Milk-Compositional Study of Metabolites and Pathogens in the Milk of Bovine Animals Affected with Subclinical Mastitis.

Bovine milk is an important food component in the human diet due to its nutrient-rich metabolites. However, bovine subclinical mastitis alters the composition and quality of milk. In present study, California mastitis testing, somatic cell count, pH, and electrical conductivity were used as confirmatory tests to detect subclinical mastitis. The primary goal was to study metabolome and identify major pathogens in cows with subclinical mastitis. In this study, 29 metabolites were detected in milk using gas chromatography−mass spectrometry. Volatile acidic compounds, such as hexanoic acid, hexadecanoic acid, lauric acid, octanoic acid, n-decanoic acid, tricosanoic acid, tetradecanoic acid, and hypogeic acid were found in milk samples, and these impart good flavor to the milk. Metaboanalyst tool was used for metabolic pathway analysis and principal component estimation. In this study, EC and pH values in milk were significantly increased (p < 0.0001), whereas fat (p < 0.04) and protein (p < 0.0002) significantly decreased in animals with subclinical mastitis in comparison to healthy animals. Staphylococcus aureus was the predominant pathogen found (n = 54), followed by Escherichia coli (n = 30). Furthermore, antibiotic sensitivity revealed that Staphylococcus aureus was more sensitive to gentamicin (79.6%), whereas Escherichia coli showed more sensitivity to doxycycline hydrochloride (80%).

Prevalence and antibiotic susceptibility of Uropathogens from cases of urinary tract infections (UTI) in Shashemene referral hospital, Ethiopia.

BACKGROUND: Urinary tract infection (UTI) remains to be one of the most common infectious diseases diagnosed in developing countries. And a widespread use of antibiotics against uropathogens has led to the emergence of antibiotic resistant species. A laboratory based cross-sectional survey was conducted in Shashemene referral hospital to determine the prevalence and antibiotic susceptibility of uropathogens. METHODS: We have collected 384 clean catch mid-stream urine samples from all suspected UTI outpatients using sterile screw capped container. The urine samples were cultured and processed for subsequent uropathogens isolation. The isolated pure cultures were grown on BiOLOG Universal Growth agar (BUG) and identified using GEN III OmniLog® Plus ID System identification protocols. The identified species were then exposed to selected antibiotics to test for their susceptibility. RESULTS: The overall prevalence of urinary tract infection in the area was 90.1%. Most frequently isolated uropathogen in our study was Escherichia coli (39.3%). While, Staphylococcus species (20.2%), Leuconostoc species (11.4%), Raoultella terrigena/Klebsiella spp./ (8.4%), Salmonella typhimurium (6.3%), Dermacoccus nishinomiyaensis (6.3%), Citerobacter freundii (5.2%) and Issatchenkia orientalis/Candida krusei/ (2.7%) were the other isolates. We find that the relationship between uropathogens and some of UTI risk factors was statistically significant (P < 0.05). Gentamicin was the most effective drug against most of the isolates followed by chloramphenicol and nitrofurantoin. In contrast, amoxicillin, vancomycin and cephalexin were the antibiotics to which most of the isolates developed resistance. CONCLUSION: Urinary tract infection was highly prevalent in the study area and all uropathogens isolated developed a resistance against mostly used antibiotics.

Bacteria in the apical root canals of teeth with apical periodontitis.

BACKGROUND/PURPOSE: Bacteria in the tooth root canal may cause apical periodontitis. This study examined the bacterial species present in the apical root canal of teeth with apical periodontitis. Antibiotic sensitivity tests were performed to evaluate whether these identified bacterial species were susceptible to specific kinds of antibiotics. METHODS: Selective media plating and biochemical tests were used first to detect the bacterial species in samples taken from the apical portion of root canals of 62 teeth with apical periodontitis. The isolated bacterial species were further confirmed by matrix-assisted laser desorption ionization-time of flight mass spectrometry. RESULTS: We found concomitant presence of two (32 teeth) or three species (18 teeth) of bacteria in 50 (80.6%) out of 62 tested teeth. However, only 34 bacterial species were identified. Of a total of 118 bacterial isolates (83 anaerobes and 35 aerobes), Prophyromonas endodontalis was detected in 10; Bacteroides, Dialister invisus or Fusobacterium nucleatum in 9; Treponema denticola or Enterococcus faecalis in 8; Peptostreptococcus or Olsenella uli in 6; and Veillonella in 5 teeth. The other 25 bacterial species were detected in fewer than five teeth. Approximately 80-95% of bacterial isolates of anaerobes were sensitive to ampicillin/sulbactam (Unasyn), amoxicillin/clavulanate (Augmentin), cefoxitin, and clindamycin. For E. faecalis, 85-90% of bacterial isolates were sensitive to gentamicin and linezolid. CONCLUSION: Root canal infections are usually caused by a mixture of two or three species of bacteria. Specific kinds of antibiotic can be selected to control these bacterial infections after antibiotic sensitivity testing.

Bacterial keratitis in a tertiary hospital in São Paulo: a 21-year review of the epidemiological, laboratory, and clinical data.

Infectious keratitis is a sight-threatening condition that is usually an ocular emergency. The visual outcome depends on prompt and accurate clinical management as well as geographic and epidemiological awareness. We conducted a retrospective observational study to define the epidemiological and laboratory profile, as well as the clinical course of bacterial keratitis in a tertiary hospital in São Paulo over 21 years. Information about age, sex, predisposing factors, topical and surgical treatment, visual acuity, ulcers' classification, bacterioscopy, culture, and antibiotic sensitivity tests were collected. This study included 160 patients. The mean age was 65.1 ± 18.4 years and risk factors were identified in 83.1 % of the patients. Empirical topical fortified cephalosporin with an aminoglycoside or fourth-generation fluoroquinolone was curative for 66.2 % of the cases. The mean treatment duration was 22.5 ± 9 days. The mean variation of visual acuity was -0.25 logMAR, p < 0.001. Culture revealed 64 % of Gram-positive bacteria. All Gram-positive bacteria were sensitive to cephalothin, vancomycin, and quinolones. All Gram-negative bacteria were sensitive to gentamicin, tobramycin, amikacin, and ciprofloxacin. These findings reinforce the importance of prompt empirical treatment of severe corneal ulcers with a fortified cephalosporin and aminoglycoside or a fourth-generation fluoroquinolone as there are equally effective. Collected data was insufficient to evaluate resistance of ocular infections over time in this population.

Identification of Virulence Markers and Phylogenetic Groups' Association, and Antimicrobial Susceptibility of Uropathogenic Escherichia coli Isolates.

BACKGROUND: Urinary tract infections represent a world public health problem, which is caused mainly by Uropathogenic Escherichia coli. Although they are originally found in the intestinal microbiota in the majority of the cases, urinary tract infections can also be caused by intra-intestinal pathogenic E. coli. OBJECTIVE: The main objective of our research is to identify the virulence factors generally associated with different pathotypes across phylogenetic groups. METHODS: E. coli were isolated from patients with urinary tract infections. Antimicrobial susceptibility tests, virulence genes and phylogroups were prospected. The data analysis were performed using the chi-square and Fisher exact test. RESULTS: In total, 72.2% of isolates showed multidrug resistant. We have also depicted an important association between E. coli from inpatients with UTIs and pap and hlyA genes (p-0.041 and p-0.019 respectively). The predominant phylogenetic group in our isolates is B2 (45.4%) followed by D (12.4%). Our results showed that 9.3% of isolates have an unknown phylogroup which shows a significant association with astA gene (p-0.008). We have as well found a significant association between B2 and three virulence genes namely pap, hlyA and invE (p-0.002, p-0.001, p-0.025 respectively); B1 and pap, hlyA genes (p-0.049 and p-0.021 respectively); E and afa gene (p-0.024). CONCLUSION: Certain virulence factors have been shown to be potential targets for drug design and therapeutic pathways in order to deal with the antimicrobial resistance problem enhanced by antibiotic therapy.

Six-year analysis of key monitoring for bacterial strain distribution and antibiotic sensitivity in a hospital.

BACKGROUND: With the widespread use of antimicrobial drugs, bacterial resistance has become a significant problem, posing a serious threat to public health. The prevalence of clinical infection strains in hospitals and their drug sensitivities are key to the appropriate use of antibiotics in clinical practice. AIM: To identify prevalent bacteria and their antibiotic resistance profiles in a hospital setting, thereby guiding effective antibiotic usage by clinicians. METHODS: Specimens from across the institution were collected by the microbiology laboratory. The VITEK 2 compact fully automatic analyzer was used for bacterial identification and antibiotic sensitivity testing, and the WHONET5.6 software was utilized for statistical analysis. RESULTS: A total of 12062 bacterial strains of key monitoring significance were detected. Staphylococcus aureus demonstrated widespread resistance to penicillin, but none of the strains were resistant to vancomycin or linezolid. Moreover, 219 strains of methicillin-resistant coagulase-negative staphylococci and 110 strains of methicillin-resistant Staphylococcus aureus were detected. Enterococcus faecalis showed moderate resistance to the third-generation quinolones ciprofloxacin and levofloxacin, but its resistance to nitrofurantoin and tetracycline was low. Enterococcus faecium displayed significantly lower resistance to third- and fourth-generation quinolones than Enterococcus faecalis. The resistance of two key monitoring strains, Escherichia coli and Klebsiella pneumoniae, to piperacillin/tazobactam was 5%-8%. However, none of the Escherichia coli and Klebsiella pneumoniae strains were resistant to meropenem. The resistance of Acinetobacter baumannii to piperacillin/sulbactam was nearly 90%. Nonetheless, the resistance to tigecycline was low, and Pseudomonas aeruginosa demonstrated minimal resistance in the antibiotic sensitivity test, maintaining a resistance of < 10% to the cephalosporin antibiotics cefotetan and cefoperazone over the last 6 years. The resistance to amikacin remained at 0.2% over the past 3 years. CONCLUSION: Our hospital's overall antibiotic resistance rate was relatively stable from 2017 to 2022. The detection rates of key monitoring strains are reported quarterly and their resistance dynamics are monitored and communicated to the entire hospital, which can guide clinical antibiotic selection.

Understanding Knowledge and Attitude of Farmers towards Antibiotic Use and Antimicrobial Resistance in Jhunjhunu District, Rajasthan India.

The misuse of antibiotics in veterinary practices by farmers is harming livestock production and food safety and leading to the rise of antibiotic resistance (AMR). This can also transfer resistant bacteria from animals to humans, posing a serious public health threat. However, we have not paid enough attention to understanding how farmers behave in this regard. Our study aims to explore farmers' behaviors and identify the factors that influence their choices. To conduct this study, we used a questionnaire with 40 questions and surveyed 208 farmers in Jhunjhunu district, Rajasthan. We analyzed the data using SPSS. Here are the key findings: About 58.3% of the farmers have some awareness of antibiotics, and 49.5% are aware of antimicrobial resistance (AMR). Notably, as the level of education increases, so does awareness of antibiotics. Unfortunately, 63.9% of the farmers are not aware of the withdrawal time, and 64% have no idea about the presence of antibiotic residues during this period. Around 75% of farmers vaccinate their animals, but approximately 56.9% of individuals have never undergone an antibiotic sensitivity test (ABST) for milk. Around 48.6% of farmers are unaware of government testing centers. Several factors hinder farmers from implementing proper animal management practices, such as the high fees of veterinarians. When their animals become sick, their first choice is home remedies, followed by using old prescriptions. Additionally, 63.9% stop treatment once the animal looks better. A significant portion (83.8%) of farmers rely on local pharmacists for medicine. It has been determined that there is no significant correlation between education, experience, age, and the level of awareness concerning withdrawal periods, the existence of government antibiotic sensitivity test (ABST) centers, and entities responsible for sending samples for ABST. In our qualitative analysis, focus groups identified significant barriers to following best farm practices and spreading awareness about AMR. These findings suggest that addressing AMR in livestock requires a comprehensive approach. This should include targeted education and awareness programs for farmers, as well as improved access to veterinary services.

Rapid antimicrobial susceptibility profiling using impedance spectroscopy.

The present antibiotic susceptibility testing (AST) techniques based on bacterial culture, gene amplification and mass spectrometry are highly time consuming, labour intensive or expensive. Impedance spectroscopy is an emerging tool for rapid bacterial analysis as it is label-free, real-time, affordable and high-throughput. The over-reliance of this technique on complex chip designs and cell enrichment strategies has, however, slowed its foray into clinical AST. We demonstrate a label-free approach in which a low conductivity zwitterionic buffer is used for boosting impedance sensitivity in simple interdigitated electrodes (IDEs) allowing rapid AST in just 20 min without any liquid flow, biofunctionalization or cell enrichment steps. The detection principle relies on measuring changes in solution resistance due to antibiotic-induced bacterial cell death or growth. While the death-based approach is faster (20 min), it's restricted to surface-acting bactericidal antibiotics. The cell growth approach is longer (60-80 min) but more versatile as it applies to all drug types. Results for antibiotic sensitivity analysis and minimum inhibitory concentration (MIC) determination are illustrated for Escherichia coli, Klebsiella pneumoniae and Staphylococcus aureus against a wide class of antibiotics (penicillins, cephalosporins, polymyxins, carbapenems etc.).

Urinary Tract Infection and Antimicrobial Susceptibility of Bacterial Isolates in Saint Joseph Kitgum Hospital, Kitgum, Uganda.

A cross-sectional study of microorganisms isolated from mid-stream urine samples obtained from 139 patients with suspected urinary tract infection (UTI) who presented leukocyturia was conducted from April to June 2019 at Saint Joseph Kitgum Hospital (Uganda). All microorganisms were identified by MALDI-TOF mass spectrometry in a laboratory in Spain. Antimicrobial susceptibility was determined on site using the disc diffusion method (Kirby-Bauer test) and these results were subsequently compared with those obtained in Spain using the Becton Dickinson Phoenix M50 device. The overall prevalence of UTI with bacterial growth was 64.0% (n = 89) (95% CI, 56.1-72.0), and 11 presented mixed infection. As a result, 100 microorganisms were isolated. The most common uropathogens were Enterococcus spp. (57%) and Escherichia coli (28%). Nitrofurantoin was the most effective drug (81.7% in Gram-positive and 87.3% in Gram-negative bacteria), followed by imipenem (94.2% and 74.5%, respectively). The highest resistance rates were observed for amoxicillin and ciprofloxacin (66.2% and 44.6%, respectively). Given the increasing trend toward antibiotic resistance, there is a need for bacteriological cultures and continuous surveillance of uropathogen antibiotic susceptibility. Use of amoxicillin and ciprofloxacin as empirical treatments for UTIs should be discontinued in Uganda. The findings of this study may be useful for clinicians, as they may improve empirical treatment.

Bacterial keratitis in a tertiary hospital in São Paulo: a 21-year review of the epidemiological, laboratory, and clinical data

ABSTRACT Infectious keratitis is a sight-threatening condition that is usually an ocular emergency. The visual outcome depends on prompt and accurate clinical management as well as geographic and epidemiological awareness. We conducted a retrospective observational study to define the epidemiological and laboratory profile, as well as the clinical course of bacterial keratitis in a tertiary hospital in São Paulo over 21 years. Information about age, sex, predisposing factors, topical and surgical treatment, visual acuity, ulcers' classification, bacterioscopy, culture, and antibiotic sensitivity tests were collected. This study included 160 patients. The mean age was 65.1 ± 18.4 years and risk factors were identified in 83.1 % of the patients. Empirical topical fortified cephalosporin with an aminoglycoside or fourth-generation fluoroquinolone was curative for 66.2 % of the cases. The mean treatment duration was 22.5 ± 9 days. The mean variation of visual acuity was −0.25 logMAR, p < 0.001. Culture revealed 64 % of Gram-positive bacteria. All Gram-positive bacteria were sensitive to cephalothin, vancomycin, and quinolones. All Gram-negative bacteria were sensitive to gentamicin, tobramycin, amikacin, and ciprofloxacin. These findings reinforce the importance of prompt empirical treatment of severe corneal ulcers with a fortified cephalosporin and aminoglycoside or a fourth-generation fluoroquinolone as there are equally effective. Collected data was insufficient to evaluate resistance of ocular infections over time in this population.

Isolation, identification, and serotyping of Avibacterium paragallinarum from quails in Indonesia with typical infectious coryza disease symptoms.

BACKGROUND AND AIM: Infectious coryza (IC) or snot is an infectious upper respiratory disease affecting chickens and birds, including quails, and it is caused by Avibacterium paragallinarum. The symptoms of IC are facial swelling, malodorous nasal discharge, and lacrimation. This study aimed to isolate, identify, and serotype the A. paragallinarum of snot in quails and to determine the sensitivity and resistance to several antibiotics. MATERIALS AND METHODS: Nine quails from Yogyakarta, Indonesia with typical snot disease symptoms were used in this study. The nasal swab was obtained and directly streaked onto a chocolate agar plate and blood agar plate (BAP), then incubated in 5% CO2 at 37°C for 24-48 h. Staphylococcus spp. was cross-streaked onto the BAP to show the satellite growth. The observation of the morphology of the suspected colony, Gram staining, and biochemical tests (catalase test, oxidase test, urease test, peptone test, and carbohydrate fermentation such as maltose, mannitol, lactose, and sorbitol) are done to identify the species of bacteria. This research also detects the serovar of A. paragallinarum using hemagglutination inhibition test.The antibiotic sensitivity tests were also performed using several antibiotics against five A. paragallinarum isolates that were cultured on Mueller-Hinton Agar and added with antibiotic discs, then incubated in 5% CO2 at 37°C for 24-48 h. RESULTS: Five isolates out of nine suspected isolates (55.5%) were A. paragallinarum. The growth of isolates from quails did not depend on the nicotinamide adenine dinucleotide (NAD) (NAD-independent). Sensitivity test was done using the five identified A. paragallinarum isolates, results showed that they were 100% sensitive to amoxicillin (AMC) and ampicillin (AMP); 100% resistant toward amikacin (AK), erythromycin (E), gentamycin (CN), and tetracycline (TE); 80% resistant toward kanamycin (K) and trimethoprim (W); 60% resistant toward chloramphenicol (C); and 20% toward enrofloxacin (ENR). The antibiotics that have an intermediate sensitivity (in between sensitive and resistant) were ENR and K, 80% and 20%, respectively. Three out of five A. paragallinarum isolates were identified as serovar B of A. paragallinarum using HI test. CONCLUSION: Five out of nine isolates (55.5%) from quails with typical IC disease symptoms identified as A. paragallinarum and sensitive toward AMC and AMP. Three out of five A. paragallinarum isolates were identified as serovar B.

16S Ribosomal RNA Gene Sequencing to Evaluate the Effects of 6 Commonly Prescribed Antibiotics.

INTRODUCTION: The rapid antibiotic sensitivity test (RAST) is a novel in-office culture and sensitivity system for endodontic infections. The purpose of this research was to validate the RAST system as a viable, in-office alternative to antibiotic sensitivity testing using turbidity to determine antibiotic sensitivities of endodontic infections. METHODS: Aspirates were taken from the root canals of 9 necrotic human teeth at the initiation of root canal therapy. These samples were cultured in the RAST medium, and antibiotic sensitivity to 6 antibiotics was tested. Further analysis was performed using 16S ribosomal RNA (rRNA) gene sequencing. RESULTS: Thirty-one bacterial phyla were identified as well as 2 phyla of the kingdom Archaea. Augmentin (Dr. Reddy's Laboratories Ltd, Hyderabad, India) and ampicillin performed identically at 24 hours, inhibiting turbidity in 100% of the samples. At 48 hours in anaerobic conditions, Augmentin outperformed ampicillin by 13%. Ciprofloxacin was the least efficacious antibiotic. At 48 hours, only 22% of anaerobic ciprofloxacin cultures affectively inhibited bacterial growth. CONCLUSIONS: The RAST medium is a viable in-office alternative to antibiotic susceptibility testing in an off-site laboratory. It is able to support the growth of a wide variety of microorganisms in both aerobic and anaerobic environments, and, in combination with 16S rRNA gene sequencing, it led to the identification of a new archaebacterial phylum, Crenarchaeota, as part of the endodontic infection microbiome.

Prevalence of enteropathogens and their antibiotic sensitivity pattern in puppies with hemorrhagic gastroenteritis.

AIM: Hemorrhagic gastroenteritis (HGE) ranging from mild to severe forms is commonly encountered in puppies. The aim of the study was to identify the prevalence of common enteropathogens and the antibiotic sensitivity pattern in puppies reported with HGE. MATERIALS AND METHODS: The canine HGE activity index, with little modification, was adopted to identify Grade III/severely affected puppies below 6 months of age. Fecal polymerase chain reaction (PCR) assay was employed to screen and compare the enteropathogens in puppies with hemorrhagic diarrhea and healthy control. RESULTS: Canine parvovirus 2b was identified in 90.3% of the diarrheic and 10% of the non-diarrheic healthy puppies. Clostridium difficile was identified in all the diarrheic puppies and in 80% of the healthy puppies. Among the diarrheic puppies, 17.7% were positive for Clostridium perfringens enterotoxin, 9.7% were positive for C. perfringens alpha toxin, 6.4% were positive for Escherichia coli shiga toxin, 6.4% were positive for E. coli enterotoxin (LT), and 3.2% were positive for canine distemper virus. Whereas, none of the healthy puppies were positive for these bacteria and toxins. Fecal antibiotic sensitivity test pattern revealed gentamicin to be sensitive in 95% of the cases, azithromycin in 50%, enrofloxacin in 25%, cefotaxime in 20%, and tetracycline in 5% of the cases. CONCLUSION: Parvoviral enteritis is predominant among puppies. Yet, bacteria and their toxins also play an important role in HGE. Gentamicin has higher sensitivity against the enteropathogens associated with the condition.