Presumptive Identification Research Articles

Thermonuclease test accuracy is preserved in methicillin-resistant Staphylococcus aureus isolates.

Introduction The nuc gene encodes a thermonuclease which is present in Staphylococcus aureus but not in coagulase-negative staphylococci (CoNS) and is the target of the rapid phenotypic thermonuclease test. The effect of nuc gene variation in methicillin-resistant S. aureus (MRSA) on the performance of PCR testing has been noted, although there are no reports about the effect of MRSA on the activity of the thermonuclease enzyme.Aim Our goals were to examine the sensitivity and specificity of the thermonuclease test used to distinguish S. aureus from CoNS cultured from blood. In addition, we aimed to assess differences in the sensitivity, specificity and accuracy of the thermonuclease test between methicillin-sensitive S. aureus (MSSA) and MRSA isolates.Methodology We performed a retrospective analysis of 1404 isolates. Each isolate from a positive blood culture was identified as a Gram-positive coccus by microscopy then analysed with the thermonuclease test (Southern Group Laboratory) prior to confirmatory identification using VITEK microbial identification platforms (bioMérieux) and cefoxitin disc diffusion testing.Results Of 1331 samples included in the final analysis, 189 were thermonuclease-positive, of which 176 were identified as S. aureus . Of the 1142 thermonuclease-negative samples, 13 were finally identified as S. aureus , giving a sensitivity of 93.1 % (95 % confidence interval [CI] 88.5–96.3) and specificity of 98.9 % (95 % CI 98.1–99.4). Of the nine proven MRSA samples, eight were thermonuclease-positive, giving a sensitivity of 88.9 % (95 % CI 51.8–99.7). Thermonuclease test accuracy for MSSA and MRSA isolates was 98.1 % (95 % CI 97.2–98.8) versus 98.8 % (95 % CI 98.0–99.3), respectively.Conclusions In the era of increasing use of molecular-based microbiology assays, the thermonuclease test remains a simple, inexpensive and robust test for the presumptive identification of S. aureus cultured from blood, irrespective of methicillin sensitivity.

The effects of three disinfection protocols on Candida spp., denture stomatitis, and biofilm: A parallel group randomized controlled trial

Statement of problemAntifungals are used to treat Candida infections. However, because of increased antifungal resistance and the length of antifungal therapy, Candida spp. infections can be prevented using the prosthesis hygiene method. Therefore, establishing efficient, safe, and low-cost hygiene protocols for complete denture wearers is necessary. PurposeThe purpose of this clinical trial was to compare 10% Ricinus communis (RC10%) and 0.5% chloramine-T (CT0.5%) with negative (water) and positive (0.25% sodium hypochlorite [SH0.25%]) controls to establish a protocol to treat denture stomatitis (DS), remove denture biofilm, reduce overall microbiota, and decrease Candida spp. on the palate and denture bases. Material and methodsThis randomized, double blind, controlled clinical trial allocated 60 DS-positive participants in parallel groups: RC10%, CT0.5%, negative control, and SH0.25%. All participants brushed their palate and dentures and applied 1 of the solutions only to the denture. The following outcomes were assessed at baseline and after 7 and 37 days: Candida spp. counts, frequency of species by presumptive identification, DS severity, and photographic quantification of biofilm. The Kruskal-Wallis and Friedman tests with stepwise step-down post hoc test compared the anticandidal effect and the DS score (between groups and time). ANOVA and the Tukey post hoc test were used for biofilm removal comparison (α=.05). ResultsMicrobial counts were solution- and time-dependent for dentures, with C. albicans, C. tropicalis, and C. glabrata being the most prevalent species. RC10% presented similar results to baseline and control after 7 and 37 days. CT0.5% reduced the CFU/mL compared with the baseline. SH0.25% was the most effective. DS reduced in all groups, independent of the solution. SH0.25% reduced biofilm the most, followed by RC10%. CT0.5% was similar to the control. ConclusionsSH0.25% demonstrated potential for Candida spp. control in denture wearers with DS. The other protocols showed intermediate activity and might be more suitable for longer immersion periods.

Prevalence and Antimicrobial Profile of Shigella Isolates in a Tertiary Care Hospital of North Karnataka: A 12-Year Study

AbstractContext:Shigella is a common cause of bacillary dysentery. Although it is reported worldwide, the majority of the infections are seen in developing countries with Shigella flexneri being the most common isolate. Prevalence of Shigella species and their antibiotic susceptibility profiles vary according to geographic area and season. Aims: In the present study, the epidemiology and antimicrobial profile of Shigella from stool samples received at our hospital for a period of 12 years (January 2006 to December 2017) was evaluated. Subjects and Methods: A total of 4578 stool samples were collected from the cases of acute gastroenteritis and diarrhoea. Samples were processed for culture and sensitivity according to standard microbiological techniques. The presumptive identification of Shigella species was done using standard conventional biochemical tests and confirmed using antisera. Results: A total of 189 (4.2%) samples yielded Shigella spp. Isolation of Shigella spp. were more frequent from males (58.2%). S. flexneri was the commonest species isolated (47.6%) followed by Shigella sonnei (11.6%), Shigella dysenteriae (4.2%) and Shigella boydii (2.1%). Non-typeable Shigella was commonly recovered. The isolates showed high resistance to ampicillin (76.7%) and co-trimoxazole (75%) while highest susceptibility was observed to ceftriaxone (79.2%). Conclusions:S. flexneri was the most prevalent species isolated at this centre. Shigella isolates from the study showed alarming resistance to recommended antibiotics. Non-typeable Shigella accounted for 34.4% isolates. Molecular discrimination between Shigella and Escherichia coli is essential.

Analysis of Powders Containing Illicit Drugs Using Magnetic Levitation

AbstractMagneto‐Archimedes levitation (MagLev) enables the separation of powdered mixtures of illicit drugs (cocaine, methamphetamine, heroin, fentanyl, and its analogues), adulterants, and diluents based on density, and allows the presumptive identification of individual components. Small samples (mass <50 mg), with low concentrations of illicit drugs, present a particular challenge to analysis for forensic chemists. The MagLev device, a cuvette containing a solution of paramagnetic gadolinium(III) chelate in a non‐polar solvent, placed between two like‐poles‐facing NdFeB magnets, allowed separation of seven relevant compounds simultaneously. In particular, initial separation with MagLev, followed by characterization by FTIR‐ATR, enabled identification of fentanyl in a sample of fentanyl‐laced heroin (1.3 wt % fentanyl, 2.6 wt % heroin, and 96.1 wt % lactose). MagLev allows identification of unknown powders in mixtures and enables confirmatory identification based on structure‐specific techniques.

Analysis of Powders Containing Illicit Drugs Using Magnetic Levitation.

Magneto-Archimedes levitation (MagLev) enables the separation of powdered mixtures of illicit drugs (cocaine, methamphetamine, heroin, fentanyl, and its analogues), adulterants, and diluents based on density, and allows the presumptive identification of individual components. Small samples (mass <50 mg), with low concentrations of illicit drugs, present a particular challenge to analysis for forensic chemists. The MagLev device, a cuvette containing a solution of paramagnetic gadolinium(III) chelate in a non-polar solvent, placed between two like-poles-facing NdFeB magnets, allowed separation of seven relevant compounds simultaneously. In particular, initial separation with MagLev, followed by characterization by FTIR-ATR, enabled identification of fentanyl in a sample of fentanyl-laced heroin (1.3 wt % fentanyl, 2.6 wt % heroin, and 96.1 wt % lactose). MagLev allows identification of unknown powders in mixtures and enables confirmatory identification based on structure-specific techniques.

Differentiation between Streptococcus pneumoniae and other viridans group streptococci by matrix-assisted laser desorption/ionization time of flight mass spectrometry

ObjectivesMatrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) is becoming the method of choice for bacterial identification. However, correct identification by MALDI-TOF of closely related microorganisms such as viridans streptococci is still cumbersome, especially in the identification of S. pneumoniae. By making use of additional spectra peaks for S. pneumoniae and other viridans group streptococci (VGS). We re-identified viridans streptococci that had been identified and characterized by molecular and phenotypic techniques by MALDI-TOF. MethodsVGS isolates (n = 579), 496 S. pneumoniae and 83 non-S. pneumoniae were analysed using MALDI-TOF MS and the sensitivity and specificity of MALDI-TOF MS was assessed. Hereafter, mass spectra analysis was performed. Presumptive identification of proteins represented by discriminatory peaks was performed by molecular weight matching and the corresponding nucleotides sequences against different protein databases. ResultsUsing the Bruker reference library, 495 of 496 S. pneumoniae isolates were identified as S. pneumoniae and one isolate was identified as non-S. pneumoniae. Of the 83 non-S. pneumoniae isolates, 37 were correctly identified as non-S. pneumoniae, and 46 isolates as S. pneumoniae. The sensitivity of the MALDI-TOF MS was 99.8% (95% confidence interval (CI) 98.9–100) and the specificity was 44.6% (95% CI 33.7–55.9). Eight spectra peaks were mostly present in one category (S. pneumoniae or other VGS) and absent in the other category and inversely. Two spectra peaks of these (m/z 3420 and 3436) were selected by logistic regression to generate three identification profiles. These profiles could differentiate between S. pneumoniae and other VGS with high sensitivity and specificity (99.4% and 98.8%, respectively). ConclusionsSpectral peaks analysis based identification is a powerful tool to differentiate S. pneumoniae from other VGS species with high specificity and sensitivity and is a useful method for pneumococcal identification in carriage studies. More research is needed to further confirm our findings. Extrapolation of these results to clinical strains need to be deeply investigated.

Outdoor Air Quality and Antibiogram Characteristics of Bacterial Isolates of Akure City Abattoirs, Nigeria

Air sampling of Onyearugbulem and FUTA abattoirs was carried out to evaluate their quality. Air sampling was done using the open-settling method on general and selective agar media of the gutter, sleeping bench, slaughtering floor dumpsite, slaughtering table and roof of the abattoirs. Using standard methods, Presumptive identification of the bacterial isolates was carried out. An array of ten (10) conventional antibiotics was used to assay the antibiotic susceptibility characteristics of the bacterial isolates. Multiple antibiotic sensitivity index (MARI) was determined for isolated bacteria. At Onyearugbulem abattoir, the total Escherichia coli count ranged from 3.6×10 cfu/ml from the gutter and slaughtering table to 6.5×10 cfu/ml from the slaughtering floor, while the total bacterial count ranged from 1.34×102 cfu/ml from dumpsite to 2.55×102 cfu/ml from the gutter, the total coliform count ranged from 2.8× 10 cfu/ml from slaughtering floor to 4.1×10 cfu/ml from sleeping bench. Several bacteria were presumptively isolated. These include: Micrococcus sp., Bacillus sp., Staphylococcus aureus, Escherichia coli, Proteus sp., Enterobacter aerogenes and Aeromonas sp. Ciprofloxacin (10 µg) had an inhibitory effect on all the Gram positive bacterial isolates with the highest inhibitory activity on S. aureus at 24.00 mm, and the Gram negative bacteria with the highest inhibitory activity on E. coli at 21.00 mm. Micrococcus sp. had the highest MARI of 0.8. These findings reveal the presence of multiple-antibiotic resistant bacteria in Onyearugbulem and FUTA abattoirs’ atmosphere. There is therefore need for routine environmental sanitation of the slaughterhouses.

Prevalence and Antibiogram Characteristics of Bacteriuria and Cadidiuria among Indigenes of Selected Parts of Akure North, Ondo State

Aims: The aim of this study was to evaluate the prevalence and antibiogram characteristics of bacteria and fungi in urine samples of some selected towns in Ondo State, Nigeria.&#x0D; Study Design: Experimental design.&#x0D; Study Location: Urine samples were collected from patients visiting University of Medical Science Teaching Hospital Akure and indigenes of Ogbese, Ipogun-Ayo and Ita-oniyan community in Ondo North, Nigeria.&#x0D; Methodology: Pre-survey of the study participants was carried out using randomly distributed questionnaires. Two hundred and forty-three (243) urine samples were randomly collected from apparently healthy male and female participants cutting across different age groups. Presumptive identification of isolated bacteria and fungi were cultured on general and differential media (cysteine lactose electrolyte deficient agar, blood agar, nutrient agar, and Potato dextrose agar, for fungal isolates) were done using standard methods. The clinical and relative typed isolates were collected from Federal Institute of Industrial Research Oshodi (FIIRO) and were subjected to antibiotic sensitivity pattern using array of ten (10) conventional antibiotics according to Clinical Laboratory Standard Instruction.&#x0D; Results: Stapylococcus aureus was the most frequently occurred (27.2%) bacteria isolated from the urine samples while, Candida albicans had the least (5.6%) occurrence. These findings revealed that bacteriuria and candidiuria were prevalent among the indigenes of Ipogun-Ayo, Ondo State, with antibiotic sensitivity to few conventional antibiotics. This is an indication of UTI among the indigenes and urgent need for medical attention.

Bacteriological Evaluation of Some Automated Teller Machine In Akure Metropolis

Background: Automated Teller Machines (ATMs) represent a good transient environment for development of pathogenic microbes. Investigation on the level of bacterial contaminants on keypads of ATMs was carried out in Akure, Nigeria. Methods: A total of sixty ATMs keypads were sampled in two major locations (main market and Federal University of Technology, campus) in Akure. Samples were collected from key-pad and screen parts of the ATM devices with sterile swab sticks and was cultured immediately on Nutrient agar, Manitol salt agar, and MacConkey agar mediums for Microbial examination. Standard bacteriological methods were employed in the analysis of the sample. Presumptive identification of bacterial isolates was by cellular morphology, Gram staining reaction, motility, catalase, coagulase test, oxidase strip test and MICROBACT Biochemical Identification system was used to identify the species of the oxidase negative Gram negative bacteria. Results: Out of 30 ATMs keypads examined in the two locations, 28 and 22 yeilded bacteria growth in Akure main Market and FUTA Campus respectively. The organisms isolated were Staphylococcus aureus, Klebsiella species, Escherichia coli, Pseudomonas and Bacillus species. All the bacterial isolates showed high resistance to Ampicillin but low resistance to Ciprofloxacin. Conclusion: The study confirmed the presence of pathogenic bacteria species on ATM key-pads with possible health implications in Akure, Nigeria. Daily and regular cleaning regimen of the key-pads with sanitizers, and public enlightenment on the hygienic usage of the keypads is necessary to reduce health risks to the users.

Histopathological features of subcutaneous mycosis: a retrospective study

Background: Subcutaneous fungal infection is commonly seen in tropical and subtropical countries particularly in India. The lesions are usually presented as swelling, in the localized form. Subcutaneous mycosis is caused by penetration of causative fungi into the subcutaneous tissue. Histopathology is one of the main tools of diagnosis in mycology as it has the advantages of early diagnosis compared to culture, low cost and it provides a presumptive identification of the infecting fungus. It also gives information about tissue reaction and supplement culture report. It gives the idea about the fungus isolated in culture is contaminant or pathogenic. Many a times tissue might be sent in formalin as fungal infection is not suspected clinically. Role of histopathologist is very important in diagnosing these subcutaneous fungal lesions. Material &amp;Methods: A retrospective study was conducted on subcutaneous fungal infections seen between January 2014 to December 2018 in the Department of pathology, IGMC&amp;RI, Puducherry. Twenty-two patients with biopsy proven subcutaneous fungal infections were included in the study. In the present study, the varied histopathological features like type of inflammatory response, presence of granulomas, necrosis, eosinophils and abscess formation were seen in the tissue sections. Results: The most common histopathology feature was giant cell reaction seen in 21 (95.5%) cases, followed by epithelioid cell granuloma in 13(59%) cases and areas of necrosis in 13 (59%) cases. Eight (36%) cases showed numerous eosinophilic infiltration 10(46%) cases of scant eosinophils and 4 (18%) cases showed absent eosinophils. These fungal structures were identified in H&amp;E stain itself in 13 (59%) cases. Conclusion: Histopathological study is most important in subcutaneous mycosis as most of the time diagnosis is unsuspected and delayed because of its rarity and varied presentation. Fungal infection should be suspected in cases with localized cystic swelling. High index of suspicion, careful microscopic examination and special stains are important for accurate diagnosis.

2152. Detection of Uropathogens Using BD Kiestra™ Total Laboratory Automation with Urine Culture Application

BackgroundUrine is the most frequently cultured specimen type for the majority of clinical microbiology laboratories. Typically, around 30% of cultures are positive for uropathogens with 70% yielding insignificant or mixed growth. BD is developing a software Urine Culture Application (UCA) for the BD Kiestra Total Laboratory Automation (TLA) system to screen images of urine culture plates, sort them based on growth vs. insignificant growth and also allow for presumptive pathogen identification.MethodsDe-identified urine specimens were inoculated onto BD BBL™ CHROMagar™ Orientation Media (CHROM; BD, Sparks, MD), CHROM/Trypticase™ Soy Agar II with 5% Sheep Blood (TSA) biplate, BD BBL MacConkey II agar, and TSA using the BD Kiestra TLA system. Plates were imaged at 24 hours using the BD Kiestra™ ReadA Compact imaging acquisition software and an algorithm was applied to the images using the UCA (Version 2.0). Semi-quantitative measurements of <100, 100–1,000, 1,000–10,000, 10,000–100,000, and >100,000 cfu/mL growth were determined by UCA for all media types and presumptive ID was determined using CHROM. Manual reading of the images by two technologists was the gold standard for comparison. For discrepant results, a third manual reader was used as an arbitrator.ResultsTesting between 877 and 934 urine specimens on each of five media types using UCA resulted in an exact semi-quantitative agreement with manual reading for 85.5–95.0% of specimens (Table 1). If semi-quantitative values ± one category of agreement are included, the number rises to 98.2–99.4% agreement. Using CHROM for presumptive identification of pure or predominant organisms, UCA was in agreement with manual identification in 251 of 272 cultures (92.3%). Of the 21 discrepant organisms, 19 were classified as “other” by manual reading but were identified as specific organisms by UCA. Definitive organism identification was not performed.ConclusionUCA was able to accurately categorize bacterial growth into five semi-quantitative categories using five media types. Pure and predominant uropathogens were accurately identified from CHROM using UCA. The use of UCA software application may enable laboratories to save time screening urine cultures by allowing more efficient use of technologist time. Disclosures All authors: No reported disclosures.

Rapid Presumptive Identification of Bacillus anthracis Isolates Using the Tetracore RedLine Alert™ Test.

A comprehensive laboratory evaluation of the Tetracore RedLine Alert test, a lateral flow immunoassay (LFA) for the rapid presumptive identification of Bacillus anthracis, was conducted at 2 different test sites. The study evaluated the sensitivity of this assay using 16 diverse strains of B. anthracis grown on sheep blood agar (SBA) plates. In addition, 83 clinically relevant microorganisms were tested to assess the specificity of the RedLine Alert test. The results indicated that the RedLine Alert test for the presumptive identification of B. anthracis is highly robust, specific, and sensitive. RedLine Alert is a rapid test that has applicability for use in a clinical setting for ruling-in or ruling-out nonhemolytic colonies of Bacillus spp. grown on SBA medium as presumptive isolates of B. anthracis.

Microbiological analysis of ready to eat foods collected from different places of Dhaka city, Bangladesh

The current study was attempted to observe the microbiological quality of ready to eat foods collected from different places of Dhaka city, Bangladesh. Ten food samples that include five fried items-fried chicken, fish fillet, shrimp fries, French fries, potato chop and five sweet items-yoghurt, pudding, ice cream, mango drink, lemon drink were analyzed during the period from September 2017 to November 2017. Conventional cultural, microscopic and biochemical tests were followed for the detection and enumeration of bacterial isolates associated with these food samples. The investigation encompassed detection of total viable bacterial count (TVBC) and presumptive identification of other pathogenic bacteria from these samples. The higher counts of TVBC, Escherichia coli, Staphylococcus aureus and Bacillus spp. were recorded as 1.94×106 cfu/g (potato chop), 3.1×104 cfu/g (fried shrimp), 7.5×103 cfu/g (fried chicken) and 1.0 x 103 cfu/g (French fries) respectively. Based on the specifications by International Commission for Microbiological Specification for Foods (ICMSF), the level of contaminations was within acceptable microbiological limits except for potato chop.&#x0D; Stamford Journal of Microbiology, Vol.8(1) 2018: 30-33

Early first-trimester transvaginal ultrasound is indicated in pregnancy after previous Cesarean delivery: should it be mandatory?

Early first-trimester transvaginal ultrasound is indicated in pregnancy after previous Cesarean delivery: should it be mandatory?

Study of vancomycin and high-level aminoglycoside-resistant Enterococcus species and evaluation of a rapid spot test for enterococci from Central Referral Hospital, Sikkim, India.

BACKGROUND:Enterococcus is an important pathogen, and with its emergence of resistance to multiple antimicrobials, the management of infection is becoming increasingly difficult.AIM:The aim of the study is to determine the prevalence, antibiotic resistance, and risk factors associated with enterococcal infection or colonization.MATERIALS AND METHODS:In this prospective study, samples from inpatients were screened for resistant enterococci. Antibiotic susceptibility testing was performed using the disc diffusion method and minimum inhibitory concentration by the agar dilution method. A modification of a test tube method of sodium chloride-esculin hydrolysis to a spot test was evaluated for its rapidity and reliability in the presumptive diagnosis of enterococci.STATISTICAL ANALYSIS USED:Fisher's exact test was used for continuous (Student's t-test) and categorical variables. Multivariate analysis was performed with logistic regression using IBM SPSS 20.0 software (Armonk, NY, USA).RESULTS:Enterococcus species were isolated from 182 samples: Enterococcus faecalis (68.7%), Enterococcus faecium (20.9%), Enterococcus gallinarum (6%), and Enterococcus durans (4.4%). Maximum resistance was to ciprofloxacin (59.3%) and least to linezolid (0.5%). The isolation rate of vancomycin-resistant enterococci (VRE) was 13.7%; 30.2% and 20.9% were of high-level gentamicin and streptomycin, respectively. All 182 Enterococcus species gave positive results within 30–60 min by the rapid spot test.CONCLUSIONS:Overall, high-level aminoglycoside resistance (HLAR) was observed more than glycopeptide resistance. Surveillance strategies need to be upgraded and implemented in order to prevent the emergence and further spread of not only VRE but also HLAR enterococci in the hospital. The spot test gave reliable and rapid results in presumptive identification of enterococci.

HiCrome Bacillus agar for presumptive identification of Bacillus and related species isolated from honey samples

This study aimed to evaluate the performance of Hicrome Bacillus™ agar for isolation and rapid identification of the aerobic spore-forming bacteria most frequently found in honey samples. A collection of 197 bacterial isolates of Bacillus, Brevibacillus, Lysinibacillus, Paenibacillus, and Rummeliibacillus belonging to different species that have been reported in honey were screened for their abilities to grow and for their colony colors and medium appearance in HiCrome Bacillus agar. Also, 21 strains from culture collections were used for comparison and quality controls. A flowchart utilizing a combination of colony and media characteristics in the chromogenic medium and a set of simple biochemical and morphological tests were elaborated for quick presumptive identification. A procedure for direct isolation from honey samples was developed. In conclusion, HiCrome Bacillus agar in combination with simple microbiological tests was highly useful for rapid and reliable identification of most Bacillus, Brevibacillus, Lysinibacillus and Paenibacillus species commonly found in honey samples facilitating isolation from polymicrobial honey.

Isolation of some gram-negative bacterial species from naturally infected silver European eel fish (Anguilla anguilla linnaeus, 1758)

The aim of this study is to inform isolations of Shewanella putrefaciens, Aeromonas sobria and A. veronii from the silver European eel fish (Anguilla anguilla) caught from the Gulf of Antalya, Turkey. Three silver eel fish samples as freshly dead (mean weight 75 g) were obtained by a local fisherman. For bacteriological study, inoculations from the head kidney, spleen and liver were made and inoculated onto Brain Hearth Infusion Agar (BHIA-S) supplemented with 1.5 % NaCl. The inoculated petri dishes were incubated at 24 ± 2°C for 72 hours. The end of the incubation period, bacterial colonies were subcultured. For presumptive identification of the bacterial strains, the strains were identified using a variety of phenotypic characteristics. The further identification of the strains was performed with PCR. Clinically, the silver eel samples showed pale gills, losses of scale, necrotic lesions on dorsal, hemorrhagies on the jaws, lateral and ventral sides of the body and around the anus and hemoragic anal fin. At necropsy, the fish had enlarged and pale spleen and the gastrointestinal tract was empty. After 72 hours, 10 bacterial strains were isolated. 6 strains as Shewanella putrefaciens, 2 strains as Aeromonas sobria and 2 strains as A. veronii were identified according to the phenotypical characteristics and PCR study results. The eel fish stocks in natural conditions are decreasing and this problem may be multifactorial such as disease, migration and overhunting. As a result of this study, A. veronii was first isolated from A. anguilla.

CONCORDÂNCIA INTEROBSERVADORES DA DETECÇÃO DO FATOR CORDA PARA A IDENTIFICAÇÃO PRESUNTIVA DO MYCOBACTERIUM TUBERCULOSIS

Detection of cord factor may favor the definitive diagnosis and initiation of antituberculosistreatment. The aim of this study was to evaluate the interobserver concordance of mycobacterial smears stained with Ziehl-Neelsen technique for the presumptive identification of the Mycobacterium tuberculosis (M. tuberculosis).Three observers, healthcare students, conducted a blind reading of 30 slides stained with Ziehl-Neelsen and their results were compared to the results of an experient reader. Kappa coefficient with 95% interval was used to evaluate the concordance betweenthe readings. No excellent interobserver concordance(κ&gt;0.75) was found. The interpretation and use of the cord factor for the presumptive identification of the M. tuberculosisis a technique that requires training for the interpretation of the variables of the smear stained with Ziehl-Neelsen technique and theoretical knowledge of the observer to deliver a correct result.

PCR based detection of tcdCΔ117 in Clostridium difficile infection identifies patients at risk for recurrence – A hospital-based prospective observational study

ObjectivesIncreasing incidence and severity of Clostridium difficile infection (CDI) in the last decades has been attributed to the emergence of hypervirulent C. difficile strain PCR-ribotype 027 (RT027). Commercial multiplex real-time PCR tests allow the presumptive identification of RT027 by detecting a single-base deletion at nt117 in the tcdC gene (tcdCΔ117). The clinical usefulness of the detection of tcdCΔ117 is unclear.Therefore, we evaluated test performance and clinical association of the detection of tcdCΔ117 in patients with CDI in a prospective observational study conducted in a tertiary care hospital in Germany. MethodsFrom June to October 2015, stool from all patients with suspected CDI was tested for C. difficile according to ESCMID guidelines. C. difficile was cultured from positive samples and ribotyping was performed. Clinical data were collected prospectively from all C. difficile positive patients. ResultsFrom 1121 tested stool samples 107 patients with CDI were included in the study. TcdCΔ117 was detected in 18 (16.8%) of these patients. Multivariable logistic regression analysis revealed an independent association of detection of tcdCΔ117 with a further episode of CDI (OR 14.6; 95% CI 3.6–58.3; p < 0.001) and death within 30 days of the positive test (OR 5.1; 95% CI 1.0–25.7; p = 0.046). As follow up data are limited, it remains unclear, whether the further episode of CDI was due to tcdCΔ117 (recurrence) or another type. ConclusionIn our setting, PCR-based detection of tcdCΔ117 identified patients at risk for recurrent CDI and increased mortality and thus may guide therapeutic choices in CDI patients at the time of diagnosis.

Isolation, identification and antibiogram of Pasteurella multocida isolated from apparently healthy rabbits in Al-Bayda, Libya

The aim of this study was to investigate the prevalence of Pasteurella multocida in upper respiratory tract (nasal cavities) of apparently healthy rabbits. Twenty adult and young rabbits were sampled, using sterile cotton swabs for sample collection from the nasal cavities of test animals. Swabs were put into nutrient broth and incubated at 37° C for 24 hrs. nasal swabs from 14 of the 40 samples rabbits (35%) showed visible growth suggestive of P. multocida, eleven (78.6%) of the isolates were males, and three isolates (21.4%) were females, whereas 26 samples (65%) were negative. Presumptive identification of the isolates was performed according to conventional bacteriological methods of gross colonial morphology, Gram staining reaction, microscopic characteristics and biochemical tests. Results achieved in this study indicate that P. multocida is harbored in the upper respiratory tract of nearly third of the rabbit population in the laboratory animal unit of the faculty of veterinary Medicine, Omer Al- Muktar university. Antimicrobial sensitivity of P. multocida isolated were tested against seven antibacterial antibiotics. 100% of the isolates were found to be sensitivity to Neomycin and 71.4% were sensitivity to Tetracycline; Erythromycin and Streptomycin should sensitivity of 50% and 14.3%, respectively. One of the hand all isolates found to be resistant to Ofloxacin, Amoxicillin and Cephalexin.