Triple Sugar Iron Research Articles

Antimicrobial Resistance Pattern of <i>Shigella</i> species Over Five Years at a Tertiary-care Teaching Hospital in North India

Sir, Shigellosis is a global human health problem, especially in developing countries where there are inadequate hygiene and unsafe water supplies (1,2). Shigella accounts for a significant proportion of cases of bacillary dysentery in many tropical and subtropical countries (3,4). Shigella is one of the most important causes of gastroenteritis-induced deaths in 3-5 million children aged less than five years in developing countries (5). The emergence of multiple drug resistance to cost-effective antimicrobials against Shigella is a matter of concern in developing countries (6). It is also well-documented that pandemic strains often exhibit multiple antimicrobial resistance and induce severe illness with high case fatality in all age-groups (7). In the tropics, most infections occur due to S. flexneri and infections that occur primarily due to Shigella sonnei are less common (8,9). Over a decade ago, results of active surveillance studies showed that, in most endemic countries, especially in Asia and sub-Saharan Africa, there was an emergence of multidrug resistance to different antimicrobials, including ampicillin, co-trimoxazole, and nalidixic acid (9). Besides the temporal changes in the antibiogram of Shigella species, it is well-known that antimicrobial susceptibility patterns in Shigella may differ between geographical areas. Such differences are never stable and may change rapidly, especially in places where antimicrobials are used excessively, particularly in developing countries (1). This warrants for robust monitoring of the pattern of antibiogram for this organism. Hence, the present study was conducted to determine the incidence and prevalence of Shigella serogroups isolated from cases of dysentery during a five-year period from 2004 to 2008 and also to determine the drug resistance pattern of Shigella in a tertiary-care hospital in North India. In this cross-sectional study, patients presenting with acute, bloody diarrhoea, suggestive of shigellosis, during 2004-2008, were included. Stool samples were collected from all patients presenting with symptoms of diarrhoea, blood or mucus in stool, fever, and abdominal pain. In total, 12,983 stool samples were received for culture and routine microscopy during 2004-2008. All the samples were subjected to routine microscopy and were examined for pus cells, trophozoites, and cysts. Only those samples which showed pus cells and/or red blood cells were cultured. The stool specimens from such symptomatic patients were cultured on differential and selective media, including MacConkey agar, Xylose-lysine desoxycholate agar, and deoxycholate citrate agar as primary plates. Loop-full sample was also inoculated into selenite-F broth. The plates were incubated at 37 [degrees]C for 24 hours. Suspected colonies on solid plates, which were Gram-negative upon Gram-staining and oxidase test-negative, were biochemically characterized as per standard procedures. Presumptive Shigella isolates on Triple Sugar Iron agar were serotyped by slide agglutination tests with Shigella polyvalent grouping and monovalent antisera (Denka Seiken Co. Ltd., Tokyo, Japan). Antimicrobial susceptibility testing and resistance patterns of the Shigella isolates to various antimicrobials were determined by the disc-diffusion technique (10). Every inoculum was prepared by inoculating 5 mL of Mueller-Hinton broth with five colonies of an 18-hour old pure Shigella culture, followed by incubation in ambient air and at 37 [degrees]C for 16 hours. Turbidity of the broth was equilibrated to match with 0.5 McFarland standards. A sterile cotton swab was dipped into the standardized suspension, drained, and used for inoculating 25 mL of Mueller-Hinton agar in a 90-mm plate. The inoculating plates were air-dried, and antimicrobial discs, included ampicillin (10 [micro]g), chloramphenicol (30 [micro]g), co-trimoxazole (25 [micro]g), tetracycline (30 [micro]g), gentamicin (10 [micro]g), nalidixic acid (30 [micro]g), and ciprofloxacin (5 [micro]g). …

SALMONELLA TYPHIMURIUM EM LINFONODOS MESENTÉRICOS DE OVINOS AO ABATE

RESUMO O rebanho de ovinos no Brasil está estimado em mais de 16 milhões de cabeças. Embora o consumo da carne desta espécie ainda seja pequeno, comparado ao de outros países, o consumo de carne, inclusive ovina, tem sido associado às doenças transmitidas por alimentos, em especial a salmonelose. No presente estudo, investigou-se a ocorrência de salmonelas em linfonodos mesentéricos e conteúdo intestinal de 175 ovinos ao abate. “Pools” constituído por cinco amostras de contéudo fecal ou 5 amostras de linfonodos de 25 g foram pre-enriquecidos em 250 mL de água peptonada tamponada e incubados a 37° C por 18-24 horas. Uma alíquota de 0,1 mL do pré-enriquecimento foi transferida para 9,9 mL de caldo de enriquecimento Rappaport-Vassiliadis e 1,0 mL do pré-enriquecimento foi transferido para 10 mL de caldo tetrationato Muller-Kaufmann, incubados a 42° C for 24h. 10 μL do caldo de enriquecimento foi semeado superfície de placas de ágar BPLS e ágar XLT4 incubadas a 37º C for 24-48h. Colônias suspeitas de salmonela foram testadas por provas bioquímicas e serologicas. Os testes bioquímicos utilizados para identificação de Salmonella foram TSI (triple sugar iron àgar), LIA (lysine iron àgar) e ágar ureia. Sorotipagem foi realizada no Laboratório de Enterobactérias do Instituto Osvaldo Cruz. Isolou-se Salmonella Tiphymurium de um pool de linfonodos mesentéricos, provenientes de cinco animais. O fato de se observar a ocorrência de salmonela em ovino portador sadio alerta para necessidade de monitorar este micro-organismo também nesta espécie, especialmente quando destinada ao abate, com vistas à produção de alimentos seguros.

ISOLATION AND BIOCHEMICALCHARACTERIZATION OF SALMONELLA & E.COLI FROM BOVINE MILK COLLECTED FROM SALE SHOPS, GOVERNAMENTAL & PRIVATE DAIRY FARMS AT QUETTA PAKISTAN

A total of 100 raw milk samples of bovine were collected in sterile screw capped tubes(each 5ml) from Government Dairy Farm(45 samples) , Private Dairy Farms (45 samples) and Private Milk Shops(10 samples) from different locations in Quetta city to isolate milk borne microbial pathogens like Salmonella and E Coli to estimate their share in the environmental pollution. All these samples were subjected to grow on Salmonella Shigella agar (SS Agar) and Xylose Lysine Deoxycholate (XLD) agar. The isolated colonies were further identified biochemically through triple sugar iron test (TSI) and Gram’s staining. From Government Dairy Farm out of forty five (45) raw milk samples six (6) samples (13%) were found positive for E Coli whereas no salmonella (0%) colony was detected. While from Private Dairy Farms out of Forty Five (45) samples ten(10) were positive for E Coli(22%) and only one(1) sample was positive(2%) for Salmonella. From private milk shops out of ten (10) samples four (4) were positive for (40%) E coli and three (3) for salmonella (30%). It is concluded from the above study that there is 4% Salmonella and 20% E.coli contamination in raw milk samples being supplied to the people in Quetta city. The high proportion of contamination (30%) of Salmonella and (40%) of E.coli was detected from Private Milk Shops due to Exposure of milk to open Environment where these both pathogens are present. which is a serious public health issue and need to create a public awareness to use treated (Pasteurized) milk and proper disposal of the waste from dairy farms as well as shops and as well as to avoid exposure of milk to open contaminated Environment of the city.

Characterisation of <i>Shigella</i> species isolated from river catchments in the North West province of South Africa

The occurrence and distribution of Shigella species in water from the five river catchments in the North West province of South Africa were investigated. Shigella is a Gram-negative, non-motile, facultative anaerobic bacillus that causes shigellosis, an important cause of morbidity and mortality in high-risk populations (such as children, the elderly and immuno-compromised individuals) that depend on river water. A total of 54 water samples collected in winter (April 2007 to July 2007) and summer (December 2007 to March 2008) were cultured on Salmonella-Shigella agar by the spread-plate method. Suspected Shigella isolates obtained were characterised by primary biochemical (Triple Sugar Iron agar and agglutination) and molecular (polymerase chain reactions, PCR) tests. Amplification of the invasion plasmid gene ( ipaH ) by PCR was done to confirm the presence of Shigella spp. in water. In total, 214 Shigella boydii , 15 Shigella dysenteriae , 11 Shigella flexneri and 2 Shigella sonnei were confirmed by serotyping in both winter and summer samples. The ipaH gene (606 bp) was present in 176 and 49 of the winter and summer isolates, respectively. The presence of Shigella spp. in water was confirmed with over 90% specificity. The need for more effective management of these river catchments and the provision of potable water and sanitation facilities is needed to minimise the occurrence and transmission of water-borne diseases caused by these and other pathogenic bacteria.

Studies on the microbial diversity of salted fishes under aerobic conditions

A total of 93 isolates of halophilic and moderately halophilic bacteria were isolated from salted fishes, Scoliodon sp. and Thrissina thryssa obtained from retail fish outlets in Cochin, Kerala, India. The viable count of the bacteria in Scoliodon sp. and Thrissina thryssa ranged from 103-106 per g. The morphological, biochemical, and physiological investigations were done to characterize the isolates, which showed good growth in medium containing 3-10% (w/v) NaCl at 37°C. In Scoliodon sp. the maximum of 87.5% isolates were Gram-positive cocci and in Thrissina thryssa 95.5% of the isolates were Gram-positive rods. The optimum conditions such as temperature, pH, and salts (NaCl) were determined. The utilization of organic compounds such as fructose, lactose, glucose, sucrose, arabinose, trehalose, maltose, salicin, mannose, cellobiose, rhamnose, dulcitol, xylose, raffinose, sorbitol, adonitol, inulin, galactose, inositol, mannitol, hydrogen sulfide, nitrate, citrate utilization, MR-VP, triple sugar iron, as well as the hydrolysis of organic compounds such as casein, gelatin, aesculin, starch, Tween 20, and Tween 80 were also investigated. Moreover, an alkalophilic bacterial strain Bacillus halodurans (B. halodurans) was isolated from Thrissina thryssa and confirmed by 16S rDNA analysis. These studies revealed that salted fishes offer an optimal environment for the viable, diverse, potentially and industrially important bacterial community.

Selective testing of β-galactosidase activity in the laboratory identification of Salmonella and Shigella species

BackgroundSalmonella and Shigella species are pathogens of great medical and public health importance. However, laboratory identification of these organisms is time consuming. Using current standard laboratory algorithms, the vast majority...

Emerging Infections: Shewanella – A Series of Five Cases

Background:Shewanella spp. are unusual cause of disease in humans; however, reports of Shewanella infections have been increasing. Shewanella is a ubiquitous organism that has been isolated from many foods, sewage, and both from fresh and salt water. Earlier it was named as Pseudomonas putrefaciens or Shewanella putrefaciens. There are several reports describing this organism causing human infections such as cellulitis, abscesses, bacteremia, wound infection, etc. It is oxidase and catalase-positive non-fermenter gram-negative rod that produces hydrogen sulfide.Aims:The study was conducted to identify Shewanella spp., which was wrongly reported as Pseudomonas spp.Materials and Methods:Clinical samples were cultured as per standard clinical laboratory procedure. We tested the non-lactose-fermenting colonies for oxidase positivity. Oxidase-positive colony was inoculated in triple sugar iron slant (TSI) to know the hydrogen sulfide production. Hydrogen sulfide positive colonies were further tested for citrate, urease, indole, and amino acid decarboxylation and acid and gas production from sugars.Results:Five isolates identified as Pseudomonas spp. during preliminary testing were proved to be Shewanella spp. on further testing.Conclusions:It will help in better understanding the epidemiology, pathogenesis and risk factors associated with these and prevention of the rare pathogenic organisms.

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The yellow maize is the third most important crop in Peru and part of the chain maize-poultry-pig, significant impact on the national economic and social activity, however, in 2011, only 40% of the corn offered corresponded to the domestic industry. Looking for alternatives to reduce the use of chemical fertilizers, have performed investigations with plant growth promoting rhizobacteria. Bacteria were isolated from the rhizosphere of maize from districts Monsefu and Reque, Lambayeque. Dilution was made into sterile saline 0.87% NaCl w/v and plated on MacConkey agar, incubating at 30°C for 48 hours. 269 pure cultures of bacteria were obtained, the biochemical reaction was investigated in agar Triple sugar iron agar Iron Lysine, agar Citrate Simons, peptone broth, red broth methyl Voges-Proskauer and nitrate broth, was identified 66% as Enterobacteriaceae of genera Pantoea (49%), Klebsiella (17%), Kluyvera (16%), Serratia (11%), Citrobacter (4%) and Hafnia (3%). The native enterobacteria were quantified to 31.67 ppm of fixed nitrogen as ammonia; 54.25 ppm indole acetic acid and 4,78 ppm solubilized phosphorus, activity proteolytic and chitinolytic and antagonistic activity of Fusarium verticillioides were also determined. 16% of native enterobacteria did not affect the emergence of hard yellow maize, 77% affected positively and 7% affected negatively. In turn, none bacteria affected survival. Was demonstrated the potential plant growth promoter of enterobacteria isolated from field crops in the region of Lambayeque.

Brodie's Abscess Caused bySalmonella enteriticaserovar Senftenberg in a Healthy Child

A 13-year-old girl presented in a local clinic with intermittent left ankle pain for 3 months without any history of trauma. She often had discomfort in walking but with no altered sensation or swelling. The patient was transferred to our hospital for the persistent pain in her left ankle. Fever and other constitutional symptoms were absent at presentation. There was no family history of specific illness and no evidence of any underlying diseases. A physical examination revealed swelling and mild tenderness of her left ankle. She had no systolic murmur or other specific findings. She undertook simple radiography and magnetic resonance imaging (MRI). Preoperatively, blood examination revealed a leukocyte count of 6.0×10/L (reference range, 4.0∼10.0×10/L) with neutrophils in 54%, a hemoglobin level of 12.5 g/dL (reference range, 12∼14 g/dL) and a platelet count of 398×10/L (reference range, 140∼ 450×10/L). C-reactive protein level was at 0.08 mg/dL (reference range, 0∼0.5 mg/dL), and an erythrocyte sedimentation rate at 6 mm/hr (reference range, <25 mm/hr). A preoperative simple radiography of lower extremity showed well-defined lytic lesion in the metadiaphyseal region of the left distal tibia (Fig. 1). MRI of the left ankle using T1-weighted and T2-weighted MRI showed a well-defined and bilobed intramedullary cystic lesion in metadiaphysis of left distal tibia about 18 mm in diameter and 40 mm in length. This lesion revealed uniform rim enhancement, marrow edema and thin periosteal reaction, and no definite cortical disruption nor soft tissue mass was noted (Fig. 2). A percutaneous needle biopsy of the lesion showed an intracortical lytic lesion with a tiny, hyperdense focus at its center and revealed chronic inflammatory tissue reaction. She underwent a surgery for the debridement of Brodie’s abscess. Aspirates of abscess during the operation were cultured sequentially and yielded Salmonealla spp., group E by performing Gram stain, Salmonella/Shigella and triple sugar iron agar findings, and antisera grouping with no other pathogenic colonies. S. enterica serovar Senftenberg was finally identified by conventional and molecular identification methods at the Institute of Health and Environment in Daegu. Antimicrobial susceptibility test was done by VITEK system (bioMerieux VITEK, Hazelwood, MO, USA) and revealed susceptible to ampicillin, cefotaxime and ciprofloxacin except trimethoprim-sulfamethoxazole. The infection was successfully treated with operational curettage and intravenous cefotaxime. After 2 weeks of the treatment, cefotaxime was changed to per oral and she returned to outpatient clinic.

A modified culture-based study of bacterial community composition in a tannery wastewater treatment plant

Though culture-independent methods have been used in preference to traditional isolation techniques for characterization of microbial community of wastewater treatment plants, it is difficult to widely apply this approach in resource-poor countries. The present study aimed to develop a test to identify the culturable portion of bacterial community in a high-strength wastewater. Wastewater samples were collected from nitrification-denitrification and settling tanks of the treatment plant of Elmo Leather AB tannery located in Borås, Sweden. After cultivating on nutrient agar with the optimal dilution (10⁻²), phenotypic and biochemical identification of the bacteria were done with colony morphology, Gram reaction, growth on MacConkey, phenylethanol media, triple sugar Iron agar slants, catalase and oxidase tests. Biochemical grouping of the isolates was done based on their test results for MacConkey, phenylethanol media, triple sugar Iron agar and oxidase test reaction. From the biochemical groups, isolates were randomly selected for API test and 16SrRNA gene sequencing. The isolates from the denitrification, nitrification tank were identified to be Paracoccus denitrificans (67%), Azoarcus spp (3%) and Spingomonas wittichii (1%). From the settling tank, Paracoccus denitrificans (22%), Corynebacterium freneyi (20%) and Bacillus cereus (1%) were identified. The grouping based on biochemical test results as well as the identification based on sequencing has shown coherence except for discrepancies with the API test. The preliminary implications of the grouping based on culture-based characteristics and its potential application for resource-limited environmental microbial studies is discussed.

A study of Bacterial Contamination of Ghanaian Currency Notes in Circulation

A study of Bacterial Contamination of Ghanaian Currency Notes in Circulation

ISOLASI, KARAKTERISASI DAN PENGELOMPOKAN STRAIN Salmonella typhi ASAL KABUPATEN SUMBA BARAT DAYA NUSA TENGGARA TIMUR BERDASARKAN SIFAT-SIFAT FENOTIP:

Typhoid fever is highly endemic in the South-West Sumba Regency, East Nusa Tenggara. The incidence rate of the diseases is high estimated at 725/100,000. It is an acute systemic infection caused by Salmonella typhi. The clinical symptoms of the disease are extremely diverse, starting from the mild form to severe ones with the most feared complication being perforation within the small intestine. Therefore, it is important to perform isolation, characterization, and grouping of S. typhi strains from the blood culture in order to determined definitely diagnosis and the different phenotypic characteristics in the community. Isolation was done in selective and differential media: BacT/ALERT FA culture media, Selenite Cystine Broth, Chromocult Coliform Agar, MacConkey Agar, and Salmonella Shigella Agar. The typical colony of Salmonella was confirmed on Triple Sugar Iron Agar, Urea agar, and L-Lysine decarboxylation media. Phenotypic characteristics of all isolates were identified using API 20E and API 50CHE diagnostics. Based on biochemical characteristics the result showed that 18 strains obtained from different geographical origins were diverse. Four strains have similarity value 100% while the remained strains have similarity value 86.3–98.4%. All of the strains were categorized in the species of S. typhi.

Isolation and characterization of Shewanella spp. from patients of food poisoning

To identify the isolates of Shewanella spp. from specimens of food poisoning based on biological and biochemical analysis. Strains were obtained from the investigation on two food poisoning episodes in September and October, 2007 in Ma'anshan city, Anhui province. In accordance with the national standard protocol (GB/T 4789), all specimens were enriched and isolated on selective medium, and the suspected strains were identified by the VITEK-32 and API20E systems. For Shewanella spp. identified by the biochemical system, more characteristics were analyzed using auxiliary biochemical, growth, hemolytic and drug-resistance tests. DNAs of Shewanella spp. were extracted, 16S rDNA was PCR amplified and sequenced with universal 16S rDNA primers. Phylogenetic tree was constructed with MEGA 4.0. After enrichment, all specimens were inoculated to selective medium and Shewanella spp. strains were isolated from 8 samples with single colony on both TCBS and BP media. The characteristics of growth in the Triple Sugar Iron (TSI) agar appeared to have had hydrogen sulfide production but no gas production or positive oxidase. No Shewanella spp. strain was detected in WS, SS and EMB media. The 8 strains were identified as Shewanella algae (S. algae) or Shewanella putrefaciens (S. putrefaciens) by VITEK-32, as S. putrefaciens by API20E system. No other enteropathogenic bacteria, including Vibrio cholerae, Salmonella, Vibrio parahaemolyticus, Proteus vulgaris or Staphylococcus aureus, were detected from those 8 samples. From 16S rDNA phylogenetic trees, 7 out of 8 Shewanella spp. were identified as S. algae, 1 as S. putrefaciens. Strains of Shewanella spp. were isolated from samples of the food poisoning episodes, providing a possible clue to investigate the role of Shewanella spp. on food poisoning.

Avaliação in vitro da atividade inibitória de Lactobacillus spp., isolados do inglúvio e cecos de aves sobre Salmonella

Inglúvios e cecos de reprodutoras comerciais de frangos de corte foram utilizados para o isolamento de Lactobacillus spp. As estirpes isoladas foram caracterizadas como Gram-positivo, catalase negativo, produtoras de gás em glicose, não produtoras de H2S em triple sugar iron e identificadas pela reação em cadeia da polimerase como Lactobacillus reuteri e Lactobacillus salivarius. A utilização da técnica spot-on-the-lawn para avaliação da inibição in vitro permitiu a análise de vários microrganismos simultaneamente. Todas as estirpes isoladas inibiram in vitro S. Enteritidis fagotipo 4, S. Enteritidis fagotipo 28, S. Typhimurium, S. Pullorum, S. Agona, S. Anatum, S. Dublin e S. Senftenberg.

Lactobacillus as a Probiotic Feed for Chickens

The present study was carried out to isolate and identify microorganisms as a probiotic feed for chicken. The strains were isolated from gut region of the chick and was inoculated in to the MRS Medium and incubated at 37 C for 48 h under anaerobiosis. The identity of culture was based on the characteristics of o Lactobacilli as presented in the Bergey’s Manual of Determinative Bacteriology using Gram Staining, Motility, Triple Sugar Iron and Fermentation of different carbon sources. Based on the criteria Lactobacillus fermentum tolerated on Inhibitory substances, Temperature and agitation were identified and tested for probiotic use for chickens. Lactobacillus fermentum shows antimicrobial activity and shows the similar effects to antibiotic in the feed. It could be a suitable strain as a probiotic feed for chickens.

ISOLASI, KARAKTERISASI DAN PENGELOMPOKAN STRAIN Salmonella typhi ASAL KABUPATEN SUMBA BARAT DAYA NUSA TENGGARA TIMUR BERDASARKAN SIFAT-SIFAT FENOTIP

Typhoid fever is highly endemic in the South-West Sumba Regency, East Nusa Tenggara. The incidence rate of the diseases is high estimated at 725/100,000. It is an acute systemic infection caused by Salmonella typhi. The clinical symptoms of the disease are extremely diverse, starting from the mild form to severe ones with the most feared complication being perforation within the small intestine. Therefore, it is important to perform isolation, characterization, and grouping of S. typhi strains from the blood culture in order to determined definitely diagnosis and the different phenotypic characteristics in the community. Isolation was done in selective and differential media: BacT/ALERT FA culture media, Selenite Cystine Broth, Chromocult Coliform Agar, MacConkey Agar, and Salmonella Shigella Agar. The typical colony of Salmonella was confirmed on Triple Sugar Iron Agar, Urea agar, and L-Lysine decarboxylation media. Phenotypic characteristics of all isolates were identified using API 20E and API 50CHE diagnostics. Based on biochemical characteristics the result showed that 18 strains obtained from different geographical origins were diverse. Four strains have similarity value 100% while the remained strains have similarity value 86.3–98.4%. All of the strains were categorized in the species of S. typhi.

Comparação entre método bioquímico e reação em cadeia de polimerase para identificação de Lactobacillus spp., isolados de aves

Lactobacilos foram isolados do inglúvio e cecos de reprodutoras pesadas e caracterizados como Gram-positivo, catalase negativo, produtores de gás em glicose e não produtores de H2S em triple sugar iron e pela fermentação de carboidratos. Utilizaram-se os iniciadores: Lac 1/23-10C para detecção de Lactobacillus acidophilus, L. crispatus, L. amylovorus, L. gasseri, L. helveticus e L. jensenii; Lac 2/LU-1' para L. acidophilus; Fer 3/Fer 4 para L. fermentum; Reu 1/Reu 2 para L. reuteri e Sal 1 e Sal 2 para L. salivarius. L. reuteri e L. salivarius foram identificados pela reação em cadeia de polimerase (PCR) e pelo teste bioquímico, enquanto L. acidophilus, L. fermentum e Lactobacillus sp. somente pelo teste bioquímico. Os resultados obtidos na PCR foram mais precisos quando comparados aos obtidos com o método bioquímico, que demonstrou ser subjetivo devido às variações na fermentação de carboidratos, principalmente na diferenciação entre L. fermentum e L. reuteri.

A Case of Catheter-Related Bacteremia of Tsukamurella pulmonis

Tsukamurella pulmonis is an aerobic actinomycete. We report a catheter-related bacteremia of T. pulmonis. A 39 yr-old male with ALL was hospitalized to receive bone marrow transplantation (BMT). Although the patient developed a high fever at the 7th hospital day (HD), it subsided with vancomycin treatment, and he received BMT at 9th HD. Fever resurged at 16th HD despite sustained treatment with vancomycin, meropenem, and amphotericin B, but subsided with removal of Hickman catheter (HC) at 19th HD. Three sets of blood cultures comprising one from the HC and two from venipunctures were taken at 7th, 16th, and 19th HD, and the distal tip of the HC was also cultured. The aerobic vials of all 3 HC-withdrawn blood cultures and one peripheral blood culture taken at 19HD and the HC tip culture grew long, straight, thin gram-positive rods that were positive on modified Kinyoun stain. This organism showed tiny, rough, grey colonies after 3-day incubation and grew to large flat colonies when incubation was extended. It was catalase-positive, urease-positive, and alkaline-slant/alkaline-deep on triple sugar iron agar, and hydrolyzed hypoxanthine. The sequence of 1,296 base pairs of 16S rRNA of this organism showed a 100.0% homology with the published sequence of T. pulmonis DSM 44142T. To our knowledge, this is the first report of T. pulmonis bacteremia in Korea.

Biofilm Production and Molecular Epidemiology of P. aeruginosa by RAPD Isolated from Intravascular Catheters in Tertiary Care Hospital

P. aeuroginosa, various culture media such as Brain Heart Infusion Agar, Citrimide Agar and Triple Sugar Iron Agar were used. Antibiotic sensitivity of the isolated strains was determined by disk diffusion test using Muller-hinton Agar medium. Moreover, antiseptic sensitivity of the strains to Decosept and Povidone-Iodine as the main antiseptic agents was identified by determining Minimal Bactericidal Concentration and Minimal Inhibitory Concentration using Nutrient Agar and Nutrient Broth media. Results: Fifty-five of the total 292 samples were positive for P. aeuroginosa. Based on the ratio of the number of positive samples to the total samples, Thalassemia ward (38.5%) was the most infected ward and taps (61.1%) was the most infected sampling place. None of the samples from room atmosphere and intensive care equipments were positive for P. aeuroginosa. Twenty percent of the samples were resistant to ceftazidim, 32.7% resistant to piperacillin, and 10.9% to both ceftazidim and piperacillin. Conclusion: Results of the study showed that contamination with P. aeuroginosa and antibiotic resistance of the isolated bacteria are the main challenges that this medical center is facing. Taking appropriate preventive measures such as using suitable antiseptic materials and installing pedals for tap waters could have major impacts on reducing hospital contamination with P.aeuroginosa and possibly other bacteria.

A Single-Tube Screen forSalmonellaandShigella

Salmonella and Shigella species are routinely sought in stool specimens submitted for culture. It is a common practice to screen lactose-negative colonies by using triple sugar iron agar, lysine iron agar, and Christensen urea agar to determine if further identification is necessary. We designed and evaluated a novel combination of media, which are layered in a single tube, for screening isolates suspected to possibly represent Salmonella or Shigella. We tested this media combination with 106 Salmonella, 56 Shigella, and 56 other gram-negative bacilli. All Salmonella and Shigella isolates tested were appropriately characterized as possible Salmonella or Shigella by using an algorithm developed for use with this media combination. Similarly, 53 (95%) of 56 other gram-negative bacilli were appropriately screened as non -Salmonella and non -Shigella isolates. This unique media combination provides the most important biochemical reactions needed to screen for Salmonella and Shigella in a single-tube format, which decreases labor by two thirds (ie, 1 tube is inoculated vs 3).