Occurrence of enterotoxigenic staphylococci in frozen fishery products

Sixty six samples of frozen fishery products consisting of frozen crab meat, frozen prawns, frozen cuttlefish and frozen mussel collected from nine cold stores situated in and around Cochin city were examined for coagulase positive staphylococci. Forty eight samples (72.72%) revealed the presence of coagulase positive staphylococci, which were then tested for enterotoxigenity, employing cellophane-over-agar method for toxin production and optimum-sensitivity-plate (OSP) method for toxin detection. Seventy seven out of one hundred and two (75.49%) coagulase positive staphylococcal isolates from the above products produced enterotoxins A, B, C, D and E either singly or in combinations. Enterotoxin A and D were detected more often than others. The occurrence of enterotoxigenic staphylococci in frozen fishery product is reported for the first time in this country.

Drug delivery strategies for photodynamic antimicrobial chemotherapy: From benchtop to clinical practice

Light and photosensitizer-mediated killing of many pathogens, termed photodynamic antimicrobial chemotherapy (PACT), has been extensively investigated in vitro. A wide range of organisms from the Gram-positive Staphylococcus aureus to the Gram-negative Pseudomonas aeruginosa have been proven to be susceptible to PACT. Multidrug-resistant strains are just as susceptible to this treatment as their naive counterparts. Both enveloped and non-enveloped viruses have demonstrated susceptibility in vitro, in addition to fungi and protozoa. Significantly, however, no clinical treatments based on PACT are currently licensed. This paper provides a comprehensive review of work carried out to date on delivery of photosensitizers for use in PACT, including topical, intranasal and oral/buccal delivery, as well as targeted delivery. We have also reviewed photo-antimicrobial surfaces. It is hoped that, through a rational approach to formulation design and subsequent success in small-scale clinical trials, more widespread use will be made of PACT in the clinic, to the benefit of patients worldwide. (C) 2009 Elsevier B.V. All rights reserved.

A potent novel antimicrobial peptide related to caerin 1.12 from the skin secretion of the Australian frog, Litoria aurea

Skin secretions from Australian frogs of the genus Litoria have been extensively studied for many years and are known to contain a large array of antimicrobial peptides that often bear their specific names — caerins (L. caerulea), aureins (L. aurea), citropins (L. citropa) and maculatins (L. genimaculata) — and each group displays distinct primary structural attributes. During a systematic transcriptome cloning study using a cDNA library derived from skin secretion of L. aurea, a series of identical clones were identified that encoded a novel 25-mer antimicrobial peptide that displayed 92% structural identity with caerin 1.12 from L. caerulea, differing in amino acid sequence at only two positions — Arg for Gly at position 7 and Leu amide for Ser amide at the C-terminus. The novel peptide had conserved Pro residues at positions 15 and 19 that flank a flexible hinge region which previous studies have suggested are important for effective orientation of the two alpha-helices within the bacterial membrane resulting in lysis of cells. As the two substitutions in the novel peptide serve to increase both positive charge and hydrophobicity, we synthesised a replicate and determined its minimal inhibitory concentration (MIC) against Gram positive Staphylococcus aureus and Gram negative Escherichia coli. The MICs for these organisms were 3 µM and 4 µM, respectively, indicating a high potency and haemolysis was

Antibacterial Effect of Human Mesenchymal Stem Cells is Mediated in Part from Secretion of the Antimicrobial Peptide LL-37

Recent in vivo studies indicate that mesenchymal stem cells (MSCs) may have beneficial effects in the treatment of sepsis induced by bacterial infection. Administration of MSCs in these studies improved survival and enhanced bacterial clearance. The primary objective of this study was to test the hypothesis that human MSCs possessed intrinsic antimicrobial properties. We studied the effect of human MSCs derived from bone marrow on the bacterial growth of Gram-negative (Escherichia coli and Pseudomonas aeruginosa) and Gram-positive (Staphylococcus aureus) bacteria. MSCs as well as their conditioned medium (CM) demonstrated marked inhibition of bacterial growth in comparison with control medium or normal human lung fibroblasts (NHLF). Analysis of expression of major antimicrobial peptides indicated that one of the factors responsible for the antimicrobial activity of MSC CM against Gram-negative bacteria was the human cathelicidin antimicrobial peptide, hCAP-18/LL-37. Both m-RNA and protein expression data showed that the expression of LL-37 in MSCs increased after bacterial challenge. Using an in vivo mouse model of E. coli pneumonia, intratracheal administration of MSCs reduced bacterial growth (in colony-forming unit) in the lung homogenates and in the bronchoalveolar lavage (BAL) fluid, and administration of MSCs simultaneously with a neutralizing antibody to LL-37 resulted in a decrease in bacterial clearance. In addition, the BAL itself from MSC-treated mice had a greater antimicrobial activity in comparison with the BAL of phosphate buffered saline (PBS)-treated mice. Human bone marrow-derived MSCs possess direct antimicrobial activity, which is mediated in part by the secretion of human cathelicidin hCAP-18/ LL-37.

Hylaranins: prototypes of a new class of amphibian antimicrobial peptide from the skin secretion of the oriental broad-folded frog, Hylarana latouchii

Amphibian skin secretions contain a broad spectrum of biologically active compounds, particularly antimicrobial peptides, which are considered to constitute a first line of defence against bacterial infection. Here we describe the identification of two prototype peptides representing a novel structural class of antimicrobial peptide from the skin secretion of the oriental broad-folded frog, Hylarana latouchii. Named hylaranin-L1 (GVLSAFKNALPGIMKIIVamide) and hylaranin-L2 (GVLSVIKNALPGIMRFIAamide), both peptides consist of 18 amino acid residues, are C-terminally amidated and are of unique primary structures. Their primary structures were initially deduced by MS/MS fragmentation sequencing from reverse-phase HPLC fractions of skin secretion that demonstrated antimicrobial activity. Subsequently, their precursor-encoding cDNAs were cloned from a skin secretion-derived cDNA library and their primary structures were confirmed unequivocally. Synthetic replicates of both peptides exhibited broad-spectrum antimicrobial activity with mean inhibitory concentrations (MICs) of 34 µM against Gram-negative Escherichia coli, 4.3 µM against Gram-positive Staphylococcus aureus and 4–9 µM against the yeast, Candida albicans. Both peptides exhibited little haemolytic activity (<6 %) at the MICs for S. aureus and C. albicans. Amphibian skin secretions thus continue to provide novel antimicrobial peptide structures that may prove to be lead compounds in the design of new classes of anti-infection therapeutics.

Aplicação de filmes de suberina na produção de materiais bio-funcionais

Trabalho Final de Mestrado para obtenção do grau de Mestre em Engenharia Química e Biológica

Bacteriological quality of individually quick-frozen (IQF) raw and cooked readyto- eat shrimp produced from farm raised black tiger shrimp (Penaeus monodon

One thousand, two hundred and sixty four samples of individually quick-frozen (IQF) peeled and deveined raw and 914 samples of cooked ready to eat shrimp samples produced from farm raised black tiger (Penaeus monodon) obtained from a seafood unit working under HACCP concept were analysed for total aerobic plate count (APC), coliform count, Escherichia coli, coagulase positive Staphylococci and Salmonella. The overall bacteriological quality of the product was found to be good. Of the frozen raw shrimp, 96% of samples showed APC below 105 while 99% of the frozen cooked ready-to-eat samples showed APC less than 104. The APC ranged from 1·0´102 to 4·2´106 cfu/gm in frozen raw shrimp and from 1·0´102 to 6·4´104 cfu/gm in the frozen cooked shrimp. Prevalences of coliforms in raw shrimp and cooked shrimp samples were 14·4% and 2·9% respectively. The coliform count in raw products ranged from 1·0´101 to 2·5´103 cfu/gm and in the cooked products, from 1·0 ´101 to 1·8´102 cfu/gm. Although all the cooked shrimp samples were free of coagulase positive staphylococci, E. coli and Salmonella, 1·0, 2·0 and 0·1% of the frozen raw shrimp samples tested positive for coagulase positive Staphylococci, E. coli and Salmonella respectively. The Salmonella strain was identified as Salmonella typhimurium. The results of the present study highlight the importance of implementation of HACCP system in the seafood industry to ensure consistent quality of frozen seafood

Effect of Delayed Icing on the Microbiological Quality of Hilsa Toli

Effect of delayed icing on the microbial quality and shelf-life of Hilsa toll was studied . Fish iced in rigor condition had a shelf-life of 11 days irrespective of the state of rigor . Fish procured from the landing centre had a shelf-life of only 8 days . It showed the presence of coagulase positive staphylococci, faecal streptococci and E. coli. Total bacterial count was low in all the samples and it increased after spoilage

Antimicrobial and Cytotoxic Activity of Fruit Extract from Syzygium cumini (L.) Skeels

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Prevalência e susceptibilidade antimicrobiana de Staphylococcusspp. em quadros de saúde e doença periodontal

The aim of this study was determine the prevalence and antimicrobial susceptibility of Staphylococcus spp. from patients with periodontal disease and periodontally healthy, correlate them with factors to host, local environment and traits of the diseases. To this, thirty adults from 19 to 55 years old were selected. They had not periodontal treatment and no antibiotic or antimicrobial was administered during three previous months. From these individuals, sites periodontally healthy, with chronic gingivitis and/or periodontitis were analyzed. Eighteen subgingival dental biofilm samples were collected through sterile paper points being six from each tooth randomly selected, representing conditions mentioned. They were transported to Oral Microbiology laboratory, plated onto Mannitol Salt Agar (MSA) and incubated at 370C in air for 48 h. Staphylococcus spp. were identified by colonial morphology, Gram stain, catalase reaction, susceptibility to bacitracin and coagulase activity. After identification, strains were submitted to the antibiotic susceptibility test with 12 antimicrobials, based on Kirby-Bauer technique. To establish the relation between coagulase-negative Staphylococcus (CSN) presence and their infection levels and host factors, local environment and traits of diseases were used Chi-square, Mann-Whitney and Kruskal-Wallis tests to a confidence level of 95%. 86,7% subjects harbored CSN in 11,7% periodontal sites. These prevalence were 12,1% in healthy sites, 11,7% in chronic gingivitis, 13,5% in slight chronic periodontitis, 6,75% in moderate chronic periodontitis and in sites with advance chronic periodontitis was not isolated CSN, without difference among them (p = 0,672). There was no significant difference to presence and infection levels of CSN as related to host factors, local environm ent and traits of the diseases. Amongst the 74 samples of CSN isolated, the biggest resistance was observed to penicillin (55,4%), erythromycin (32,4%), tetracycline (12,16%) and clindamycin (9,4%). 5,3% of the isolates were resistant to oxacilin and methicillin. No resistance was observed to ciprofloxacin, rifampicin and vancomycin. It was concluded that staphylococci are found in low numbers in healthy or sick periodontal sites in a similar ratio. However, a trend was observed to a reduction in staphylococci occurrence toward more advanced stages of the disease. This low prevalence was not related to any variables analyzed. Susceptibility profile to antibiotics demonstrates a raised resistance to penicillin and a low one to methicillin. To erythromycin, tetracycline and clindamycin was observed a significant resistance

Peritonitis in Latin America

Peritoneal dialysis has a high acceptance rate in Latin America, thus the knowledge concerning complication patterns is of great relevance. This work reviews Latin American data on peritonitis, the most serious complication of peritoneal dialysis.The incidence of peritonitis has been reduced over time, concomitantly with the incorporation of safer exchange systems and the use of prophylactic measurements. Today, rates tower than 1 episode per 24 patient-months are commonly reported. Furthermore, changes in causative organisms have been observed, with predominance of Staphylococcus aureus up through the mid-1990s, as welt as increases in coagulase-negative staphylococcus and participation of gram negatives. However, the prevalence of S. aureus is still high, due possibly to climatic conditions and the elevated prevalence of carriers. Resolution rate varies from 55% to 78%, transfer to hemodialysis from 10.9% to 15.4%, and death in 3% to 9.9% of cases. Outcome is worse in S. aureus episodes compared to those with coagulase-negative staphylococcus, despite the higher percentage of oxacillin-resistant strains among the former. In general, despite socioeconomic or climatic conditions, our results are similar to those in developed countries, perhaps as a consequence of technological improvements and/or center expertise.

Essential Oils Against Foodborne Pathogens and Spoilage Bacteria in Minced Meat

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Chemical Constituents of the Methanolic Extract of Leaves of Leiothrix spiralis Ruhland and Their Antimicrobial Activity

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Ação de antibióticos e quimioterápicos sobre alguns agentes baxterianos da mastite bovina, isolados da água utilizada no processo de obtenção do leite

Three strains of Staphylococcus aureus, 73 coagulase negative Staphylococcus and 28 Escherichia coli strains,isolated from water samples from 10 dairy farms, were tested for ''in vitro'' sensitivity to antibiotics and chemotherapeutics. The results showed that all samples isolated presented resistance to at least one active drug tested. The percentage of S. aureus, coagulase negative Staphylococcus and E. coli strains that exhibited resistance to the three active drugs were 100.00%, 84.93% and 71.43%, respectively. These results are important mainly due to the role of water as a vehicle for transmission of mastitis bacterial agents during the milking process.

Avaliação microbiológica da superfície dos implantes ortopédicos metálicos usados para fixar fraturas ósseas

In the majority of cases of bone fracture requiring surgery, orthopedic implants (screw-plate and screw) are used for osteosynthesis and the infections associated with such implants are due to the growth of microorganisms in biofilms. The objective of this study was to identify microorganisms recovered from osteosynthesis implants used to fix bone fractures, to assess the viability of the cells and the ability of staphylococci to adhere to a substrate and to determine their sensitivity/resistance to antimicrobials. After surgical removal, the metal parts of austenitic stainless steel (ASTM F138/F139 or ISO NBR 5832-1/9) were transported to the Laboratory of Clinical Microbiology, washed in buffer and subjected to ultrasonic bath at 40±2 kHz for 5 minutes. The sonicated fluid was used to seed solid culture media and cell viability was assessed under the microscope by with the aid of a fluorescent marker. The production of extracellular polysaccharide by Staphylococcus spp. was investigated by means of adhesion to a polystyrene plate. The profile of susceptibility to antimicrobials was determined by the disk diffusion assay. The most frequently isolated bacteria included coagulase-negative Staphylococcus resistant to erythromycin, clindamycin and oxacillin. Less frequent were Pseudomonas aeruginosa resistant to trimethoprim/sulfamethoxazole and ampicillin, Acinetobacter baumannii resistant to ceftazidime, Enterobacter cloacae resistant to cephalothin, cefoxitin, cefazolin, levofloxacin and ciprofloxacin, Bacillus spp. and Candida tropicalis. The observation of slides by fluorescence microscope showed clusters of living cells embedded in a transparent matrix. The test for adherence of coagulase-negative Staphylococcus to a polystyrene plate showed that these microorganisms produce extracellular polysaccharide. In conclusion, the metal parts were colonized by bacteria related to orthopedic implant infection, which were resistant to multiple antibiotics.