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PubMed1996

Microbiological analysis of 'black pudding', a Trinidadian delicacy and health risk to consumers.

The prevalences of Salmonella, Escherichia coli and Staphylococcus aureus in black pudding which originated from local vendors and supermarkets in Trinidad were determined. The enterotoxigenicity of S. aureus strains and occurrence of O157:H7 strains amongst E. coli isolates were also investigated. For the 100 black puddings each sampled from supermarkets and vendors, the mean total aerobic plate count (TAPC) per g was 1.8 x 10(7) +/- 1.5 x 10(7) and 1.5 x 10(8) +/- 2.3 x 10(8), respectively. E. coli was isolated from 56 (56.0%) black pudding samples from supermarkets with a mean count per g of 9.2 x 10(6) +/- 7.9 x 10(6) compared to a prevalence of 79% (79 of 100) and mean count per g of 3.2 x 10(7) +/- 4.7 x 10(7) for samples from local vendors. The difference between prevalences was statistically significant (P < or = 0.001; chi 2). Only 1 (2.2%) of 45 strains of E. coli from supermarket-purchased pudding tested, was an 0157:H7 strain compared to 9 (13.6%) of 66 strains of E. coli from vendor-sold black pudding. The difference was not statistically significant (P > or = 0.05; chi 2). Five (5.0%) of 100 black pudding samples from supermarkets yielded Salmonella, with S. ohio being the predominant serotype. For vendor-sold black pudding however, 11 (11.0%) samples were positive for Salmonella with a new serotype, S. unnamed (4,12:d-) being responsible for 50% (6 of 12) of isolates from this source. Forty samples each of black pudding from supermarkets and vendors were all (100.0%) positive for S. aureus with mean counts per g being 3.1 x 10(5) +/- 8.8 x 10(5) and 3.3 x 10(6) +/- 7.7 x 10(6), respectively. Overall, 27 (33.8%) of 80 strains of S. aureus tested were enterotoxigenic producing staphylococcal enterotoxins A(SEA), SEB, SEC, SED or a combination. It was concluded that black pudding poses a high risk to consumers based on the prevalence, microbial load and toxigenicity of the pathogens detected.

Culture Media↗

PubMed1995

The use of immunological methods to detect and identify bacteria in the environment.

Immunological detection methods have become increasingly important in microbial ecology for the tracking of specific microorganisms and for community analysis. For a reliable application of these techniques, the monoclonal antibodies or polyclonal antisera used have to fulfill several quality criteria. Cross reactivity, cellular localization of the antigenic determinant, affinity characteristics and the expression of the antigenic determinant at environmental conditions have to be determined. Immunological methods can be used for the identification, quantification and enrichment of specific bacteria in extracts as well as for the visualization of cells in situ. The sensitivity of advanced immunological methods can be compared to PCR techniques. Using image processing of epifluorescence micrographs or confocal laser scanning microscopy, the immunofluorescence approach can now be applied to study complex environmental samples.

Journal Article↗

PubMed2005

Inflammation and atherosclerosis.

PURPOSE: The aim of this article is to discuss the role of inflammation in atherosclerosis. SUMMARY: An initial chemical, mechanical or immunological insult induces endothelial dysfunction. This triggers a cascade of inflammatory reactions, in which monocytes, macrophages, T lymphocytes and vascular smooth muscle cells participate. Leukocyte adhesion molecules, cytokines, growth factors and metalloproteinases participate in all stages of atherogenesis. Almost all of the traditional risk factors for atherosclerosis are associated with and participate in the inflammatory process. Many infectious agents, mainly Chlamydia pneumoniae, have been proposed as potential triggers of the cascade. The immune system has been implicated in plaque formation, through the activation of cellular and humoral immunity against innate or microbial heat shock protein 60. Methods of detection of systemic or local plaque inflammation have been developed and research is being conducted on the potential use of anti-inflammatory and antibiotic drugs in atherosclerosis.

Atherosclerosis↗

PubMed2002

A multivariate statistical analysis to follow the course of disease after infection of cats with different strains of the feline immunodeficiency virus (FIV).

A descriptive multivariate assay is described which is suitable to analyze results of a biological experiment with small sample size but high qualitative and quantitative complexity of variables. This type of assay allows evaluation of multiple variables observed in the course of an experimental virus infection (e.g. viremia, nucleic acid detection, antibody titers, clinical parameters, anti-microbial treatments or vaccination) in a single graph. In our study, a multiple correspondence analysis (MCA) was used to correlate a total of 145 measurements from each of a dozen of variables measured in five groups of three cats infected by five isolates of feline immunodeficiency virus (FIV). Three groups of virus isolates with distinct virulence were defined and correlation between dynamics of lymphocyte subset counts and viral virulence was established. Comparison between the primary stages of illness and follow-up examinations were of prognostic value and are thus helpful for development and monitoring of therapeutic strategies.

Acute Disease↗

PubMed1997

Microbial production of a novel trihydroxy unsaturated fatty acid from linoleic acid.

A bacterium isolated from a dry soil sample collected from McCalla, AL, USA, converted linoleic acid to a novel compound, 12,13,17-trihydroxy-9 (Z)-octadecenoic acid (THOA). The organism is a Gram-positive, non-motile rod (0.5 microns x 2 microns). It was identified as a species of Clavibacter ALA2. The product was purified by high pressure liquid chromatography, and its structure was determined by 1H and 13C nuclear magnetic resonance and Fourier transform infrared spectroscopies, and by mass spectrometer. Maximum production of THOA with 25% conversion of the substrate was reached after 5-6 days of reaction. THOA was not further metabolized by strain ALA2. This is the first report of a 12,13,17-trihydroxy unsaturated fatty acid and its production by microbial transformation. Some dihydroxy intermediates were also detected. THOA has a structure similar to those of known plant self-defense substances.

Bacteria↗

PubMed2000

Evolution of a degradative bacterial consortium during the enrichment of naphtha solvent.

A microbial mixed culture able to degrade naphtha solvent, a model of hydrocarbon aromatic mixture, was isolated from a hydrocarbon-polluted soil. Composition of the population was monitored by phenotypic and molecular methods applied on soil DNA, on whole enrichment culture DNA, and on 85 isolated strains. Strains were characterized for their 16S rDNA restriction profiles and for their random amplified polymorphic DNA profiles. Catabolic capabilities were monitored by phenotypic traits and by PCR assays for the presence of the catabolic genes methyl mono-oxygenase ( xylA, M), catechol 2,3 dioxygenase (xylE) and toluene dioxygenase (todC1) of TOL and TOD pathways. Different haplotypes belonging to Pseudomonas putida, Ps. aureofaciens and Ps. aeruginosa were found to degrade aromatic compounds and naphtha solvent. The intrinsic catabolic activity of the microbial population of the polluted site was detected by PCR amplification of the xylE gene directly from soil DNA.

Bacteria↗

PubMed2004

Toll-like receptor 2 and Toll-like receptor 4 expression in human adrenals.

Toll-like receptors (TLRs) are key elements in the innate immune response, functioning as pattern-recognition receptors for the detection and response to endotoxins and other microbial ligands. Inflammatory cytokines play an important role in the activation of the hypothalamic-pituitary-adrenal HPA axis during inflammation and sepsis. The newly recognized major role of TLR2 and TLR4 and the adrenal stress response during critical illnesses such as inflammation and sepsis demand comprehensive analysis of their interactions. Therefore, we analyzed TLR2 and TLR4 expression in human adrenal glands. Western blot analysis demonstrated the expression of TLR2 and TLR4 in the human adrenocortical cell line NCI-H295. Immunohistochemical analysis of normal human adrenal glands revealed TLR2 and TLR4 expression in the adrenal cortex, but not in the adrenal medulla. Considering the crucial role of the HPA axis and the innate immune response during acute sepsis or septic shock, elucidating the functional interaction of these systems should be of great clinical relevance.

Adrenal Cortex↗

PubMed2002

Transcriptional modulation of bacterial gene expression by subinhibitory concentrations of antibiotics.

Antibiotics such as erythromycin and rifampicin, at low concentrations, alter global bacterial transcription patterns as measured by the stimulation or inhibition of a variety of promoter-lux reporter constructs in a Salmonella typhimurium library. Analysis of a 6,500-clone library indicated that as many as 5% of the promoters may be affected, comprising genes for a variety of functions, as well as a significant fraction of genes with no known function. Studies of a selection of the reporter clones showed that stimulation varied depending on the nature of the antibiotic, the promoter, and what culture medium was used; the response differed on solid as compared with liquid media. Transcription was markedly reduced in antibiotic-resistant hosts, but the presence of mutations deficient in stress responses such as SOS or universal stress did not prevent antibiotic-induced modulation. The results show that small molecules may have contrasting effects on bacteria depending on their concentration: either the modulation of bacterial metabolism by altering transcription patterns or the inhibition of growth by the inhibition of specific target functions. Both activities could play important roles in the regulation of microbial communities. These studies indicate that the detection of pharmaceutically useful natural product inhibitors could be effectively achieved by measuring activation of transcription at low concentrations in high-throughput assays using appropriate bacterial promoter-reporter constructs.

Anti-Bacterial Agents↗

PubMed1992

Characterization and growth response of bacteria in soil following application of carbofuran.

Enhanced biodegradation of carbofuran (2, 3-dihydro-2, 2 dimethyl-7-benzofuranyl methyl carbamate) is an economically significant, but poorly understood, microbial phenomenon in soil. A series of experiments was conducted to examine short term changes in soil bacterial populations stimulated by carbofuran application at field rates. In the field experiment, commercially formulated carbofuran and butylate (S-ethyl diisobutyl carbamothioate) were applied at 5.6 kg ai ha-1 and 8.4 kg ai ha-1, respectively, on a soil (Putnam silt loam) exhibiting enhanced degradation of carbofuran. In laboratory studies, technical grade carbofuran (20 mg kg-1 soil) was applied to samples of the field soil. Bacterial populations were estimated using non-selective (tryptic soy agar) and selective media containing carbofuran or butylate. Largest population increases in pesticide-treated soil were observed between 7 and 15 days after treatment (DAT) compared to populations in non-treated soil. Significant increases (P less than 0.05) in total bacterial populations and presumed carbofuran-degraders due to carbofuran application were associated with increased populations of Pseudomonas spp. and Flavobacterium spp. Application of carbofuran appeared to provide a competitive advantage to these species over actinomycetes persisting beyond 20 DAT. Growth responses of bacteria to carbofuran in the Putnam soil were compared to those in a native prairie soil (Mexico silt loam), which exhibited a much slower rate of carbofuran degradation. Bacterial population response to carbofuran was measurable, but small and short-lived. Perpetuation of the enhanced degradation phenomenon may lie in a persistent pesticide-induced competitive advantage given to a very small segment of the microbial population. This advantage may not be detectable after 20 days using conventional plating techniques.

Bacteria↗

PubMed2006

Effect of dry honey on the shelf life of packaged turkey slices.

The development of off-flavors from oxidation reactions in cooked turkey products is a common problem and results in a less desirable, rancid flavor. Various strategies have been evaluated to minimize this off-flavor development, including vacuum and modified atmosphere packaging, feeding antioxidants to animals, and use of antioxidants in the final product. A natural protein-sugar reaction called the Maillard reaction produces a brown pigment, flavors, and antioxidants. This research tested the addition of honey to turkey breast meat before processing to retard production of oxidation products related to off-flavor. Three levels (0, 5, 15%) of dry honey were mixed with raw turkey breast meat pieces, then the mixture was stuffed into casing and cooked. The cooking process facilitated the Maillard reaction and the development of an antioxidative effect. The cooked chubs were then cooled, sliced, and vacuum-packaged as individual slices. The slices were refrigerated and tested for color, flavor, oxidative rancidity, and microbial growth over 11 wk. Sensory panelists detected increased sweetness and no negative flavor impact on acceptability for turkey with added honey. The addition of honey enhanced the oxidative stability of the meat, as indicated by lower TBA values, hexanal content, and oxidative stability index. Honey did impart a slightly darker color with lower lightness values but had no effect of redness and yellowness values.

Animals↗

PubMed2000

Diagnostic particle agglutination using ultrasound: a new technology to rejuvenate old microbiological methods.

Microbial antigen in clinical specimens can be detected rapidly by commercial test-card latex agglutination, but poor sensitivity is a potential difficulty. Antigen detection by immuno-agglutination of coated latex micro-particles can be enhanced in comparison with the conventional test-card method in both rate and sensitivity by the application of a non-cavitating ultrasonic standing wave. Antibody-coated micro-particles suspended in the acoustic field are subjected to physical forces that promote the formation of agglutinates by increasing particle-particle contact. This report reviews the application of ultrasound to immuno-agglutination testing with several commercial antibody-coated diagnostic micro-particles. This technique is more sensitive than commercial card-based agglutination tests by a factor of up to 500 for fungal cell-wall antigen, 64 for bacterial polysaccharide and 16 for viral antigen (in buffer). The detection sensitivity of meningococcal capsular polysaccharide in patient serum or CSF has been increased to a stage where serotyping by ultrasound-enhanced agglutination is comparable to that achievable with the PCR, but is available more rapidly. Serum antigen concentration as measured by ultrasonic agglutination has prognostic value. Increasing the sensitivity of antigen detection by increasing the acoustic forces that act on suspended particles is considered. Employing turbidimetry to measure agglutination as part of an integrated ultrasonic system would enable the turnover of large numbers of specimens. Ultrasound-enhanced latex agglutination offers a rapid, economical alternative to molecular diagnostic methods and may be useful in situations where microbiological and molecular methods are impracticable.

Agglutination Tests↗

PubMed2002

A strategy to retrieve the whole set of protein modules in microbial proteomes.

Protein homology is often limited to long structural segments that we have previously called modules. We describe here a suite of programs used to catalog the whole set of modules present in microbial proteomes. First, the Darwin AllAll program detects homologous segments using thresholds for evolutionary distance and alignment length, and another program classifies these modules. After assembling these homologous modules in families, we further group families which are related by a chain of neighboring unrelated homologous modules. With the automatic analysis of these groups of families sharing homologous modules in independent multimodular proteins, one can split into their component parts many fused modules and/or deduce by logic more distant modules. All detected and inferred modules are reassembled in refined families. These two last steps are made by a unique program. Eventually, the soundness of the data obtained by this experimental approach is checked using independent tests. To illustrate this modular approach, we compared four proteobacterial proteomes (Campylobacter jejuni, Escherichia coli, Haemophilus influenzae, and Helicobacter pylori). It appears that this method might retrieve from present-day proteins many of the modules which can help to trace back ancient events of gene duplication and/or fusion.

Bacterial Proteins↗

PubMed2006

In vitro calculus detection with a moved smart ultrasonic device.

STUDY OBJECTIVE: The objective of subgingival instrumentation of periodontally diseased root surfaces is to remove the adhering microbial biofilm and calcified deposits. Recently, we have described an automated calculus detection system under static conditions. Clinically however, the tip of the system has to be moved over tooth surfaces. It was thus necessary to study the entire system in motion. METHODS: The detection device is based on a conventional dental piezoelectric ultrasonic handpiece with a conventional scaler insert. The impulse response of the mechanical oscillation system is analysed by a fuzzy logic-based computerized algorithm, which classifies various surfaces. The present study investigates dental surface recognition properties of the new system with the tip being moved over teeth surfaces in vitro. Following a training set of 7977 measurements (3960 calculus, 4017 cement) on 200 extracted teeth, 1363 measurements were conducted on 34 teeth unknown to the system. RESULTS: The surfaces cementum and calculus were correctly classified in 78% within the training set and in 81% within the set unknown, with a kappa value of 0.68. CONCLUSION: It was shown that this method of automatic recognition of tooth surfaces is able to distinguish between different tooth surfaces in vitro independently from tip movements.

Dental Calculus↗

PubMed1970

Limits of microbial existence: temperature and pH.

A microscopic survey made to detect the presence of bacteria in hot springs of varying temperature and pH characteristics revealed that in neutral and alkaline hot springs bacteria are found at temperatures up to the boiling point of water (92 degrees to 100 degrees C, depending on the altitude). In hot springs of increasing acidity the upper temperature limit at which bacteria are found decreases; at pH 2 to 3 the upper temperature limit is 75 degrees to 80 degrees C. Bacteria have thus been able to evolve with the ability to grow at either high temperature or high acidity, but not at both high temperature and high acidity. These results suggest that there are physicochemical limitations of the environment beyond which life is impossible.

Bacteria↗

PubMed1981

Scanning electron microscope evidence for bacterial colonization of a drinking-water distribution system.

The surfaces of water distribution mains and suspended particulate matter from drinking water were examined by using scanning electron microscopy to investigate the nature and extent of association of microorganisms with these surfaces. In addition, X-ray energy-dispersive microanalysis was used to determine the elemental constitution of the pipe surface. Though distributed sparsely and randomly along the pipe surface, a variety of morphologically distinguishable bacteria-like structures and microcolonies were observed. The morphologies of the individual cells varied form chain-forming cocci to filamentous and prosthecate cell types. The iron-oxidizing bacterium Gallionella, recognized by its characteristic helical stalks, was observed both in water samples and attached to pipe surfaces. Attachment of some microbes to the pipe surface was apparently mediated by extracellular fibrillar appendages. Large numbers of rod-shaped bacteria were also evident adhering to the surfaces of suspended detritus or silt particles recovered from water samples by filtration. X-ray energy scans of the pipe surface revealed the presence of five major elemental constituents including silicon, phosphorous, sulfur, calcium, and iron. Smaller quantities of the elements zinc, magnesium, aluminum, potassium, and manganese were also detected. The public health significance of sessile microbial communities in drinking-water distribution systems is discussed.

Bacteria↗

PubMed1997

Emerging issues in microbiological food safety.

Many microorganisms previously unrecognized as food-borne or harmful are emerging as human pathogens transmitted by food. This is a result of recent acquisition of key virulence factors, detection by newly developed isolation procedures, or astute detective-like laboratory skills of microbiologists. Six microbial pathogens, including Shiga toxin-producing Escherichia coli, Listeria monocytogenes, Arcobacter butzleri, Helicobacter pylori, Cryptosporidium parvum, and Cyclospora, have become recognized as significant causes of human illness. Although the ecology and epidemiology of illness caused by some of these pathogens have not been fully elucidated, food has the potential of being an important vehicle in their dissemination. Existing technologies and new approaches such as irradiation and hazard analysis critical control point (HACCP) programs are useful tools in the control of food-borne hazards. However, because of ever-changing products, processes, food-handling practices, societal habits, and pathogens, emerging food-borne diseases will continue to be an important public health concern.

Campylobacter Infections↗

PubMed2001

Sexual and enteric bacterial infections eliciting reactive arthritis.

In this review synonymous definitions of reactive arthritis are discussed first. Major clinical symptoms, their infectious etiology and epidemiology define post-dysenteric and post-veneral forms of reactive arthritis. Classical (smear, culture, biochemistry, antigen detection) and molecular (DNA and RNA detections) techniques are used in the routine microbial diagnosis that is retrospective in the majority of cases. In the pathomechanism of this disorder, HLA-B27 antigen positivity of patients is a frequent risk factor. Molecular mimicry between microbial and self-antigens, abnormal antigen presentation leading to incomplete CD8+ T lymphocyte activation might contribute to the persistence of microbial antigens that elicit clinical symptoms. Treatment is rarely successful with antimicrobial chemotherapy.

Anti-Infective Agents↗

PubMedDate not supplied

Sequence-based source tracking of Escherichia coli based on genetic diversity of beta-glucuronidase.

High levels of fecal bacteria are a concern for recreational waters; however, the source of contamination is often unknown. This study investigated whether direct sequencing of a bacterial gene could be utilized for detecting genetic differences between bacterial strains for microbial source tracking. A 525-nucleotide segment of the gene for beta-glucuronidase (uidA) was sequenced in 941 Escherichia coli isolates from the Clinton River-Lake St. Clair watershed, 182 E. coli isolates from human and animal feces, and 34 E. coli isolates from a combined sewer. Environmental isolates exhibited 114 alleles in 11 groups on a genetic tree. Frequency of strains from different genetic groups differed significantly (p < 0.03) between upstream reaches (Bear Creek-Red Run), downstream reaches, and Lake St. Clair beaches. Fecal E. coli uidA sequences exhibited 81 alleles that overlapped with the environmental set. An algorithm to assign alleles to different host sources averaged approximately 75% correct classification with the fecal data set. Using the same algorithm, the percent of environmental isolates assignable to humans decreased significantly between Bear Creek-Red Run (30 +/- 3%) and the beaches (17 +/- 2%) (p < 0.05). Birds accounted for approximately 50% of assignable environmental isolates. For combined sewer isolates, the same algorithm assigned 51% to humans. These experiments demonstrate differences in the frequency of different E. coli strains at different locations in a watershed, and provide a "proof in principle" that sequence-based data can be used for microbial source tracking.

DNA, Bacterial↗