniae (Fisher’s Exact test p < 0.001). A McNemar test did not reveal a statistically significant association. Although the antibacterial resistance profile was not broad, five out of nine S. aureus isolates were biofilm producers and four out of five K. pneumoniae isolates were biofilm producers. Conclusions: Surface and core cultures of tonsils are comparable with a differing incidence between the surface and the punch biopsy cultures for S. aureus and K. pneumoniae. A larger quantity of bacteria exist in surface samples suggesting that a biopsy sample may be less challenging in evaluating recurrent tonsillitis. We recommend that antibacterial susceptibility results are considered alongside the biofilm-forming potential of isolated bacteria.

EVALUATION OF SOME ANTIBIOTIC RESISTANCE IN STAPHYLOCOCCUS AUREUS ISOLATED BY MEDICAL LABORATORIES ADEN, YEMEN

The study was carried out to determine antibacterial resistance profiles of methicillin-resistant Staphylococcus aureus (MRSA) isolates from clinical samples of patients during the duration, from January 2019 to July 2020 in five governmental and private medical laboratories of Aden governorate, Yemen. A total of 267 MRSA isolates were tested with a disc diffusion method from 4 clinical specimens. The isolates were examined for methicillin resistance using vancomycin, erythromycin, amikacin, and gentamicin.&#x0D; The results revealed a large number of MRSA isolates showed resistance to erythromycin (43.5%) and Vancomycin (28%). A lower number of MRSA isolates showed resistance to amikacin (6.25%), gentamicin (11.75%). A majority of MRSA isolates were from urine specimen (162) followed by (56) wounds, (34) pus, and (25) semen specimens. The highest proportion of MRSA isolates were obtained from Al-medina medical laboratory (31%) and Alborg laboratory (24%).

Source and antibiotic susceptibility of gram-negative bacteria causing superficial incisional surgical site infections

BackgroundSurgical site infections (SSIs) are a major public health problem worldwide and the second-most frequently reported type of nosocomial infection. The presence of Gram-negative bacteria (GNB) in the hospital environment is significantly associated with SSIs. As such, this study aimed to explore the rate, antibiotic profile and common sources of bacteria in SSI. MethodsA cross-sectional study was conducted between August 2018 and July 2019 in three surgical centers in Kalar city, Kurdistan region, Iraq. A total of 512 patients, who underwent surgery were followed up with for superficial incisional SSI. Pre-, intra- and postoperative swabs were collected from patients and the surrounding hospital environment and processed for isolation and identification of GNB by microbiological and molecular methods. The isolates were typed by sequencing and tested for antibiotic resistance using the disc-diffusion technique. ResultsSSI developed in 113 (22.07%) of the patients studied. GNB were involved in 48 (9.38%) cases; among these, Klebsiella spp. were the predominant cause 21/48 (43.75%), followed by Escherichia coli 14/48 (29.17%), Pseudomonas aeruginosa 7/48 (14.58%), Enterobacter spp. 3/48 (6.25%) and A. baumannii 2/48 (4.17%). Citrobacter koseri showed the lowest rate of infection 1/48 (2.08%). Sequencing analysis and the antibacterial resistance profile revealed that 25 (74.28%) of the 48 SSI isolates were from the hospital environment, whereas the rest were exogenous, with an undetermined source. The involved GNB were highly resistant to most antibiotics tested. ConclusionSSIs caused by GNB were mostly exogenous or from the hospital environment, not from endogenous sources. All of the bacterial isolates detected from SSI patients were multidrug resistant.

Occurrence and antibiogram of Listeria species in raw pork, beef, and chicken meats marketed in Enugu State, Southeast Nigeria.

Aim:This study was undertaken to isolate Listeria (L.) species from raw meats sold in markets in Enugu State, Southeast Nigeria, and to determine the antibacterial resistance profile.Materials and Methods:Twenty-five grams of beef (n=144), chicken meat (n=144), and pork (n=144) were collected randomly from supermarkets and general markets in Enugu State. Isolation of Listeria was done using half and full Fraser broths, and polymyxin acriflavine lithium chloride ceftazidime aesculin mannitol agar. Identification of isolates was done using an analytical profile index kit specific for Listeria. Confirmation of the genus Listeria was done by a polymerase chain reaction. The resistance of the isolates was determined using the disk diffusion method.Results:Listeria was isolated from 39/144 (27.1%) chicken meat, 19/144 (13.2%) pork, and 66/144 (45.8%) beef samples cultured. Listeria innocua was the predominant species in chicken meat (52.6%) and beef (81.8%) samples. Listeria grayi, Listeria welshimeri, and Listeria ivanovii were also isolated from the beef and chicken meat samples. More than 65% of the isolates were resistant to penicillin, rifampicin, ciprofloxacin, sulfamethoxazole-trimethoprim, and cephalothin. All the isolates from beef and pork samples and 23 (92%) from chicken meat samples, were resistant to ≥3 classes of antibacterial agents. Mean multiple antibiotic resistance index (MARI) was 0.77 (range=0.42-1.00), 0.58 (range=0.25-0.83), and 0.79 (range=0.58-0.92) for the isolates from beef, chicken meat, and pork samples, respectively. All the isolates had MARI >0.2.Conclusion:Multidrug-resistant Listeria strains contaminate raw beef, pork, and chicken meats marketed in Enugu State, Southeast Nigeria.

Prosthetic valve endocarditis caused by Pseudomonas aeruginosa with variable antibacterial resistance profiles: a diagnostic challenge

BackgroundInfective endocarditis (IE) caused by gram-negative bacilli is rare. However, the incidence of this severe infection is rising because of the increasing number of persons at risk, such as patients with immunosuppression or with cardiac implantable devices and prosthetic valves. The diagnosis of IE is often difficult, particularly when microorganisms such as Pseudomonas aeruginosa, which rarely cause this infection, are involved. One of the mainstays for the diagnosis of IE are persistently positive blood cultures with the same bacteria, while polymicrobial bacteremia usually points to another cause, e.g. an abscess. The antimicrobial resistance profile of some P. aeruginosa strains may change, falsely suggesting an infection with several strains, thus further increasing the diagnostic difficulties.Case presentationA 66-year old male patient who had a transcatheter aortic valve implantation (TAVI) one year previously developed fever seven days after an elective inguinal hernia repair. During the following four weeks, P. aeruginosa with different antibiotic resistance profiles was repeatedly isolated from blood cultures. Repeated trans-esophageal echocardiograms (TEE) were negative and an infection by different P. aeruginosa strains was suspected. Extensive diagnostic workup for an infectious focus was performed with no results. Finally, an oscillating mass on the aortic valve was detected by TEE five weeks after the initial positive blood cultures. P. aeruginosa endocarditis was confirmed by culture of the surgically removed valve. Whole genome sequencing of the last two P. aeruginosa isolates (valve and blood culture) revealed identical strains, with genome mutations for AmpR, AmpD and OprD.ConclusionsThe diagnosis of prosthetic valve endocarditis is particularly difficult for several reasons. The modified Duke criteria have a lower sensitivity for patients with prosthetic valve endocarditis and the infection may be caused by “unusual” pathogens such as P. aeruginosa. Patients with repeatedly positive blood cultures should make clinicians suspicious for endocarditis even if imaging studies are negative and if isolated pathogens are “unusual”. Repeatedly positive blood cultures for P. aeruginosa should be considered as “persistent bacteremia” (suspicious for IE) even in the presence of different antibiotic susceptibility patterns, since P. aeruginosa might rapidly activate or deactivate resistance mechanisms depending on antibiotic exposition.

Occurrence and Antibiogram of Extended-Spectrum Cephalosporin- and Cephamycin-Resistant Escherichia coli in Asymptomatic University Students

Background: Apparently healthy individuals could serve as reservoirs and disseminators of extendedspectrum cephalosporin (ESC)- and cephamycin (cefoxitin, FOX)-resistant, and extended-spectrum β-lactamase-producing (ESBL-P) Escherichia coli which jeopardizes antibacterial therapy thereby posing a threat to the health of infected individuals/carriers. Objectives: This study aimed to screen healthy asymptomatic students in the University of Nigeria, Nsukka (UNN) as potential reservoirs of ESC- and FOX-resistant and ESBL-P E. coli and to determine the antibacterial resistance profile. Materials and Methods: Anal swabs were collected from 190 randomly selected healthy asymptomatic students of both genders in UNN between March and July 2018. ESC-resistant E. coli was isolated using MacConkey agar with 2 µg/mL ceftazidime. ESBL production was assessed by combination disc method while cephamycin resistance was determined using cefoxitin disc screening. Phenotypic resistance of the isolates was determined using disc diffusion method. Results: Out of 190 samples, 20 (10.2%) demonstrated growth. Of these, 6 (30%) were FOX resistant (putative AmpC-producers) but none produced ESBL. The resistance of the isolates was 100% to ampicillin (AMP), 95% to ceftazidime (CAZ), tetracycline (TET) and sulfamethoxazole-trimethoprim (SXT), 30% to FOX and chloramphenicol (CHL), 85% to ciprofloxacin (CIP), enrofloxacin (ENR) and streptomycin (STR), and 65% to kanamycin (KAN). All the isolates were susceptible to meropenem (MEM). Among the 20 isolates, 1 (5%) was resistant to 2 classes of antibacterial agents while 19 (95%), including all the FOX-resistant strains, were resistant to ≥ 3 classes of antibacterial agents. The isolates exhibited 11 multiple antibacterial resistance patterns with AMP, CAZ, FOX, TET, CIP, ENR, STR, KAN, SXT being predominant. Conclusion: Healthy asymptomatic students in UNN are potential reservoirs and disseminators of ESC- and cephamycin (FOX)-resistant E. coli.

Multidrug-resistant enterobacteria colonize commercial day-old broiler chicks in Nigeria.

Aim:This study was conducted to isolate generic enterobacteria from day-old broiler chicks in Nigeria, determine the antibacterial resistance profile, and assess multidrug resistance.Materials and Methods:The birds were sourced from five purposively-selected hatcheries (identified as A, B, C, D and E) in Southwest Nigeria. Non-duplicate cloacal swabs were collected from a total of 75 (15 birds per hatchery) randomly selected apparently healthy birds. Sampling was done in three batches of five chicks per batch at 2-week interval. Isolation of enterobacteria was done using MacConkey agar. The resistance of the isolates was determined using the disk diffusion method.Results:Of 15 processed samples of birds from each hatchery, all samples from hatcheries B, D, and E, 10 (66.7%) and 14 (93.3%) samples from hatcheries A and C, respectively, yielded pure cultures of Escherichia coli. Klebsiella was also isolated from 1 (7.1%) of the 14 and 2 (13.2%) of the 15 growth-positive samples from hatcheries C and D, respectively. The range of resistance among E. coli isolates was tetracycline (86.7-100%), ampicillin (80-100%), gentamicin (60-85.7%), sulfamethoxazole-trimethoprim (46.7-92.9%), enrofloxacin (40-100%), ciprofloxacin (26.7-86.7%), streptomycin (10-80%), cefotaxime (26.7-73.3%), amoxicillin-clavulanic acid (13.3-60%), and ceftazidime (6.7-40%). Klebsiella and all E. coli isolate from chicks of hatcheries B, C, and E, 80 and 93.3% of those from chicks of hatcheries A and D, respectively, exhibited resistance to three or more classes of antibacterial agents.Conclusion:Commercial day-old broiler chicks in Nigeria are colonized by multidrug-resistant coliforms (E. coli and Klebsiella) and are potential reservoirs and disseminators of these organisms.

Microbiological and antibacterial resistance profile in canine otitis externa – a comparative analysis

The aim of the present study was to compare the prevalence of microbial agents involved in canine otitis externa and their sensitivity to antibacterial drugs in two periods: 2007–2011 and 2013–2017. For 2013–2017, coagulase-positive staphylococci were the dominating bacterial species (186 isolates), followed by Pseudomonas aeruginosa (82 strains). The rate of isolated yeasts (mainly Malassezia pachydermatis) was substantially high (152 isolates). Compared to the earlier period (2007–2011) a tendency to more frequent occurrence of co-infections was noted – 61.7% and more than 80% of co-infections involved yeasts. Antibacterial resistance patterns showed a clear trend to increased resistance of coagulase-positive staphylococci and β-haemolytic streptococci to amoxicillin/clavulanic acid (42% and 50% respectively) and gentamicin (29%, 40%). Increased resistance of P. aeruginosa was established to gentamicin (16%) and amikacin (18%). The prevalence of pseudomonads resistant to enrofloxacin was lower (27%).

Occurrence and Antibiogram of Generic Extended-Spectrum Cephalosporin-Resistant and Extended-Spectrum β-Lactamase-Producing Enterobacteria In Horses

Abstract This study was conducted to isolate generic extended-spectrum cephalosporin (ESC)-resistant and extended-spectrum β-lactamase (ESBL)-producing enterobacteria from horses in Nigeria, and to determine the antibacterial resistance profile. Rectal swabs were collected from 155, systematic randomly selected, apparently-healthy horses. Isolation of ESC-resistant enterobacteria was done using Mac Conkey agar with ceftazidime. ESBL production was assessed by combination disc method. Resistance of the isolates was determined using disc diffusion method. Out of 155 samples, 5.2% gave positive growth. From these, 11 ESC-resistant enterobacteria comprising of 36.4% E. coli, 36.4% Salmonella spp. and 27.2% Proteus spp., were obtained. From 11 isolates, 45.5% consisting of all the 4 E. coli and 1 Proteus isolates, were ESBL-producers, these were recovered from 4 (2.6%) out of the 155 horses sampled. Resistance of the E. coli isolates was 25% to aztreonam (AZT), 75% to amoxicillin-clavulanic acid (AMC), gentamicin (GEN), perfloxacin (PEF), and sulphamethoxazoletrimethoprim (SXT-TRI), 50% to ofloxacin (OFL) and 100% to ampicillin (AMP), ceftazidime (CTZ), cefotaxime (CTX), chloramphenicol (CHL), streptomycin (STR), tetracycline (TET), sparfloxacin (SPA), ciprofloxacin (CIP), norfloxacin (NOR) and enrofloxacin (ENR). Resistance of the Salmonella isolates was 50% to PEF and 100% to CTZ, CTX, AMP, AZT, AMC, CHL, GEN, STR, TET, SPA, CIP, OFL, NOR and ENR. Resistance of the Proteus isolates was 25% to AMC, CHL, STR, TET, SPA and NOR, and 100% to CTZ, CTX, AZT and AMP. Resistance of the isolates to more than 3 classes of antibacterial agents tested was 75% for Proteus and 100% for E. coli and Salmonella, respectively. This study showed that horses in Nigeria are potential reservoirs and disseminators of ESC-resistant and ESBL-producing Enterobacteriaceae.

Occurrence, Antibiotic Resistance, and Population Diversity of Listeria monocytogenes Isolated From Fresh Aquatic Products in China.

Listeria monocytogenes is an important Gram-positive foodborne pathogen. However, limited information is available on the comprehensive investigation and potential risk of L. monocytogenes in fresh aquatic products, which are popular to consumers in China. This study aimed to determine the occurrence, virulence profiles, and population diversity of L. monocytogenes isolated from aquatic products in China. In total, 846 aquatic product samples were collected between July 2011 and April 2016 from 43 cities in China. Approximately 7.92% (67/846) aquatic product samples were positive for L. monocytogenes, 86.57% positive samples ranged from 0.3 to 10 MPN/g, whereas 5.97% showed over 110 MPN/g by the Most Probable Number method, which included two samples of products intended to be eaten raw. Serogroups I.1 (serotype 1/2a), I.2 (serotype 1/2b), and III (serotype 4c) were the predominant serogroups isolated, whereas serogroup II.1 (serotype 4b) was detected at much lower frequencies. Examination of antibacterial resistance showed that nine antibacterial resistance profiles were exhibited in 72 isolates, a high level susceptibility of 16 tested antibiotics against L. monocytogenes were observed, indicating these common antibacterial agents are still effective for treating L. monocytogenes infection. Multilocus sequence typing revealed that ST299, ST87, and ST8 are predominant in aquatic products, indicating that the rare ST299 (serotype 4c) may have a special ecological niche in aquatic products and associated environments. Except llsX and ptsA, the 72 isolates harbor nine virulence genes (prfA, actA, hly, plcA, plcB, iap, mpl, inlA, and inlB), premature stop codons (PMSCs) in inlA were found in four isolates, three of which belonged to ST9. A novel PMSC was found in 2929-1LM with a nonsense mutation at position 1605 (TGG→TGA). All ST87 isolates harbored the ptsA gene, whereas 8 isolates (11.11%) carried the llsX gene, and mainly belonged to ST1, ST3, ST308, ST323, ST330, and ST619. Taken together, these results first reported potential virulent L. monocytogenes isolates (ST8 and ST87) were predominant in aquatic products which may have implications for public health in China. It is thus necessary to perform continuous surveillance for L. monocytogenes in aquatic products in China.