Human Pus Research Articles

Resilient Staphylococcus aureus in Mastitis Milk: Insights into Antibiotic Resistance and Biofilm Formation

Background: Staphylococcus aureus is capable of producing a wide range of virulence factors. It is a deadly bacterium that is resistant to the majority of antibiotics that are typically provided. The ability of S. aureus to establish chronic, implant associated infections and our inability to cure them, is directly associated with its biofilm formation, creating an environment where bacteria can grow and persist while protected from the host immune response and antibiotic therapy. Antibiotic resistance of S. aureus is widely spreading with low cure rate. Methods: A total of 62 S. aureus strains were isolated from animal pus, human pus, animal skin, human skin, mastitic milk, normal milk and unprocessed meat samples. The isolates were genotypically confirmed by 23S rRNA ribotyping for S. aureus. Using 14 antibiotics belonging to different generations were tested for their resistance pattern from various sources. Result: All the mastitic milk, animal pus, human pus and unprocessed meat S. aureus isolates from all the places of sampling showed 100% sensitivity towards imipenem while isolates obtained from mastitic milk, human skin, animal skin showed 100% resistance towards ampicillin and ampicillin+sulbactum combination.

Characterization of the microorganisms of human pus samples- a tertiary care hospital based study

The term "pyogenic infection" describes an infectious agent-induced localized inflammation and pus production caused by a buildup of dead leukocytes. 120 patients' specimens were gathered and grown on blood agar and MacCokey agar. Biochemical tests were used to identify colonies grown in these conditions, and subsequently their antibiotic susceptibility was examined. When combined with the patient's medical history, these data showed how common pyogenic infections were in BIHS General hospital, Dhaka. Within 86.67% of the Positive pyogenic infection were due to bacterial agents, with Staphylococcus aureus (23.07%), Escherichia coli (15.39%), Klebsiella (15.39%), Proteus (15.39%), Pseudomonas (7.69%), Enterococci (7.69%), Citrobacter (7.69%), Enterobacter (7.69%) being the major pathogens. Female was predominant than male which was 72 (60%) cases and 48 (40%) cases respectively. The most common age group for both the male and female groups was 40–60 years old, with 16 (13.33%) and 64 (53.33%) instances, respectively. In conclusion, Staphylococcus aureus is the most often isolated bacterium following aerobic culture of pus. Nonetheless, there are fewer gram-positive cocci than gram-negative bacilli.

Phenotypic and Molecular Characterization of Pseudomonas aeruginosa Isolated from Human specimens

Pseudomonas aeruginosa (P. aeruginosa) is considered an opportunistic pathogen that causes high morbidity and mortality and is responsible for a high proportion of hospital infections pathogen that can cause several disease syndromes in the community and hospitals. Recently, P. aeruginosa infections having antibiotic resistance have been increasing and serious problems are being experienced in treatment. This study aimed to determine the prevalence, antibiogram pattern, and molecular identification of oprL virulence gene of P. aeruginosa recovered from human pus. One hundred samples from human pus were collected for bacteriological identification before being subjected to molecular identification. The results revealed that 13/100 (13%) were positive for P. aeruginosa on Citramide ager. All isolates showed high sensitivity to Imipenem, Apramycin, Amikacin, Norfloxacin, Sulphamethoxazol, with 100%, 92.31%, 84.61%, 9.23%, and 61.53% respectively. For the molecular characterization of P. aeruginosa (5 isolates) for detection of oprL virulence gene. The results indicated that oprL genes were the detected genes in all tested samples with (100%).In conclusion; the obtained results provide important data about the importance of the oprL virulence gene and pathogenicity of P. aeruginosa as well as the widespread of MDR strains that complicated the treatment options in human community

Historic nucleic acids isolated by Friedrich Miescher contain RNA besides DNA.

One hundred fifty years ago, Friedrich Miescher discovered DNA when he isolated "Nuclein"-as he named it-from nuclei of human pus cells. Miescher recognized his isolate as a new type of molecule equal in importance to proteins. He realised that it is an acid of large molecular weight and high phosphorus content. Subsequently, he discovered Nuclein also in the nuclei of other cell types, realised that it chemically defines the nucleus, and speculated on its role in proliferation, heredity and fertilisation. While now universally recognised as the discoverer of DNA, whether Miescher also discovered RNA has not yet been addressed. To determine whether his isolation also yielded RNA, we first reproduced his historic protocols. Our resulting modern Nuclein contained a significant percentage of RNA. Encouraged by this result, we then analysed a sample of Nuclein isolated by Miescher from salmon sperm. Assuming that the RNA present in this sample had degraded to nucleobases, we tested for the presence of uracil in the historic Nuclein. Detection of significant levels of uracil by LC-UV-MS demonstrates that Miescher isolated both forms of nucleic acid-DNA and RNA-and underlines the fundamental nature of his discovery for the field of molecular genetics.

Antibacterial activity of Indian propolis and its lead compounds against multi-drug resistant clinical isolates

IntroductionThe limited anti-infectious drugs and growing antibiotic resistance among pathogenic bacteria underscore the urgent need to explore novel antimicrobial agents, preferably from a natural source. Propolis is the potent natural antimicrobial agent, which produced by honeybees using various plant exudates. However, it has been minimally studied against multidrug-resistant (MDR) microorganisms. In the present study, the authors have investigated the antibacterial activity of ethanolic extracts of Indian melifera propolis (IMP) samples and combinations of their lead compounds against three human clinical isolates. MethodsThree multidrug-resistant microorganisms (MDRMs) were isolated from a human pus sample, and their molecular identification was carried out by 16S rRNA sequencing. The antibacterial activity of the IMP extracts and different combinations of chrysin, galangin, and phenethyl caffeate was determined by minimum inhibitory concentrations (MIC) using the 96-well plate microdilution method. ResultsAmongst, 19 IMP studied samples, IMP-5, IMP-14, and IMP-16 samples had the most potent antimicrobial activity against three MDR isolates. These samples had antimicrobial efficacy in the order of IMP16 > IMP14 > IMP5. The combinations of chrysin, galangin, and phenethyl caffeate had the lowest MIC values than individual components and above potent IMP extracts. ConclusionsCertain IMP samples and combinations of chrysin, galangin, and phenethyl caffeate could be the best natural therapeutic agents to control the pathogenicity of MDRMs.

Investigation on antibiotic resistance property of bacteria enumerated from human pus samples

This study was focused on evaluating the presence of commercial antibiotic resistant pathogenic bacteria present in the pus samples of patients from secondary care hospital Namakkal district of Tamil Nadu. About 494 pus samples from a total of 729 samples contained antibiotic resistant pathogenic bacteria. Among several bacterial species, the strains of Pseudomonas, Staphylococcus, Klebsiella, Escherchia, Enterococcus, and Micrococcus were found as the most predominant bacteria from pus samples. About 4 isolates showed resistance to more commercial antibiotics. These 4 strains were identified as P. aeroginosa CM1 to CM4. These strains showed resistance up to 98% for more number of commercial antibiotics than other strains. These strains were identified through molecular characterization (16S rRNA) and phylogenetic tree analysis. Fortunately, these 4 numbers of P. aeruginosa strains were susceptible to meropenem and amikacin antibiotics. These results declared that the pathogenic bacteria present in the soft tissue of wounds and others could create resistance to the commercial antibiotic, which is frequently administered to the patients.

Frequency and Antimicrobial Susceptibility Pattern of Pseudomonas Aeruginosa in Human Pus Samples at Holy Family Hospital Rawalpindi

Background: Nosocomial infections are great threat for hospitalized patients and Pseudomonas aeruginosa has emerged as one of the most potent nosocomial pathogens along with its diverse mechanisms to counter the various antimicrobial agents such as aminoglycosides, fluoroquinolones, monobactems, third generation cephalosporins, carbapenams and broadspectrum penicillins. P. aeruginosa is one of the well-known pyogenic bacteria and is 3rd leading cause of pyogenic infections with the variable frequency depending on geographical region and clinical setting. P. aeruginosa is intimately associated with pyogenic nosocomial infections. Objectives: Since multidrug resistant strains of P. aeruginosa have posed serious threats and are frequently implicated in nosocomial infections. Methods: Pus swab were sampled under aseptic conditions and cultured on blood and Muller Hinton agar. Gram reaction, pigment production, Oxidase, indole reaction and citrate test were used to confirm isolate. Antibiotic susceptibility was performed b Kirby Bauer technique. Results compiled by us in this cross sectional study, showed 58 cases of P. aeruginosa out of 289 cases. This included 43% males and 57% females. Majority of the patients were of young age, with mean age 38 years. Antibiotic sensitivity revealed resistance to gentamicin was 50%, amikacin was 64%, ciprofloxacin and Aztronem 66%, Cefaparazone 69%, Tzaocin 71% and meropenem and sulzone was 79%. While Colistin and Ceftazidime were the most effective in 85% and 89% of cases respectively. The multidrug resistant strains of P. aeruginosa infections accounted for 32.76% of total P. aeruginosa infections. This study reveals high prevalence of multidrug resistant organisms at the set of our study. Based on this study, we suggest adopting the strategies to minimize the risk of nosocomial infections to slow down the rapidly growing multidrug resistance. These strategies may include, stricter antiseptic measures, fastening the recovery process and reducing the hospital stay and considering other alternates. Besides this, we would like to suggest the precise use of antibiotic susceptibility facility to reduce the nosocomial infection associated complications.

Targeting polyelectrolyte networks in purulent body fluids to modulate bactericidal properties of some antibiotics.

The response of the human immune system to most bacterial infections results in accumulation of neutrophils at infection sites that release a significant quantity of DNA and F-actin. Both are negatively charged polyelectrolytes that can interact with positively charged host defense molecules such as cathelicidin-delivered LL-37 peptide or other cationic antibiotic agents. Evaluation of the ability of bacterial outgrowth (using luminescence measurements or counting colony-forming units) to form a biofilm (quantified by crystal violet staining) and analysis of the structure of DNA/F-actin network by optical microscopy in human pus samples treated with different antibiotics in combination with plasma gelsolin, DNAse 1, and/or poly-aspartic acid revealed that bactericidal activity of most tested antibacterial agents increases in the presence of DNA/F-actin depolymerizing factors.

Evaluation of small-subunit rRNA touchdown polymerase chain reaction for direct detection of Entamoeba histolytica in human pus samples from patients with amoebic liver abscess

Purpose: The aim of the present study was to evaluate the use of touchdown polymerase chain reaction (TD-PCR) for the detection of Entamoeba histolytica in liver pus samples obtained from patients with a clinical diagnosis of amoebic liver abscess (ALA) using small-subunit rRNA (SSU rRNA) as the target gene. Materials and Methods: Microscopic examination in vitro culture and serological test for the detection of E. histolytica in 67 pus samples obtained from ALA patients was performed. Molecular studies were carried out by both conventional PCR and TD-PCR targeting the SSU rRNA gene using the same sets of primers and the results were compared. Results: TD-PCR detected the presence of E. histolytica in 86.5% of the liver pus samples within 2.5 h as compared with 82.08% by conventional PCR within 3.5– 4 h. Conclusion: TD-PCR assay may serve as a relatively better detection method for E. histolytica over conventional PCR with respect to the turnaround time, increased sensitivity, specificity and yield.

Isolation of Carnobacterium sp. from a human blood culture

Carnobacterium species have been isolated from the environment and are not regarded as human pathogens, although they are known to cause disease in fish. Only two reports describing isolation of Carnobacterium species from human pus were found in the literature. We report what we believe to be the first isolation of Carnobacterium sp. from a human blood culture.

Inflammatory sites as a source of plasma neopterin: Measurement of high levels of neopterin and markers of oxidative stress in pus drained from human abscesses

Objectives Plasma neopterin is a clinical marker of inflammation. Interferon-γ triggers 7,8-dihydroneopterin and its oxidation product, neopterin, to be released from macrophages. 7,8-dihydroneopterin is a potent antioxidant which can protect macrophages from oxidative damage in vitro. This study examined whether 7,8-dihydroneopterin/neopterin levels reach sufficient concentrations in human pus to provide antioxidant activity and be the source of plasma neopterin. Design and methods Pus was removed by needle aspiration from 19 patients and examined for total neopterin, protein-bound DOPA, dityrosine, α-tocopherol, lipid oxidation and protein carbonyls. Results Total neopterin was detected between 50 nM and 1.2 μM, with an average concentration of 0.51 μM. Significant quantities of oxidized proteins and lipids were detected. α-Tocopherol concentrations positively correlate with total neopterin levels. Conclusions Total neopterin levels found in the pus was up to 100 times higher than that reported in plasma, suggesting plasma neopterin originates from inflammatory sites. The concentration of total neopterin suggests that 7,8-dihydroneopterin could act as an antioxidant during inflammation.

Zinc, Iron, Copper, Selenium, Lactoferrin, and Ferritin in Human Pus

Restriction of zinc and iron available for microbial growth in tissues are well-recognized host defense mechanisms. The present studies were performed to characterize some constituents of human pus that may affect these important host defenses. Zinc, iron, copper, calcium, and magnesium in pus were measured using an atomic absorption spectrophotometer; selenium was measured fluorometrically. Ferritin was measured with a fluorometric enzyme immunoassay, and lactoferrin was measured with a radial diffusion assay. The growth of Escherichia coli at 37 degrees C was measured in pus supernate adjusted to pH 5.5 or 7.4, in boiled supernate, or in supernate adjusted with 1.3 mM iron or 0.9 mM zinc singly or together. Zinc and iron concentrations in pus exceeded normal serum. Calcium and magnesium levels were 2- to 3-fold lower and higher, respectively, than normal serum values. Lactoferrin concentrations of were 880 +/- 48 microg/mL and ferritin levels were 20,726 +/- 2,667 ng/mL. Growth of an E coli strain was inhibited in pus at pH 5.5 but not at pH 7.4, and growth was enhanced by addition of iron or zinc to E coli suspended in pus at pH 6.7. To our knowledge, this is the first report of the zinc, iron, copper, selenium, lactoferrin, and ferritin levels of human pus. These studies provide additional insight into host defense mechanisms mediated by the restriction of the bioavailability of zinc and iron in suppurative infection.

Hyaluronidase activity in human pus from which Streptococcus intermedius was isolated.

Hyaluronidase (HAase) activity was detected in both a human pus sample and the culture supernatant of the only bacterial isolate from the pus, Streptococcus intermedius, using a zymographic technique. The optimum pH range for HAase activity was similar for both samples. Although the bands showing the strongest HAase activity of these samples differed from each other with respect to molecular size, both samples were equally inhibited by an antiserum raised against HAase of S. intermedius. These results suggest that S. intermedius may produce HAase in vivo as well as in vitro, and that this enzyme and/or its fragments may play an important role in host tissue degradation.

Comparison of human polymorphonuclear leukocytes from peripheral blood and purulent exudates by high resolution 1H MRS

Human pus samples from various sites, including soft tissue and pleural cavity, as well as sputum from patients with cystic fibrosis have been analyzed by 1H magnetic resonance spectroscopy (MRS) and found to generate high resolution spectra. Assignments have been made for neutral lipid, amino acids, taurine, and lactate. Human peripheral blood polymorphonuclear leukocytes have also been examined and a similar spectrum containing these lipid and metabolite resonances was detected in cells stimulated with lipopolysaccharide. In unstimulated cells, only resonances from taurine and lactate were observed, suggesting that the presence of the lipid and metabolite resonances is an indicator of cellular activation of polymorphonuclear leukocytes. This is the first report of 1H MRS applied to intact human polymorphonuclear leukocytes and extends available documentation as to which types of normal and/or diseased tissue contain MR visible molecules.

Effect of the abscess environment on the antimicrobial activity of ciprofloxacin

The present studies were conducted to identify factors in human purulent material that might limit or enhance the activity of ciprofloxacin against bacteria causing suppurative infection. Ciprofloxacin, imipenem, and ampicillin were tested with regard to binding or inactivation by pus. The bactericidal activity of ciprofloxacin and imipenem were tested against Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, or Staphylococcus aureus in human pus with a pH of 6.0 incubated at 37°C under aerobic or anaerobic conditions. The effect of single or combination drug therapy with 20 mg/kg of ciprofloxacin, imipenem, or rifampin given every 12 hours was tested against E. coli or P. aeruginosa in polymicrobic murine abscesses that had been produced by subcutaneous injection of either of those organisms mixed with Bacteroides fragilis and autoclaved human stool. Antibiotic levels and the number of bacteria surviving in pus were quantitated. Therapy of subcutaneous abscesses was delayed 72 hours to test drug efficacy against organisms in well-established infections. Levels of ampicillin, imipenem, or ciprofloxacin were reduced from 10 μg/ml to 3.1 ± 4.0, 2.7 ± 3, or 5.8 ± 2 μg/ml, respectively, after incubation in eight pus specimens for 24 hours at 37°C. Ampicillin levels were reduced to less than 1 μg/ml in four pus specimens containing beta-lactamase. Imipenem levels were undetectable in two specimens and were 0.2 μg/ml in one specimen. Ciprofloxacin binding to pus supernate or sediment appeared to be explained by its binding to the deoxyribonucleic acid (DNA) present in pus. Activity of 5 μg/ml of ciprofloxacin against four E. coli or K. pneumoniae strains in pus in vitro was greater than that of twofold higher concentrations of imipenem. The bactericidal activity of ciprofloxacin and imipenem were comparable but substantially reduced against S. aureus and P. aeruginosa in pus. Ciprofloxacin alone or regimens combining ciprofloxacin with rifampin or rifampin plus imipenem reduced the number of E. coli in polymicrobic subcutaneous abscesses but had little effect on P. aeruginosa in polymicrobic abscesses. The anaerobic abscess milieu appeared to inhibit the growth of P. aeruginosa. Ciprofloxacin activity in abscess fluid did not appear to be adversely affected by acid pH, aerobic or anaerobic conditions of incubation, the abscess constituents, or the binding of ciprofloxacin to the DNA in pus. Ciprofloxacin was bound to DNA of bacterial or human origin. Binding by pus was reversible, and binding to DNA extracts of pus was blocked by pretreatment of extracts with deoxyribonuclease but not by pretreatment with ribonuclease. The ability of ciprofloxacin to kill organisms growing in the adverse conditions of the abscess milieu may contribute to its success as a chemotherapeutic agent for treatment of patients with suppurative infection.

Ｂａｃａｍｐｉｃｉｌｌｉｎ経口投与後のヒト血清，歯根嚢胞および膿中へのａｍｐｉｃｉｌｌｉｎの移行に関する研究

Ｂａｃａｍｐｉｃｉｌｌｉｎ経口投与後のヒト血清，歯根嚢胞および膿中へのａｍｐｉｃｉｌｌｉｎの移行に関する研究

Effect of abscess milieu on bactericidal activity of LY146032 against staphylococci.

The antistaphylococcal activity of LY146032 was tested in human pus in vitro and in a murine abscess model in vivo. The drug was not degraded by pus containing beta-lactamase and had equally good or better activity than nafcillin or vancomycin against Staphylococcus aureus or Staphylococcus epidermidis in vitro and in vivo. Its activity was slightly enhanced when used in combination with rifampin and tobramycin.

Granulocyte collagenase and elastase and the plasma protease inhibitors in human pus.

Granulocyte collagenase and elastase was demonstrated in human pus using specific antisera. The enzymes were complexed by alpha1-antitrypsin and alpha2-macroglobulin but also were present as free proteases. All purulent exudates showed free elastolytic and collagenolytic activity.

Inactivation of penicillin by purulent exudates.

Four of 22 specimens of human pus inactivated up to 90% of added penicillin within one hour in vitro. Ampicillin and cephaloridine were also inactivated, but streptomycin and fusidic acid were not. The effect was not related to the protein content of the pus, nor to its pH value. Microbes that may produce beta-lactamase in small quantities were isolated from three of the four specimens, but the enzyme was not detected in the pus by physical methods nor by microbiological inhibition assay. The inactivating effect was shown to be a property of the solid portion of the pus, and was absent from the filtrate. We suggest that the effect may be an intrinsic property of the host, which should be investigated further as it has important implications for clinical practice.

THE CHICK EMBRYO ANTIGEN (LYGRANUM) TEST FOR LYMPHOGRANULOMA VENEREUM

The purpose of this communication is to report on our experience with a relatively new antigen introduced in 1940 by Grace, Rake and Shaffer 1 as being superior to mouse brain antigen for the performance of the Frei test. This antigen (known as lygranum) is made by growing the lymphogranuloma venereum virus in the yolk sac of the developing chick embryo, while lygranum control is made from the yolk sac itself. In 1941 Sulkin and co-workers, 2 using as did the originators of the test a 7 mm. papule as a minimal positive reaction, found in 42 patients with lymphogranuloma venereum 40 with a positive reaction to lygranum and 38 with a positive reaction to human bubo pus. There were easily recognized nonspecific papules in some of the control subjects. The Robinsons 3 in 1942 performed simultaneous tests with lygranum and bubo pus on 127 patients. They found 91 per