Evaluation of Pathogens Causing Urinary Tract Infection and Their Sensitivity Pattern

Background: UTI constitute a major public health problem in India accounting 2nd most common infection next to respiratory tract infection. They are responsible for increasing treatment cost and significant morbidity.Aim:-To determine the incidence of UTI, evaluation of pathogens responsible and their antimicrobial susceptibility pattern in the population.Methods:Urine samples were collected from 300 patients attending the OPD Patna medical college, Patna during the period of 18 months (January 2017 to June 2018) Antimicrobial sensitivity testing was done for the bacterial isolates present in the sample by Kirby- Bauer disc diffusion method. Only those samples were taken into consideration which develops count equal to or greater than 1*105CFU/ml as indicated by Kass.Results:Out of 300 samples collected 146 (48.66%)) yielded bacterial growth. Out of 146 culture isolates E.Coli was the most common pathogen followed by klebsiella, CoNS and staphylococcus. Antibiotic sensitivity was performed on all the isolates. It was observed that highest sensitivity was 49.31% to amikacin, gentamycin (45.89%), nitrofurantoin (38.35%) meropenem (27.39%).Conclusions:It was observed that high grade of resistance to ampicillin, cotrimoxazole, ciprofloxacin, cefuroxime, chloramphenicol, cefotaxime, cefazolin, amoxicillin + clavulanic acid and gentamycin is present as a result of misuse or improper use of antibiotic in the community. Hence urine culture is necessary for the diagnostic screening of UTI before the treatment.

Probiotics in the Prevention of the Calcium Oxalate Urolithiasis

Nephrolithiasis ranks third among urological diseases in terms of prevalence, making up about 15% of cases. The continued increase in the incidence of nephrolithiasis is most probably due to changes in eating habits (high protein, sodium, and sugar diets) and lifestyle (reduced physical activity) in all developed countries. Some 80% of all kidney stones cases are oxalate urolithiasis, which is also characterized by the highest risk of recurrence. Frequent relapses of nephrolithiasis contribute to severe complications and high treatment costs. Unfortunately, there is no known effective way to prevent urolithiasis at present. In cases of diet-related urolithiasis, dietary changes may prevent recurrence. However, in some patients, the condition is unrelated to diet; in such cases, there is evidence to support the use of stone-related medications. Interestingly, a growing body of evidence indicates the potential of the microbiome to reduce the risk of developing renal colic. Previous studies have primarily focused on the use of Oxalobacterformigenes in patients with urolithiasis. Unfortunately, this bacterium is not an ideal probiotic due to its antibiotic sensitivity and low pH. Therefore, subsequent studies sought to find bacteria which are capable of oxalate degradation, focusing on well-known probiotics including Lactobacillus and Bifidobacterium strains, Eubacterium lentum, Enterococcus faecalis, and Escherichia coli.

Nanocluster Mediated Photothermia Improves Eradication Efficiency and Antibiotic Sensitivity of Helicobacter Pylori

Background: Helicobacter pylori (H. pylori) eradication plays a crucial role in gastric cancer prevention, but the antimicrobial resistance of H. pylori is obstructing this elimination process. In this study, we developed nanoclusters (NCs) from Zn0.3Fe2.7O4 nanoparticles using a poly(ethylene glycol)-b-poly(ε-caprolactone)-based nanocarrier as an innovative antibiotic-independent H. pylori management.Results: The nanocluster showed minimal toxicity and maximal biocompatibility. With a low concentration (50 µg/mL) of NCs under a short time period (～2 min) of near-infrared (808nm) irradiation, we kept the culture medium temperature to 41 °C for 20 minutes with continuous irradiation. The heated NCs exhibited efficient photothermal effects and resulted in an excellent inhibition of H. pylori growth, adhesion ability and cell vacuolization ability in in vitro investigation. Transmission electron microscopy showed a dramatic morphologic change after NCs photothermia on H. pylori, including cell wall and membrane rupture, as well as ribosome damage. Besides, levofloxacin and clarithromycin resistance were improved after photothermal treatment in H. pylori NCTC 11637 and/or clinical strains, however metronidazole resistance was unchanged. We also discovered a significant decrease in the biofilm formation of H. pylori under the NCs-based photothermal application, while efflux pump function was unchanged.Conclusions: Based on this novel NCs-based photothermal approach, we were able to demonstrate in vitro a significant inhibition of both H. pylori growth and molecular toxicity, and its improvement in antibiotic resistance alone with the eradication of H. pylori biofilms previously believed to be resistant to conventional antibiotics.

Detection of Nocardia by 16S Ribosomal RNA Gene PCR and Metagenomic Next-Generation Sequencing (mNGS)

In this study, the aim was to investigate the discriminatory power of molecular diagnostics based on mNGS and traditional 16S ribosomal RNA PCR among Nocardia species. A total of fourteen clinical isolates from patients with positive Nocardia cultures and clinical evidence were included between January 2017 and June 2020 in HeNan Provincial People’s Hospital. DNA extraction and 16S rRNA PCR were performed on positive cultures, and pathogens were detected by mNGS in these same samples directly. Among the 14 Nocardia isolates, four species were identified, and N. cyriacigeorgica (8 cases) is the most common species. Twelve of the 14 Nocardia spp. isolates were identified by the two methods, while two strains of N. cyriacigeorgica were not identified by mNGS. All tested isolates showed susceptibility to trimethoprim-sulfamethoxazole (SXT), amikacin and linezolid. Apart from Nocardia species, other pathogens such as Acinetobacter baumannii, Klebsiella pneumonia, Aspergillus, Enterococcus faecalis, Human herpesvirus, etc., were detected from the same clinical samples by mNGS. However, these different pathogens were considered as colonization or contamination. We found that it is essential to accurately identify species for determining antibiotic sensitivity and, consequently, choosing antibiotic treatment. 16S rRNA PCR was useful for identification of nocardial infection among species, while this technique needs the clinicians to make the pre-considerations of nocardiosis. However, mNGS may be a putative tool for rapid and accurate detection and identification of Nocardia, beneficial for applications of antimicrobial drugs and timely adjustments of medication.

A Wake-Up Call: Determination of Antibiotics Residue Level in Raw Meat in Abattoir and Selected Slaughterhouses in Five Local Government in Kano State, Nigeria.

The frequent use of antibiotics may result in drug residues that can be discovered at varying quantities in animal products such as milk or meat. The presence of pharmaceutical residues in food above the MRLs has been perceived globally by various persons. Antibiotics residues are present in food, which can endanger human health by causing antibiotic sensitivity, allergic reactions, microflora imbalance, bacterial resistance to antibiotics in microorganisms, and financial loss to the food industry. Farmers around the world utilize them on a sporadic basis for both preventative and curative purposes. This study assessed the antibiotics residues in raw meat sold in 6 slaughter houses in Kano States. The study is a descriptive cross-sectional study involving six (6) major slaughter house in Kano state. Muscle, Kidney and liver samples were collected from each slaughterhouse. The antibiotic residues in the meat samples were analysed using high performance liquid chromatography (HPLC) for tetracycline, ciprofloxacin and oxytetracycline residue results were presented in charts and tables. Out of a total of 18 beef samples analyzed during this study, 15 (83%) of the total samples had detectable levels of tetracycline residues from which 6(33.3%) had tetracycline residues at violative levels above the WHO/FAO maximum residue limits (MRLs), out of those 18 beef samples analyzed during this study, 6(33%) of the total samples had detectable levels of oxytetracycline residues from which 3(17%) had oxytetracycline residues at violative levels above the WHO/FAO maximum residue limits (MRLs) and out of those 18 beef samples analyzed during this study, 12(67%) of the total samples had detectable levels of ciprofloxacin, all levels are below the WHO/FAO maximum residue limits (MRLs). This high level of tetracycline and oxytetracycline residues in greater proportion of meat destined for human consumption at violative levels could be as a result of the indiscriminate use and misuse of veterinary drugs as commonly practiced among livestock producers and marketers without observing withdrawal period prior to slaughter. These results indicate that consumers may be predisposed to health hazards and hinder international meat trade from Nigeria. Regulatory authorities should therefore ensure compliance with good agricultural practices including withdrawal period of drugs used for treatment of food animals.

Systematic whole-genome sequencing reveals an unexpected diversity among actinomycetoma pathogens and provides insights into their antibacterial susceptibilities

Mycetoma is a neglected tropical chronic granulomatous inflammatory disease of the skin and subcutaneous tissues. More than 70 species with a broad taxonomic diversity have been implicated as agents of mycetoma. Understanding the full range of causative organisms and their antibiotic sensitivity profiles are essential for the appropriate treatment of infections. The present study focuses on the analysis of full genome sequences and antibiotic resistance profiles of actinomycetoma strains from patients seen at the Mycetoma Research Centre in Sudan with a view to developing rapid diagnostic tests. Seventeen pathogenic isolates obtained by surgical biopsies were sequenced using MinION and Illumina methods, and their antibiotic resistance profiles determined. The results highlight an unexpected diversity of actinomycetoma causing pathogens, including three Streptomyces isolates assigned to species not previously associated with human actinomycetoma and one new Streptomyces species. Thus, current approaches for clinical and histopathological classification of mycetoma may need to be updated. The standard treatment for actinomycetoma is a combination of sulfamethoxazole/trimethoprim and amoxicillin/clavulanic acid. Most tested isolates were not susceptible to sulfamethoxazole/trimethoprim or to amoxicillin alone. However, the addition of the β-lactamase inhibitor clavulanic acid to amoxicillin increased susceptibility, particularly for Streptomyces somaliensis and Streptomyces sudanensis . Actinomadura madurae isolates appear to be particularly resistant under laboratory conditions, suggesting that alternative agents, such as amikacin, should be considered for more effective treatment. The results obtained will inform future diagnostic methods for the identification of actinomycetoma and treatment.

Resistansi Antibiotik Secara Fenotip Dan Deteksi Gen TetA pada Sampel Hati Ayam di Pasar Dukuh Kupang Surabaya

The purpose of this study was to determine the resistance of several antibiotics phenotypically and genotypically to detect the tetA gene from broiler chicken liver samples at Dukuh Kupang market, Surabaya. A total of 30 samples were taken and then prepared aseptically and sterile. Isolation on Eosin Methylene Blue Agar (EMBA) media, then microscopic examination using gram staining and biochemical tests of Triple Sugar Iron Agar (TSIA), Sulfide Indole Motility (SIM), Methyl Red (MR), Voges Prouskauers (VP) and Simons Citrate Agar (SCA). The identified Escherichia coli colonies were tested for antibiotic sensitivity using the Kirby Bauer method, then isolates that were proven to be resistant to tetracycline antibiotics were followed by genetic testing using the Polymerase Chain Reaction (PCR) method. The results showed that 20 of the 30 samples were positive for Escherichia coli. Escherichia coli isolates from chicken liver samples showed resistance to 30 µg tetracycline antibiotics by 85% (17 of 20 samples) Researchers also compared with other antibiotics, the highest resistance to ampicillin 10 µg was 90% (18 out of 20 samples), gentamicin resistance was 10 µg by 50% (10 of 20 samples) and 30 µg chloramphenicol antibiotic resistance by 30% (6 of 20 samples). The isolates that were resistant to tetracycline were confirmed by Polymerase Chain Reaction to detect the tetA gene with the final product in the form of a band with a length of 210 bp. Bacterial isolates resistant to Tetracycline antibiotics did not always show TetA gene expression in the PCR test. Keywords: Antibiotic Resistance; Escherichia coli; Market; TetA gene Abstrak Tujuan dari penelitian ini yaitu untuk mengetahui resistansi beberapa antibiotik secara fenotip dan secara genotip mendeteksi gen tetA dari sampel hati ayam broiler di pasar Dukuh Kupang Surabaya. Sebanyak 30 sampel diambil kemudian dipreparasi secara aseptis dan steril. Isolasi pada media Eosin Methilen Blue Agar (EMBA), selanjutnya dilakukan pemeriksaan mikroskopis menggunakan pewarnaan gram dan uji biokimiawi Triple Sugar Iron Agar (TSIA), Sulfide Indol Motility (SIM), Methyl Red (MR), Voges Prouskauers (VP), dan Simons Citrat Agar (SCA). Koloni Escherichia coli yang teridentifikasi dilakukan uji sensitifitas antibiotik dengan metode Kirby bauer, selanjutnya isolat yang terbukti resistan terhadap antibiotik tetrasiklin dilanjutkan pemeriksaan genetik dengan metode Polymerase Chain Reaction (PCR). Hasil penelitian menunjukkan bahwa 20 dari 30 sampel positif Escherichia coli. Isolat Escherichia coli asal sampel hati ayam menunjukkan resistansi terhadap antibiotik Tetrasiklin 30 µg sebesar 85 % (17 dari 20 sampel) Peneliti juga melakukan perbandingan dengan antibiotik lainnya, resistensi tertinggi pada antibiotik ampisilin 10 µg sebesar 90 % (18 dari 20 sampel), resistensi gentamisin 10 µg sebesar 50 % (10 dari 20 sampel) dan resistensi antibiotik kloramfenikol 30 µg sebesar 30 % (6 dari 20 sampel). Isolat yang resisten terhadap tetrasiklin dikonfirmasi dengan Polymerase Chain Reaction untuk mendeteksi gen tetA dengan produk akhir berupa band dengan panjang 210 bp. Isolat bakteri yang resistan terhadap antibiotik Tetrasiklin tidak selalu menunjukkan ekspresi gen tetA pada uji PCR. Kata kunci: Escherichia coli; Gen TetA; Pasar; Resistansi Antibiotik.

Correlation of biofilm production with antibiotic susceptibility pattern of Pseudomonas aeruginosa from various clinical specimens

Background: Pseudomonas aeruginosa is one of the most prevalent nosocomial pathogens that cause a life-threatening infection. One of the important characteristics of P. aeruginosa is biofilm formation which leads to antibiotic resistance. Aims and Objectives: The aim of the study was to study the antibiotic resistance pattern of P. aeruginosa isolates and correlation with their biofilm-production. Materials and Methods: A total of 87 P. aeruginosa isolates from different clinical specimens were processed and confirmed by conventional microbiological methods as per standard methodology. Antibiotic sensitivity testing was done for all isolates. Biofilm producing isolates were identified by the microtiter plate method (MTPM). Results: Of 87 P. aeruginosa isolates, majority were from pus 33 (38%), followed by urine 26 (30%), sputum 19 (22%), body fluids 7 (8%), and blood 2 (2%). Biofilm producing isolates showed more resistance in comparison to non-biofilm producers. The observed difference between biofilm formation for multidrug resistant and susceptible isolates was found to be statistically significant. Conclusion: MTPM method was an effective test for detection of biofilm formation and was also able to verify biofilm production by P. aeruginosa. This indicated a higher propensity among the clinical isolates of P. aeruginosa to form biofilm and revealed a positive correlation between biofilm formation and antibiotic resistance. This indicates the need for testing of even susceptible isolates for virulence factors such as biofilm production.

Tertiary hospital sewage as reservoir of bacteria expressing MDR phenotype in Brazil

High doses of antibiotics used in hospitals can affect the microbial composition of sewers, selecting resistant bacteria. In this sense, we evaluated the antibiotic resistance profile and the multiresistant phenotype of bacteria isolated in sewage from a tertiary hospital in the interior São Paulo state, Brazil. For bacteria isolation, 10 µL of sewage samples were sown in selective culture media and the isolates were identified using VITEK-2 automatized system. The antibiotic sensitivity test was performed by disk diffusion. High percentages of resistance were found for amoxicillin, ampicillin, ceftazidime, clindamycin, vancomycin and the multidrug-resistant phenotype (MDR) was attributed to 60.7% of the isolates. Our results show bacteria classified as critical/high priority by WHO List of Priority Pathogens (Enterococcus and Staphylococcus aureus resistant to vancomycin and Enterobacteriaceae resistant to carbapenems) in hospital sewage. Therefore, the implementation of disinfection technologies for hospital sewage would reduce the bacterial load in the sewage that will reach urban wastewater treatment plants, minimizing superficial water contamination and bacterial resistance spread in the environment.

Bacteriological profile, antibiotic susceptibility pattern and other factors related to neonatal meningitis: A cross-sectional hospital-based study from West Bengal

Background: Neonatal sepsis and meningitis is an important cause of neonatal mortality and morbidity especially in the developing countries. Bacteriological profile of meningitis and antibiotic sensitivity pattern may vary from one region to another. Aims and Objectives: We have planned this study to know the etiological agent of neonatal meningitis with its antibiotic sensitivity profile and to evaluate some other associated risk factors of meningitis. Materials and Methods: This observational, cross-sectional study was done for a period of 1 year in the SNCU and NICU of a district Medical College of West Bengal in neonates presented with clinical sepsis and meningitis. Sepsis screen, blood culture, cerebrospinal fluid (CSF) study, and culture sensitivity was done and recorded along with demographic data, clinical presentation, outcome, and other associated factors. Results: We found meningitis in 55 neonates out of 250 clinical sepsis. CSF culture was positive in 42 cases with Escherichia coli (30.9%), Klebsiella (26.1%), Staphylococcus aureus (16.6%), Acinetobacter (14.2%) and Coagulase negative Staphylococcus (CoNS 11.9%) as prevalent organism. E. coli and Klebsiella were mostly sensitive to Amikacin, Levofloxacin, and Colistin whereas less sensitive to Cefotaxime, Pipercilin-tazobactam or Meropenem and Acinetobacter showed good sensitivity only to Levofloxacin. Among the gram-positive organism, S. aureus and CoNS were only sensitive to Linezolid, Vancomycin, and Teicoplanin. Conclusion: This type of study should help to make a proper antibiotic policy for an institution so that the empirical first-line antibiotic can be started with good effect in cases of neonatal sepsis and meningitis before the arrival of culture sensitivity report.