Isolation Of Organism Research Articles

Isolation and identification of specific spoilage organism from Tilapia wastes enzymatic hydrolysates

Specific spoilage organism of Tilapia Wastes enzymatic hydrolysates during chilled storage was studied by PCR- DGGE and PCR- RFLP.Results indicated that Tilapia enzymatic hydrolysates spoilaged about 20 days at 4℃,and the total number of bacteria increased during 0 ~ 12 days( 8.71 × 102~ 8.13 × 107 cfu / g) gradually,became stable during 12~20 days.There were three types of bacteria by PCR- RFLP analysis of 16 S r RNA from 18 strains of bacteria selected randomly. Type one which dominated 61% of the total bacteria was highly similar with Pseudomonas.The maps of PCR- DGGE showed that the bacteria reduced gradually before spoilaged,and there was only one single band of bacteria at last.Sequencing analysis results of DGGE bands showed that there were three types of bacteria Pseudomonas sp.,Aeromonas and Acinetobacter in the Tilapia Wastes enzymatic hydrolysates,Pseudomonas fluorescens was the predominant bacteria when spoilaged,this conclusion was consistent with RFLP.

Epidemiological Survey of Tonsillitis Caused by Streptococcuspyogenes Among Children in Awka Metropolis (A Case Study of Hospitals in Awka Community, Anambra State)

The epidemiological survey of tonsillitis caused by Streptococcus pyogenes among children in Awka metropolis was carried out using samples from three different hospitals. A total number of 73 throat swabs were collected from children aged between 5 weeks - 5 years. Swab sticks were streaked onto Blood agar and MacConkey agar for isolation of the test organism. Organisms recovered were Streptococcus pyogenes, Streptococcus pneumoniae, Escherichia coli, Staphylococcus species, Proteus species, Pseudomonas species and Klebsiella species. Streptococcus pyogenes presented the highest number; 19 out of the 39 bacterial isolates recovered. The results showed that the prevalence of bacterial tonsillitis, specifically Streptococcus pyogenes, is 8% in infants aged between 5weeks to 11months and 17% in children aged between 1-5 years. From the results, the higher percentage of the prevalence of bacterial tonsillitis in children aged 1-5 years showed that school children are quite susceptible to Tonsillitis. This is because tonsillitis can be transmitted directly from person to person and indirectly via formites as well as respiratory secretions.This research work has shown that the drugs, Cefuroxime, Azithromycin, Ceftazidine and Gentamicin were 100% sensitive to Streptococcus pyogenes, hence can be recommended for the control of this organism.

Observed variations of Plasma Procalcitonin Amonsgt Sexually Active Males With Suspected Bacterial-Urogenital Infection In Yola Metropolis

Background: Male urogenital infection in Nigeria has increased due to alarming rise of unprotected sex amongst the sexually active males in Nigeria. Most of these case leads to a limiting advanced disease state such as kidney infections, bacteropsermia/infertility and reinal failure. AIM: To study the elevated serum Procalcitonin levels on patients with bacterial urogenital infections as an adjunct of bacterial-fungi-viral urogenital infections for diagnosis/prognosis and antimicrobial monitoring. PATIENTS and METHODS: The study group cut across adults from the ages of 20 years and above, 40 males who were suspected to have urogenital infection and 40 who were confirmed cases of urogenital infections those with suspected urogenital infections and those with confirmed urogenital infections. Using standard microbiological techniques urine and urethral swabs were taking and proper cultural techniques were carried out to isolate the bacterial/Fungi origin of infection, blood samples were collected from patients with suspected and confirmed urogenital infections for further analysis of the Procalcitonin ratio. RESULT: a total of 40 suspected males with signs and symptoms of urogenital infections where included in this study, a microbiological analysis was carried out on MSU(midstream urine clean catch) and urethral swab, and seminal fluid analysis, 40% yielded growth of gram Staphylococcus aureus and CoNs, 22.5% yielded growths of Escherichia coli, Proteus mirabilis, Acinectobacter spp, Klebseilla oxytoca while 15% had fungi/Parasitic origins,( Candida albicans and Trichomonas vaginalis) 22.5% showed no significant growth. The mean PCT ratio across the infectious agents were, bacterial origin ( 0.4-1.1 CI 0.34-0.62) fungi/parasitic origin (0.3-1.0 CI 0.28-0.56) Conclussion: although there was no remarkable difference between the confirmed cases of urogenital infection and suspected cases this showed the accuracy of PCT and its use. The reasonable increased levels of PCT in urogenital infections may serve as prognosis for asypmtomatic cases which often result to infertility, although most of the suspected cases where in the hospital wards either with prolonged chronic reinal failure or urinary tract infection this poses an interest to link clinical isolation of pathogenic organism and a symbiotic relationship of serologic PCT monitoring.

Detection of ESBL Producing Bacteria in Cases of Urinary Tract Infection in Pediatric Department at Benha University Hospital

Background: Extended-spectrum beta-lactamases (ESBLs) are rapidly evolving group of beta-lactamase enzymes commonly produced by the Gram negative bacteria. Objectives: This work aimed to detect the bacterial pathogens that cause urinary tract infection and to detect ESBL producing bacteria among them in Pediatric Department at Benha University Hospital. Methodology: This study was conducted on 118 children patients suffering from UTI collected from Outpatient and Inpatient of pediatric department at Benha University during the period from January 2015 to the end of July 2015. They were 77 girls and 41 boys and their ages ranged from 1 to 13 years.118 Urine samples were subjected to isolation and identification of causative organisms, screened for ESBL production by the pattern of antimicrobial susceptibilities. Phenotypic identification for ESBL production was confirmed by double disc diffusion test, bla SHV and bla CTX-M genes in ESBL, producing isolates were detected by using multiplex PCR. Results: This study showed that the most common cause of children UTI was E.coli (55.1%) followed by klebsiella pneumoniae (21.2%). Thirty seven (31.4%) of 118 bacterial isolates that tested by double disc confirmatory test were ESBL positive. Among the ESBL producing bacteria, 16 out of 37 (43.24%) have CTX-m and SHV genes together while 3 out of 37 (8.1%) have SHV gene only. Conclusion: Our findings showed that majority of ESBL isolates were positive for both CTXm and SHV genes (43.24%) but SHV gene alone was positive only in (8.1%) of isolates, So studies should be done in different regions of Egypt to find the common ESBL genes present in that geographical area for epidemiological purposes. Also ESBL producing isolates have been increasingly recognized in hospitals in Egypt and are associated with multiple drug resistance. Thus, ESBL producing organisms should be identified quickly so that appropriate antibiotic usage and infection control measures can be implemented.

Resident microbial flora amongst nursing staff as a source of nosocomial infection

Background: Infections due to hospital-acquired microbes is an evolving problem worldwide. Prevalence as high as 19%, of these infections in developing countries poses a challenge to health care providers. A major source of horizontal transmission of the nosocomial infections are thought to be bacterial contamination of palm of hand of health care providers like doctors and nurses. Hand hygiene is the single most cost effective preventive measure against hospital acquired infections and education is an important tool to ensure its implementation. The present study was undertaken, to demonstrate the presence of resident microbial flora on the hands of nursing staff after duty hours working at various critical areas in a tertiary care hospital with their antibiogram. Materials and Methods: This cross sectional analytical study was carried out after ethical clearance from June to August 2013on 100 nursing staff of either sex working in critical areas of tertiary care hospital after due informed consent from the nurses. A sterile swab moistened with sterile normal saline was used to collect the swabs from dominant(working) hands after working hours. The collected swabs were processed for isolation, identification and antibiotic sensitivity of organisms. Results: Out of 100 swabs collected from the dominant hands of nurses working in critical areas, 83 showed the growth. All the samples cultured shows growth of more than 2 organisms. The major Gram Positive pathogenic organisms grown were Coagulase negative Staphylococcus spp. 40 (48.19%) followed by Staphylococcus aureus 30 (36.14%) whereas amongst Gram Negative Klebsiella spp. 5(6.025%) and Pseudomonas aeruginosa 4(4.81%) predominated.. Conclusions: In the present study, the level of contamination of nursing staff was found to be high. Resident flora consisting of multi drug resistant pathogenic organisms can be greatly reduced by an increase in hand-hygiene awareness coupled with organizational interventions. We recom

Potential Bacterial Zoonotic Pathogens Isolated from a Major Abattoir and its Receiving Surface Water in Abeokuta, Nigeria

The rapid pace and scale of urbanization in Nigeria has led to large volumes of domestic and industrial wastes, which pose threats to the environment and public health. Currently, in Nigeria, there seems to be neither sufficient measures nor facilities to treat abattoir effluents for environmental safety or to recover usable energy and material from abattoir by-products. This study was conducted to assess the microbial status of the Lafenwa abattoir effluent and its receiving surface water. The Total viable and Coliform counts (TBC and TCC) using surface plating and multiple tube test techniques, respectively were performed. Bacterial contaminants were isolated and identified by standard microbiological procedures. Furthermore, various methods for waste disposal at the abattoir were assessed. The mean TBC and TCC for waste water during and after slaughtering were 5.2x107, 4.9x107 and 4.26x107, 3.06x107 cfu/ml respectively. The receiving surface water had mean TBC and TCC of 4.15x107, 3.83x107, and 3.89x107, 2.87x107 cfu/ml respectively. Waste disposal in the abattoir was by open dumping of solid wastes while effluent was discharged into the nearby Ogun River, which is also used by butchers for meat processing. Bacterial organisms isolated from the effluent include Enterobacter aerogenes, Hafnia alvei, Erwinia mallotivora, Edwardsiella ictaluri, Enterobacter amnigenus and Escherichia coli O157strains while Proteus miriabilis, Staphylococcus spp, Pseudomonas aeruginosa, Enterobacter intermedius, Yersinia aleksiciae, Serratia odorifera, Enterobacter cloacae, Enterobacter aerogens and Eschericia coli O157 strains were isolated from the surface water. The high microbial load and the isolation of pathogenic organisms of public health importance especially Escherichia coli O157 strains further demonstrates the need for adequate waste disposal and treatment. Also, the current methods of waste disposal at the abattoir greatly reduces the quality of the surrounding environment and portends a risk to public health and food safety.

Comparative assessment of antimicrobial efficacy of different hand sanitizers: An in vitro study.

To evaluate the antimicrobial efficacy of four different hand sanitizers against Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, Escherichia coli, and Enterococcus faecalis as well as to assess and compare the antimicrobial effectiveness among four different hand sanitizers. The present study is an in vitro study to evaluate antimicrobial efficacy of Dettol, Lifebuoy, PureHands, and Sterillium hand sanitizers against clinical isolates of the aforementioned test organisms. The well variant of agar disk diffusion test using Mueller-Hinton agar was used for evaluating the antimicrobial efficacy of hand sanitizers. McFarland 0.5 turbidity standard was taken as reference to adjust the turbidity of bacterial suspensions. Fifty microliters of the hand sanitizer was introduced into each of the 4 wells while the 5th well incorporated with sterile water served as a control. This was done for all the test organisms and plates were incubated in an incubator for 24 h at 37΀C. After incubation, antimicrobial effectiveness was determined using digital caliper (mm) by measuring the zone of inhibition. The mean diameters of zones of inhibition (in mm) observed in Group A (Sterillium), Group B (PureHands), Group C (Lifebuoy), and Group D (Dettol) were 22 ± 6, 7.5 ± 0.5, 9.5 ± 1.5, and 8 ± 1, respectively. Maximum inhibition was found with Group A against all the tested organisms. Data were statistically analyzed using analysis of variance, followed by post hoc test for group-wise comparisons. The difference in the values of different sanitizers was statistically significant at P < 0.001. Sterillium was the most effective hand sanitizer to maintain the hand hygiene.

Utility of septic screen in early diagnosis of neonatal sepsis

Background and Objectives: (Signs of neonatal sepsis are subtle) and common to various illnesses making the diagnosis of sepsis difficult. Results of isolation of organism and culture are available only after 48 – 72 hours. Hence the need to find out different laboratory determinants helpful in detecting neonatal sepsis. A ‘septic screen’ of the following investigations was evaluated as a reliable indicator of neonatal sepsis – total leukocyte count, absolute neutrophil count, immature to total neutrophil ratio, micro erythrocyte sedimentation rate, C reactive protein. Methods: This study was carried over a period of 18 months in a tertiary care hospital. A total of 250 neonates admitted with a clinical diagnosis of neonatal sepsis were included. Total leukocyte count was performed using an automated cell counter, differential count was done on peripheral smears stained with Wright stain, micro ESR was measured using capillary tubes with an internal diameter of 1.1 mm and length 75mm, CRP was estimated by commercially available latex agglutination kit. The cut off values used were Leukopenia 0.2, microESR >15mm, CRP positive (>1mg/dl) Results: Blood culture was positive in 75 cases (30%) only and the commonest organism cultured was Acinetobacter. On comparing various tests with blood culture as gold standard they were found to be statistically significant (p

Principles of antibiotic therapy in pediatric emergency department

Infectious diseases are among the most common conditions seen in pediatric emergency departments. Of these, in bacterial infections, appropriate antibiotic therapy should be started quickly. Specific antibiotic therapy is optimally driven by microbiologic diagnosis, predicated on isolation of the pathogenic organism, and supported by antimicrobial susceptibility testing. However, given the inherent difficulties that can arise in collecting specimens from pediatric patients, and given the high risk of mortality and disability associated with serious bacterial infections in very young infants, much of pediatric infectious disease practice is based on a clinical diagnosis with empirical use of antibacterial agents. Therefore, it is important to know the age-appropriate differential diagnosis with respect to likely pathogens. This information affects the choice of antimicrobial agent and also the dose, dosing interval, and route of administration. The patterns of antimicrobial resistance in the community and for the potential causative pathogen being empirically treated must also be considered. Antimicrobial resistance has reached crisis proportions, driven by the emergence of new resistance mechanisms and by overuse of antibiotics. It is important for practitioners to use antibiotics only as necessary, with the narrowest feasible antimicrobial spectrum, to help thwart emergence of resistance. Key Words: Bacterial Infections; Child; Anti-Bacterial Agents; Drug Resistance, Microbial; Emergency Service, Hospital

Description of Respiratory Microbiology of Children With Long-Term Tracheostomies.

There is little evidence in the medical literature to guide empiric treatment of pediatric patients with long-term tracheostomies who present with signs and symptoms of a bacterial respiratory infection. The overall goal of this study was to describe the respiratory microbiology in this study population at our institution. This study was a retrospective chart review of all subjects with tracheostomies currently receiving care at the Arkansas Center for Respiratory Technology Dependent Children. Descriptive statistics were used to describe the respiratory microbiology of the full study group. Several subgroup analyses were conducted, including description of microbiology according to time with tracheostomy, mean time to isolation of specific organisms after the tracheostomy tube was placed, association between Pseudomonas aeruginosa or methicillin-resistant Staphylococcus aureus isolation and prescribed antibiotic courses, and description of microbiology according to level of chronic respiratory support. Available respiratory culture results up to July 2011 were collected for all eligible subjects. Descriptive statistics were used to describe subject characteristics, and chi-square analysis was used to analyze associations between categorical data. P < .05 was considered statistically significant. A total of 93 subjects met inclusion criteria for the study. The median (interquartile range) age at time of tracheotomy was 0.84 (0.36-3.25) y, and the median (interquartile range) time with tracheostomy was 4.29 (2.77-9.49) y. The most common organism isolated was P. aeruginosa (90.3%), with Gram-negative organisms predominating. However, 55.9% of the study population had a respiratory culture positive for methicillin-resistant S. aureus. The first organism isolated after tracheostomy placement was Methiciliin-sensitive S. aureus was isolated the soonest after tracheostomy placement. Specific organisms were not related to level of chronic respiratory support or likelihood of receiving antibiotics. This study provides an updated overview of the variety of potential pathogens isolated from respiratory cultures of pediatric subjects with long-term tracheostomies.

Vaccines Through Centuries: Major Cornerstones of Global Health.

Multiple cornerstones have shaped the history of vaccines, which may contain live-attenuated viruses, inactivated organisms/viruses, inactivated toxins, or merely segments of the pathogen that could elicit an immune response. The story began with Hippocrates 400 B.C. with his description of mumps and diphtheria. No further discoveries were recorded until 1100 A.D. when the smallpox vaccine was described. During the eighteenth century, vaccines for cholera and yellow fever were reported and Edward Jenner, the father of vaccination and immunology, published his work on smallpox. The nineteenth century was a major landmark, with the “Germ Theory of disease” of Louis Pasteur, the discovery of the germ tubercle bacillus for tuberculosis by Robert Koch, and the isolation of pneumococcus organism by George Miller Sternberg. Another landmark was the discovery of diphtheria toxin by Emile Roux and its serological treatment by Emil Von Behring and Paul Ehrlih. In addition, Pasteur was able to generate the first live-attenuated viral vaccine against rabies. Typhoid vaccines were then developed, followed by the plague vaccine of Yersin. At the beginning of World War I, the tetanus toxoid was introduced, followed in 1915 by the pertussis vaccine. In 1974, The Expanded Program of Immunization was established within the WHO for bacille Calmette–Guerin, Polio, DTP, measles, yellow fever, and hepatitis B. The year 1996 witnessed the launching of the International AIDS Vaccine Initiative. In 1988, the WHO passed a resolution to eradicate polio by the year 2000 and in 2006; the first vaccine to prevent cervical cancer was developed. In 2010, “The Decade of vaccines” was launched, and on April 1st 2012, the United Nations launched the “shot@Life” campaign. In brief, the armamentarium of vaccines continues to grow with more emphasis on safety, availability, and accessibility. This mini review highlights the major historical events and pioneers in the course of development of vaccines, which have eradicated so many life-threatening diseases, despite the vaccination attitudes and waves appearing through history.

Diagnosis of spacer-associated infection using quantitative cultures from sonicated antibiotics-loaded spacers: implications for the clinical outcome.

Recent studies showed that a positive microbiological result from sonication of the PMMA spacer was associated with poor outcome of patients, but no quantitative analysis has yet been performed. For this purpose, a prospective analysis of 50 spacers (46 patients) was performed. All spacers were processed according to a previously described protocol, including centrifugation and quantitative culture. Clinical data and outcome were also analysed. A statistical relationship between the results of the cultures and the outcome of the patient was assessed. Sixteen patients were diagnosed with spacer-associated infection. Thirteen out of 50 spacers gave a positive culture. Nine of 13 presented with growth of an organism not isolated in the first-stage cultures, and in 7 out of 13 the organisms count was high (>10,000 CFU/ml). We have detected a significant statistical relationship between poor outcome and positive cultures, high colony counts, isolation of different organisms, positive periprosthetic cultures and spacer-associated infection. The detection in a sonicated, antibiotic-loaded PMMA spacer of organisms other than those isolated in the first surgical samples or high colony counts of any organisms is diagnostic with regard to spacer-associated infection.

Septic shock in chronic dialysis patients: clinical characteristics, antimicrobial therapy and mortality.

To describe the clinical characteristics and in-hospital mortality of chronic dialysis-dependent end-stage kidney disease patients with septic shock in comparison to septic shock patients not receiving chronic dialysis. Using an international, multicenter database, we conducted a retrospective analysis of data collected from 10,414 patients admitted to the intensive care unit (ICU) with septic shock from 1989 to 2013, of which 800 (7.7 %) were chronic dialysis patients. Data on demographic characteristics, sites of infection, microbial pathogens, antimicrobial usage patterns, and in-hospital mortality were aggregated and compared for chronic dialysis and non-dialysis patients. Multivariate time-varying Cox models with and without propensity score matching were constructed to determine the association between dialysis and in-hospital death. Septic shock secondary to central venous catheter infection, peritonitis, ischemic bowel, and cellulitis was more frequent in chronic dialysis patients. The isolation of resistant organisms (10.7 vs. 7.1 %; p = 0.005) and delays in receiving antimicrobials (6.0 vs. 5.0 h) were more common in chronic dialysis patients than in non-dialysis patients. Delayed appropriate antimicrobial therapy was associated with an increased risk of death in chronic dialysis patients (p < 0.0001). In-hospital death occurred in 54.8 and 49.0 % of chronic dialysis and non-dialysis patients, respectively. After propensity score matching, there was no difference in overall survival between chronic dialysis and non-dialysis patients, but survival in chronic dialysis patients decreased over time compared to non-dialysis patients. The demographic and clinical characteristics of chronic dialysis patients with septic shock differ from those of similar non-dialysis patients. However, there was no significant difference in mortality between the chronic dialysis and non-dialysis patients with septic shock enrolled in this analysis.

Evaluation of Infection In First Three Months of Renal Transplant – A Hospital Based Study

Background:The success of renal transplantation depends on a compromise between achieving sufficient immunosuppression to avoid rejection of the graft and maintaining a sufficient level of immuno competence to protect the recipient from infection. This study was undertaken to findout the prevalence of infection of renal allograft, to determine cause of infection, isolation of organism, antibiotic sensitivity and to evaluation of fate of infection within first three months of post transplant period.Methods: All the subjects of this study had received live related renal transplantation. Triple immunosuppressive therapy was started day before transplantation. Before enrollment in this study, all base line investigations of recipient and donor were done. All the patients were investigated at 3rd, 7th, 14th, 21st, 28th day, end of 2nd and 3rd month and whenever infection was suspected any time within first 3 months of transplantation. Patients of primary graft failure were excluded from this study. Results were expressed as mean ± standard deviation (mean ± SD).Fisher exact test or chi-square tests were performed for data analysis. All analysis were done using SPSS windows package and p value &lt;0.05 were considered significant.Results:In the present study, 71% patients had suffered from infection within first 3 months of post transplant period. Among them 71% occurred during hospital stay. Infection rate was 62% at the first month followed by 25% in 2nd month and 13% at 3rd month. Ninty six percent causes were bacterial infection and 4% were viral infection. Bacterial infection included urinary tract infection (79%), respiratory tract infection (6%), septicaemia (7%) and wound infection (4%). During follow up, most of the UTI were of bacterial origin, male and female had suffered from similar incidence of UTI (72% vs 69%), that was statistically insignificant (p=NS). Most of the episodes of UTI (90%) were associated with bacteriuria caused by E.coli.Conclusion: In the present study showed that, infection was the most common complication within first 3 months of transplant patient. UTI was the most common and remains major cause of morbidity of renal transplant recipients.J Dhaka Medical College, Vol. 23, No.2, October, 2014, Page 153-156

Molecular characterization of clinical isolates of Rhodococcus equi with PCR assay based on virulence plasmid marker

Rhodococcus equi is one of the most important pathogens of foals, in which it causes a disease manifesting in pyogranulomatous bronchopneumonia, abscesses, lymphadenitis or ulcerative enterocolitis. R. equi can be pathogenic to other domestic and wild animals and humans as well. Although, R. equi is prevalent in India, the work carried out in our country has not gone much beyond isolation of organism from clinical cases of foal pneumonia. Therefore, the present study was carried out for characterization of R. equi strains isolated from clinical cases based on plasmid markers (traA, vapA and vapB genes) and antibiotic sensitivity. In the present study, 298 samples (nasal swab, 136; fecal sample, 130; soil, 28; tissue, 4) were collected and processed for isolation, identification, and characterization of R. equi via biochemical test, antimicrobial susceptibility test and PCR. A total of 28 R. equi isolates could be recovered from clinical samples. All the 28 isolates were found sensitive to chloramphenicol, erythromycin, oxytetracycline, ciprofloxacine, neomycin and rifampin while resistant to ampicillin, trimethoprim, sulphadiazine, cloxacin, amikacin, cephalexin, and kanamycin in in vitro antimicrobial assay. PCR typing based on plasmid gene markers: traA, vapA, and vapB revealed that vapA plasmid was present in 26 isolates whereas it was absent in 2 isolates. Periodic monitoring of horse farm before and after foaling season is recommended for diagnosis of R. equi and initiating requisite bio-security and therapeutic measures.

Foodborne Illness Among School Children in Ga East, Accra.

A food borne illness was reported in Ga-East district of Greater Accra Region among school children in May, 2007 after eating food provided at school. The objective of the investigation was to determine the source, mode of contamination and the causative agent. A case-control study was conducted, cases were schoolchildren with abdominal symptoms and controls were children of the same sex and class without any symptom during the same period. The school children were selected by systematic sampling. Food handlers and the children were interviewed by a structured questionnaire. Food handlers were physically examined and their stools and blood examined. The kitchen for food preparation was inspected. Risks of food borne infection from the foods eaten were determined using attack rates. The minimum, peak and maximum incubation periods were 2, 11 and 61 hours respectively. The source was rice and groundnut soup (with the highest attack rate difference). Stool and blood samples of food handlers were not infective. Storage facility for food items was poor. No food samples were available for organism isolation. A protocol to prevent such outbreaks was nonexistent. The short incubation period and symptoms presented suggest an infective origin. The storage of the meat may potentially have been the point of contamination. The study showed that the schoolchildren ate contaminated food although the investigation could not determine the causative agent. Protocols to prevent such outbreaks need to be developed for the schools.

Predictive value of blood culture in acute bacterial meningitis

To evaluate predictive value of blood culture as an effective alternative laboratory tool for isolation of causative organism in acute pyogenic meningitis. It is a hospital based prospective longitudinal hospital based study performed from May 2007 to May 2009. A total number of 310 children of age group 2 months-14 years who presented to the hospital with probability of bacterial meningitis were screened in the study. Eighty cases excluded by preset exclusion criteria and rest 230 formed the analytic sample. Pretreatment blood culture with concomitant lumbar puncture or lumbar puncture within 2 h of antibiotic administration for cerebrospinal fluid analysis was obtained. The main outcome parameter was the predictive value of blood culture for isolation of causative organism in acute bacterial meningitis. Blood culture revealed a sensitivity of 73.28% (95% confidence interval [CI]: 64.85%–80.63%), specificity of 87.88% (95% CI: 79.78%–93.58%), positive predictive value of 88.9% (95% CI: 81.40%–94.13%), negative predictive value of 71.31% (62.42%–79.14%), positive likelihood ratio of 5.97 and negative likelihood ratio of 0.30. Probability of isolation of causative organism by blood culture in bacterial meningitis was found to be highest among infants (82%) and least in children aged above 5 years (54.17%). When compared to gold standard cerebrospinal fluid diagnostic criteria, blood culture has a high sensitivity, specificity, positive predictive value and likelihood ratio to be an effective alternative laboratory tool for isolation of causative organism in acute bacterial meningitis especially in infants and younger children.

Mining environmental high-throughput sequence data sets to identify divergent amplicon clusters for phylogenetic reconstruction and morphotype visualization.

Environmental high-throughput sequencing (envHTS) is a very powerful tool, which in protistan ecology is predominantly used for the exploration of diversity and its geographic and local patterns. We here used a pyrosequenced V4-SSU rDNA data set from a solar saltern pond as test case to exploit such massive protistan amplicon data sets beyond this descriptive purpose. Therefore, we combined a Swarm-based blastn network including 11 579 ciliate V4 amplicons to identify divergent amplicon clusters with targeted polymerase chain reaction (PCR) primer design for full-length small subunit of the ribosomal DNA retrieval and probe design for fluorescence in situ hybridization (FISH). This powerful strategy allows to benefit from envHTS data sets to (i) reveal the phylogenetic position of the taxon behind divergent amplicons; (ii) improve phylogenetic resolution and evolutionary history of specific taxon groups; (iii) solidly assess an amplicons (species') degree of similarity to its closest described relative; (iv) visualize the morphotype behind a divergent amplicons cluster; (v) rapidly FISH screen many environmental samples for geographic/habitat distribution and abundances of the respective organism and (vi) to monitor the success of enrichment strategies in live samples for cultivation and isolation of the respective organisms.

High-risk Oncogenic Human Papilloma Virus Infection of the Foreskin and Microbiology of Smegma in Prepubertal Boys

To evaluate the prevalence and genotypes of human papillomavirus (HPV) infection and microbiological status of the foreskin in asymptomatic prepubertal boys. A total of 100 prepubertal healthy boys who underwent a standard circumcision procedure were included in the study. High-risk HPV status was determined by real-time polymerase chain reaction for the genotypes 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68. The swabs were immediately sent to microbiology laboratory for culture and sensitivity tests. The median age at the time of surgery was 5.7years (range, 2 months-9 years). High-risk HPV was detected in 9 foreskins (9%). Positive samples showed are HPV16 (n= 3), 31 (n= 2), 39 (n= 3), and 51 (n= 1). There were a total of 72 organism isolates: 54 gram-positive (75%) and 17 gram-negative (23.6%) bacteria and 1 (1.4%) Candida. The most commonly isolated gram-negative bacterium was Escherichia coli (41.2%), whereas the commonly isolated gram-positive bacterium was Enterecoccus sp. (57.4%). Our results showed that subclinical high-risk HPV infections are found in the foreskin, which could be a reservoir for HPV-associated diseases, and smegma seems not to be a risk factor for drug-resistant infection in children.

Prevalence of Vibrio spp. in raw shrimps (Parapenaeus longirostris ) and performance of a chromogenic medium for the isolation of Vibrio strains

Vibrios are natural inhabitants of estuarine ecosystems and some species may pose public health problem as agents of sporadic or collective food-borne infections associated with the consumption of fish or shellfish. Samples of raw shrimp (n=299), fished in coastal areas of the city of Agadir, Morocco, and collected from its fish marketplace, were examined for the presence of pathogenic vibrios. Microbiological analysis was carried out according to a protocol using thiosulphate citrate bile sucrose (TCBS) agar and CHROMagar Vibrio (CV) as selective media. Presumptive-positive colonies were identified by biochemical and species-specific PCR systems, and further tested by PCR for the presence of pathogenicity factors. The overall prevalence of Vibrio spp. in shrimps was 55·8%. Vibrio parahaemolyticus was recovered from 25 of 299 samples (8·4%), Vibrio cholerae from six samples (2%) and Vibrio alginolyticus from 161 samples (53·8%). No virulence genes were found among V.parahaemolyticus and V.cholerae isolates. The CHROMagar Vibrio plating medium was found to be more efficient than the Thiosulphate Citrate Bile salt Sucrose (TCBS) Agar in the isolation of Vibrio organisms. A significant proportion of shrimps marketed and consumed in Morocco are caught in the coastal region of the city of Agadir. This study provides interesting data of prevalence of Vibrio spp. in raw shrimps as well as better understanding of their potential virulence. It is apparent from this study that genes and primers used in multiplex PCR for identification and detection of virulence factors, can be used to monitor shrimps for the presence of potentially pathogenic strains of Vibrio cholerae and Vibrio parahaemolyticus. The results highlight the added value of using a chromogenic medium for research and isolation of pathogenic Vibrio in seafood, more specific and accurate than TCBS.