scholarly journals The threat of persistent bacteria and fungi contamination in tuberculosis sputum cultures

2021 ◽  
Vol 21 (2) ◽  
pp. 628-632
Author(s):  
Grace Muzanyi ◽  
Aber Peace ◽  
Bonny Wamuntu ◽  
Akol Joseph ◽  
Joanita Nassali
Keyword(s):  
Polymyxin B ◽  
Sputum Culture ◽  
Standard Laboratory ◽  
Indicator Tube ◽  
Tb Treatment ◽  
Significant Difference ◽  
Bacteria And Fungi ◽  
Mgit Culture ◽  
Culture Positive ◽  
Culture Sample

Background: Tuberculosis (TB) sputum culture contaminants make it difficult to obtain pure TB isolates.We aimed to study and identify persistent TB sputum culture contaminants post the standard laboratory pre-culture sample decontamination techniques. Methods: This was a longitudinal study of TB sputum culture contamination for a cohort of TB patients on standard treatment at: baseline, during TB treatment and post TB treatment. Sputum samples were decontaminated with 1.5%NaOH and neutralized using 6.8 Phosphate buffer solution.Sputum was then inoculated into MGIT (mycobactrial growth indicator tube) supplemented with 0.8ml PANTA. A drop of each positive MGIT culture was sub cultured onto blood agar and incu- bated for 48 hours at 35 -37OC.Any growth was identified using growth characteristics and colony morphology. Results: From October 2017 through May 2019;we collected 8645 sputum samples of which 8624(99.8%) were eligible and inoculated into MGIT where 2444(28.3%)samples were TB culture positive and 255(10.4%)were positive for contam- inants:237 none-tuberculosis bacteria, 12 fungi and 6 mixed(none-tuberculous bacteria+fungi).There was no statistically significant difference between none tuberculosis bacteria and fungi in the treatment (OR=1.4,95%CI:0.26–7.47,p=0.690) and the post treatment TB phases(OR=2.02,95%CI:0.38–10.79,p=0.411)Vs baseline. Conclusion: None-tuberculous bacteria and fungi dominate the plethora of TB sputum culture contamination and persist beyond the standard laboratory pre-culture decontamination algorithm. Keywords: Bacteria; Fungi; Inoculation; PANTA (Polymyxin B; Amphotericin B; Nalidixic acid; Trimethoprim; Azlocillin).

scholarly journals Accuracy of Xpert Ultra in Diagnosis of Pulmonary Tuberculosis among Children in Uganda: a Substudy from the SHINE Trial

2020 ◽  
Vol 58 (9) ◽  
Author(s):  
Willy Ssengooba ◽  
Jean de Dieu Iragena ◽  
Lydia Nakiyingi ◽  
Serestine Mujumbi ◽  
Eric Wobudeya ◽  
...  
Keyword(s):  
Positive Result ◽  
Diagnostic Challenge ◽  
Indicator Tube ◽  
Lowenstein Jensen ◽  
Mgit Culture ◽  
Mycobacterial Growth ◽  
Culture Positive ◽  
Mycobacterial Growth Indicator Tube ◽  
Higher Sensitivity ◽  
Growth Indicator

ABSTRACT Childhood tuberculosis (TB) presents significant diagnostic challenges associated with paucibacillary disease and requires a more sensitive test. We evaluated the diagnostic accuracy of Xpert MTB/RIF Ultra (Ultra) compared to other microbiological tests using respiratory samples from Ugandan children in the SHINE trial. SHINE is a randomized trial evaluating shorter treatment in 1,204 children with minimal TB disease in Africa and India. Among 352 samples and one cervical lymph node fine needle aspirate, one sample was randomly selected per patient and tested with the Xpert MTB/RIF assay (Xpert) and with Lowenstein-Jensen medium (LJ) and liquid mycobacterial growth indicator tube (MGIT) cultures. We selected only uncontaminated stored sample pellets for Ultra testing. We estimated the sensitivity of Xpert and Ultra against culture and a composite microbiological reference standard (any positive result). Of 398 children, 353 (89%) had culture, Xpert, and Ultra results. The median age was 2.8 years (interquartile range [IQR], 1.3 to 5.3); 8.5% (30/353) were HIV infected, and 54.4% (192/353) were male. Of the 353, 31 (9%) were positive by LJ and/or MGIT culture, 36 (10%) by Ultra, and 16 (5%) by Xpert. Sensitivities (95% confidence intervals [CI]) were 58% (39 to 65% [18/31]) for Ultra and 45% (27 to 64% [14/31]) for Xpert against any culture-positive result, with false positives of <1% and 5.5% for Xpert and Ultra. Against a composite microbiological reference, sensitivities were 72% (58 to 84% [36/50]) for Ultra and 32% (20 to 47% [16/50]) for Xpert. However, there were 17 samples that were positive only with Ultra (majority trace). Among children screened for minimal TB in Uganda, Ultra has higher sensitivity than Xpert. This represents an important advance for a condition which has posed a diagnostic challenge for decades.

The MPB64 immunochromatography assay: an analysis of doubtful results

2020 ◽  
Vol 50 (4) ◽  
pp. 340-343
Author(s):  
Chanchal Kumar ◽  
Kamal Shrivastava ◽  
Anupriya Singh ◽  
Naresh Kumar Sharma ◽  
Jitender Yadav ◽  
...  
Keyword(s):  
Positive Culture ◽  
World Health ◽  
Indicator Tube ◽  
Polymerase Chain ◽  
Probe Assay ◽  
Mgit Culture ◽  
Health Organization ◽  
Culture Positive ◽  
Positive Results ◽  
Growth Indicator

Culture remains the gold standard for tuberculosis (TB) diagnosis, and the mycobacteria growth indicator tube (MGIT), endorsed by the World Health Organization (WHO), is widely used. Further identification of a positive culture is done with the help of an immunochromatography assay, which often shows faint bands that are difficult to interpret. We analysed 125 BACTEC MGIT culture positive results, of which 11/16 (68.7%) of the doubtful assays, analysed by MGIT™ TBc Identification test (TBcId), were positive for Mycobacterium tuberculosis complex (MTBC), the remaining being non-tuberculous mycobacteria as determined by an in-house duplex polymerase chain reaction and line probe assay. Guidelines on faint or doubtful bands in immunochromatography assays are important so as not to overlook true-positive cases of TB.

scholarly journals 1476. Impact of an Educational Campaign on Rates of Sputum Culture Acquisition as an Opportunity for Antibiotic De-escalation

2020 ◽  
Vol 7 (Supplement_1) ◽  
pp. S739-S739
Author(s):  
Jessica Snawerdt ◽  
Derek N Bremmer ◽  
Dustin R Carr ◽  
Thomas L Walsh ◽  
Tamara Trienski ◽  
...  
Keyword(s):  
Length Of Stay ◽  
Hospital Mortality ◽  
Community Acquired Pneumonia ◽  
Sputum Culture ◽  
Post Intervention ◽  
Significant Difference ◽  
Educational Campaign ◽  
The Difference ◽  
Hospital Acquired ◽  
Culture Acquisition

Abstract Background The 2019 community-acquired pneumonia (CAP) guidelines recommend obtaining a sputum culture in patients who are empirically treated for methicillin-resistant Staphylococcus aureus (MRSA) or Pseudomonas aeruginosa to assist clinicians in optimizing antimicrobial therapy. A previous study at our institution found respiratory cultures were rarely obtained in patients with CAP. As a result of these findings, an educational campaign was implemented to promote the use of an induced sputum protocol. Methods This was a multicenter, retrospective cohort study that included patients who were ≥18 years of age, had a diagnosis of CAP, and received ≥48 hours of anti-pseudomonal antibiotics. Patients were excluded if mechanically ventilated within 48 hours of admission or diagnosed with hospital-acquired or ventilator-associated pneumonia. Patients were grouped into pre- and post-intervention time periods. The intervention involved education on obtaining respiratory cultures including technique on induced sputums and updates to CAP order sets. The primary outcome was the rate of sputum culture acquisition. Secondary outcomes included duration of anti-pseudomonal and anti-MRSA therapy, in-hospital mortality, and length of stay. Results A total of 143 patients met inclusion criteria, 72 in the pre-implementation group and 71 in the post-implementation group. Baseline characteristics were similar between the two groups. More patients in the post-implementation group had a sputum culture obtained but the difference was not statistically significant (38.9% vs 53.5%; p=0.08). Anti-pseudomonal therapy was continued for an average of 5.6 days pre-implementation and 5.2 days post-implementation (p=0.499). There was also not a significant difference in anti-MRSA duration between the two groups (3.4 days vs 3.2 days; p=0.606). In-hospital mortality and length of stay were similar between the two groups. Conclusion An educational campaign focusing on the acquisition of induced sputums led to an increase in rates of sputum cultures collected. However, this did not correlate with a decrease in duration of anti-MRSA or anti-pseudomonal therapy. Further interventions should be made to optimize de-escalation of broad spectrum antibiotics based on sputum culture results. Disclosures All Authors: No reported disclosures

scholarly journals The Nutritive Value of Commonly Consumed Processed and Unprocessed Vegetables in South-Southern Nigeria

2020 ◽  
pp. 263-273
Author(s):  
Lilian C. Aburime ◽  
Onot O. Ekpe ◽  
Gift O. Okpuruwu ◽  
Chidera T. Nweke
Keyword(s):  
Social Science ◽  
Dietary Fibre ◽  
Nutrient Intake ◽  
Nutritive Value ◽  
Chemical Analyses ◽  
Standard Laboratory ◽  
Laboratory Methods ◽  
Significant Difference ◽  
Southern Nigeria ◽  
Mineral Compositions

Aim: The nutritive value, proximate, vitamin and mineral compositions of seven commonly consumed vegetables in South-Southern Nigeria were determined. Methodology: Pods and leaves of vegetables were divided into two parts. One part was analyzed raw and the remaining part was cooked by boiling. The raw vegetable leaves and okro pods were separately washed and dried using a food dehydrator (40ºC) for 24hrs. After drying, it was milled into fine flour using an electric blender. The second part of the vegetables were processed by boiling using variable time, after boiling, they were separately drained and dried using a food dehydrator at a temperature of about 50ºC for 48 hrs. Chemical analyses were carried out using standard laboratory methods. Means and standard deviations were calculated using the statistical package for social science. The least significant difference (LSD) was accepted at P = .05 significance. Results: For the raw vegetables the results were as follows: Moisture 71.2 – 91.2%, ash 0.9- 2.9%, dietary fibre 9.2 – 13.1%, fat 0.3 -1.4%, protein 2.1 – 8.2%, available Carbohydrate 1.0 – 8.7%, phosphorus 8 -52 mg/100 g, potassium 263- 1152 mg/100 g, sodium 3-23 mg/100 g, calcium 183-815 mg/100 g, magnesium 67-217 mg/100 g, iron 0.26-1.27 mg/100 g, zinc 0.26-1.10 mg,  folate 3-13 mcg/100 g. The results of the boiled vegetables were: moisture 80.9– 93.8%, protein 1.7 – 3.2%, fat 0.2 – 0.5%, ash 0.5– 1.1%, dietary fibre 3.9 – 9.3%, available carbohydrate 3.8 – 9.9%, phosphorus 7 – 35 mg/100 g, Potassium 0.33 – 300 mg/100 g, sodium 0.3 – 20 mg/100 g, magnesium 45 – 132 mg/100 g, calcium 82 – 200 mg/100 g, iron 0.21 -0.60 mg/100 g, zinc 0.13 – 0.30mg/100g. Folate 3 – 6 mcg/100 g. The range of the percentage contributions of the vegetables that are consumed raw to the recommended nutrient intake or recommended dietary allowance of adults are as follows: protein 5-18%, fat 1-3%, carbohydrates 1-6%, dietary fibre 3-52%, iron 2-14%, zinc 6-36%, calcium 26 -116%, phosphorus 8-31%, sodium 0.3-1%, Potassium 8-32%, Magnesium 26-99%, Folate 3-4%. Conclusion: Boiling significantly reduced most nutrient studied. Bitter leaves are more nutrient dense than other studied vegetables. Knowing the food compositions of these vegetables will promote their use.

scholarly journals Treatment outcomes and influencing factors on patients with Pulmonary Tuberculosis: a retrospective study

2021 ◽  
Vol 3 (2) ◽  
pp. 105-108
Author(s):  
Jamshid Ayatollahi ◽  
◽  
Abolhasan Halvani ◽  
Mohammadhesam Gharaei Khezri ◽  
Hossein Shahcheraghi ◽  
...  
Keyword(s):  
Side Effects ◽  
Frequency Distribution ◽  
Cross Sectional Study ◽  
The Body ◽  
Sputum Culture ◽  
Response To Treatment ◽  
Sputum Smear ◽  
Drug Side Effects ◽  
Cross Sectional ◽  
Significant Difference

Introduction: Tuberculosis infection caused by Mycobacterium tuberculosis is one of the most common infectious diseases, especially in countries such as Iran. The course of treatment and the number of drugs used vary depending on the severity of the disease and the parts of the body involved. The resistant tuberculosis to treatment has increased in recent years. Thus, this study was conducted to investigate the frequency distribution of response to treatment of patients with tuberculosis in Sirjan, Iran. Methods: This descriptive cross-sectional study investigated all patients with tuberculosis in Sirjan city who had referred to health centers during the years 2011-2019. The data collection tool was a pre-prepared checklist that included information on age, sex, sputum smear results, sputum culture results, diabetes, patients' nationality, drug side effects, and response to treatment. Finally, data was entered into SPSS version 22, and analyzed. Results: In this study, the overall response rate was 83% and the mortality rate was 10%. Between the frequency distribution of response to treatment in terms of gender, age, sputum smear results, sputum culture results, patients' nationality and diabetes was not statistically significant difference. Also, no statistically significant difference was found between the frequency distribution of pulmonary TB treatment response in terms of drug allergy, drug hepatitis and other drug side effects. Conclusion: According to results, can be concluded that none of the variables: age, sex, smear and culture result, and history of diabetes have no an effect on response to treatment and mortality of tuberculosis.

scholarly journals The role of honey in healing of bedsores in cancer patients

2012 ◽  
Vol 01 (02) ◽  
pp. 66-71 ◽  
Author(s):  
Aramita Saha ◽  
Subrata Chattopadhyay ◽  
Md. Azam ◽  
Prabir Kr. Sur
Keyword(s):  
Wound Healing ◽  
Cancer Patients ◽  
Assessment Tool ◽  
Low Cost ◽  
Critical Difference ◽  
Visual Analogue Pain ◽  
Wound Assessment ◽  
Significant Difference ◽  
Bacteria And Fungi ◽  
And Control

Abstract Bacground: Honey was used to treat infected wounds as long as 2000 years before bacteria were discovered. It has been reported to have inhibitory action to around 50 species of bacteria and fungi (aspergillus, penicillium). Usually, Metronidazole powder is used in our palliative clinic for wound healing due to low cost & effectivity. Honey is cheap, easily available ingredient with high astringent activity. Objective: Objectives of the study were to find out the effectiveness of Honey in terms of rate of wound healing & pain control in bedsores of cancer patients. Materials and Methods: 40 cancer patients with bedsore wounds were randomly assigned (1:1 ratio i.e. 20 in each arm) for Study Arm (Honey plus Metronidazole powder) and Control Arm (only Metronidazole powder), attending Palliative clinic of our department in between July 2010 to September 2011. Washing of the wound with normal saline done daily before application of above medicaments. Change of posture & soft bed were encouraged in both groups. A pre designed interview proforma, standardised Bates Jensen Wound Assessment Tool and Visual Analogue Pain assessment scale were used to collect and assess data. Results: There was significant difference in wound healing status (F value = 6.523; Critical Difference =14.03, P&nit;0.05) from day 10 and pain reduction also (F value = 6.638 and Critical Difference = 1.667, P&nit;0.05) from day 7 in study arm. Conclusion: Application of honey dressing provides a better wound healing, rapid pain relief in cancer patients with bedsores in palliative settings.

Experimental cardiac necrosis in hypobaric and anemic hypoxia

1975 ◽  
Vol 39 (2) ◽  
pp. 205-208 ◽  
Author(s):  
J. J. McGrath ◽  
B. Ostadal ◽  
J. Prochazka ◽  
M. Wachtlova ◽  
V. Rychterova
Keyword(s):  
White Male ◽  
Male Rats ◽  
Standard Laboratory ◽  
Simulated Altitude ◽  
Subcutaneous Injections ◽  
The Third ◽  
Infant Rats ◽  
Significant Difference ◽  
Standard Laboratory Diet ◽  
Cardiac Necrosis

Resistance to isoproterenol-induced cardiac necrosis (IPRO) was compared in rats exposed to two types of hypoxia (i.e., hypobaric and anemic). IPRO was induced by two consecutive, subcutaneous injections of isoproterenol (80 mg/kg) at 24-h intervals. The animals were killed on the third day and the severity of the lesion was evaluated on a 0 (no damage) to 4 (severely damaged) scale. White male rats (HA) were exposed in a barometric chamber to a simulated altitude of 7,000 m (307 mmHg) for 4 h/day for 24 days. Two groups of control rats were kept at sea level; one group (SLA) was the same age and one group (SLW) was the same weight as the altitude-exposed rats. The HA rats were significantly more resistant to IPRO with a mean necrogenic rating of 1.8 compared to 3.3 for the SLA and SLW rats. Infant rats (AA) were made anemic by feeding full-cream milk and glucose for 100 days after weaning. Two groups of control animals were fed a standard laboratory diet; one group (AC) was the same age and one group (AW) was the same weight as the AA rats. There was no significant difference in the necrogenic ratings of the AA (3.3), AC (3.5), or WC (3.7) hearts. Thus, hypobaric hypoxia affords some protection against IPRO which is not afforded by anemic hypoxia. Similarities and differences in the two hypoxias are discussed.

4 SUPPRESSION OF ASH2L ALTERS DNA METHYLATION AND HISTONE PATTERNS DURING BOVINE EMBRYONIC DEVELOPMENT

2016 ◽  
Vol 28 (2) ◽  
pp. 131
Author(s):  
M. D. Snyder ◽  
J. H. Pryor ◽  
M. D. Peoples ◽  
G. L. Williamson ◽  
M. C. Golding ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Embryonic Development ◽  
Blastocyst Stage ◽  
Early Embryonic Development ◽  
Standard Laboratory ◽  
Significant Difference ◽  
Commercial Supplier ◽  
Secondary Antibodies ◽  
Laboratory Protocol

Epigenetic patterns established during early bovine embryogenesis via DNA methylation and histone modification patterns are essential for proper gene expression and embryonic development. We have previously discovered that suppression of absent, small, or homeotic-like (ASH2L) with small interfering RNA (siRNA) had no significant effect during in vitro embryo development when compared with its respective control (31.3 ± 2.0% standard error of the mean, n = 466 v. 34.8 ± 1.9%, n = 418). Analysing DNA methylation and histone modifications via immunocytochemistry will further explain the role of ASH2L during embryonic development, specifically at the blastocyst stage. In this experiment, we obtained mature bovine oocytes from a commercial supplier (De Soto Biosciences, Seymour, TN) and preformed IVF following standard laboratory protocol. Eighteen hours after IVF, presumptive zygotes were divided into 3 treatments: noninjected controls, nontargeting siRNA injected controls (siNULL), and injection with siRNA targeting ASH2L (siASH2L). Each embryo was injected with ~100 pL of 20 nM siRNA previously verified to suppress expression of ASH2L by ~79%. Embryos were cultured in Bovine Evolve (Zenith Biotech, Guilford, CT) supplemented with 4 mg mL–1 of BSA (Probumin, Millipore) for 7 days. Blastocysts from each treatment (N = 601) were fixed and prepared for immunocytochemistry following standard laboratory protocol. The following primary antibodies were used to target specific DNA and histone methylation marks: 5mc mAb (Epigentek, Farmingdale, NY), 5hmc pAb, H3K4me3 pAb (Active Motif, Carlsbad, CA), H3K4me2 pAb, H3K9me2–3 mAb, and H3K27me3 mAb (Abcam, Cambridge, MA). Embryos were fluorescently labelled with the following secondary antibodies: Alexa Flour 488 Goat Anti-Rabbit, Alexa 488 Donkey Anti-Goat, and Alexa Flour 594 Goat Anti-Mouse (Invitrogen, Carlsbad, CA). The DNA was stained with Hoechst 33342 (Invitrogen). Fluorescent images were captured using the Zeiss Stallion digital imaging work station. Ratio averages (targeting mark/DNA) were calculated and statistical analysis performed using one-way ANOVA and Tukey’s honestly significant difference to assess treatment effects. The ratio of DNA methylation to total DNA increased in siASH2L as compared with control and siNULL embryos (0.35 ± 0.01, 0.26 ± 0.02, and 0.30 ± 0.01, respectively; P < 0.01). The 5hmC was inversely related to 5mC levels and decreased in siASH2L embryos (0.75 ± 0.01, 0.93 ± 0.02, 0.87 ± 0.02, respectively; P < 0.0001). The H3K4me3 and H3K27me3 are also inversely related with decreased H3K4me3 in siASH2L versus control and siNULL embryos (0.48 ± 0.02, 0.57 ± 0.02, 0.58 ± 0.02, respectively; P < 0.001) and increased H3K27me3 (0.62 ± 0.02, 0.053 ± 0.01, 0.54 ± 0.02, respectively; P < 0.001). No differences were observed in H3K9me2–3 or H3K4me2 labelling across treatments. These results indicate that ASH2L may play a role in DNA methylation by decreasing 5mc and 5hmc conversion, which is a key event during early embryonic development. Suppression of ASH2L also alters global levels of H3H4me3 and H3K27me3, which may lead to transcription aberrations. Further analysis of siASH2L embryos via RNA-seq will help define its role during early embryonic development.

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