In Vitro Assessment on Designing Novel Antibiofilms of Pseudomonas aeruginosa Using a Computational Approach.

An anti-biofilm that can inhibit the matrix of biofilm formation is necessary to prevent recurrent and chronic Pseudomonas aeruginosa infection. This study aimed to design compounds with a new mechanism through competitive inhibitory activity against phosphomannomutase/phosphoglucomutase (PMM/PGM), using in vitro assessment and a computational (in silico) approach. The active site of PMM/PGM was assessed through molecular redocking using L-tartaric acid as the native ligand and other small molecules, such as glucaric acid, D-sorbitol, and ascorbic acid. The docking program set the small molecules to the active site, showing a stable complex formation. Analysis of structural similarity, bioavailability, absorption, distribution, metabolism, excretion, and toxicity properties proved the potential application of ligands as an anti-biofilm. In vitro assessment with crystal violet showed that the ligands could reach up to 95.87% inhibition at different concentrations. The nitrocellulose membrane and scanning electron microscopic visualization showed that the untreated P. aeruginosa biofilm was denser than the ligand-treated biofilm.

Odontogenic brain abscess due to Anaerococcus prevotii infections: A case report and review article.

BACKGROUND: Odontogenic brain abscess is a rare case primarily caused by normal flora such as Anaerococcus prevotii. CASE PRESENTATION: A 60-years-old Indonesian female complained of severe left side headaches, hearing loss, a decrease of consciousness, several episodes of nausea and vomiting, and hemiparesis dextra for 5 days. Three months previously, she performed dental operative procedures on the left side of the first and second lower molar and debridement of phlegmon on the left side of the mouth. Head CT scan suggests multiple brain abscesses or high-grade glioma, non-communicating hydrocephalus and suggestive mastoiditis. The patient underwent excision surgery and abscess culture, which resulted in Anaerococcus prevotii. The patient received a metronidazole antibiotic, and on the seventh day, his condition improved. DISCUSSION: Identifying bacterial infection in the brain abscess is crucial for effective treatment. Abscess removal in the brain and antibiotics are treatments for brain abscesses. CONCLUSION: Odontogenic brain abscess caused by Anaerococcus prevotii infection effectiveness with surgical excision and antibiotics.

Isolation and Identification of Escherichia coli O157:H7 Lytic Bacteriophage from Environment Sewage.

Escherichia coli O157:H7 is one of the pathogenic bacteria causing foodborne disease. The use of lytic bacteriophages can be a good solution to overcome the disease. This study is aimed at isolating lytic bacteriophages from environmental sewage with E. coli O157:H7 bacterial cells. The sample used in this study was eight bacteriophages, and the technique used in identifying E. coli O157:H7 carriers of the stx1 and stx2 genes was PCR. The double layer plaque technique was used to classify bacteriophages. Plaque morphology, host specificity, and electron micrograph were used to identify the bacteriophages. The result obtained plaque morphology as a clear zone with the largest diameter size of 3.5 mm. Lytic bacteriophage could infect E. coli O157:H7 at the highest titer of 10 × 108 PFU/mL. Bacteriophages have been identified as Siphoviridae and Myoviridae. Phage 3, phage 4, and phage 8 could infect Atypical Diarrheagenic E. coli 1 (aDEC1) due to their host specificity. The Friedman statistical tests indicate that lytic bacteriophage can significantly lyse E. coli O157:H7 (p = 0.012). The lysis of E. coli O157:H7 by phage 1, phage 2, phage 3, and phage 5 bacteriophages was statistically significant, according to Conover's posthoc test (p < 0.05). The conclusion obtained from this study is that lytic bacteriophages from environmental sewage could lyse E. coli O157:H7. Therefore, it could be an alternative biocontrol agent against E. coli O157:H7 that contaminates food causing foodborne disease.

The Analysis of OmpA and Rz/Rz1 of Lytic Bacteriophage from Surabaya, Indonesia.

A good strategy to conquer the Escherichia coli-cause food-borne disease could be bacteriophages. Porins are a type of ß-barrel proteins with diffuse channels and OmpA, which has a role in hydrophilic transport, is the most frequent porin in E. coli; it was also chosen as the potential receptor of the phage. And the Rz/Rz1 was engaged in the breakup of the host bacterial external membrane. This study aimed to analyze the amino acid of OmpA and Rz/Rz1 of lytic bacteriophage from Surabaya, Indonesia. This study employed a sample of 8 bacteriophages from the previous study. The OmpA analysis method was mass spectrometry. Rz/Rz1 was analyzed using PCR, DNA sequencing, Expasy Translation, and Expasy ProtParam. The result obtained 10% to 29% sequence coverage of OmpA, carrying the ligand-binding site. The Rz/Rz1 gene shares a high percentage of 97.04% to 98.89% identities with the Siphoviridae isolate ctTwQ4, partial genome, and Myoviridae isolate cthRA4, partial genome. The Mann-Whitney statistical tests indicate the significant differences between Alanine, Aspartate, Glycine, Proline, Serine (p=0.011), Asparagine, Cysteine (p=0.009), Isoleucine (p=0.043), Lysine (p=0.034), Methionine (p=0.001), Threonine (p=0.018), and Tryptophan (p=0.007) of OmpA and Rz/Rz1. The conclusion obtained from this study is the fact that OmpA acts as Phage 1, Phage 2, Phage 3, Phage 5, and Phage 6 receptors for its peptide composition comprising the ligand binding site, and Rz/Rz1 participates in host bacteria lysis.

Association of blood isolate's multi antibiotic resistance-index on laboratory-confirmed bloodstream infection: A cross-sectional study.

BACKGROUND: A not optimal way of the insertion of the intravenous catheter can be one of the factors that cause bloodstream infection (BSI) that should be confirmed with blood culture, and if positive it is called Laboratory-Confirmed Bloodstream Infection (LCBI). One of the surveillance methods of nosocomial infection that is commonly used is the Multi Antibiotic Resistance (MAR)-Index. The aimed of study was association of MAR-index from blood isolates on LCBI category. METHOD: This study used a cross-sectional study with a consecutive sampling method. Data collection for this study includes identification of micromaterial profile, antimicrobial test, MAR-Index, and LCBI category. The analysis used is the Mann Whitney test with p < 0.05. RESULT: There were 43 isolates of LCBI 1, 26 isolates of LCBI 2, and none of the LCBI 3. Microorganisms in the LCBI category 1 were Staphylococcus aureus (53.4%), Acinetobacter baumannii (20.9%), Escherichia coli (9.3%), Klebsiella pneumonia (7.0%), Pseudomonas aeruginosa (4.7%), and Enterococcus faecalis (4.7%) with the MAR-Index ranged from 0.22 to 0.91. Microorganisms in the LCBI category 2 were Staphylococcus haemolyticus (69.3%), Staphylococcus epidermidis (19.3%), Staphylococcus hominis (3.8%), Streptococcus viridans (3.8%), and Corynebacterium jeikeium (3.8%) with the MAR-Index ranging between 0.11 and 0.79. There is no significant difference of MAR-index between LCBI 1 and 2 (p = 0.424) and no association of MAR-index on LCBI (p = 0.571). CONCLUSION: Most LCBI type 1 is Staphylococcus aureus and LCBI type 2 is Staphylococcus haemolyticus which there is no significant association of MAR-index on LCBIs.

Frequency of diarrheagenic Escherichia coli among children in Surabaya, Indonesia.

Diarrheagenic Escherichia coli (DEC) is a major etiologic agent of childhood diarrhea in developing countries. We investigated the frequency of DEC in stool samples from 125 diarrheal children (age, 1-10 years) and 92 non-diarrheal children in Surabaya, Indonesia. The non-diarrheal children served as healthy controls. DEC was detected in 23 of 125 (18.4%) and 47 of 92 (51.1%) samples in the diarrheal and non-diarrheal children, respectively. Enteropathogenic E. coli was the most prevalent in the non-diarrheal children (25.0%), and its prevalence was significantly higher than that in the diarrheal children (0.8%) (P < 0.0001). Interestingly, Shiga toxin-producing E. coli (4.3%) was detected only in the non-diarrheal children (P = 0.031). This is the first study comparing between diarrheal children with non-diarrheal or healthy children to investigate the role of DEC in pediatric diarrheal diseases in Indonesia.

ß-D-xylosidase from Geobacillus thermoleovorans IT-08: biochemical characterization and bioinformatics of the enzyme.

The gene encoding a thermostable ß-D-xylosidase (GbtXyl43B) from Geobacillus thermoleovorans IT-08 was cloned in pET30a and expressed in Escherichia coli; additionally, characterization and kinetic analysis of GbtXyl43B were carried out. The gene product was purified to apparent homogeneity showing M r of 72 by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The enzyme exhibited an optimum temperature and pH of 60 °C and 6.0, respectively. In terms of stability, GbtXyl43B was stable at 60 °C at pH 6.0 for 1 h as well as at pH 6-8 at 4 °C for 24 h. The enzyme had a catalytic efficiency (k cat/K M) of 0.0048 ± 0.0010 s(-1) mM(-1) on p-nitrophenyl-ß-D-xylopyranoside substrate. Thin layer chromatography product analysis indicated that GbtXyl43B was exoglycosidase cleaving single xylose units from the nonreducing end of xylan. The activity of GbtXyl43B on insoluble xylan was eightfold higher than on soluble xylan. Bioinformatics analysis showed that GbtXyl43B belonging to glycoside hydrolase family 43 contained carbohydrate-binding module (CBM; residues 15 to 149 forming eight antiparallel ß-strands) and catalytic module (residues 157 to 604 forming five-bladed ß-propeller fold with predicted catalytic residues to be Asp287 and Glu476). CBM of GbtXyl43B dominated by the Phe residues which grip the carbohydrate is proposed as a novel CBM36 subfamily.

Phenotypic and genotypic characterization of Vibrio cholerae clinically isolated in Surabaya, Indonesia.

The phenotypic and genotypic characteristics of 6 clinical strains of Vibrio cholerae isolated in Surabaya, Indonesia in 2009 were examined. The DNA fingerprints obtained suggested that these isolates were not from a single clone. Furthermore, all isolates produced cholera toxin and possessed the classical type of toxin B subunit gene, thus meaning that this is the first report of the occurrence of El Tor variants of V. cholerae in Indonesia. Although all isolates were sensitive to almost all antibiotics tested, including ampicillin, chloramphenicol, ciprofloxacin, gentamicin, levofloxacin, kanamycin, nalidixic acid, norfloxacin, streptomycin, trimethoprim-sulfamethoxazole, and tetracycline, and had no mutation in the gyrA and parC genes, they nevertheless possessed the class 1 integron that is a molecular vehicle for the acquisition of antibiotic resistance genes, suggesting that they have the potential to acquire the genetic element for drug resistance.

Emergence of fluoroquinolone-resistant strains of Salmonella enterica in Surabaya, Indonesia.

Typhoid fever remains a major health problem in developing countries. Fluoroquinolones such as ciprofloxacin emerged as the 1st-choice treatment of enteric fever, including typhoid, in the 1990s. Recently, Salmonella typhi strains with resistance to ciprofloxacin have been increasingly reported in several countries, although the fluoroquinolone-resistant clinical strain has not been reported in Indonesia. In the present study, we examined the drug susceptibility and the presence of gyrA mutations in 17 clinical strains of S. typhi isolated from Surabaya, Indonesia, in 2006 (9 strains) and 2008 (8 strains). Although all 9 isolates from 2006 were sensitive to all tested antibiotics and had no mutation in the gyrA gene, all 8 isolates from 2008 were resistant to nalidixic acid and ampicillin and had a gyrA mutation at codon 87. In addition, 3 of 8 strains from 2008 showed multiple drug resistance, including resistance to chloramphenicol, trimethoprim-sulfamethoxazole, and ciprofloxacin. Therefore, newer drugs, such as ceftriaxone, cefixime, and azithromycin, might be effective in this situation. This is the 1st report of the emergence of fluoroquinolone-resistant clinical strains of S. typhi with a gyrA mutation, and it reveals a health risk due to multidrug-resistant strains in Indonesia.

Induction of thymus-derived gammadelta T Cells by Escherichia coli enterotoxin b subunit in peritoneal cavities of mice.

We examined the activation of intraperitoneal T cells in BALB/c mice by the Escherichia coli enterotoxin B subunit, which induced a specific Th2 type of T-cell response to intraperitoneally coadministered bovine immunoglobulin G. The numbers of both gammadelta and alphabeta T cells increased significantly after intraperitoneal administration of the B subunit in a time-dependent manner; these numbers were not affected by the B-subunit G33D mutant, which is defective in GM1 ganglioside-binding ability. Early after administration a small number of gammadelta T cells produced either interleukin-4 (IL-4) or gamma interferon, while late after administration primarily IL-10-producing gammadelta T cells were detected. gammadelta T cells induced by the B subunit did not express a characteristic V gene over the time course of the study. The induction of gammadelta T cells did not occur in athymic nu/nu mice but could be induced upon transplantation of fetal AKR thymus-like alphabeta T cells. gammadelta T cells in athymic nu/nu mice with a fetal thymic graft predominantly expressed the donor Thy-1.1 antigen but not the host Thy-1.2 antigen. The induction of these T cells, however, could not be restored by coadministration of the B subunit with peritoneal cells from normal mice. These results suggest that the B subunit activates intraperitoneal gammadelta and alphabeta T cells in a manner dependent upon its ability to bind to GM1 ganglioside. gammadelta T cells induced by the B subunit are Th2-type cells derived from the thymus. These gammadelta T cells may be functionally involved in specific Th2 responses to the B subunit, which possibly acts as an adjuvant through the influence of alphabeta T cells.