NMR studies of the interaction between Bacillis subtilis neutral proteinase and inorganic metal compounds

The interaction between Bacillis subtilis neutral proteinase (B.S.NP) and inorganic metal compounds (CoCI2, NiCI2) was investigated by 1H NMR spectroscopy. It has been shown that the Zn(Ⅱ) ion in the active center of the native enzyme may directly interact with external CoCI2 and NiCI2, producing Co(Ⅱ)- and Ni(Ⅱ)-substituted derivatives, and their 1H NMR spectra were obtained for the first time. From the 1H NMR spectra, the coordinated structure of the active center in the native enzyme was described.

Treatment of primary nasal tuberculosis with anti-tumor necrosis factor immunotherapy:A case report

BACKGROUND Primary nasal tuberculosis(TB)is a rare form of extrapulmonary TB,particularly in patients receiving anti-tumor necrosis factor(TNF)immunotherapy.As a result,its diagnosis remains challenging.CASE SUMMARY A 58-year-old male patient presented to the ear,nose,and throat department with right-sided nasal obstruction and bloody discharge for 1 month.He was diagnosed with psoriatic arthritis and received anti-TNF immunotherapy for 3 years prior to presentation.Biopsy findings revealed chronic granulomatous inammation and a few acid-fast bacilli,suggestive of primary nasal TB.He was referred to our TB management department for treatment with oral anti-TB agents.After 9 months,the nasal lesions had disappeared.No recurrence was noted during follow-up.CONCLUSION The diagnosis of primary nasal TB should be considered in patients receiving TNF antagonists who exhibit thickening and crusting of the nasal septum mucosa or inferior turbinate,particularly when pathological findings suggest granulomatous inflammation.

油脂下脚料降解菌的选育

从油污土样等样品分离获得两种高产蛋白酶的菌株,命名为M1、DB2,其酶活力分别213.04,206.95 U/mL。利用该两株菌分别对大豆粕,芝麻粕,菜籽粕作为发酵基质进行了初步的降解蛋白能力比较研究,结果表明M1、DB2均能很好的降解三种粕中的粗蛋白。M1、DB2的最适生长温度分别为30、35℃,最适生长pH分别为7、8,最佳生长时间分别为12、32 h。初步鉴定M1、DB2为芽孢杆菌属。

Dynamic of Soil Microorganisms from Root Region of Ginseng with Different Growing Years

Objective To see the dynamic of fungi, bacilli and actinomyces communities from root region of ginseng with different growing years.Method With ginseng root region soils from several sampling sites of Jilin Province as materials, concentrations of fungi, bacilli and antinomyces were evaluated by spread-plate method. Result Though there are differences on statistic data among soil samples, commonly with the increasing of growing years, concentration of fungi in ginseng root region increased, which were on the contrary for bacilli and antinomyces, and bacilli changed even more significant than antinomyces. Conclusion Concentrations of soil microorganisms can be influenced by soil type, planting mode and growing years simultaneously, but growing years influenced even more significantly.

Diagnosis of Crohn's disease in India where tuberculosis is widely prevalent

AIM:To define the parameters that positively predict diagnosis of Crohn's disease (CD) and differentiate it from gastrointestinal tuberculosis (GITB). METHODS:This prospective study over 3 years was carried out in the consecutive Indian patients with definite diagnosis of CD and equal numbers of patients with definite diagnosis of GITB. Demographic, clinical, laboratory, morphological and histological features were noted in all the patients. Serological tests such as p-ANCA, c-ANCA, IgA ASCA and IgG ASCA, were performed. Endoscopic biopsy and/or surgical tissue specimens were subjected to smear and culture for acid-fast bacilli (AFB) and tissue polymerase chain reaction for tuberculosis (TB PCR). Diagnosis of CD and GITB was based on the standard criteria. Data were analyzed using univariate Chi-square test and multiple logistic regression (MLR). RESULTS:The study is comprised of 26 patients with CD (age 36.6 ± 8.6 year, male:female, 16:10) and 26 patients with GITB (age 37.2 ± 9.6 year, male:female, 15:11). The following clinical variables between the two groups (CD vs TB) were significant in univariate analysis:duration of symptoms (58.1 ± 9.8 vs 7.2 ± 3.4 mo), diarrhoea (69.2% vs 34.6%), bleeding per rectum (30.7% vs 3.8%), fever (23.1% vs 69.2%), ascites (7.7% vs 34.6%) and extra-intestinal manifestations of inflammatory bowel disease (61.5% vs 23.1%). Of these, all except ascites and extra-colonic manifestations were found statistically significant by MLR. Accuracy of predicting CD was 84.62% based on the fever, bleeding P/R, diarrhoea and duration of symptoms while it was 63.4% when histology was reported as inflammatory bowel disease and 42.3% when there was recurrence of disease after surgery. Accuracy of predicting GITB was 73.1% when there was co-existing pulmonary lesions and/or abdominal lymphadenopathy;75% when tuberculosis was reported in histology;63.4% when granuloma was found in histology;82.6% when TB PCR was positive;and 61.5% when smear and/ or culture was positive for AFB. Serological test was not useful in differentiation of CD from GITB. Positivity rates for CD and GITB were:p-ANCA-3.8% and 3.8%, c-ANCA-3.8% and 0%, IgA ASCA-38.4% and 23.1%, and IgG ASCA-38.4% and 42.3%, respectively. CONCLUSION:Simple clinical parameters like fever, bleeding P/R, diarrhoea and duration of symptoms have the highest accuracy in differentiating CD from GITB.

A middle-aged lady with a pyogenic liver abscess caused by Clostridium perfringens

The pyogenic liver abscess caused by Clostridium perfringens (C. perfringens ) is a rare, but rapidly fatal infection. It is usually associated with malignancy and immunosuppression. We report the case of 50-year-old lady with the secondary liver metastases from rectal cancer presented with fever and epigastric pain. The identification of Grampositive bacilli septicaemia, the presence of gas-forming liver abscess and massive intravascular hemolysis should lead to the suspicion of C. perfringens infection. Here we review twenty cases published since 1990 and their clinical features are discussed. The importance of "an aggressive treatment policy" with multidisciplinary team approach is emphasized.

Diagnostic efficacy of Ziehl-Neelsen method against fluorescent microscopy in detection of acid fast bacilli

Objective:To investigate the application of Ziehl-Neelsen(Z-N) and fluorescent microscopy in detection of acid fast bacilli(AFB).Methods:Duplicate smears were prepared from 260 sputum samples and stained with Z-N and fluorescent staining(FS) methods.The efficiency of both methods in primary diagnosis of tuberculosis were evaluated.Results:The smears were positive for AFB in 15(5.77%) samples by Z-N staining method and in 16(6.15%) samples by FS method.The sensitivity and specificity of Z-N staining method against FS method were 93.75% and 100%respectively.Conclusions:Though lesser cost-effective than Z-N,FS method is a more sensitive and better case finding tool in detection of AFB.

The characteristics of pulmonary tuberculosis amongst patients attending chest clinic, including age, sex, occupation and hemoglobin concentration in Benin City, Nigeria

Aim: To study characteristics of pulmonary tuberculosis amongst patients attending chest clinics in Benin City. Materials and Methods: A total of 816 subjects comprising of 563 (69%) males and 253 (31%) females were enlisted for this study. Sputum specimen and blood were collected from all subjects. Acid alcohol fast bacilli from sputa were examined using the Ziehl Nelson staning technique. Hemoglobin concentration was determined using the method described by Dacis and Lewis. Result: Pulmonary tuberculosis infection was significantly affected by age among the male and female subjects studied (P - 30 years and 41 - 50 years had the highest risk of acquiring TB amongst the male and female population respectively. Although the unemployed subjects had the highest prevalence of TB (35%), occupation did not significantly affect the prevalence of TB (P > 0.05). Irrespective of gender, anemia was significantly associated with pulmonary tuberculosis infection (P female subjects. Anemia was associated with TB infection. Intervention effort at curbing the prevalence of TB and anemia is advocated.

Clinical and laboratory features of PCR-confirmed periocular tuberculosis in China

Experts lack knowledge of periocular tuberculosis（TB） in China. Nested polymerase chain reaction（PCR） shows advantages in diagnosis of extrapulmonary TB. Our study aims to explore the clinical and laboratory features of PCR-confirmed periocular TB. We retrospectively reviewed medical records of presumptive periocular TB and performed nested PCR test to confirm diagnosis. Nine cases were recruited. Clinical symptoms were chronic and insidious. Eight cases achieved favorable visual acuity, while one underwent enucleation due to fungalTB panophthalmitis. Sensitivity of caseous necrosis, acidfast bacilli（AFB） staining and interferon γ release assay（T-SPOT） test are 33.3%, 44.4% and 85.7% respectively. Low lymphocyte percentage（P=0.019） and high monocytelymphocyte ratio（P=0.042） positively correlate with AFB staining. Male gender（P=0.048） and Langhans giant cell（P=0.048） positively correlate with caseous necrosis. To conclude, traditional TB ancillary tests are not as sensitive as nested PCR technique. Several factors facilitate diagnosis including male gender, decreased lymphocytes, and typical Langhans giant cells.

Correlation of Serum C-Reactive Protein with Disease Severity in Tuberculosis Patients

Purpose: To study the factors influencing sputum smear conversion including Serum C-Reactive Protein (CRP) and its correlation with disease severity in tuberculosis patients. Method: Levels of Serum-CRP concentrations were deter-mined in 60 patients with pulmonary tuberculosis, 30 healthy volunteers and patients in follow-up after completion of antitubercular treatment (DOTS therapy). Results: Serum-CRP levels were found to be significantly higher in smear-positive group as compared with the follow-up patients and smear-negative control group. The values were 43.65 ± 23.68, 9.88 ± 5.23 and 4.04 ± 3.85 mg/L respectively (P Conclusion: Serum-CRP levels are significantly correlated with disease severity in patients with active pulmonary tuberculosis. Thus these findings from the present study would certainly add new criteria for early diagnosis of TB, which may lead to development of new strategies to treat TB.

Tuberculosis:an experience from Mycobacterium smears and culture analysis

Objective:Simple tests like direct smear of the acid fast bacilli(AFB) and Mycobacterium culture could assist the diagnosis of tuberculosis.This study is aimed at reviewing the outcome of smears,culture results and contamination rate among specimens requested for AFB smear and Mycobacterium culture.Methods:Retrospective laboratory data analysis requesting for Mycobacterium culture from January 2005 till December 2006 was done in a tertiary teaching hospital of Universiti Sains Malaysia,Kubang Kerian,Kelantan,Malaysia.Results: Four hundred and sixty seven(36.6%) isolates grew from 1 277 specimens.Of these isolates,314 (67.2%) grew Mycobacterium tuberculosis,23(4.9%) grew Mycobacterium other than tuberculosis and 38 (8.1%) grew contaminants.Among the M.tuberculosis cultures,165(52.5%) had growth of more than 100 confluent colonies,whereas 39 cultures(12.4%) had growth of less than 19 colonies.Direct smear for AFB among smear positive cases showed presence of more than 50 bacilli/line in 231(49.5%) cases and smear negative cases accounted for 63(13.5%).Among smear positive cases,291(94.5%) cultures grew Mycobacterium species and another 17(5.5%) cultures grew contaminants.In smear negative cases,32(62.7%) cultures grew Mycobacterium species and 19(37.3%) cultures grew contaminants.Conclusion:The results from data analysis of the Mycobacterium cultures should be critically utilized in order to review the laboratory performance and to improve its services in the future.Some of the data is also useful to the administrators of the hospital in terms of estimating the risk of occupational hazard faced by the health care workers.

Metallo-β-lactamase producing nonfermentative gram-negative bacteria:An increasing clinical threat among hospitalized patients

Objective:To detect and evaluate the various methods for metallo-β-lactamases(MBL) production in Pseudomonas aeruginosa(P.aeruginosa) and Acinetobacter species.Methods:A total of 109 P.aeruginosa and 85 Acinetobacter species were screened for imipenem resistance by Kirby- Bauer disc diffusion methods.Detection of MBL production was(lone by imipenem-EDTA combined disc test,double disc synerygy test(DDST) and imipenem-EDTA MBL E test.Results: A total of 63(57.8%) strains of P.aeruginosa and 46(54.1%) strains of Acinetobacter spp.were found to be resistant to imipenem.Of the 63 imipenem resistant P.aeruginosa tested for MBL production.44(69.89;) were found to be positive and among 46 imipenem resistant Acinetobacter. 19(41.3%) were shown to be the MBL producers.Conclusions:Imipenem-EDTA combined disc test and MBL E test are equally effective for MBL detection in both P.aeruginosa and Acinetobacter spp.,but given the cost-constraints,combined disc can be used as a convenient screening method in the clinical microbiology laboratory.

Poncet's disease: An unusual presentation of tuberculosis in a diabetic lady

Authors describe a 53-year-old woman who presentedto their diabetes clinic with a three week history of multiple painful and swollen joints. She had been diagnosed with type 2 diabetes 5 years back. On examination, both knee joints and left ankle were swollen. A soft tissue swelling appeared over the medial end of the left clavicle few days later. Rheumatoid arthritis, collagen vascular diseases and other common causes of polyarthritis were ruled out by appropriate investigations. Non steroidal anti-inflammatory drugs failed to give satisfactory pain relief and the arthritis persisted. Conventional cultures of synovial fluid samples including cultures for tuberculosis were negative. Computed tomography showed a space occupying lesion involving the left sternoclavicular joint. Fine needle aspiration from the lesion was performed and acidfast bacilli were demonstrated in the smear using ZiehlNeelsen stain. The explanation of her arthritis was therefore tuberculous arthritis in left sternoclavicular joint and reactive arthritis in the rest of the joints. A diagnosis of Poncet's disease was considered in her case. We treated her with standard anti-tuberculosis drugs and the arthritis resolved within a few days. She remained symptom-free at her 2 years' follow-up.

Detection of oqxA and oqxB efflux pump genes among nosocomial coliform bacilli:An observational cross-sectional study

Objectives:To identify and test the antibiotic susceptibility of nosocomial coliform bacilli and investigate the presence of oqxA and oqxB genes among the multidrug-resistant(MDR)phenotypes.Methods:One hundred and twenty different healthcare-associated infection samples were collected.Coliform bacilli were isolated,identified by conventional methods,and then antibiotic susceptibility tests were done using the VITEK2 system and disk diffusion methods.OqxAB operon was identified using a conventional PCR-based technique.oqxA and oqxB genes were compared between MDR Klebsiella pneumonia(K.pneumonia)phenotypes and MDR Escherichia coli(E.coli)phenotypes.Besides,oqxAB operons were compared between phenotypes of K.pneumonia and E.coli isolates.Results:Seventy coliform bacilli were isolated with the predominance of K.pneumonia and E.coli.Besides,82.1%of K.pneumonia strains and 53.3%of E.coli isolates were MDR phenotypes.Significant more oqxB genes alone were found in MDR E.coli than that in MDR K.pneumoniae phenotypes(χ^(2)=10.160,P=0.003).OqxAB operon was significantly more in MDR phenotypes of E.coli than that in the susceptible phenotypes(P<0.001).There was significantly less of this operon in susceptible E.coli isolates than that in susceptible K.pneumoniae isolates(P<0.001).OqxAB positive isolates that were also resistant to fluoroquinolones,tetracycline,trimethoprim,and chloramphenicol,most probably suggested functional pumps.Conclusions:MDR coliform bacilli are strongly implicated in healthcare-associated infection.Attention should be paid to the presence of oqxAB pump,as an important mechanism in the development of resistance against many antimicrobials because it contributes to co-resistance with other categories;therefore,this pump could be a good target for efflux pump inhibitors.

Experimental Study on the Antibacterial Effect of Origanum Volatile Oilon Dysentery Bacilli In Vivo and In Vitro

To observe the germistatic and germicidal effects of origanum volatile oil (OVI) on the dysentery bacteria, the abdominal cavity of mice was infected with Shigella sonne (Sh. sonnei) and Shigella flexneri (Sh. flexneri) F 2a. After OVI was given to the mice via gastric lavage, the effects of OVI on the infected mice were observed. The minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) for dysentery bacteria were determined in vitro. The results showed that origanum volatile oil showed obvious protective effect on mice infected with Sh. sonnei and Sh. flexneri F 2a. and it had germistatic and germicidal effects on dysentry bacteria. We are led to conclude that origanum volatile oil is an effective medicine against the infection of dysentery bacteria.

Detection of Acid Fast Bacilli in Tuberculous Lymph Node Tissue and Touch Preparations—A Comparative Study

Background: The most common form of extrapulmonary tuberculosis is lymphadenitis. Lymphadenitis is considered mainly a disease of young children and has a peak age of onset at 20 to 40 years. Objectives: The purpose of this study was to detect and compare the presence of acid fast bacilli in lymph node biopsies and their touch preparations using Ziehl Neelsen and fluorescence staining. Materials and Methods: The lymph node biopsies and their touch preparations were prepared and stained with Haematoxylin and eosin to determine their morphology. Slides were stained with ZN staining and fluorescent staining technique using Auramine O and Rohdamine B to demonstrate the presence of acid fast bacilli in tissue and their touch preparations. Results: AFB was detected in 56% of biopsies and 54% of touch preparations. In addition in these cases we observed that fluorescent staining technique using Auramine O and Rohdamine B is superior to ZN stain because low magnification used in fluorescent staining technique makes it possible to scan the smear rapidly and detects the AFB as glowing spots even if it is present in small numbers. Conclusion: The results of AFB in tissue biopsies are in equivalent in their touch preparation.

Obligate aerobic, gram-positive, weak acid-fast, nonmotile bacilli, Tsukamurella tyrosinosolvens: Minireview of a rare opportunistic pathogen

Tsukamurella species are obligate aerobic,gram-positive,weak acid-fast,nonmotile bacilli.They are found in various environments,such as soil,water,sludge,and petroleum reservoir wastewater,and belong to the order Actinomycetales.In 2016,there was a reclassification of species within the genus Tsukamurella,merging the species Tsukamurella tyrosinosolvens(T.tyrosinosolvens)and Tsukamurella carboxydivorans.Tsukamurella species are clinically considered to be a rare opportunistic pathogen,because most reported cases have been related to bacteremia and intravascular prosthetic devices and immunosuppression.To date,it has been isolated only from human specimens,and has always been associated with clinical disease;human infections are very rare.Reported infections have included pneumonia,brain abscesses,catheter-related bloodstream infections,ocular infections,bacteremia,and sepsis presenting with septic pulmonary emboli in patients who are immunocompromised.To date,there is no commercially available test for identification.On the other hand,sequence-based identification,including matrix-assisted laser desorption ionization time-of-flight mass spectrometry,is an alternative method for identifying clinical isolates that are either slow growers or difficult to identify through biochemical profiling.The golden standards for diagnosis and optimal management still remain to be determined.However,newer molecular biological techniques can provide accurate identification,and contribute to the appropriate selection of definitive therapy for infections caused by this organism.Combinations of several antimicrobial agents have been proposed for treatment,though the length of treatment for infections has yet to be determined,and should be individualized according to clinical response.Immunocompromised patients often experience severe cases due to infection,and life-threatening T.tyrosinosolvens events associated with dissemination and/or failure of source control have occurred.Favorable prognoses can be achieved through earlier identification of the cause of infection,as well as successful management,including appropriate antibiotic therapy together with source control.Further analyses of similar cases are required to establish the most adequate diagnostic methods and treatment regimens for infections.

Status of ESBL Producing Bacteria Isolated from Skin Wound at a Tertiary Care Hospital in Bangladesh

Background: ESBL producing bacteria are increasing with an alarming rate with a wide range of infections. Objective: The purpose of the present study was to see the status of ESBL producing bacteria isolated from skin wounds. Methodology: This cross sectional study was conducted in the Department of Microbiology at Mymensingh Medical College, Bangladesh from January 2011 to June 2011 for a period of 6 months. All the patients, at any age with both sexes presented with skin wound infection, were taken as study population. Wound swab was taken from all patients. Specimens were processed and bacteria were isolated and identified according to standard procedure. The ESBL status was confirmed by double disc diffusion test (DDDT) and minimum inhibitory concentration (MIC) by agar dilution method by standard procedure according to Clinical Laboratory Standard Institute (CLSI). Antimicrobial resistance was done by disc diffusion method. Result: A total number of 84 wound swabs were taken of which the most common ESBL producing bacteria were Esch. coli (61.5%),?Proteus species (78.3%) and Klebsiella species (88.9%). All the isolates were sensitive to imipenem and nitrofurantoin followed by amikacin (92.9%). Conclusion: In conclusion, ESBL producing E. coli is the most common bacteria causing skin wound infection followed by Proteus species with a reduced sensitivity towards antibiotics.

Serratia marcescens and other non-AACEK GNB endocarditis: A case report and review of literature

BACKGROUND Non-Aggregatibacter aphrophilus,Aggregatibacter actinomycetemcomitans,Cardiobacterium hominis,Eikenella corrodens,Kingella spp.(non-AACEK)gramnegative bacilli(GNBs)are an infrequent and challenging cause of endocarditis associated previously with mainly intravenous drug use.Currently,this pathology has increasingly become a healthcare-associated issue.Current guidelines do not clearly define the management of non-AACEK GNB endocarditis due to a lack of prospective trials.We review characteristics,outcomes and treatment of non-AACEK GNB endocarditis,in particular Serratia marcescens endocarditis.CASE SUMMARY We describe the case report of a 46-year-old man who presented to the emergency department with high-grade fever and a purulent exudate on an intracardiac device site.Serratia marcescens mitral valve endocarditis as a consequence of complicated generator pocket infection was diagnosed.The patient was treated with complete device removal and a long course of broadspectrum antibiotics for 6 wk after surgery with intravenous piperacillintazobactam and ciprofloxacin,which was later switched to oral ciprofloxacin and sulfamethoxazole-trimethoprim.The patient had complete resolution of symptoms and inflammatory parameters at the end of the treatment and at follow-up.CONCLUSION Long-term dual-antibiotic therapy containing a beta-lactam is indicated for most non-AACEK GNB endocarditis, whereas valve surgery may not be necessary inall patients.

Detection of 16S rRNA Methylase Genes in Gram-Negative Bacilli Isolated from Hospitals in Changchun, China

Methylation of 16S rRNA is an important mechanism of aminoglycoside resistance among gram-negative pathogens. In this report, 16S rRNA methylase genes were amplified using PCR among gram-negative bacillus isolates from hospitals in the Changchun area of China and 16S rRNA methylase genotypes (armA, rmtB, rmtA, rmtC, rmtD, and npmA) were identified by direct sequencing. Fifty of the isolates (43.1%) harbored 16S rRNA methylase genes. The common 16S rRNA methylase genes were armA and rmtB (12.1% and 31.0%, respectively), whereas the rmtA, rmtC, rmtD, and npmA genes were absent from the sample. It suggests that the predominant 16S rRNA methylase genes among gramnegative bacilli in the Changchun area are armA and rmtB.