Culture Tubes Research Articles

Comparison of different diagnostic modalities for isolation of Mycobacterium Tuberculosis among suspected tuberculous lymphadenitis patients

Tuberculosis is a communicable disease with high morbidity and mortality rates in developing countries. The study's primary objective is to compare conventional methods such as acid-fast bacillus (AFB) culture and microscopy with rapid diagnostic methods. The secondary objective is to compare histopathological and microbiological findings in suspected patients with tubercular lymphadenitis. A total of 111 samples (August 2018 to September 2019) of lymph nodes were processed for AFB microscopy, AFB cultures, drug-susceptibility testing (DST), histopathology, and Xpert Mycobacterium Tuberculosis (MTB)/resistance to Rifampin (RIF) assays. Out of 111 lymph node samples, 6 (5.4%) were positive for AFB smear microscopy, 84 (75.6%) were positive for AFB culture, 80 (70.7%) were positive on Gene Xpert, and 102 (91.8%) were indicative of tuberculosis for histopathology studies. Mycobacteria growth indicator tube (MGIT) culture positivity was 84 (75.6%) higher than solid Lowenstein-Jensen (LJ) culture 74 (66.6%). Positive cultures underwent phenotypic DST. Two cases were Multidrug-resistant (MDR) on DST, while three cases were Rifampicin resistant on Gene Xpert. The sensitivity of Genexpert was (62%) against the conventional AFB culture method. The poor performance of conventional lymphadenitis diagnostic methods requires early and accurate diagnostic methodology. Xpert MTB/RIF test can help in the treatment of multidrug-resistant TB cases. Nonetheless, rapid and conventional methods should be used for complete isolation of Mycobacterium tuberculosis.

Synthesis of Gingerol-Metals Complex and in-vitro Cytotoxic Activity on Human Colon Cancer Cell Line.

Herbs are excellent sources of medicinal substances, and their curative abilities have been recognized to treat many ailments and are used for example as antioxidants, analgesics, anti-inflammatories, antipyretics, and many other medicinal uses. The properties of natural compounds and their health effects have been studied extensively, especially those that originate from plant sources such as ginger. The ginger plant contains many chemical compounds, such as 6-gingerol, which is characterized by containing active groups such as carbonyl and hydroxide, which can be attached to metal molecules. This is what was done in this study, where the formation of complexes with a group of metals was studied and their effect on cancer cells was investigated. These complexes will open new horizons for further study of medicinal uses. The synthesis of gingerol-metal complexes was carried out by conjugating gingerol molecules with Ag, Au, Cd, Co, Cu, Ni, and Zn metal ions. The extracted gingerol was transferred to culture tubes and deionized water-DMSO were added followed by sonication. The tubes were incubated at 90°C for two days as well as the control sample. The samples were then filtered and the complex solutions were transferred into new tubes for further studies. Different characterization techniques such as FT-IR, UV-visspectroscopy, FESEM, and EDX are used to confirm the formation of the complexes. The in vitro of the complexes was tested by the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay against the human colorectal cancer cell lines HCT116 and HT29 which exhibited strong cytotoxicity. The gingerol-metal complexes showed an enhancement as an anticancer agent compared to the control. The in vitro anticancer activity showed that the Ag-gingerol complex showed the most activity among the other complexes. Gingerol-metal complexes can inhibit cancer cells, noting that the potency of the complex depends on the type of metal used.

Isolation of biofilm formers from aquaculture and study, the effect of antibiofilm activity of chitosan mediated silver nanoparticle

Bacterial biofilms are the root cause of biofouling and are due to the agglomeration of microbes and microorganisms on a surface that is surrounded or held together by extracellular polymeric substances. Bacteria produce extracellular polymeric substances (EPS), lipopeptides, and other extracellular matrix proteins, which enable them to adhere on to help them attach to surfaces and bind to one another. These extracellular substances can adhere to metals and varied surfaces, possibly producing toxic compounds such as. sulfhydryl groups, In Aquaculture fam and Ponds such Biofilm forms are of greatest threat by paving the way to settle down for broad diversity of epibiotic organisms. They together can contribute economic impact of Aquaculture and farms and ponds.Chitin, the second most abundant polymer on earth, is a commercially valuable product. The deacetylated form of chitin, chitosan possesses more attractive and useful properties. In this study, the raw material used was the waste of the Spp of shrimp, Penaeus indicus. Chitosan-Ag Nano complex was synthesized and used for its antibiofilm-forming properties. Microbes capable to form biofilm isolated from contaminated sites were isolated and characterized for their identity using microtiter plate assay, tube culture assay, and slide culture assay biofilm formers were observed from two different environments such as paddy fields and ponds. Antibiofilm-forming properties of chitosan nanoparticles were assessed using microtiter plate assay.

Modification of root architecture of rice by guard cell and rhizosphere halotolerant bacteria under saline stress

A significant global issue for agriculture is soil salinity. Rice, being one of the most salt-sensitive cereals, is greatly affected by salinity. High salt concentrations have a detrimental impact on root growth of rice. Halotolerant bacteria are known to alleviate salinity stress and have been shown to alter the root morphology of some plants. This possible alteration effect of halotolerant bacteria on rice is not well known. In this study, the halotolerant bacterial strains Enterobacter cloacae GCH3, Acinetobacter sp. GCH4, Acinetobacter calcoaceticus RSH6 and Bacillus tequilensis RSH8 were used to evaluate the early alterations of rice root architecture. These bacteria were inoculated into rice seedlings growing in culture tubes under gnotobiotic conditions, and root architecture variables of developing roots were assessed periodically using non-destructive 2-dimensional imaging and processing in both normal and salt stress conditions. The images were analyzed and processed to obtain 20 root architecture variables in which, the variables such as maximum number of roots, network solidity, number of lateral roots, network surface area, root thickness, area, volume and bushiness were significantly increased with time course due to halotolerant bacterial inoculation than uninoculated control. The root and shoot biomass was also found to be increased by bacterial inoculation. Number of connecting components, median number of roots, specific root length and major ellipse axes were negatively impacted by halotolerant bacteria. Enterobacter cloacae GCH3 and Bacillus tequilensis RSH8 excelled in most of the traits in improving the root architecture of rice under saline conditions which can promote the early growth stages of root and protect the plant from saline stress effects.

3D‐Printed Centrifugal Pump Driven by Magnetic Force in Applications for Microfluidics in Biological Analysis (Adv. Healthcare Mater. 24/2022)

Dynamic Cell Cultures Article 2200593 by Shoji Takeuchi and co-workers proposes a 3D-printed centrifugal pump driven by magnetic force. Precise control of the flow rate and a wide range and variety of flow profiles are achieved by controlling the rotational speed of the motor. Compact size enables simple setups within commercially available culture dishes and tubes. Moreover, demonstrations of laminar flow production and perfusion culture are presented for biological applications in microfluidic devices.

Characterization of biofilm formation by Mycobacterium smegmatis during different environmental stress conditions: An in-vitro study

Aim: In-vitro characterisation of biofilm produced by Mycobacterium smegmatis (M. smegmatis), a surrogate model for biofilm production by Mycobacteria, and to evaluate the impact of different environmental stress on mycobacterial growth and biofilm formation. Methodology: M. smegmatis biofilms were studied using tissue culture plate and tube adherence methods. Confocal Laser scanning microscopy (CLSM) and scanning electron microscopy (SEM) were used to study the 3D structure and surface morphology, respectively. Additionally, the effect of different environmental stress, such as the absence of essential ions, exposure to harsh environmental conditions, such as acidic environment or oxidative stress, on mycobacterial biofilm formation and mycobacterial growth was assessed. Results: All the exposures, except for carbon supplemented media had a detrimental effect on the number of viable counts and on biofilm formation by mycobacteria (p<0.001). Growth in low pH and oxidative stress was found to be maximum showing reduction by 98% when compared with control. Interpretation: Our findings present various environmental conditions that profoundly affect biofilm formation and thus, may find practical implications in future as effective mycobacterial control strategies having attributes of mycobacterial growth as well as biofilm inhibition. Key words: Biofilm, Environmental stress, Multi-drug resistance, Mycobacterium

Pulmonary non-tuberculous mycobacteria in colonisation and disease in The Gambia

The clinical relevance of pulmonary non-tuberculous mycobacteria (PNTM) in The Gambia is unknown. The aim of this study was to estimate the prevalence of non-tuberculous mycobacteria (NTM) in colonisation, and the burden of clinically relevant pulmonary NTM (PNTM) disease in The Gambia. This was a cross-sectional study of the prevalence of NTM in participants aged ≥ 15 years, in a nationwide tuberculosis (TB) prevalence survey between December 2011 and January 2013. We enrolled 903 participants with suspected NTM and NTM cultures were confirmed by 16S rRNA gene sequencing analyses. We applied the American Thoracic Society/Infectious Disease Society of America (ATS/IDSA) diagnostic criteria to determine clinical relevance of NTM. A total of 575 participants had acid-fast bacilli (AFB) positive Mycobacterial Growth Indicator Tube (MGIT) cultures and 229 (39.8%) were NTM. M. avium complex was by far the most isolated NTM (71.0%), followed by M. fortuitum (9.5%) and M. nonchromogenicum (2.9%). Older participants (> 24 years old) were four times more likely to have NTM in their sputa. Only 20.5% (9/44) NTM cases met the ATS/IDSA criteria for NTM disease. This study provides important data on the prevalence of NTM in pulmonary samples of suspected TB cases with AFB positive cultures from a nationally representative population in The Gambia. Enhanced PNTM surveillance is recommended to better understand the contribution of NTM to pulmonary disease.

Role of endotracheal tube culture or aspirate culture in identifying mechanically ventilated patients at risk for ventilator-associated pneumonia in neonatal intensive care unit at a tertiary care centre

Background: Ventilator-associated pneumonia (VAP) is the second most common hospital-acquired infection in Neonatal intensive care units (NICU). The standard methods of diagnosis are not popular because of their invasive nature and cost. Endotracheal aspirate (ETA) is a relatively easy and non-invasive method. The aim of the study was to document the role of routine endotracheal aspirate culture performed in predicting causative organism and selecting effective antibiotic therapy in the event of subsequent ventilator-associated pneumonia.Methods: This study was conducted in the NICU of a medical college over ten months (September 2019 to June 2020). Newborns up to 28 days of life admitted in NICU and mechanically ventilated for more than 48 hours were enrolled in this prospective observational study (cross-sectional study). After 48 hours of ventilation, ET aspirate is collected and sent for culture and sensitivity testing in the microbiology lab. A Chi-square test was applied. P value<0.05 was considered statistically significant.Results: A total of 50 patients were included in this study, of which 15 patients developed VAP. Among the 6 VAP pathogens in this study, 94.12% were gram negative bacilli and 5.88% were gram positive cocci. Acinetobacter baumanii (52.80%) was the most predominating causative organism, followed by Klebsiella, Pseudomonas and E. coli (11.76%), followed by coagulase-negative staphylococcus and Enterobacter cloacae (5.88%).Conclusions: Acinetobacter baumanii was the most common organism isolated. Endotracheal tube aspirate cultures help in the early identification of patients at risk for developing VAP.

Generation of good manufacturing practice grade virus-specific T cells for the management of post-transplant CMV infections

Adoptive transfer of antigen-specific T cells has recently emerged as a successful strategy to treat viral infections following hematopoietic cell transplantation (HCT). Ex-vivo expanded donor-derived virus-specific T cells (VSTs) can be safe and effective, devoid of all the drug-related adverse effects. The study aimed to manufacture cGMP grade VSTs from healthy donors, characterize the VST product and demonstrate its safety and efficacy. Peripheral blood mononuclear cells (PBMCs) collected from six healthy donors were stimulated with pepmix that mimics the pp65 antigenic epitope of CMV and cultured for 14 days in G-Rex culture tubes. Post pepmix exposure and expansion the median CD3% was 98.8% (range:95.5% to 99.9%) while the median CD4% and CD8% were 49.1% (range:21.3% to 86.6%) and 43.9% (range:12.7% to 75.5%) respectively. The percentage of IFNγ+ cells was much higher among the CD8+ T cells (median - 18.47%; range 6.50% - 45.82%) when compared to CD4+ T cells (median − 2.74%; range 0.47% - 18.58%) and there was a switch from the CD45RA+ naive phenotype to CD45RA- effector memory phenotype in the 4 samples that achieved a >5 fold expansion. The VSTs were cytotoxic to the pepmix pulsed lymphoblasts (efficacy) while they did not induce cytolysis in the lymphoblasts that were not exposed to the pepmix (safety). This feasibility exercise helped us optimize the starting cell dose for the culture and clinical grade culture strategies, subset characterization and cytotoxicity assays. The approach could be applied to the clinical practice where virus-specific T cell infusions could be given for post-transplant viral infections.

P245 Arthrinium species, a filamentous ascomycetes isolated from samples of human cutaneous infections-report from a medical mycology laboratory of Assam, North-East India

Poster session 2, September 22, 2022, 12:30 PM - 1:30 PMObjectivesThis study aims to report the isolation of closely related Arthrinium species from superficial skin lesions of five cases from a medical mycology laboratory of Assam, North East India.MethodsThe lesions were decontaminated with 70% ethanol and skin scrapings were collected on a sterilized glass plate. Direct mounts were prepared in 10%-20% KOH and cultures were put in Saboraud's Dextrose Agar with antibiotics, 5% sheep blood agar, and dermatophyte test medium (Himedia, India). Plates and tubes were incubated as per standard mycological techniques described. Molecular identification was done using ITS sequence analysis using ITS1 and ITS4 universal primers.ResultsDirect mount showed presence of hyphae with arthrospores in 3/5 cases. In one case, fungal hyphae was seen along with spore-like oval or round structures of about 3-4 μm diameter. Pure growth was seen after 7-14 days in multiple culture tubes in all five cases. Colonies were white, downy initially becoming white, and floccose on further incubation. Subculture on PDA in all the cases for 15-20 days revealed black, round, and oval spores of 3-5 μm suggesting Arthrinium spp.The taxonomical identification was done by constructing a phylogenetic tree of the ITS sequences of the Arthrinium isolates of this study along with reference Arthrinium strains and Seiridium phylicae as the outgroup taxa.The phylogenetic analysis clustered the isolates of this study into closely related Arthrinium species.ConclusionThe genus Arthrinium belonging to the family Apiosporaceae, class Sordariomycetes which comprises of a group of filamentous ascomycetes fungi is rarely reported from human infections. We are reporting closely related Arthrinium spp from five cases of skin lesions from Assam, North East India. Three of the 5 cases hailed from tea garden areas of Assam. Arthrinium isolation in clinically significant cases and in multiple tubes may not be disregarded as a contaminant.

Reduction of blood C-reactive protein concentration complements the resolution of sputum bacillary load in patients on anti-tuberculosis therapy.

BackgroundTuberculosis (TB) is a difficult-to-treat disease requiring the combination of four antibiotics for a minimum of 6 months. Rapid and quantitative biomarkers to monitor treatment response are urgently needed for individual patient management and clinical trials. C-reactive protein (CRP) is often used clinically as a rapid marker of inflammation caused by infection. We assessed the relationship of TB bacillary load and CRP as biomarkers of treatment response.MethodsXpert MTB/RIF-confirmed pulmonary TB cases were enrolled for treatment response assessment in Mozambique. Treatment response was measured using the Tuberculosis Molecular Bacterial Load Assay (TB-MBLA) in comparison with standard-of-care Mycobacterium Growth Indicator Tube (MGIT) culture at baseline and at weeks 1, 2, 4, 8, 12, 17, and 26 of treatment. Blood CRP concentration was measured at baseline, week 8, and week 26. Treatment response was defined as increase in MGIT culture time to positivity (TTP), and reduction in TB-MBLA-measured bacillary load and blood CRP concentration.ResultsOut of the 81 screened presumptive TB cases, 69 were enrolled for 6-month treatment follow-up resulting in 94% treatment completion rate. Four participants did not complete TB treatment and 22 participants had missing CRP or TB-MBLA results and were excluded from TB-MBLA-CRP analysis. The remaining 43 participants—median age, 31 years old [interquartile range (IQR): 18–56]; 70% (30/43) male; and 70% (30/43) infected with HIV—were considered for analysis. Culture TTP and bacillary load were inversely correlated, Spearman’s r = −0.67, p < 0.0001. Resolution of sputum bacillary load concurred with reduction of blood CRP, r = 0.70, p < 0.0001. At baseline, bacillary load had a median (IQR) of 6.4 (5.5–7.2), which reduced to 2.4 (0.0–2.9) and 0.0 (0.0–0.0) log10 CFU/ml at months 2 and 6 of treatment, respectively. Correspondingly, blood CRP reduced from 1.9 (1.6–2.1) at baseline to 1.3 (0.9–1.7) and 0.4 (0.1–0.8) log10 mg/dl at months 2 and 6 of treatment, respectively. CRP reduction trialed bacteriological resolution at a rate of −0.06 log10 mg/dl compared to a bacillary load of 0.23 log10 CFU/ml per week. Consequently, 14 (33%) and 37 (88%) patients had reduced CRP to normal concentration and bacillary load to zero by the end of treatment, respectively. Pre-treatment CRP concentration and bacillary load, and resolution during treatment were slightly lower in HIV co-infected patients but not significantly different from HIV-uninfected TB patients.ConclusionTB-MBLA-measured bacillary load and blood CRP complement each other in response to anti-TB therapy. Slow CRP reduction probably reflects residual TB bacilli in the lung not expectorated in sputum. Combining both measures can improve the accuracy of these biomarkers for monitoring TB treatment response and shorten turnaround time since the results of both assays could be available in 24 h.

How to simply and efficiently screen microbial strains capable of anaerobic biosynthesis of biosurfactants: Method establishment, influencing factors and application example evaluation.

Microbial resources capable of anaerobic biosynthesis of biosurfactants are increasingly interested for their application in oxygen-deficient environments, such as in-situ microbial enhanced oil recovery and anaerobic bioremediation. How to simply and efficiently screen microbial strains capable of anaerobic biosynthesis of biosurfactants need be further studied in depth. In this study, an efficient and simple screening method was established based on the oil displacement characteristic of biosurfactants combined with the anaerobic culture technology using microplate assays. Strains whose anaerobic culture in microwells can form oil displacement circles with diameters larger than 10 mm were screened for scale-up culture in anaerobic tubes. The screened strains which can reduce the surface tension of anaerobic culture to lower than 45 mN/m were verified as positive strains. Using this screening method, eight positive strains and thirteen positive strains were screened from oil reservoir produced water and oily sludge, respectively. Through phylogenetic analysis, some screened strains were identified as Pseudomonas sp., Bacillus sp., and Enterobacter sp. This study also found that more microbial strains might be isolated after enrichment culture of environmental samples, whereas more microbial species would be isolated without enrichment. Suspension of environmental samples prepared with distilled water or normal saline had no significant effect. The established screening method is highly targeted and efficient for microbial strains capable of anaerobic biosynthesis of biosurfactants. The diameter of oil displacement circle is a reliable screening indicator. This study will contribute to explore more microbial resources which can anaerobically biosynthesize biosurfactants.

The associative effects of sunflower straw, sunflower plate, sunflower seed shells associated with concentrate and alfalfa evaluated by using an in vitro gas production technique

This study aimed to evaluate the multiple-factor associative effects (AEs) of concentrate (C) to sunflower straw (SS) to alfalfa (A) ratios, concentrate to sunflower plate (SP) to alfalfa ratios, concentrate to sunflower seed shell (SSS) to alfalfa ratios at 40 : 60 : 0, 40 : 45 : 15, 40 : 30 : 30, 40 : 15 : 45, 40 : 0 : 60 and 30 : 70 : 0, 30 : 55 : 15, 30 : 40 : 30, 30 : 25 : 45, 30 : 10 : 60, 30 : 0 : 70, respectively, by using an in vitro gas production (GP) method. Thirty-three feed combinations and nine single feeds C, SS, A; C, SP, A; C, SSS, A were incubated respectively for 72 h in a GP tube. A total of 42 treatment combinations were tested, including 33 feed mix combinations [(5 + 6) × 3] and nine single raw materials, a total of 42 sample culture tubes. Each sample culture tube was repeated 20 times, totalling 840 (42 × 20) sample culture tubes. In this study, a total of 12 batches of in vitro culture tests were carried out. Seventy sample tubes and six blank tubes were cultured in each batch (eliminating systematic errors). A total of 912 sample culture tubes were used. The GP values of 0, 2, 4, 6, 9, 12, 24, 36, 48, 72 h were recorded. The GP parameters a, b, c, a + b were calculated by a single exponential equation. The AE values were calculated based on 72 h GP and weighted estimation value of 33 feed combinations. In the incubation fluid pH, ammonia nitrogen (NH&lt;sub&gt;3&lt;/sub&gt;–N) and volatile fatty acids (VFA) after 72 h incubation were determined and dry matter digestibility (DMD), organic matter digestibility (OMD) were measured in the residue. The single-factor associative effects index (SFAEI) and multiple-factor associative effects index were calculated. The results indicated that groups 40 : 45 : 15 (C:SS:A), 30 : 25 : 45 (C : SS : A), 40 : 60 : 0 (C : SP : A), 40 : 45 : 15 (C : SP : A), 30 : 40 : 30 (C : SP : A), 40 : 45 : 15 (C : SSS : A), 40 : 30 : 30 (C : SSS : A), 30 : 25 : 45 (C : SSS : A) and 30 : 10 : 60 (C : SSS : A) had higher a, b, GP&lt;sub&gt;72h&lt;/sub&gt;, NH&lt;sub&gt;3&lt;/sub&gt;–N, DMD, OMD, moreover, higher SFAEI (the AE of GP&lt;sub&gt;72h&lt;/sub&gt;, NH&lt;sub&gt;3&lt;/sub&gt;–N, total VFA, DMD and OMD) than other groups (P &amp;lt; 0.05). It was concluded that the optimal feed combinations occurred when concentrate/sunflower straw/alfalfa ratios were 40 : 45 : 15 and 30 : 25 : 45; concentrate/sunflower plate/alfalfa ratios were 40 : 60 : 0, 40 : 45 : 15 and 30 : 40 : 30; concentrate/sunflower seed shell/alfalfa ratios were 40 : 45 : 15, 40 : 30 : 30, 30 : 25 : 45 and 30 : 10 : 60.

Combining Metabolic Engineering and Multiplexed Screening Methods for 3-Hydroxypropionic Acid Production in Pichia pastoris.

Production of 3-hydroxypropionic acid (3-HP) in Pichia pastoris (syn. Komagataella phaffii) via the malonyl-CoA pathway has been recently demonstrated using glycerol as a carbon source, but the reported metrics were not commercially relevant. The flux through the heterologous pathway from malonyl-CoA to 3-HP was hypothesized as the main bottleneck. In the present study, different metabolic engineering approaches have been combined to improve the productivity of the original 3-HP producing strains. To do so, an additional copy of the gene encoding for the potential rate-limiting step of the pathway, i.e., the C-terminal domain of the malonyl-CoA reductase, was introduced. In addition, a variant of the endogenous acetyl-CoA carboxylase (ACC1 S1132A ) was overexpressed with the aim to increase the delivery of malonyl-CoA. Furthermore, the genes encoding for the pyruvate decarboxylase, aldehyde dehydrogenase and acetyl-CoA synthase, respectively, were overexpressed to enhance conversion of pyruvate into cytosolic acetyl-CoA, and the main gene responsible for the production of the by-product D-arabitol was deleted. Three different screening conditions were used to classify the performance of the different strains: 24-deep-well plates batch cultures, small-scale cultures in falcon tubes using FeedBeads® (i.e., slow release of glycerol over time), and mini bioreactor batch cultures. The best two strains from the FeedBeads® screening, PpHP8 and PpHP18, were tested in bioreactor fed-batch cultures using a pre-fixed exponentially increasing feeding rate. The strain PpHP18 produced up to 37.05 g L−1 of 3-HP at 0.712 g L−1 h−1 with a final product yield on glycerol of 0.194 Cmol−1 in fed-batch cultures. Remarkably, PpHP18 did not rank among the 2-top producer strains in small scale batch cultivations in deep-well plates and mini bioreactors, highlighting the importance of multiplexed screening conditions for adequate assessment of metabolic engineering strategies. These results represent a 50% increase in the product yield and final concentration, as well as over 30% increase in volumetric productivity compared to the previously obtained metrics for P. pastoris. Overall, the combination of glycerol as carbon source and a metabolically engineered P. pastoris strain resulted in the highest 3-HP concentration and productivity reported so far in yeast.

Comparison of TrueNat polymerase chain reaction and mycobacterium growth indicator tube culture in the diagnosis of pulmonary and extrapulmonary tuberculosis

Comparison of TrueNat polymerase chain reaction and mycobacterium growth indicator tube culture in the diagnosis of pulmonary and extrapulmonary tuberculosis

POTENSI DAN ANALISA BIAYA PENYEDIAAN BIBIT TEBU (Saccharum officinarum L.) UNGGUL SECARA IN VITRO DI PT. GUNUNG MADU PLANTATIONS

&lt;p&gt;&lt;em&gt;Sugarcane productivity included cane yield and sugar extracted rate (rendemen) fluctuated annually. The main causes of low sugarcane productivity is quality of planting material with no high sugar extracted rate (sugar rendemen) potential, resistance to pests and diseases, stability production, tolerance to climate changed and early age of harvest. For increasing of the yield productivity and producing a superior sugarcane planting material with competitiveness cost, the vegetative propagation through in vitro culture technique via shoot tip culture was carried out. The objective of this research is to produce a superior sugarcane planting in vitro and analyze the costs required. The research was carried out at the Tissue Culture Laboratory, PT. Gunung Madu Plantations (GMP) in March-May 2021. The research stages started from the preparation of explant source plants, sterilization of explants and tools, manufacture of stock solutions and planting media, subculture, and analysis of the costs needed to provide superior sugarcane seeds. The results showed that the basal swelling of explants, increasing the number of shoots, root elongation, and the method of providing superior sugarcane seeds from apical shoot explants in vitro were relatively more competitive with the price of plantlets per culture tube contained 10 clones costly were Rp. 20,328, compared with conventional propagation methods. with the price of sugar cane seedlings material were Rp 1,603/stem.&lt;/em&gt;&lt;/p&gt;

Repeated-batch fermentation of Scheffersomyces stipitis in biofilm reactor for ethanol production from the detoxified and glucose- or xylose-enriched rice husk hydrolysate and its kinetic modeling

The present study aims to determine the best biofilm support material (BSM) via culture tube fermentation, produce ethanol from the detoxified and glucose- or xylose-enriched rice husk hydrolysate (RHH) by using Scheffersomyces stipitis (ATCC 58784) in the biofilm reactor, and model kinetically the ethanol fermentation performed in culture-tubes and the biofilm reactor. The activated charcoal was employed to detoxify the acid-pretreated RHH. As a result of the culture-tube fermentation performed in both glucose- and xylose-based media, a BSM that includes polypropylene (50%), soybean hulls (35%), soybean flours (5%), yeast extract (5%), bovine albumin (5%), and some minerals (w/w) was chosen. With this BSM, ethanol yield (YP/S) and production (P) were 38.83% and 3.61 g/L for glucose-based medium and 33.06% and 3.80 g/L for xylose-based medium. Ethanol fermentations were performed at 30 °C temperature, medium pH 6.2, and 150 rpm agitation rate without aeration. Based on the results of the fermentation in the repeated-batch biofilm reactor established with the selected BSM, maximum YP/S and P were 35.01% and 2.57 g/L when glucose was used as the sole carbon source (Medium A). Regarding the fermentation in which the RHH was used, YP/S and P were 23.50% and 1.98 g/L when 75% RHH (1.125-L) and 25% glucose (0.375-L) were used as carbon sources, respectively (Medium D). When the RHH (50%, 0.75-L) was supplemented with xylose (50%, 0.75-L), YP/S and P were 22.65% and 1.96 g/L, respectively (Medium E). Additionally, modeling results demonstrated that ethanol production was growth-dependent since α ≠ 0 and β≌0 and sugars were utilized for ethanol production and microbial growth because γ≫m. Consequently, the RHH is a promising carbon source to produce ethanol in the repeated-batch biofilm reactor. Also, the biofilm reactor constructed was operated for 648 h without contamination. Therefore, the biofilm reactor system can be used for long-term production of value-added products using microorganisms.

Organogenesis from Leaf-derived Calli of S. potatorum L. f.: A Vulnerable Medicinal Tree Species

An efficient protocol for in vitro regeneration of Strychnos potatorum L. f. through indirect organogenesis was developed using mature leaf disc explants. The explants were selected from twenty years old tree. Cultured on Murashige and Skoog’s medium supplemented with 2,4-dichloro phenoxy acetic acid (2,4-D) was used for the induction of organogenic calli. A pale yellow coloured friable callus developed at 2 mg/L-1 2,4-D where showed higher morphogenic potential. Further de-novo shoots formation and multiplication in optimal concentrations of cytokinins with different combinations of auxins. An average mean shoots number (14.2±0.42) and shoots height (3.5±0.27cm) was obtained from piece of calli in 6-benzylaminopurine (BAP) 1 mg/L-1 with 0.5 mg/L-1 α-naphthalene acetic acid (NAA). BAP in combinations with NAA was found to be superior to shoot development than other combination used. The well developed shoots were transferred to root induction medium for effective rooting. The rooting medium consisted of in half-strength MS medium augmented with 0.5 mg/L-1 NAA was used. The maximal mean number of roots per shoots 6.5±0.53 was recorded in 4 weeks. The rooted plantlets were removed from the culture tubes, gently washed with sterile double distilled water to remove agar adhering on the plant surface. The regenerated plants were transferred to plastic cups filled with a potting mixture containing sterile sand and soilrite (1:1 ratio) and allowed to grow in the greenhouse. The slowly acclimatized plants were further transferred to substrate containing red soil and farmyard manure (1:1) for effective hardening. 75% survival rate was recorded. This protocol can be useful for the ex-situ conservation and rehabilitation of this vulnerable medicinal tree S. potatorum.&#x0D; Keywords: Indirect organogenesis, friable callus, shoot development, 6-benzylaminopurine, α-naphthalene acetic acid

3D-Printed Centrifugal Pump Driven by Magnetic Force in Applications for Microfluidics in Biological Analysis.

In recent years, microfluidic systems have been extensively utilized for biological analysis. The integration of pumps in microfluidic systems requires precise control of liquids and effort-intensive set-ups for multiplexed experiments. In this study, a 3D-printed centrifugal pump driven by magnetic force is presented for microfluidics and biological analysis. The permanent magnets implemented in the centrifugal pump synchronized the rotation of the driving and operating parts. Precise control of the flow rate and a wide range and variety of flow profiles are achieved by controlling the rotational speed of the motor in the driving part. The compact size and contactless driving part allow simple set-ups within commercially available culture dishes and tubes. It is demonstrated that the fabricated 3D-printed centrifugal pump can induce laminar flow in a microfluidic device, perfusion culture of in vitro tissues, and alignment of cells under shear stress. This device has a high potential for applications in microfluidic devices and perfusion culture of cells.

Diagnostic Performance of Molecular Rapid Test in Establishing Diagnosis of MDR-TB

Tuberculosis (TB) is a global health problem and is the leading cause of morbidity and mortality in many developing countries. Multi Drug Resistant Tuberculosis (MDR TB) becomes one burden of health problems considering the high morbidity and mortality rates. Establishment of MDR TB diagnosis is still a challenge, related to the tools and methods used, while cultural examination as gold standard is expensive and takes a long time. This study aims to evaluate diagnostic performance of Molecular Rapid Test or MRT (GeneXpert) in establishing diagnosis of MDR TB using Mycobacteria Growth Indicator Tube (MGIT) culture as gold standard. Using a cross sectional design, this study involved 51 subjects, a total of 26 (51%) male and 25 (49%) female, adult patients suspected TB, treated at dr. Ario Wirawan Lung Hospital (RSPAW) Salatiga. Mean age 48.2 + 16.35 years, (17-79 years). The MRT for TB and MDR TB detection showed 13 and 7 true positives, 32 and 43 true negatives; obtained 68.4% and 87.5% Sensitivity, 100% and 100% Specificity, 100% and 100% Positive Predictive Value (PPV), 84.2% and 97.7% Negative Predictive Value (NPV), respectively. A specific analysis to diagnose MDR TB by MRT on TB patient groups showed 7 true positives, 0 false positives, 11 true negatives and 1 false negative; obtained 87.5% Sensitivity, 100% Specificity, 100% PPV and 91.7% NPV. For establishing diagnosis of MDR TB, MRT provides perfect specificity. One false negative MDR TB in MRT results is likely resistant to other than rifampicin.