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PubMed2004

Anthrax in Wabessa village in the Dessie Zuria district of Ethiopia.

In 2002 an investigation of sudden death in a goat in Wabessa village in the Dessie Zuria district of Ethiopia was undertaken using fresh blood brought to the Kombolcha Regional Veterinary Laboratory. The sample was examined using standard bacteriological techniques and animal pathogenicity tests were also performed. The laboratory investigation revealed Bacillus anthracis as the cause of sudden death. Information gathered from stockowners in the same village revealed other similar recent cases and deaths, both in animals and humans, with farmers clearly describing the clinical signs and necropsy findings of anthrax. The disease occurs annually in this area in May and June, and in the 2002 outbreak mortality rates of 7.7%, 32.7% and 47.1% were observed in cattle, goats and donkeys, respectively. This study indicates that the community of this particular village neither knows of, nor practises, any of the conventional methods for anthrax control. The cutaneous form of the disease in humans and the environmental contamination associated with the practise of opening cadavers are briefly described and the findings are discussed with reference to the epidemiology of anthrax in both Ethiopia and elsewhere. Control strategies are also recommended.

Adult↗
PubMed2001

Molecular epidemiologic features and antimicrobial susceptibility profiles of various ribotypes of Pseudomonas aeruginosa isolated from humans and ruminants.

OBJECTIVES: To assess automated ribotyping for characterization of Pseudomonas aeruginosa isolates and to identify their type prevalence and geographic distribution. SAMPLE POPULATION: 39 human and 56 ruminant P aeruginosa isolates. PROCEDURES: Isolates were identified by use of bacteriologic techniques and automated Pvull-based ribotyping. Susceptibility to antimicrobials was tested in vitro. Data were analyzed for index of discrimination; prevalence ratio; geographic distribution of ribotypes found only in humans, only in cows, or only in goats (single-host ribotypes); and geographic distribution of ribotypes found in humans and ruminants (multihost ribotypes). RESULTS: All isolates were typeable (45 ribotypes, 35 single-host ribotypes). Ribotyping index of discrimination was 0.976. More isolates (45.3%) than expected yielded multihost ribotypes (22% of all ribotypes). Although 8.6% of single-host ribotypes were found in 4 or more isolates, 60% of multihost ribotypes were found in 4 or more isolates. Ninety percent of multihost ribotypes were isolated from different geographic areas, whereas 3.0% of single-host ribotypes were isolated from different geographic areas. All ruminant isolates were susceptible to gentamicin and polymyxin B. In contrast, antibiogram profiles differed for human isolates from different geographic areas. Susceptibility to antimicrobials differentiated 6 isolates not distinguished by ribotyping. CONCLUSIONS AND CLINICAL RELEVANCE: Automated ribotyping with Pvull discriminated more isolates than in vitro antimicrobial susceptibility. In combination, both tests provided more information than either test alone. Given the greater prevalence and geographic distribution of multihost ribotypes, immunocompromised humans and lactating ruminants may have a greater risk for disease if exposed to multihost P aeruginosa ribotypes, compared with single-host ribotypes.

Animals↗
PubMed1985

The effect of diet on the mammalian gut flora and its metabolic activities.

The review will encompass the following points: A brief introduction to the role of the gut flora in the toxicology of ingested food components, contaminants, and additives, including known pathways of activation and detoxication of foreign compounds and the implication of the flora in enterohepatic circulation of xenobiotics. The advantages and disadvantages of the various methods of studying the gut flora (classical bacteriological techniques, metabolic and enzymological methods) will be critically discussed with special reference to their relevance to dietary, toxicological, and biochemical studies. Sources of nutrients available to the gut flora will be described including host products (mucus, sloughed mucosal cells, hormones, proteins) and exogenous nutrients derived from diet. An account of the problems involved in studies of dietary modification with special reference to the use of stock laboratory animal diets, purified diets, and human dietary studies. The influence of dietary modification on the flora will be assessed on the basis of changes in numbers and types of bacteria and their metabolic activity, drawing on data from human and animal studies. The effects of manipulation of the quantity and quality of protein, fat, and indigestible residues (fiber) of the diet will be described together with their possible implications for toxicity of ingested compounds.

Amino Acids↗
PubMed1976

Microbiological aspects on peritonsillar abscesses.

In 37 patients with unilateral quinsy put was collected with a syringe technique and bacteriologically examined. Beta-hemolytic streptococci were isolated from 17 abscesses; in 8 of these, however, together with other bacteria, mainly anaerobes. Anaerobic bacteria, often more than one species, were found in 28 abscesses. Streptococcal serology including AST-O, Streptozyme and separate ADNAse test showed high titres or titre rises in 22 out of 30 examined cases. There was one case with beta-hemolytic streptococci in the abscess but negative serology. In the remaining cases a possible primary etiological role of anaerobes is suggested. The effect of combined surgical drainage and treatment with antibiotics (ampicillin or penicillin V) was good.

Bacteriological Techniques↗
PubMedDate not supplied

Urine culture collected from gel-based diapers: developing a novel experimental laboratory method.

BACKGROUND: Urinary tract infection diagnosis is based on urine culture, taken from a midstream collection. Obtaining samples in this manner is difficult in elderly patients suffering from incontinence and in infants. OBJECTIVES: (1) Develop a method for urine collection using gel-based diapers and (2) compare culture results from gel-based diapers with those of the same urine, examined by accepted methods. METHODS: Urine was collected and cultured by standard bacteriologic techniques at the microbiology laboratory of Soroka Medical Center, Beer-Sheva, Israel. Gel-based diapers were manually dampened with the same urine samples, and samples from diapers were cultured 0 to 3 hours after dampening. RESULTS: Comparison showed good correlation between direct urine cultures and cultures obtained from gel based diapers (R = 1.000). Total sensitivity was 100% and specificity 97%. CONCLUSIONS: (1) Urine can be cultured from gel-based diapers and (2) a larger sample of in vivo trials will be needed to make this technique clinically applicable in an outpatient setting.

Cell Culture Techniques↗
PubMed2003

The effects of increased intra-abdominal pressure on bacterial translocation.

In this study, we investigated the effect of different values of intra-abdominal pressure on bacterial translocation. Twenty-four Wistar-Albino rats were divided into four groups. The animals belonging to the Control group were not subjected to any increased intra-abdominal pressure. In groups I, II and III, an intra-abdominal pressure of 14, 20, and 25 mmHg, respectively, was established by carbon dioxide pneumoperitoneum for a period of 60 minutes. Four hours after the pneumoperitoneum, all animals were sacrificed to evaluate the degree of bacterial translocation at this time. Liver, spleen and mesenteric lymph nodes were excised under sterile conditions. Bacterial growth was assessed using standard bacteriological techniques and compared statistically. The Kruskal-Wallis and Mann-Whitney U tests were used for the statistical analysis. Different amounts of bacterial growth were found in all of the animals subjected to increased intra-abdominal pressure, except for the controls. Bacterial translocation was detected at an intra-abdominal pressure of 14 mmHg but this finding was not statistically significant (p > 0.05). There was a significant increase in bacterial growth in animals subjected to an intra- abdominal pressure of 20 mmHg or above (p < 0.001). As a result, we found that bacterial translocation started when the intra-abdominal pressure reached a level of 14 mmHg. Patients should be closely monitored for septic complication risks following laparoscopic procedures in which the intra-abdominal pressure exceeds 20 mmHg.

Abdomen↗
PubMed1999

Escherichia coli O157:H7 in retail ground beef in Seattle: results of a one-year prospective study.

Escherichia coli O157:H7 was sought systematically in 1,400 samples of retail ground beef in Seattle in a 1-year prospective study. Sorbitol-nonfermenting, lactose-fermenting, indole-positive colonies isolated after enrichment culture were probed for the presence of Shiga toxin genes. Totals of 67,040 sorbitol-nonfermenting and 66,705 sorbitol-fermenting colonies were characterized, but E. coli O157:H7 was not identified. The sensitivity of this technique was usually sufficient to detect E. coli O157:H7 at a concentration below 1 CFU/g of meat. These data demonstrate that retail ground beef in Seattle is neither frequently nor heavily contaminated with E. coli O157:H7.

Animals↗
PubMed2003

Improved detection of Salmonella spp. in foods by fluorescent in situ hybridization with 23S rRNA probes: a comparison with conventional culture methods.

This report describes a new technique for the detection and identification of Salmonella species in food with the use of fluorescent in situ hybridization (FISH) with 23S rRNA-targeted oligonucleotide probes. Two species-specific 23S rRNA-targeted oligonucleotide probes (Sal-1 and Sal-3) were selected, and one (Sal-544) was newly designed. The relative specificities of these probes were compared with those of bacterial 23S rRNA sequences from the GenBank database and tested by in situ hybridization with bacterial cell smears of pure cultures. Fifty-one tested reference strains of Salmonella serovars belonging to subspecies I (enterica) hybridized with these probes. No cross-reactions with 46 other strains of the family Enterobacteriaceae or with another 14 bacterial strains from other families were observed. Storage of a Salmonella Panama test strain under various environmental conditions (2, 5, and 15% NaC1; -20 degrees C, 4 degrees C, and room temperature; pHs of 3.3 to 7.4) did not adversely affect the FISH method. No matrix effects were observed with 18 different kinds of foods. FISH was able to detect Salmonella spp. in 52 (probe Sal-1), 56 (probe Sal-3), and 35 (probe Sal-544) of 225 naturally contaminated food samples after 16 h of incubation in a preenrichment broth. When conventional culture and detection methods were used, Salmonella could be isolated from only 30 of these 225 samples. In contrast, FISH failed to identify Salmonella in only two of the culture-positive samples when Sal-1 and Sal-3 were used and in only three of the culture-positive samples when Sal-544 was used.

Animals↗
PubMed1986

Nosocomial transmission of a strain of Staphylococcus aureus causing toxic shock syndrome.

A strain of Staphylococcus aureus producing toxic shock syndrome toxin-1 was repeatedly isolated from the nares of a neurosurgeon. This strain was identical to strains cultured from two of his patients who developed toxic shock syndrome after laminectomy. The relatedness of the isolates was shown by Southern blot hybridization analyses using chromosomal transposons as probes. This approach should be considered, in addition to standard bacteriologic techniques, as an effective method to analyze the relatedness of nosocomial isolates.

Adult↗
PubMed1999

Evaluation of the MicroScan system for identification of staphylococci.

Conventional biochemical tests were compared with reactions in a multiple test system, MicroScan Walkaway (Dade Diagnostic Inc. MicroScan Divison, West Sacramento, California) in conjugation with the Combo Pos ID Panels (Dade Diagnostic Inc. MicroScan Divison, West Sacramento, California), in order to evaluate the accuracy for the identification of 99 clinical isolates of Staphylococcus spp. and five reference strains. False-negative or positive reactions were detected from Voges-Proskauer, urease and mannose tests. A good correlation was found among the two identification systems for the fermentation of trehalose, lactose, raffinose, as well as for arginine dyhydrolase, esculin hydrolisis and nitrate reduction. From the results of the present study, it is concluded that the MicroScan Walkaway system is a reliable method for identification of staphylococci (94.23%), although 8.2% could be identified to the species level only after use of additional test.

Bacterial Typing Techniques↗
PubMed2000

Characterization of a highly glycopeptide-resistant Enterococcus gallinarum isolate.

BACKGROUND AND PURPOSE: Adequate treatment of emergency infection involving antibiotic-resistant bacteria such as vancomycin-resistant Enterococcus requires a convergence of clinical and bacteriologic techniques. An isolate of Enterococcus gallinarum, designated as TSGH63, is known to be uncommonly vancomycin-resistant. This study investigated the genetic determinant for this unique characteristic. METHODS: After completing the conventional identification and sensitivity tests, the genomic content of E. gallinarum TSGH63 was extracted and analyzed by pulse-field electrophoresis. A set of specific primers for vanA, vanB, vanC1, and vanC2/C3 genes was then applied in a multiplex polymerase chain reaction (PCR) to differentiate its genetic content. To locate the determinant for high vancomycin resistance, the electrophoresis profile was further analyzed by Southern blot using the digoxigenin (DIG)-labeled vanA gene probe. Finally, interspecies transfer of the vancomycin-resistance determinant of E. gallinarum TSGH63 was tested by a conjugation experiment in vitro. RESULTS: A 50-kb plasmid was identified in the analysis of the genomic extract of E. gallinarum TSGH63 by pulse field electrophoresis. Using multiplex PCR, we demonstrated that E. gallinarum TSGH63 harbors a vanA gene in addition to a vanC1 gene. The DIG-labeled vanA gene-specific probe bound to the plasmid exclusively on the Southern blot. The plasmid-carried vanA gene, but not the vanC1 gene, was found to be transferable from TSGH63 to E. faecalis JH2-2 by conjugation in vitro. CONCLUSIONS: This is the first report of isolation of E. gallinarum with a high level of resistance to glycopeptides in Taiwan. The demonstrated interspecies transfer of the vancomycin-resistance gene highlights the importance of stringent control of the use of vancomycin.

Conjugation, Genetic↗
PubMed1998

[Pediatric Haemophilus influenzae type b meninngitis in Hefei city: an epidemiologic study].

OBJECTIVE: To obtain epidemiologic information about Hib meningitis from Hefei. METHODS: Data were collected from 1990 to 1992 by a coordinative group including 13 hospitals. All children with a clinical diagnosis of acute bacterial meningitis were enrolled and specimens were taken for etiologic studies. CSF and blood were tested by standard bacteriologic technique. CSF, blood and concentrated urine were tested directly for detecting antigen by CIE. The data were analysed by epidemiologic methods. RESULTS: Of 60 cases of bacterial meningitis clinically diagnosed, 31 (51.7%) were CIE positive for Hib antigen. Only 3 cases of CSF culture were positive for Hib. The annual incidence of Hib meningitis in Hefei was culculated as 4.81/100,000 children younger than 15 years and 10.66/100,000 children younger than 5 years. The case fatality rate was 9.7%. 21.4% of survivors suffered from neurological or psychological problems. CONCLUSIONS: Using antigen detection combined with bacterial culture, we could make etiologic diagnosis in 90% of the cases. Hib is the most common cause of bacterial meningitis, but the incidence is much lower than in most parts of the world.

Adolescent↗
PubMed1999

Isolation and typing methods for the epidemiologic investigation of thermotolerant campylobacters.

Thermotolerant campylobacters, C. jejuni, C. coli, C. lari and C. upsaliensis, are spiral bacteria involved in human enteric disease. The prevalence of these emerging pathogens, mainly C. jejuni and to a lesser extent C. coli, as etiologic agents of enteric disease in industrialized countries has increased over the last decade. The isolation and culture of these microorganisms is tedious and time-consuming mainly due to their complex nutritional and environmental requirements. This review discusses the techniques and methods developed for the selective isolation of thermotolerant campylobacters from food, environmental and clinical samples. Additionally, both traditional and newer molecular biology techniques applied to this group of thermophilic organisms for typing and taxonomic purposes are summarized.

AIDS-Related Opportunistic Infections↗
PubMed2000

Identification of Vibrio spp. (other than V. vulnificus) recovered on CPC agar from marine natural samples.

Two hundred and eighty four presumptive but not confirmed Vibrio vulnificus isolates grown on cellobiose-polymixin B-colistin agar (CPC) at 40 degrees C, recovered from sea water samples from Valencia, Spain, during a microbiological survey for V. vulnificus, were phenotypically identified. Most of the isolates (91%) corresponded to Vibrio species. V. harveyi (24%) and V. splendidus(19%) were the most abundant species identified, followed by V. navarrensis (13%), V. alginolyticus (8%) and V. parahaemolyticus (5%). The ability to grow on CPC agar and ferment cellobiose of several V. vulnificus strains from different origins and serovars, including reference strains, was tested. Most serovar E isolates and 25% of non-serovar E isolates could not grow on CPC agar.

Agar↗
PubMed2000

Throat carriage of pneumococci in healthy school children in the Union Territory of Pondicherry.

BACKGROUND & OBJECTIVES: Colonisation of Streptococcus pneumoniae in the throat is common among children the world over. Little is known about the relationship of nasopharyngeal carriage and invasive disease or the way it spreads within the households and close confines. There is a paucity of data on the colonization of Strep. pneumoniae in the throat of healthy children in India. To determine the prevalence of pneumococcal carriage in school children of urban and rural Pondicherry, a study was undertaken. METHODS: Throat swabs of healthy school-going children between 5-10 yr of age were examined for pneumococcal carriage, by standard bacteriological techniques. RESULTS: A prevalence rate of 24.3 per cent was noted. There was no difference in the carriage rate among the rural children when compared to urban children. No age, sex or geographical predilection of pneumococcal carriage was noted. A statistically significant seasonal variation, however, was seen. Carriage rate increased during the colder months and was found to be the highest in the months of March and November. INTERPRETATION & CONCLUSIONS: Strep. pneumoniae circulates in the community among healthy children. Carriage rate is influenced by seasonal variation.

Carrier State↗
PubMedDate not supplied

[Multiplication of Listeria in milk products].

The results of the evaluation of the multiplication dynamics of Listeria cells in milk and Bifidok, a lactic acid product, are presented. The samples were inoculated on thioglycol agar and studied at different exposure time after incubation at 37 degrees C, 20 degrees C and 4 degrees C. The study revealed the intensive multiplication of Listeria cells in milk, also during storage in a household refrigerator. The presence of bifidobacteria mixed with kefir-producing culture in dairy products was shown to essentially inhibit the growth of Listeria cells which were not detected by bacteriological techniques on day 7.

Bifidobacterium↗