Presence Of Gram Research Articles

Mapping of Microbial Diversity of Gautala Reserve Forest in Aurangabad (District) (M.S.), India

Microorganisms help in the fertility and endurance of soil. Additionally supporting the growth of several biological systems, soil and soil microorganisms fill in as the best mode for plant growth. Soil microbes are essential for recycling old plant material and decaying organic matter. In the upkeep of the Environment microbial diversity plays a dominant role. The current paper is focused on the seasonal variations in the microbial count, which are increased or decreased (Impact on counts) in the Gautala reserve forest. In the present study, soil samples from Gautala Reserve Forest were collected from 15 different sites during the rainy, winter, and summer seasons. The present study was attempted to cover the microbial diversity of the whole Gautala forest through covering maximum sampling spots. The average total microbial count (TMC) in the rainy season for plate 1 and plate 2 was 36.73 and 35.46 respectively and the final count was 36.4 × 106. The average TMC in the winter season for plate 1 and plate 2 was 32.4 and 31.93 respectively and the final count was 32.46 × 105. The average TMC in the summer season for plate 1 and plate 2 was 37.13 and 36.4 and the final count was 37.6 × 104. The bacterial colony at sampling points 1, 4, and 7 exhibits presence of Gram-Negative Bacteria; whereas the rest of the sampling spots showed presence of Gram - Positive Bacteria. There were three types of Gram-negative and 12 were found to be Gram-positive bacteria with three genera i.e. Pseudomonas spp., Bacillus spp., and micrococcus spp. This paper discusses seasonal fluctuations in microbial counts and associated laboratory culture techniques and statistical analysis methods.

MO702: Etiology and the Impact of Refractory Peritonitis on Clinical Outcomes of Patients on Peritoneal Dialysis—12 Years’ Single-Center Experience from Turkey

Abstract BACKGROUND AND AIMS The outcomes of refractory peritonitis in peritoneal dialysis (PD) patients have been reported to be inferior to those of solitary peritonitis. The current study aimed to examine the factors associated with treatment failure in PD patients experiencing refractory peritonitis. METHOD This single-center retrospective study included all episodes of refractory peritonitis in adult PD patients in Marmara University Hospital, Turkey, between 2009 and 2020. Patient characteristics, microbiological data, outcomes and factors associated with refractory peritonitis were analysed. The primary outcome was peritonitis-related catheter loss. Secondary outcomes were hospitalization and peritonitis-related death. RESULTS Overall, 236 episodes of refractory peritonitis occurring in 135 patients were included. Gram-positive, gram-negative and fungal infections accounted for 44.1%, 20.4% and 2.4% of all peritonitis episodes, respectively. Forty-seven patients (34.8%) needed catheter removal, 2 patients (1.5%) died due to peritonitis complications and 59 episodes (25%) needed hospitalization. Mean fifth day PD fluid cell count was significantly greater among patients who required PD catheter removal (3621.3 $ \pm $ 3144.1 versus 1589.4 $ \pm $ 2316.6 P < 0.001). Furthermore, patients with >1000/mm3 cell count on the fifth day had higher rate of catheter removal (72.3% versus 37.6%, P < 0.001) as compared to patients with cell count under 300/mm3. Treatment failure was more common in peritonitis episodes caused by gram (−) organisms (31.9% versus 14.8%, P:0.012). Pseudomonas and fungi-associated peritonitis were also significantly correlated with catheter loss (P:0.001 and P: <0.001) (Table 1). When peritonitis episodes with more and <1000 cells/mm3 on the fifth day were compared, there were more episodes with gram (−) bacteria (29.7% versus 12.9%, P:0.003) and hospitalization (41.9% versus 11.4%, P: <0.001) in the group with more than 1000 cells/mm3. In the multivariate analysis, factors associated with catheter loss were a cell count of >1000 on the fifth day and hospitalization, while presence of gram (+) bacteria related peritonitis was inversely correlated with catheter loss (Table 2). CONCLUSION Our study shows that the PD cell count on the fifth day of peritonitis can be used as a prognostic tool to determine the prognosis of refractory peritonitis episodes. Although we were unable to show the adverse prognostic effect of gram (−) bacteria related peritonitis, gram (+) bacteria related peritonitis was associated with better outcome. Prospective studies are needed to assess the risk factors for adverse outcomes of patients with refractory peritonitis, as the evidence in this area is sparse.

Antimicrobial Activity of Lactic Acid Bacteria from Kunun-Zaki on Selected Clinical Pathogens

Aims: To investigate the anti-microbial activity of lactic acid bacterial isolates from locally fermented Kunun-zaki beverage.
 Study Design: The study was designed to study the anti-microbial effect of lactic acid bacterial isolates obtained from Kunun-zaki beverage.
 Place and Duration of Study: Department of Applied Microbiology Enugu State University of Science and Technology, Agbani, June 2009 and January 2010.
 Methodology: Fresh Kunun drink was prepared from millet following standard modified procedures. Isolation of lactic acid bacteria from the beverage sample was carried out using MRS agar medium. Morphological and biochemical characterizations were performed to identify the organisms. Finally, the bacterial isolates were tested for antibacterial activity against clinical pathogens – Bacillus anthracis, Clostridium perfringens, Escherichia coli, Streptococcus faecalis, Micrococcus luteus, Klebsiella pneumonia, Pseudomonas aeruginosa, Bacillus cereus, Bacillus subtilis and Staphylococcus aureus obtained from Microbiology Department Nnamdi Azikiwe University Awka Nigeria.
 Results: Morphological and biochemical characteristics revealed the presence of Gram’s positive bacteria which were identified as Lactobacillus casei and Lactobacillus brevis. The antimicrobial activity of the isolates (L. casei and L. brevis) against various pathogenic strains recorded 90±0.002% inhibition. Only Micrococcus luteus was absolutely resistant to antimicrobial treatments with Lactobacillus casei and Lactobacillus brevis. 
 Conclusion: The study concluded that Lactobacillus brevis and Lactobacillus casei from kunun-zaki drink possess strong antimicrobial activity against a wide range of clinical pathogens. Since lactic acid bacteria are known to possess many health benefits, some of the therapeutic effects reported from the consumption of the beverage may be attributed to these organisms.

Soil microbial community and microbial residues respond positively to minimum tillage under organic farming in Southern Germany

In a field trial comprising organic farming and minimum tillage management strategies in Scheyern, Germany, we evaluated the long-term (21-year) effects of organic farming (use of a diverse crop rotation with legume cover crop and without application of synthetic fertilizer or pesticides) and minimum tillage (6–8cm depth) on the microbial community structure and microbial residues in Cambisols. Organic farming had a positive effect on microbial biomass, total phospho-lipid fatty acids (PLFA), Gram (+) bacteria, Gram (−) bacteria and the arbuscular mycorrhizal fungi (AMF) indicator PLFA 16:1ω5 and amino sugars. The increase in presence of Gram (+) bacteria when compared to integrated farming was also reflected by increased content of bacterial muramic acid (MurN), i.e. an increased formation of bacterial residues. Minimum tillage significantly increased microbial biomass N and the fungal PLFA 18:2ω6,9, averaging the values of upper (0–8cm) and deeper (12–25cm) soil, but had no effects on PLFA 16:1ω5. Minimum tillage generally resulted in a negative depth gradient of almost all microbial properties analyzed. The only important exception was fungal galactosamine (GlcN), which led to increases in the fungal C/bacterial C ratio and in the contribution of microbial residue C to SOC in the deeper soil. Significant second order tillage×management interactions indicated that minimum tillage effects on microbial biomass and PLFA indices (Gram (+) and (i15:0+i17:0)/(a15:0+a17:0)) were much stronger in the organic farming system than in the integrated farming system. Redundancy analysis (RDA) showed SOC and H2O content predominantly affected the microbial community structure in the present study. Minimum tillage in combination with organic farming appears to be an effective agricultural strategy that enhances soil microbial biomass, microbial residues and bacterial and fungal abundances. The results indicate that the positive effects of minimum tillage on microbial community can be enhanced by organic farming. Microbial residues as a fraction of SOC respond faster to farming management than to tillage.

Microbial Analysis of Biomedical Wastes from Selected Health Facilities in Parts of Edo South and its Public Health Implication

This study, aimed at the microbial analysis of biomedical waste (BMW) was carried out using 100 samples of 10 different BMW collected in duplicates from 5 busy primary healthcare centers (PHCs).The research findings showed a high prevalence of <em>E.coli</em> (39%) and <em>S.aureus</em> (32%) which were both statistically significant at P≤0.05, while the least isolated organisms were <em>K. pnuemoniae</em> (10%) and <em>B.</em><em> subtilis</em> (4%) and were statistically not significant at P≥0.05. Samples from dressings and beddings were found to contain the highest microbial loadof 25 and 13 respectively while the least number of isolates were from expired cytotoxic drugs (2) and lancets (1).The biochemical tests showed the presence of Gram positive and negative organisms with record of both aerobic and anaerobic isolates from the BMW. The investigation revealed that BMW contains mixed bacterial community with some being pathogenic and pose a public health hazard to both health workers and other community members, therefore adequate treatment measures should be given to all BMW before disposal.

Antimicrobial Resistance of ESBL Producing Coliforms Isolated from Retail Meat Samples

Extended Spectrum Beta lactamases (ESBLs) are special enzymes which are found in groups and can hydrolyze the third generation cephalosporins. Their Production is the commonest cause of resistance to beta-lactam antibacterial agents among Gram-negative bacteria. There is spread of drug resistance among food animals due to the use of antimicrobials. Pathogens develop resistance in their animal reservoirs and then these resistant strains are transmitted to humans where they may cause infections which are difficult to treat. In this study the antimicrobial resistance due to ESBL producing coliforms in humans and in edible animal meat was examined. The main objective of this study was to determine the fact that a substantial increase in antimicrobial resistance posed by ESBL producing coliforms in humans is due to edible animal meat. Various chicken, meat and beef samples were collected from different sites in Faisalabad. The samples were enriched in Trypticase Soy Broth and were streaked on Nutrient Agar then on MacConkey Agar to check the presence of Gram negative bacteria. Gram Negative isolates were isolated by gram staining and biochemical test (Triple Sugar Iron; TSI). Antibiotic susceptibility testing was performed through disc diffusion method. In which 28% isolates were resistant to Cefixime, 20% to Ceftriaxone, 24% to Clavulanic acid and 30% to Cefotaxime. As the next step, a set of PCR was optimized for amplification of major genes of ESBL conferring resistance to selected antibiotics. As a result 20% Bla (CTX-M), 4% Bla(OXA), 2% Bla(PER) and 1% Bla(Ges) were amplified through PCR. These are the major genes of ESBL which are going to become more and more resistant towards the first, second and third generation cephalosporins.

Effects of antiresorptive agents on osteomyelitis

The effects of antiresorptive agents (e.g., alendronate [Aln], osteoprotegerin [OPG]) on bone infection are unknown. Thus, their effects on implant‐associated osteomyelitis (OM) were investigated in mice using PBS (placebo), gentamycin, and etanercept (TNFR:Fc) controls. None of the drugs affected humoral immunity, angiogenesis, or chronic infection. However, the significant (P < 0.05 vs. PBS) inhibition of cortical osteolysis and decreased draining lymph node size in Aln‐ and OPG‐treated mice was associated with a significant (P < 0.05) increase in the incidence of high‐grade infections during the establishment of OM. In contrast, the high‐grade infections in TNFR:Fc‐treated mice were associated with immunosuppression, as evidenced by the absence of granulomas and presence of Gram+ biofilm in the bone marrow. Collectively, these findings indicate that although antiresorptive agents do not exacerbate chronic OM, they can increase the bacterial load during early infection by decreasing lymphatic drainage and preventing the removal of necrotic bone that harbors the bacteria.

Liquid Crystal Emulsions as the Basis of Biological Sensors for the Optical Detection of Bacteria and Viruses

AbstractA versatile sensing method based on monodisperse liquid crystal (LC) emulsion droplets detects and distinguishes between different types of bacteria (Gram +ve and −ve) and viruses (enveloped and non‐enveloped). LCs of 4‐cyano‐4'‐pentylbiphenyl transition from a bipolar to radial configuration when in contact with Gram −ve bacteria (E. coli) and lipid‐enveloped viruses (A/NWS/Tokyo/67). This transition is consistent with the transfer of lipid from the organisms to the interfaces of the micrometer‐sized LC droplets. In contrast, a transition to the radial configuration is not observed in the presence of Gram +ve bacteria (Bacillus subtilis and Micrococcus luteus) and non‐enveloped viruses (M13 helper phage). The LC droplets can detect small numbers of E. coli bacteria (1–5) and low concentrations (104 pfu mL−1) of A/NWS/Tokyo/67 virus. Monodisperse LC emulsions incubated with phosholipid liposomes (similar to the E. coli cell wall lipid) reveal that the orientational change is triggered at an area per lipid molecule of ∼46 Å2 on an LC droplet (∼1.6 × 108 lipid molecules per droplet). This approach represents a novel means to sense and differentiate between types of bacteria and viruses based on their cell‐wall/envelope structure, paving the way for the development of a new class of LC microdroplet‐based biological sensors.

Specific multigenotypic diagnosis of nosocomial pneumonia in ICU

Mortality rate of nosocomial pneumonia in ventilated ICU patients (pts) is about 20-25%. Despite advances in critical care, standard criteria for pneumonia are still inaccurate. Antibiotherapy is often started on a 'best guess' basis to anticipate frequently negative bacteriological results (Ba-cul). Main etiological pathogens are staphylococci, streptococci, pseudomonads and enterobacteriaceae. A multigenotypic and sequential molecular identification (Mol-id) was applied on bronchoalveolar lavages (BAL) from 12 pts with clinical-radiological evidence of infection with (n = 9)/without (n = 3) positive Ba-cul. DNA extraction and duplicate specific amplification of 16S rDNA from BAL was first used to identify signals corresponding to the presence of Gram + (G+), Gram - (G-), or mixed G+/G-. Any G+ signal was followed by femA and mecA multiplex (mpx)-PCR for species-specific identification of staphylococci and methicillin resistance, and by mpx-PCR for S. pneumoniae. Any G- signal was followed by mpx-PCR for species-specific identification of Pseudomonads (aeruginosa, cepacia, maltophilia) vs other G- for which the 16S rDNA amplicon was sequenced. Mol-id identified mixed G+/G- in 2/3 negative Ba-cul. Ba-cul gave G+ or G- only in 3 and 4/12 pts, respectively. Among these, Mol-id confirmed either G+ (2/3) or G- (2/4), and found a mixed signal in 3/7 pts. Mixed G+/G- was found by Ba-cul and Mol-id in 2/12 BAL, but in one case, S. epidermidis was found by Mol-id and enterococci by Ba-cul. Staphylococci (MRSA, other) and Ps. aeruginosa were found by Ba-cul and Mol-id in 2 and 3 pts, respectively. Current results suggest that Mol-id is a useful adjunct bringing more insight to standard criteria of nosocomial pneumoniae in ICU and can be a timely relevant guide for antibiotherapy.