Conventional Biochemical Tests Research Articles

First report of Kurthia gibsonii in the digestive system of Apis mellifera in an indigenous community in the south of Mexico

The insect digestive system is an outstanding niche for recognizing and understanding features and interactions with unique microbiota. Apis melliferaplay an important role in honey production and pollination, but their populations have decreased over time, so their study and comprehension are vital. Specimens were collected to study the culturable microbiota of the digestive system in an indigenous community in southern Mexico. Digestive samples of bees from four apiaries were randomly analyzed and cultured, and Gram-positive bacilli were identified through conventional biochemical tests and with molecular markers. Kurthia gibsoniiwas identified in 7.5% (6/80) of the cases, and the sequences obtained were reported in GenBank. This is the first report of K. gibsoniiin the digestive system of bees in apiaries in southern Mexico.

ISOLATION, MOLECULAR IDENTIFICATION AND ANTIBIOTIC SENSITIVITY OF SALMONELLA FROM BUFFALO FECES IN SYLHET DISTRICT OF BANGLADESH

Salmonella is a widely distributed foodborne pathogen affecting humans and animals around the globe. This cross-sectional bacteriological study was aimed at isolation of Salmonella from fecal samples of buffalos in Sylhet district (Upazilas Jaintapur and Fenchuganj) of Bangladesh, their molecular confirmation, and learning their antibiotic sensitivity patterns. A total of 334 fecal samples were collected using a simple random sampling technique. Standard conventional bacteriological culture and biochemical tests were done to isolate and confirm Salmonella. The isolates were confirmed with polymerase chain reaction (PCR). In this study, Salmonella was isolated from 56 samples (16.77%) using bacteriological culture methods and biochemical tests, and all isolates were confirmed in PCR tests. The prevalence of Salmonella was estimated to be 17.57% and 13.93% in Jaintapur and Fenchuganj Upazilas, respectively. Buffalo calves under one year of age had a higher prevalence (24.32%) than older animals aged 1 - 2 (18.62%), 2 - 4 (13.25%), and > 4 years (10.67%). Furthermore, the prevalence was substantially higher in diarrheic animals (72.22%) than in their healthy counterparts (13.60%). It did not vary significantly (p > 0.05) between animals from Jaintapur and Fenchuganj Upazilas. Likewise, no significant difference (p > 0.05) in Salmonella isolates was detected between different age groups. The results revealed that the isolation rate of Salmonella was significantly (p < 0.05) higher in female and diarrheic animals. The most effective antibiotics against the majority of Salmonella isolates were gentamycin (100%), levofloxacin (100%), and ciprofloxacin (76.67%). On the other hand, Salmonella isolates were highly resistant to tetracycline (100%), ampicillin (87.5%), and streptomycin (78.6%). Pathogenic microorganisms in feces are a potential risk to public health due to environmental and animal food contamination. Therefore, infection control and establishing strategic antibiotic therapy should be a priority.

Occurrence of Salmonella Species in Local and Commercial Chickens Slaughtered at Samaru and Sabongari Live Bird Markets, Kaduna State

Salmonellosis is a bacterial infection caused by members of the genus Salmonella. It is one of the most common and important zoonotic diseases. The route of infection from animals to humans is usually through contaminated food, water and environment. Bacteriological investigation on 303 cloacal swab samples collected from local (139) and commercial (164) chickens slaughtered at Samaru and Sabon-gari live bird markets, Kaduna State was carried out to determine the occurrence of Salmonella species. Cultural isolation and identification using conventional biochemical tests, microgen test kit™ and Polymerase Chain Reaction (PCR) amplification of invA gene of the Salmonella isolates was carried out. Isolates were subjected to antimicrobial susceptibility tests, phenotypic detection of Extended Spectrum Beta Lactamases (ESBLs) producing Salmonella isolates using the modified Clinical and Laboratory Standards Institute (CLSI) ESBL confirmatory test. The occurrence of Salmonella species based on cultural and biochemical tests was found to be 13 (4.29%), microgen kit identified 7 (53.8%) of the 13 Salmonella species while PCR based on invA gene confirmed 9 (69.2%) as the overall occurrence rate of Salmonella species. Meanwhile, the study also revealed rate of occurrence of Salmonella species in local chickens 8 (61.5%) which was higher compared to commercial chickens 1 (7.69%) with a statistical significant association (χ2 = 8.775, P = 0.003) between the occurrence of Salmonella species in local and commercial chickens. Antimicrobial susceptibility testing was carried out using panel of 12 antibiotics from 8 different antibiotic classes. Highest rate of sensitivity of the isolates to antibiotics was observed for Ofloxacin with 90%, while all the isolates (100%) were resistant to Nalidixic Acid, Cefazolin, Amoxicillin + Clavulanic Acid, Oxacillin and Penicillin. This study revealed multidrug-resistance (MDR) profile of Salmonella with a total of (25%) of Salmonella isolates being resistant to 5 antibiotics belonging to 4 different classes of antimicrobials with the highest multiple antibiotic-resistant (MAR) indices ranging from 0.58 to 0.91. All the nine (9) isolates of Salmonella tested were found to be negative for ESBLs production. Therefore, it is concluded that Salmonella species are present in local and commercial chickens slaughtered at Samaru and Sabon-gari live bird markets, Kaduna State and could pose serious public health risks to handlers and to consumers of poultry meat and its products.

First US Case of Vagococcus Lutrae Infection in a Human: A Case Report

Abstract Introduction/Objective Vagococcus Lutrae is a gram-positive, catalase-negative bacterium, that has been isolated from different animal species. It is an extremely rare pathogen in humans and has been documented in only two human cases globally. We present a case of V. Lutrae infection in humans, which is the third in the world and first in the US to be documented in literature. Methods/Case Report Our patient was a 75-year-old male with a history of hypertension and hyperlipidemia who presented with a right groin abscess with associated skin necrosis and cellulitis. V. Lutrae was identified in the culture of the abscess fluid. The diagnosis was made after conventional biochemical tests, automated methodology, and mass spectrometry (MALDI-TOF MS). The patient underwent incision and drainage of the abscess, was treated with antibiotics, and showed improvement upon follow-up. Results (if a Case Study enter NA) NA Conclusion Our case highlights the importance of considering V. Lutrae as a potential pathogen in patients with skin infections, particularly those with a history of contact with contaminated water or soil. Further studies are needed to better understand the epidemiology and clinical manifestations of V. Lutrae infections in humans.

Coagulase-Negative Staphylococci as an Etiologic Agent of Ovine Mastitis, with a Focus on Subclinical Forms.

The objective of this review is to investigate the distribution and the characteristics of coagulase-negative Staphylococci (CoNS) implicated in ovine mastitis, and especially in subclinical cases, in order to provide a global perspective of the current research data and analyze specific critical aspects of the issue. PRISMA guidelines were implemented in the search of the last 20 years of the related literature in two databases. In total, 139 studies were included in this review. Relevant data were tracked down, assembled, and compared. Regarding the geographical distribution, most studies originated from Europe (68), followed by South America (33). Lacaune was the most examined breed, while S. epidermidis was the predominantly identified species, representing approximately 39% of the obtained isolates. Antibiotic resistance in the relevant bacteria was documented mostly for Penicillin (32.8%) and Amoxicillin (32.1%), while biofilm- and toxin-associated genes were encountered in variable rates because significant inequalities were observed between different articles. Significantly higher rates of antimicrobial resistance were detected in Asia and South America compared to Europe. Finally, the diagnostic procedures carried out in the respective studies were evaluated. Conventional culture and biochemical tests were mostly performed for simple strain identification; therefore, further molecular investigation of isolates should be pursued in future studies, as this will provide important data regarding specific aspects of the implication of CoNS in ovine mastitis.

Bacterial profile of surgical site infection and antimicrobial resistance patterns in Ethiopia: a multicentre prospective cross-sectional study

BackgroundGlobally, surgical site infections (SSI) are the most commonly reported healthcare-associated infections.MethodsA multicentre study was conducted among patients who underwent surgical procedures at four hospitals located in Northern (Debre Tabor), Southern (Hawassa), Southwest (Jimma), and Central (Tikur Anbessa) parts of Ethiopia. A total of 752 patients clinically studied for surgical site infection were enrolled. The number of patients from Debre Tabor, Hawassa, Jimma, and Tikur Anbessa, hospitals was 172, 184, 193, and 203, respectively. At each study site, SSI discharge culture was performed from all patients, and positive cultures were characterized by colony characteristics, Gram stain, and conventional biochemical tests. Each bacterial species was confirmed using Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI TOF). An antimicrobial susceptibility test (AST) was done on Mueller–Hinton agar using the disk diffusion method. Logistic regression analysis was used to assess associations of dependent and independent variables. A p-value < 0.05 was considered statistically significant. Data were analysed using STATA 16 software.ResultsAmong 752 wound discharge cultures performed, 65.5% yielded growth. Among these, 57.9% and 42.1% were Gram-negative and Gram-positive isolates, respectively. In this study, a total of 494 bacteria were isolated; Staphylococcus aureus (31%), Escherichia coli (20.7%), and Klebsiella pneumoniae (9.8%) were the most common. Rare isolates (0.8% each) included Raoultella ornithinolytica, Stenotrophomonas maltophilia, Alcalignes faecalis, Pantoea ecurina, Bacillus flexus, and Paenibacillus tylopili. Enterobacteriaceae showed high levels of resistance to most of the tested antibiotics but lower levels of ertapenem (32.9%), amikacin (24.3%), imipenem (20.3%), and meropenem (17.6%) resistance. Multidrug-resistant (MDR) frequency of Enterobacteriaceae at Debre Tabor, Hawassa, Jimma, and Tikur Anbessa hospitals was 84.5%, 96.5%, 97.3%, and 94%, respectively. Ages ≥ 61 years (AOR = 2.83, 95% CI: 1.02–7.99; P 0.046), prolonged duration of hospital stay (AOR = 4.15, 95% CI: 2.87–6.01; P 0.000), history of previous antibiotics use (AOR = 2.83, 95% CI: 1.06–2.80; P 0.028), history of smoking (AOR = 2.35, 95% CI: 1.44–3.83; P 0.001), emergency surgery (AOR = 2.65, 95% CI: 1.92–3.66; P 0.000), and duration of operation (AOR = 0.27, 95% CI: 0.181–0.392; P 0.000) were significant risk factors.ConclusionThe most prevalent isolates from Gram-positive and Gram-negative bacteria across all hospitals were S. aureus and E. coli, respectively. Many newly emerging Gram-negative and Gram-positive bacteria were identified. Variation between hospitals was found for both SSI etiology type and MDR frequencies. Hence, to prevent the emergence and spread of MDR bacteria, standard bacteriological tests and their AST are indispensable for effective antimicrobial stewardship.

Evaluation of Immulex S. pneumoniae Omni test for the direct detection of S. pneumoniae from positive blood cultures

Rapid and early identification of Streptococcus pneumoniae from positive blood cultures is crucial for the management of patients with bloodstream infections (BSI). Many identification systems in microbiology laboratories have difficulty differentiating S. pneumoniae from other closely related species in the Streptococcus mitis group. To overcome this limitation, we developed a rapid workflow in our laboratory combining direct MALDI-TOF MS identification with the Immulex S. pneumoniae Omni test (SSI Diagnostica, Denmark) for rapid detection of S. pneumoniae directly from positive blood cultures. The workflow was evaluated using 51 Streptococcus isolates. Compared to conventional biochemical testing, our new workflow demonstrates 100 % specificity and sensitivity for the detection and differentiation of S. pneumoniae from other closely related species. Our new workflow is accurate, cost-effective, and can easily be implemented in microbiology laboratories that already perform direct MALDI-TOF identification from positive blood cultures to improve the management of patients with invasive pneumococcal disease. ImportanceInvasive pneumococcal disease remains a major public health problem worldwide. Reducing the time to identify Streptococcus pneumoniae in positive blood cultures allows patients to be treated sooner with more targeted and effective antibiotics. We evaluated a two-step protocol where positive blood cultures are first tested directly by MALDI-TOF MS and any samples containing Streptococcus species are tested by Immulex S. pneumoniae Omni test to both detect and differentiate S. pneumoniae from other closely related Streptococcus species. Our study results showed 100 % sensitivity and specificity, and a much faster turn-around time than conventional methods.

Development of Polymerase Chain Reaction assay protocol for assessment Salmonella sp. in cow raw milk

Salmonella sp. is a pathogenic bacterium that may associated with acute diarrhoea in human. These bacteria may be transmitted in a variety of ways, including consumption of contaminated cow raw milk. Salmonella sp. is troublesome to assessment due to methodological restrictions. The aim of this study was to development the protocol of Polymerase Chain reaction (PCR) assay for Assessment Salmonella sp. in cow raw milk. This method is comprised of (1) biochemical assay and (2) bacterial DNA purification from Selenite Cystine Broth and XLD Xylose Lysine Deoxycholate culture using Wizard® Genomic DNA Purification Kit followed by PCR detection. The biochemical assay is divided into several stages, namely bacterial isolation, Gram staining and conventional biochemical tests. The PCR optimization was done with Salmonella sp. The oligonucleotide primer invasion protein (invA) gene (F: 5’-TCGTCATTCCATTACCTACC-3’; R: 5’-AAACGTTGAAAAACTGAGGA-3’) were used for targeting the diagnosis of Salmonella at the genus level. In biochemical tests, Salmonella sp. results showed posit if result of catalase, oxidase, citrate, TSIA, lactose-sucrose-mannitol fermentation, urea, and methyl-red. Conversely, negative result for Voges-Proskauer. PCR protocol consist of 30 PCR cycles with initial denaturation at 94°C for 45 seconds, denaturation at 94°C for 20 seconds, annealing at 57°C for 15 seconds, extension at 72°C for 15 seconds and finally at 72°C for 2 minutes. The conventional method detection results obtained as many as 9 positive samples Salmonella sp. and the PCR method obtained 7 positive samples Salmonella sp. In conclusion, our results indicate that the developed protocol would be utilized as a routine analysis for monitoring cow raw milk contamination and the protocol of the PCR technique provides results at a handful of time required by the biochemical assay (24 hours compared to 2–3 days).

An Investigation of Bacterial Pathogens Associated with Diseased Nile Tilapia in Small-Scale Cage Culture Farms on Lake Kariba, Siavonga, Zambia

This study investigated disease outbreaks in farmed Oreochromis niloticus (Nile tilapia) in Siavonga among small-scale cage culture farms on Lake Kariba in order to establish bacterial etiological agents associated with fish mortality and to determine their antibiotic susceptibility. A total of 300 fish samples from 11 farms were aseptically collected and bacteria were isolated from the kidney, liver, brain, and spleen. The isolates were identified using their morphological characteristics and conventional biochemical tests. The antibiotic susceptibility of selected bacteria was determined by the Kirby–Bauer disc diffusion method. The following well-known fish pathogens were identified at a prevalence of Aeromonas spp. (13%), Pseudomonas spp. (10.3%), Micrococcus spp. (9.7%), Klebsiella spp. (8.7%), Lactococcus spp. (7.3%), Streptococcus spp. (7.0%), and Acinetobacter spp. (7.0%). All the isolates tested were susceptible to doxycycline, and complete resistance to ciprofloxacin, co-trimoxazole, and cephalothin was recorded in the Bacillus spp. The observed resistance could be attributed to bacteria from terrestrial sources as fish farmers do not administer antibiotics to fish. To our knowledge, this is the first study to establish the occurrence of several bacterial species infecting tilapia in Zambia and the first to determine the antibiotic susceptibility of fish bacteria among small-scale farms on Lake Kariba. The current study provides baseline information for future reference and fish disease management on Lake Kariba and in Zambia.

DETECTION OF CARBAPENEMASE-PRODUCING CARBAPENEM-RESISTANT IN BLOOD CULTURE ISOLATES BY MCIM AND ECIM AND ITS SUSCEPTIBILITY TO TIGECYCLINE AND MINOCYCLINE

Objective: In this study, we aimed to detect different Carbapenemase-producing carbapenem-resistant Enterobacterales (CREs) in blood isolates by phenotypic, modified carbapenem inactivation methods (mCIM) and amp; EDTA-carbapenem inactivation methods (eCIM), and also to study the susceptibility of these CREs toward Tigecycline and Minocycline. Methods: This prospective study included 100 non-duplicate Enterobacterales organisms isolated from 250 blood samples positive for Enterobacterales that showed resistance to carbapenem (Imipenem). The isolates were identified by conventional routine biochemical tests. CRE isolates were screened for Carbapenemase production by the Clinical and Laboratory Standards Institute (CLSI)-recommended, mCIM and eCIM for evidence of the production of matello-beta-lactamase. Antimicrobial susceptibility for Tigecycline and Minocycline drugs was tested by the disk diffusion method on Mueller–Hinton agar according to CLSI guidelines, and susceptibility patterns were recorded. Clinical diagnosis data were collected from the requisition forms sent to our laboratory during test procedures. Results: Out of 100 (40%) CRE isolates tested for mCIM, 34 samples showed positive results for the Carbapenemase enzyme. Among the mCIM-positive isolates, Klebsiella spp. showed the highest prevalence of 58.8% (20/34). While among mCIM-positive isolates (22/34), 64.7% were positive for eCIM (Matello beta-lactamase producer). Maximum samples had been received from the NICU ward from patients diagnosed with early-onset sepsis; 41.2% of these were Carbapenemase-producing Enterobacterales. Among CRE isolates, 86 isolates were susceptible to Tigecycline and 24 isolates were susceptible to Minocycline. 70 CRE isolates were susceptible to Tigecycline but resistant to minocycline, and all CRE isolates resistant to Tigecycline were not susceptible to Minocycline.

Phenotypic characterization and antimicrobial susceptibility profiles of &lt;i&gt;Vibrio cholerae&lt;/i&gt; isolates during the October 2022 and January 2023 outbreak in North-Kivu province, The Democratic Republic of Congo

Background: Cholera is an infectious disease characterized by severe watery diarrhea, frequently occurring in outbreaks which affects many communities in the Democratic Republic of the Congo (DRC). At the end of October 2022, a cholera outbreak was declared in the camp of internally displaced people (IDP) of Kanyaruchinya, 20 kilometers north of Goma, the provincial capital of the North-Kivu province in DRC, as well as in other IDP camps and settlements around the city of Goma. The aim of this study was to phenotypically characterize Vibrio cholerae isolates associated with this outbreak, and to determinate their antimicrobial susceptibility profiles.&#x0D; Methodology: Between October 31, 2022 and January 31, 2023, faecal swab samples were collected into Cary-Blair medium from 1604 and 538 patients with clinical signs of cholera at the Kanyaruchinya IDP, and IDPs camps and settlements around the city of Goma, respectively. After enrichment in 1% alkaline peptone water, the samples were cultured on thiosulphate-citrate-bile salt-sucrose (TCBS) agar for isolation and phenotypic characterization of V. cholerae O1 using conventional biochemical tests and serotyping technique. Antimicrobial susceptibility of selected isolates was peformed to a panel of 8 antibiotics by the disk diffusion method in accordance with EUCAST and CLSI guidelines.&#x0D; Results: Vibrio cholerae was cultured from 807 samples (50.3%) of 1604 patients from the Kanyaruchinya IDP, and from 206 samples (38.3%) of 538 patients around the city of Goma (p&lt;0.01). All the V. cholerae isolates from the Kanyaruchinya IDP (807/807, 100.0%) were serotyped as V. cholerae O1 Inaba whereas 136 (66.0%), 67 (32.5%), and 3 (1.5%) V. cholerae O1 isolates from around Goma were serotyped as V. cholerae O1 Ogawa, V. cholerae O1 Inaba, and Hikojima respectively. Antimicrobial susceptibility test on 174 and 62 isolates selected randomly from the 807 and 206 V. cholerae isolates from the Kanyaruchinya IDP camp, and from around the city of Goma respectively, showed that all the tested V. cholerae O1 isolates were resistant to polymyxin and cotrimoxazole, while being susceptible to tetracycline and azithromycin. All tested V. cholerae O1 isolates from Kanyaruchinya IDP camp displayed a unique antimicrobial susceptibility profile characterized by resistance to ampicillin, cotrimoxazole and chloramphenicol, and susceptiblility to ciprofloxacin, norfloxacin, azithromycin, tetracycline and doxycycline. Their counterparts from settlements around the city of Goma displayed a more variable antimicrobial susceptibility profile.&#x0D; Conclusion: Our results suggest that a single V. cholerae O1 Inaba clone probably caused the cholera outbreak in the Kanyaruchinya IDP camp, whereas during the same period, several V. cholerae clones (Ogawa, Inaba and Hikojima) were associated with the cholera outbreak around the city of Goma.

Emergence of clinical &lt;i&gt;vanA&lt;/i&gt;-type vancomycin-resistant &lt;i&gt;Staphylococcus aureus&lt;/i&gt; isolates in National Orthopaedic Hospital Dala, Kano, Nigeria

Background: The increasing prevalence of multi drug resistance (MDR) in strains of Staphylococcus aureus is a major challenge in the selection of an appropriate therapeutic agents, especially in persistent orthopaedic infections. This study investigated the patterns of antimicrobial resistance and identified the genetic determinants of resistance in S. aureus isolates from orthopaedic patients.&#x0D; Methodology: This was a descriptive cross-sectional study of hospitalized patients at National Orthopaedic Hospital Dala (NOHD), Kano, Nigeria from whom urine samples, and nasal and wound swabs were collected for isolation of S. aureus. Samples were cultured on standard media and S. aureus isolated and identified using both conventional biochemical tests and a standard rapid diagnostic kit. The antibiotic susceptibility was determined to a panel of 15 antibiotics using the modified Kirby-Bauer disc diffusion method. Vancomycin minimum inhibitory concentration (MIC) of each isolate was determined using vancomycin Epsilon-test strip. mecA and vanA were detected by multiplex polymerase chain reaction (PCR) assay.&#x0D; Results: From the total of 134 samples, S. aureus was isolated from 36 (26.8%); 10 (7.4%) from urine, 13 (9.7%) from nasal swab, and 13 (9.7%) from wound swab. Thirty-four (94.4%) isolates were phenotypically methicillin (cefoxitin) resistant (MRSA), while 2 (5.6%) isolates were methicillin sensitive (MSSA). Phenotypic resistance rate of the S. aureus isolates was highest to gentamicin (94.4%), followed by penicillin (88.8%), cephalosporins and fluoroquinolones (87.4%), while rate was lowest to vancomycin (11.1%, 4/36). Seventeen (47.2%) were MDR, 16 (44.4%) were extensively drug resistant (XDR), and 2 (5.6%) were pan-drug resistant (PDR) S. aureus isolates. The mecA gene was detected in 4 (11.8%) of the 34 phenotypic MRSA isolates and vanA genes in 2 (50.0%) of the 4 phenotypic VRSA isolates.&#x0D; Conclusion: The detection of vanA and mecA in clinical S. aureus isolates in this study is an indication that clinical VRSA has emerged in MRSA population in Nigeria. This emergence can pose a major threat to primary care-givers and a public health challenge among the daily inhabitants of National Orthopaedic Hospital Dala (NOHD), Kano and the community at large.

A prospective study evaluating the effect of a ‘Diagnostic Stewardship Care-Bundle’ for automated blood culture diagnostics

We prospectively implemented a diagnostic stewardship care-bundle checklist, 'Sepsis-48 DSB', with the aim of reducing intervening duration of key steps of automated blood culture diagnostics (aBCD). Sepsis-48 DSB was implemented for automated blood culture bottles (BCBs) received from adult intensive care units (AICUs) during the intervention period (P2; July 2020-June 2021) and intervening durations were compared with those during the retrospective, pre-intervention period (P1; March-June 2020). During both periods, provisional blood culture reports (pBCR) were issued wherein direct microbial identification (dID) was performed in BCBs with Gram-negatives by directly inoculating conventional biochemical tests and direct antimicrobial susceptibility testing (dAST) using EUCAST RAST method. The results were compared with the standard of care (SoC) method (i.e. full incubation followed by identification and AST by VITEKⓇ-2 Compact). During P2, significant reductions in loading time (LT) [median: 63.5 vs. 32 minutes, P < 0.001], time to dID+dAST performance (TTD) [186 vs. 115 minutes, P=0.0018] and an increase in compliance to bundle targets [LT ≤45: 44% vs. 66%, P=0.006 and TTD ≤120: 34% vs. 51.7%, P=0.03] were observed. Using dID+dAST method, results were read 694 minutes earlier than SoC method. Of 176 pBCR, 165 (94%) were concordant with SoC in microbial identification of species. Categorical agreement for any drug-bug combination was 92.7% (1079/1164) and corresponding major, very major, and minor error rates were 8.8% (19/216), 4.9% (45/921), and 1.8% (21/1164), respectively. The 'diagnostic stewardship care-bundle' strategy was successfully implemented with considerable diagnostic accuracy leading to significant reductions in duration of targeted steps of aBCD.

Extended Spectrum B-Lactamase (ESBL) and Metallo B-Lactamase (MBL) Production in Gram-Negative Bacteria isolated from Urinary Tract Infection Patients.

Background: The rate of infections caused by Morganella species is reported to be affected by difficulties in their isolation. Enterobacter, species, Citrobacter freundii, Serratia marcescens, and Morganella morganii are the major pathogens involved in hospital acquired infections. All of them are extensively reported in the infections of urinary tract, respiratory tract, cutaneous disorders, catheter associated infections and infections of the central nervous system.Objective: To investigate the frequency of ESBL producing bacteria in Urinary Tract Infection. Material and Methods: A cross-sectional study of 200 urine samples was collected from the outdoor patient departments of Khyber Teaching Hospital Peshawar through consecutive sampling technique during April, 2019 to June 2019. The urine samples were collected and then cultured on selective media i.e., Cysteine Lactose Electrolyte Deficient Agar (CLED) and on MacConkey agar plates. Isolates were identified by conventional morphological and biochemical tests while antibiotic susceptibility was analyzed by “Agar disc diffusion method” using different antibiotics and their zone of inhibition was measured. Results: Total 200 samples processed, 125 (62.5%) isolates, including 67 (53.6%) females, tested positive for the growth of Gram-negative bacteria. Among the identified species, 76 (60.8%) isolates produced ESBLs, with Escherichia coli accounting for 32.6%, Enterobacter for 23.2%, and Morganella species for 12.0%. Moreover, 77 (61.6%) of the isolates were found to be MBL positive, with 30 (24.0%) of them being E. coli, 28 (22.4%) being Enterobacter, and 19 (15.2%) being Morganella spp. The isolates formed a zone of inhibition like a clover leaf and demonstrated a sensitivity of 85 (68%) to antibiotics in the carbapenem class, such as imipenem and meropenem. Cefoparazone/Sulbactam 80 (64.0%), Ofloxacin 79 (63.2%), and Amikacin 78 (62.4%) were the next three antibiotics with high sensitivity. Ampicillin 106 (84.8%) showed the highest resistance, followed by Nitrofurantoin 84 (67.2%) and Ceftriaxone 79 (63.2%), in that order. Conclusion: The study indicated that ESBL-producing bacteria were present frequently among UTI patients. Most of the isolates exhibited strong carbapenem class antibiotic sensitivity. The findings of this study may contribute to the formulation of tactics to lower the incidence of ESBL-producing UTI.Keywords: Antibiotics resistance, Gram Negative bacteria, Plasmid-mediated enzymes, UTI.

High burden of ESBL and carbapenemase-producing gram-negative bacteria in bloodstream infection patients at a tertiary care hospital in Addis Ababa, Ethiopia.

Bloodstream infection due to beta-lactamase and carbapenemase-producing gram-negative bacteria poses a substantial challenge to the effectiveness of antimicrobial treatments. Therefore, this study aimed to investigate the magnitude of beta-lactamase, carbapenemase-producing gram-negative bacteria, and associated risk factors of bloodstream infections in patients at a tertiary care hospital, in Addis Ababa, Ethiopia. An institutional-based cross-sectional study was conducted with convenience sampling techniques from September 2018 to March 2019. Blood cultures were analyzed from 1486 bloodstream infection suspected patients across all age groups. The blood sample was collected using two BacT/ALERT blood culture bottles for each patient. Gram stain, colony characteristics, and conventional biochemical tests were used to classify the gram-negative bacteria at the species level. Antimicrobial susceptibility testing was carried out to screen beta-lactam and carbapenem drug-resistant bacteria. The E-test was conducted for extended-spectrum-beta-lactamase and AmpC-beta-lactamase-producers. A modified and EDTA-modified carbapenem inactivation method was conducted for carbapenemase and metallo-beta-lactamases producers. Data collected using structured questionnaires and medical records were reviewed, encoded, and cleaned using EpiData V3.1. software. The cleaned data were exported and analyzed using SPSS version 24 software. Descriptive statistics and multivariate logistic registration models were used to describe and assess factors associated with acquiring drug-resistant bacteria infection. A p-value <0.05 was considered statistically significant. Among 1486 samples, 231 gram-negative bacteria were identified; of these, 195(84.4%) produce drug-hydrolyzing enzymes, and 31(13.4%) produce more than one drug-hydrolyzing enzyme. We found 54.0% and 25.7% of the gram-negative bacteria to be extended-spectrum-beta-lactamase and carbapenemase-producing, respectively. The extended-spectrum-beta-lactamase plus AmpC-beta-lactamase-producing bacteria account for 6.9%. Among the different isolates Klebsiella pneumonia 83(36.7%) was the highest drug-hydrolyzing enzyme-producing bacteria. Acinetobacter spp 25(53.2%) was the most carbapenemase producer. Extended-spectrum-beta-lactamase and carbapenemase-producing bacteria were high in this study. A significant association between age groups and extended-spectrum-beta-lactamase producer bacterial infection was seen, with a high prevalence in neonates (p = <0.001). Carbapenemase showed a significant association with patients admitted to the intensive care unit (p = 0.008), general surgery (p = 0.001), and surgical intensive care unit (p = 0.007) departments. Delivery of neonates by caesarean section, and insertion of medical instruments into the body were exposing factors for carbapenem-resistant bacterial infection. Chronic illnesses were associated with an extended-spectrum-beta-lactamase-producing bacterial infection. Klebsiella pneumonia and Acinetobacter species showed the greatest rates of extensively drug-resistant (37.3%) and pan-drug-resistance (76.5%), respectively. According to the results of this study, the pan-drug-resistance prevalence was found to be alarming. Gram-negative bacteria were the main pathogens responsible for drug-resistant bloodstream infections. A high percentage of extended-spectrum-beta-lactamase and carbapenemase-producer bacteria were found in this study. Neonates were more susceptible to extended-spectrum-beta-lactamase and AmpC-beta-lactamase-producer bacteria. Patients in general surgery, caesarean section delivery, and intensive care unit were more susceptible to carbapenemase-producer bacteria. The suction machines, intravenous lines, and drainage tubes play an important role in the transmission of carbapenemase and metallo-beta-lactamase-producing bacteria. The hospital management and other stakeholders should work on infection prevention protocol implementation. Moreover, special attention should be given to all types of Klebsiella pneumoniae and pan-drug resistance Acinetobacter spp transmission dynamics, drug resistance genes, and virulence factors.

Survey of Multidrug Resistanct Bacteria in Patients with Lower Respiratory Tract Infection Attending Some Hospitals in Kebbi State, Nigeria

The rise in multidrug resistance is a growing public health concern among agents of respiratory tract infection, which is responsible for morbidity, mortality and costs in Africa. This study was designed to survey for multidrug resistant bacterial pathogens in patients with lower respiratory tract infection (LRTI) attending some hospitals in Kebbi State, Nigeria. Three hundred and fifty sputum samples were collected from patients with the symptoms of LRTI attending six different hospitals in Kebbi State. The samples were all screened for bacterial pathogens using standard microbiological techniques. The bacterial isolates were identified using conventional biochemical tests and then confirmed using commercial biochemical test kit (MICROBACT) according to manufacturer’s instruction. Antimicrobial susceptibility tests were determined using disc diffusion method to detect resistant isolates as well as multidrug-resistant (MDR) isolates. The overall incidence of multidrug-resistant isolates in this study was 39.8%. High level of MDR was observed among Staphylococcus aureus (28.0%) and then Klebsiella pneumoniae (21.0%). MDR were also detected amongst Aeromonas hydrophila (11.6%), Pseudomanas aeruginosa (11.6.0%), Klebsiella oxytoca (11.6%), Burkholderia pseudomallei (7.1%) Escherichia coli (4.7%) and Acinetobacter baumannii (4.7%). In conclusion, MDR isolates were predominately isolated among Staphylococcus aureus and Klebsiella pneumoniae. Indiscriminate use of antibiotics remains the risk factor for developing multidrug resistance isolates.

Characterization of lactic acid bacteria isolated from street pickles of Dhaka, Bangladesh

Traditionally fermented pickles are a popular street food in Bangladesh famous for their unique flavors and health benefits. Pickles are often prepared by fermentation using lactic acid bacteria (LAB) that can act as probiotics. The study was aimed to isolate and characterize lactic acid bacteria from pickle samples collected from streets of Dhaka city, as well as assess the microbial quality of pickles for food safety.A total of 30 pickle samples of different kinds were collected from streets of Dhaka city. Isolation and identification were conducted using conventional cultural and biochemical tests, followed by molecular confirmation of identity. Antibiotic susceptibility of isolates was investigated against 7 antibiotics of different groups. Antimicrobial activity of LAB isolates was analyzed by well-diffusion assay and phenotypic enterocin activity assay. Physiological characterizations of LAB were performed to determine their tolerance to temperature, salt, pH, bile, carbohydrate fermentation pattern, proteolytic activity and biofilm formation.Fifty isolates were obtained from pickle samples, of which 18% was identified as LAB, including Enterococcus faecalis (6) and Enterococcus faecium (3). The rest included S. aureus (18), E. coli (11), Klebsiella spp. (5), Salmonella (3), Shigella (3) and Pseudomonas aeruginosa (1). Antibiotic resistance pattern revealed higher occurrence of resistance against azithromycin among the non-LAB isolates, but none of the LAB isolates were found to resist any of the antibiotics used. Antimicrobial activity of LAB isolates was not observed against the foodborne isolates. All LAB isolates fermented a wide range of carbohydrates and showed adequate tolerance to salt, pH, temperature and bile. Out of 9 isolates, 5 displayed proteolytic activity, and 6 were found as strong biofilm producer.These results suggest that although the LAB isolates from street pickles collected from Dhaka does not have antimicrobial activities, they still have potential to be used as probiotics. It also shows high occurrence of antibiotic resistant foodborne pathogens in pickles, indicating that consumption of such street food can be serious health hazard.

Bacterial vaginosis, vulvovaginal candidiasis, trichomonal vaginitis and aerobic vaginitis in women from Egypt.

Infectious vaginitis is prevalent in developing countries. Most of the females suffer from vaginal infections at least once per lifetime. Due to limited resources, many infections are misdiagnosed or undiagnosed. Good diagnosis of these infections is critically important and will definitely help to guide treatment and prevent recurrence. A total of 1080 vaginal swabs were collected from symptomatic females. Nugent's score and Amsel's criteria were applied to diagnose bacterial vaginosis (BV). A rapid test was used to identify Gardnerella vaginalis. Trichomonal vaginitis (TV) was diagnosed through microscopic examination. Vulvovaginal candidiasis (VVC) was also identified microscopically and using conventional culture. Finally, aerobic vaginitis (AV) was detected using Donder's scale combined with conventional culture and biochemical tests. There was no statistically significant association between age and type of vaginal infection (p=0.130). Vulvovaginal inflammation, itching and redness were significantly associated with VVC (p≤0.012). BV was detected as single infection in 43.8%, followed by VVC 24.2%. On the contrary, AV and TV were scarcely detected among the participants; 4.9% and 0.5% respectively. Mixed infections between BV and VVC were noted in 26.6%. BV showed the highest prevalence followed by VVC. Mixed infections between BV and VVC were evidently noted, therefore good reliable diagnosis using cost-effective methods is crucial for proper treatment. Aerobic vaginitis showed low prevalence and most of the Streptococcus spp. were isolated from pregnant females. The low prevalence of Trichomonas vaginalis may be due to the dependance on conventional methods for diagnosis, and thus more advanced diagnostic tools are required.

Inhibitory activity of Mentha spicata oils on biofilms of Proteus mirabilis isolated from burns

Introduction and Aim: Proteus mirabilis is an opportunistic pathogen, infecting humans, through the release of endotoxins and enzymes such as urease, hemolysin, protease, DNase etc. One of the factors contributing to its virulence is its unusual ability to form crystalline biofilms. This study aimed to investigate the effect of Mentha spicata volatile oil on P. mirabilis biofilm formation. Materials and Methods: P. mirabilis was isolated from infected wound of burns of patients using conventional biochemical tests. Vitek 2-Compact System was used to confirm the diagnosis of bacterial isolates. The isolates were tested for their susceptibility to 11 antibiotics. Aqueous and alcoholic extracts as well as Volatile Oil and natural Menthol extracted from M. spicata were tested for their ability to inhibit biofilm formation by P. mirabilis. Results: In this study 16 out of the 45 burn injury samples were tested positive for P. mirabilis. Bacterial isolates were found to be resistant to the drugs Levofloxacin and Norfloxacin, with percentages of 10.2% and 11.8%, respectively. Majority of these isolates had the capacity to produce several virulence factors, including biofilm in variable amounts and the enzymes protease, hemolysin, DNase, and gelatinase. The volatile oil and natural component menthol extracted from M. spicata inhibited the formation of biofilm at increasing concentrations. Conclusion: The volatile oil and the natural menthol compound of M. spicata are effective in inhibiting biofilm formation by P. mirabilis.

Prevalence and Antibiotic Susceptibility Profile of Staphylococcus aureus in Commercialized Food in Oran, Algeria

Background: With regard to health-threatening infections, Staphylococcus aureus is the leading cause of polymorphic infections varying from a banal tegumentary infection to numerous lethal illnesses. Furthermore, it is the third commonest bacterial cause of food-borne infections worldwide.This study aimed to investigate contamination, prevalence, and antibiotic resistance of S. aureus isolated from several widely marketed food products (raw and processed) in the region of Oran, Algeria.&#x0D; Methods: A total of 350 food samples including prepared meals (n=110), dairy products (n=42), pastry (n=78), meat and its derivatives (n=97), and other commercially available foods (egg products, sweets, and sauces) (n=23) have been randomly purchased from diverse sale outlets and screened for S. aureus strains during the period from July 2021 to September 2022. The isolation and identication of S. aureus bacteria were preformed using conventional culture and biochemical tests such as catalase, coagulase, and DNase tests. Furthermore, the strains were screened for their resistance to five different antimicrobial drugs using the agar diffusion method.&#x0D; Results: The overall prevalence of S. aureus determined among the collecteded samples was 31.14% (109/350), where, pastry products harbored the highest contamination rate (43.59%), and prepared meals, the lowest contamination rate (18.18%). The strains presented a high level of resistance (58.71-39.44%) for gentamycin and oxacilin, respectively. Moreover, the lowest level of resistance was observed against erythromycin 16.51%, and about 83% of strains presented multidrug resistance.&#x0D; Conclusion: The significative prevalence and the high level of multidrug resistant of S. aureus highlights the seriousness to improve food contamination prevention programs and underlines that good hygiene practices at sale outlets has a major impact on the sanitary quality of commercialized food products.