A study to compare the diagnostic accuracy of GeneXpert MTB/RIF/assay and its comparison with liquid culture in clinically suspected cases of genital tuberculosis attending outpatient department of tertiary center

Background: Tuberculosis is a major health issue globally despite a declining trend in mortality with effective diagnosis and treatment, an estimated 10.4 million persons developing active TB each year with 1.33 million deaths. Objective of this study was to evaluate role of GeneXpert MTB/RIF/assay in diagnosis of female genital tuberculosis in suspected cases of tuberculosis.Methods: It was a cross sectional study done in department of obstetrics and gynecology in S. N. Medical college Agra for a period of 2 year (July 2017 to October 2019). 70 cases were selected from OPD of department of obstetrics and gynecology, S. N. Medical College Agra who met the inclusion and exclusion criteria after taking proper consent. In all selected cases endometrial biopsy sample was taken using endometrial biopsy curette in premenstrual period. All samples of endometrial biopsy were taken under all aseptic precaution from both corneal ends, anterior and posterior wall and lower part of uterus using endometrial biopsy curette and sample was collected in two separate sterile vials having normal saline and was sent for GeneXpert MTB/RIF/assay and liquid culture simultaneously.Results: Out of total 70 clinically suspected cases of female genital tuberculosis in between 20-45 years of age cough with expectoration 94% was the most common respiratory symptom followed by fever 81%, weight loss 56% and anorexia 54%. Prevalence of genital tuberculosis in active pulmonary tuberculosis patients was 30%. Irregular menstruation, vaginal discharge and pelvic pain were present in 69%, 60% and 52% patients respectively.Conclusions: The overall sensitivity of CBNAAT was 22% and specificity was 77%. The overall sensitivity of liquid culture was 28% and specificity was 71%.

Development of a Novel Spawn (Block Spawn) of an Edible Mushroom, Pleurotus ostreatus, in Liquid Culture and its Cultivation Evaluation

Mushroom cultivation has gained increased attention in recent years. Currently, only four types of spawn, including sawdust spawn, grain spawn, liquid spawn, and stick spawn, are commonly available for mushroom cultivation. This limited spawn diversity has led to difficulty in selecting suitable inoculum materials in some cultivation. In this study, three small blocks of lignocellulosic agro-wastes and one block of a synthetic matrix were prepared as support for growing Pleurotus ostreatus in liquid medium. Mycelium-adsorbed blocks were then evaluated for their potential as block spawn for fructification. Our results indicated that the edible fungus was adsorbed and abundantly grew internally and externally on loofah sponge and synthetic polyurethane foam (PUF) supports and also has the ability to attach and grow on the surface of sugarcane bagasse and corncob supports. The mycelia of P. ostreatus adhered on corncob exhibited the highest metabolic activity, while those on the PUF showed the least activity. Mycelial extension rates of block spawns made of agro-waste materials were comparable to that of sawdust spawn, but the block spawn of PUF showed a significantly lower rate. No significant differences in cropping time and yield were observed among cultivations between experimental block spawns and sawdust spawns. Moreover, the corncob block spawn maintained its fruiting potential during an examined period of 6-month storage. The developed block spawn could be practically applied in mushroom cultivation.

Evaluation of different laboratory methods for auxiliary diagnosis of pulmonary tuberculosis / 临床检验杂志

Objective@#To evaluate the effectiveness of different laboratory methods for the supplementary diagnosis of pulmonary tuberculosis（PTB）and provide reference data for the early diagnosis of PTB. @*Methods@#A total of 298 suspected PTB patients, who were diagnosed and treated in the outpatient department of Shanghai Tongren Hospital from January 2016 to December 2018, were divided into 3 groups: active PTB (138 cases)，inactive PTB (43 cases) and non-PTB (117 cases) group. Sputum acid-fast staining, MGIT liquid culture system and Xpert MTB/RIF test were performed to detect the sputum specimens. The sensitivity and specificity were compared by Chi-square test. @*Results@#The three methods showed certain significance for distinguishing active PTB, inactive PTB combined with non-PTB (χ 2 values were 89.08, 138.94 and 137.12 respectively, all P＜0.01). There was no significant difference for the positive rate of the three methods between inactive PTB and non-PTB. The sensitivities of acid-fast staining, MGIT liquid culture, Xpert MTB/RIF test and the combination of three methods in the diagnosis of active PTB were 45.7% (63/138), 63.8% (88/138), 65.4% (87/133) and 78.2% (104/133) respectively. The sensitivities of MGIT culture and Xpert MTB/RIF test were significantly higher than that of acid-fast staining (χ 2 was 35.79 and 11.26 respectively，all P＜0.01). There was no significant difference for the sensitivities between MGIT liquid culture and Xpert MTB/RIF test（χ 2 was 29.87, P＞0.05) . The sensitivity of combined detection was higher than that of single detection（χ 2 was 30.84, 64.62, 70.14, respectively, all P＜0.01）. The diagnostic specificities of the three methods and their combination were 99.1%（116/117), 98.3%（115/116）, 99.1%（113/114) and 97.3%（110/113）respectively. There was no significant difference for the specificities of the three methods. @*Conclusion@#High sensitivities of MGIT liquid culture and Xpert MTB/RIF test were shown in PTB diagnosis. Combined detection of the three methods may improve the sensitivity of detection.

Rapid detection on resistance of pyrazinamide for Mycobacterium tuberculosis by microplate allochroic silica gel assay / 国际检验医学杂志

Objective To establish a method of judge the pyrazinamide(PZA) susceptibility for Mycobacterium tuberculosis(MTB) with 24-hole micro-liquid culture silica gel color plate,and evaluate the clinical application value of this method.Methods According to the result of MGIT960,to detect PZA drug susceptibility for 30 MTB clinical isolates of whose PZA susceptibility were known with silica gel color plate.The effects of different pH value,different inoculation concentrations on the results were observed,and the optimum detection conditions were discussed.Finally,the 98 MTB clinical isolated of whose PZA susceptibility were unknown were simultaneously detected by gel color plate and MGIT960,the sensitivity,specificity and accuracy were judged by PZA drug sensitivity.Results The best pH was 5.8-5.9 and the best concentration was 2.5×10-1 mg/mL in gel color plate.The best results were read after 7-14 d.The sensitivity,specificity and accuracy were 95.50%,96.30%,and 98.21% respectively just at PZA critical concentration was 100 μg/mg.The sensitivity,specificity and accuracy were 90.90%,92.59%,and 91.84% respectively just at PZA critical concentration was 200 μg/mg.Conclusion The PZA susceptibility for MTB with 24-hole micro-liquid culture silica gel color plate is accurate,rapid,low-cost and simple.

The effects of different strength of MS media in solid and liquid media on in vitro growth of Typhonium flagelliforme

Objective: To determine the effects of different strength of Murashige and Skoog (MS) media (full, 1/2 and 1/4) in solid and liquid media on in vitro growth of Typhonium flagelliforme (T. flagelliforme), whereby an optimum media composition can be provided for mass propagation of T. flagelliforme. Methods: Rhizome bud of T. flagelliforme was obtained from the axenic in vitro established T. flagelliforme plantlets in Plant Tissue Culture Laboratory, Universiti Teknologi MARA, Shah Alam. Rhizome bud was used as explant and cultured onto shoot proliferation medium under different strength of MS media (full, 1/2, 1/4) in solid and liquid culture media. Results: After 6 weeks of culture, the number of shoot, number of leaf, number of root, height of shoot, fresh weight, dry weight and chlorophyll content of T. flagelliforme were analyzed. A comparison was made between liquid and solid culture media. The results revealed that the liquid culture media were more effective for all the growth parameters (shoot height, shoot number, leaf number, root number, fresh weight, dry weight, chlo-rophyll a and chlorophyll b content) compared to solid culture media. Apart from that, this study revealed the positive relationship between strength of MS media and type of culture media (solid and liquid media) to the growth of T. flagelliforme. Growth of T. flagelliforme was improved when MS strength was increased in liquid media. In contrast, growth of T. flagelliforme was improved when MS strength was decreased in solid media. Conclusions: Through this study, an optimum media composition for mass propagation of T. flagelliforme had been established by observing effects of MS media strength and type of culture media (solid and liquid media) on the growth of T. flagelliforme.

The effects of different strength of MS media in solid and liquid media on in vitro growth of Typhonium flagelliforme

Objective To determine the effects of different strength of Murashige and Skoog (MS) media (full, ½ and ¼) in solid and liquid media on in vitro growth of Typhonium flagelliforme (T. flagelliforme), whereby an optimum media composition can be provided for mass propagation of T. flagelliforme. Methods Rhizome bud of T. flagelliforme was obtained from the axenic in vitro established T. flagelliforme plantlets in Plant Tissue Culture Laboratory, Universiti Teknologi MARA, Shah Alam. Rhizome bud was used as explant and cultured onto shoot proliferation medium under different strength of MS media (full, ½, ¼) in solid and liquid culture media. Results After 6 weeks of culture, the number of shoot, number of leaf, number of root, height of shoot, fresh weight, dry weight and chlorophyll content of T. flagelliforme were analyzed. A comparison was made between liquid and solid culture media. The results revealed that the liquid culture media were more effective for all the growth parameters (shoot height, shoot number, leaf number, root number, fresh weight, dry weight, chlorophyll a and chlorophyll b content) compared to solid culture media. Apart from that, this study revealed the positive relationship between strength of MS media and type of culture media (solid and liquid media) to the growth of T. flagelliforme. Growth of T. flagelliforme was improved when MS strength was increased in liquid media. In contrast, growth of T. flagelliforme was improved when MS strength was decreased in solid media. Conclusions Through this study, an optimum media composition for mass propagation of T. flagelliforme had been established by observing effects of MS media strength and type of culture media (solid and liquid media) on the growth of T. flagelliforme.

Desempenho da cultura líquida MGIT após implementação em uma rede de laboratórios públicos do estado de São Paulo / Performance of liquid culture MGIT after implementation in a network of public laboratories of Sao Paulo state

A OMS, em 2007, recomendou a implementação da cultura líquida para o diagnóstico da tuberculose (TB) e teste de sensibilidade para países de baixa e média renda. Neste estudo foi avaliado odesempenho da cultura líquida MGIT em condição de rotina após dois anos de implantação em uma rede de laboratórios públicos. Foi efetuada análise retrospectiva de dados da cultura líquida,realizadas em dez laboratórios regionais do Instituto Adolfo Lutz, de janeiro a março de 2010. Foram incluídas amostras submetidas a baciloscopia, cultura líquida MGIT automatizada ou manual eidentificação presuntiva do complexo Mycobacterium tuberculosis (CMTB). Foram detectadas 1.159 culturas positivas. Destas, 113 (9,7%) contaminaram, e 1.046 foram analisadas, sendo 850 (81,3%) CMTB, 116 (11,1%) micobactérias não tuberculosas e 6 (0,6%) Nocardia sp. A taxa de contaminação foi de 2,2% e o acréscimo da cultura para o diagnóstico da TB foi de 29,9%. A média do tempo de detecção da cultura foi de 14,7 dias (DP+/- 11,7 dias). A acurácia da identificação presuntiva foide 91,3%. A cultura líquida MGIT demonstrou ser excelente alternativa para efetuar diagnóstico da TB e das micobacterioses, em razão da rapidez possibilitando uma intervenção rápida e eficaz no tratamento.

In 2007, WHO recommended the implementation of liquid culture for tuberculosis (TB) diagnosis anddrug-susceptibility test in low and middle-income countries. This study evaluated the performanceof MGIT culture in routine condition after two years of its implementation in a public laboratoriesnetwork. This is a retrospective study, which analyzed the data on the liquid culture performed in ten regional laboratories of the Institute Adolfo Lutz, from January to March 2010. The data included clinical samples submitted to microscopy, automated or manual MGIT culture and presumptive M. tuberculosis complex (MTBC) identification by analyzing the cord formation. Culture waspositive in 1,159 samples. Of these, 113 (9.7%) contaminated, and 1,046 were analyzed, of which 850 (81.3%) were identified as MTBC, 116 (11.1%) as non-tuberculous mycobacteria and 6 (0.6%)as Nocardia sp. Contamination rate was 2.2% and the contribution of culture to the TB diagnosis was 29.9%. The detection mean time was 14.7 days (SD+/-11.7 days). The accuracy of the presumptive identification of MTBC was 91.3%. MGIT liquid culture demonstrated to be an excellent alternative for diagnosing TB and mycobacterioses, because of the rapidity of diagnosis, thus allowing an immediate and effective treatment.

Desempenho da cultura líquida MGIT após implementação em uma rede de laboratórios públicos do estado de São Paulo / Performance of liquid culture MGIT after implementation in a network of public laboratories of Sao Paulo state

A OMS, em 2007, recomendou a implementação da cultura líquida para o diagnóstico da tuberculose (TB) e teste de sensibilidade para países de baixa e média renda. Neste estudo foi avaliado o desempenho da cultura líquida MGIT em condição de rotina após dois anosde implantação em uma rede de laboratórios públicos. Foi efetuada análise retrospectiva de dados da cultura líquida, realizadas em dez laboratórios regionais do Instituto Adolfo Lutz, de janeiro a março de 2010. Foram incluídas amostras submetidas a baciloscopia, cultura líquida MGIT automatizada ou manual e identificação presuntiva do complexo Mycobacterium tuberculosis (CMTB). Foram detectadas 1.159 culturas positivas. Destas, 113 (9,7%) contaminaram, e 1.046 foram analisadas, sendo 850 (81,3%) CMTB, 116 (11,1%) micobactérias não tuberculosas e 6 (0,6%) Nocardia sp A taxa de contaminação foi de 2,2% e o acréscimo da cultura para o diagnóstico da TB foi de 29,9%. A média do tempo de detecção da cultura foi de 14,7 dias (DP+/- 11,7 dias). A acurácia da identificação presuntiva foi de 91,3%. A cultura líquida MGIT demonstrou ser excelente alternativa para efetuar diagnóstico da TB e das micobacterioses, em razão da rapidez possibilitando uma intervenção rápida e eficaz no tratamento.

In 2007, WHO recommended the implementation of liquid culture for tuberculosis (TB) diagnosis and drug-susceptibility test in low and middle-income countries. This study evaluated the performance of MGIT culture in routine condition after two years of its implementation in a public laboratories network.This is a retrospective study, which analyzed the data on the liquid culture performed in ten regional laboratories of the Institute Adolfo Lutz, from January to March 2010. The data included clinical samples submitted to microscopy, automated or manual MGIT culture and presumptive M. tuberculosis complex (MTBC) identification by analyzing the cord formation. Culture was positive in 1,159 samples. Of these, 113 (9.7%) contaminated, and 1,046 were analyzed, of which 850 (81.3%) were identified as MTBC, 116 (11.1%) as non-tuberculous mycobacteria and 6 (0.6%) as Nocardia sp. Contamination rate was 2.2% and the contribution of culture to the TB diagnosis was 29.9%. The detection mean time was 14.7 days (SD+/-11.7 days). The accuracy of the presumptive identification of MTBC was 91.3%. MGIT liquid culture demonstrated to be an excellent alternative for diagnosing TB and mycobacterioses, because of the rapidity of diagnosis, thus allowing an immediate and effective treatment.

Clinical performance of automated real-time nucleic acid amplification technology for rapid and simultaneous detection of tuberculosis and rifampicin resistance assay in diagnosis of pulmonary tuberculosis in 214 cases / 中华传染病杂志

Objective To evaluate the clinical application of the automated nucleic acid amplification technology for rapid and simultaneous detection of tuberculosis and rifampicin resistance (Xpert M TB/RIF) in diagnosis of pulmonary tuberculosis in Wenzhou .Methods A total of 214 patients with suspected pulmonary tuberculosis , clinical diagnosed tuberculosis or suspected drug‐resistant tuberculosis were enrolled in this study .The patients′ sputum specimens were collected for acid‐fast smear ,liquid culture and Xpert M TB/RIF assay .The sensitivity and specificity of three tests for the detection of mycobacterium tuberculosis (M TB) were determined using the clinical final diagnosis as golden standard .The sensitivity and specificity of Xpert M TB /RIF for the detection of rifampin resistance were determined using liquid drug susceptibility as the golden standard .The sensitivity and specificity were compared using chi‐squared test .Results Using the clinical final diagnosis as golden standard ,Xpert M TB/RIF had significantly higher sensitivity than sputum acid‐fast smear (69 .5% vs 44 .1% , χ2 = 23 .31 ,P 0 .05) .The sensitivities of Xpert M TB /RIF in smear positive and smear negative M TB samples were 97 .4% and 47 .5% ,respectively ;the specificities in smear positive and smear negative M TB samples were both 100 .0% .The sensitivity and specificity of Xpert M TB /RIF for the detection of rifampin resistance were 92 .9% and 98 .8% ,respectively using the liquid drug susceptibility test as golden standard .Conclusions Xpert M TB/RIF can detect M TB and rifampin resistance rapidly and accurately with higher sensitivity ,which is highly valuable in practical application .

Effect of sucrose and potassium nitrate on biomass and saponin content of Talinum paniculatum Gaertn. hairy root in balloon-type bubble bioreactor

Objective: To increase biomass and saponin production in hairy root culture of Talinum paniculatum Gaertn. (T. paniculatum) in balloon-type bubble bioreactor (BTBB). Methods: Hairy roots which were collected from leaf explants of T. paniculatum were infected by Agrobacterium rhizogenes strain LB510. The hairy roots were cultivated at 400 mL Murashige and Skoog liquid medium without growth regulator (MS0) in 1 000 mL BTBB. Each BTBB had 2 g hairy roots as initial inoculum and these cultures were treated with various concentrations of sucrose (3%, 4%, 5%, 6%w/v) and potassium nitrate (0.5, 1.0, 1.5 and 2.0 strength of MS medium). Cultures were maintained for 14 days. Fresh and dry weights of hairy roots at the end of culture were investigated. Results: Various concentrations of sucrose influenced the biomass accumulation of hairy roots. Maximum biomass was reached by MS medium supplemented with 6% sucrose and it was approximately threefold higher than control. Culture supplemented with po-tassium nitrate at 2.0 strength of MS0 could increase biomass accumulation of hairy roots until 0.14 g dry weight and it was almost threefold higher than control. However, the maximum saponin content was obtained by MS medium supplemented with 5%sucrose and 2.0 strength potassium nitrate of MS. Conclusions: Based on this research, those conditions can be used to produce biomass and saponin of hairy root of T. paniculatum in the large scale.

Effect of sucrose and potassium nitrate on biomass and saponin content of Talinum paniculatum Gaertn. hairy root in balloon-type bubble bioreactor

Objective: To increase biomass and saponin production in hairy root culture of Talinum paniculatum Gaertn. (T. paniculatum) in balloon-type bubble bioreactor (BTBB). Methods: Hairy roots which were collected from leaf explants of T. paniculatum were infected by Agrobacterium rhizogenes strain LB510. The hairy roots were cultivated at 400 mL Murashige and Skoog liquid medium without growth regulator (MS0) in 1. 000 mL BTBB. Each BTBB had 2 g hairy roots as initial inoculum and these cultures were treated with various concentrations of sucrose (3%, 4%, 5%, 6% w/v) and potassium nitrate (0.5, 1.0, 1.5 and 2.0 strength of MS medium). Cultures were maintained for 14 days. Fresh and dry weights of hairy roots at the end of culture were investigated. Results: Various concentrations of sucrose influenced the biomass accumulation of hairy roots. Maximum biomass was reached by MS medium supplemented with 6% sucrose and it was approximately threefold higher than control. Culture supplemented with potassium nitrate at 2.0 strength of MS0 could increase biomass accumulation of hairy roots until 0.14 g dry weight and it was almost threefold higher than control. However, the maximum saponin content was obtained by MS medium supplemented with 5% sucrose and 2.0 strength potassium nitrate of MS. Conclusions: Based on this research, those conditions can be used to produce biomass and saponin of hairy root of T. paniculatum in the large scale.

Antibacterial Effect of Shenju Lotion in Vitro / 医药导报

Objective To study the antibacterial effect of Shenju lotion in vitro. Methods The diameter of inhibition zone was determined by paper-disc agar-diffusion method. Minimum inhibitory concentration ( MIC) and Minimum bactericidal concentration ( MBC ) was determined by culture medium dilution methodand agar medium plate method, respectively. Antibacterial effect was compared between Shenju lotion and city sale of the gynecological lotion. Results Inhibitory effects of Shenju lotion on 5 pathogenic strains were significantly better than that of city sale of the gynecological lotion at the same concentrations (P<0. 05). MIC of Shenju lotion on Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Bacillus subtilis and Candida albicans was 33. 75, 67. 5, 67. 5, 67. 5 and 33. 75 mg ·mL-1 , respectively. MBC of Shenju lotion on Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Bacillus subtilis and Candida albicans was 33. 75, 67. 5, 67. 5, 135 and 33. 75 mg ·mL-1 , respectively. Conclusion Shenju lotion has obvious bacteriostasis and sterilization effect.

Multiplicación in vitro de Psidium guajava L. en sistemas de inmersión temporal / In vitro multiplication of Psidium guajava L. in temporary immersion systems

La introducción de nuevos cultivares de guayabo (Psidium guajava L.) amerita su propagación masiva, lo cual solo puede ser satisfecho mediante la micropropagación. Sin embargo la micropropagación convencional dejó de ser económicamente eficiente, debido al uso de agentes gelificantes y el elevado número de operaciones manuales, por esta razón se planteó en esta investigación, generar una metodología que permita disminuir los costos de producción por la exclusión del gelificante en los medios de cultivo, evaluando los sistemas de inmersión temporal (SIT) en la multiplicación in vitro de guayabo. Para lo cual, se evaluó el efecto del cultivo en SIT, se comparó los SIT tipo BIT® y RITA® y se evaluó el tiempo (1 y 2 min) y frecuencia (3 y 4 veces/día) de inmersión. Luego de seis semanas de cultivo se evaluó: número de brotes (NB), numero de nudos (NN), longitud de brote (LB) y coeficiente de multiplicación (CM). Con el empleo de SIT se logró valores superiores para NB (2,17), NN (3,5), LB (10,7 mm) y CM (2,8). En la comparación entre SIT tipo RITA y BIT, valores superiores se obtuvieron con el RITA® para NB (3,8), NN (3,8), LB (16,6 mm) y CM (10,4). Se determinó que con 2 min de inmersión se logró los mayores valores de NB (3,7), NN (13,4), LB (15,3 mm) y con 2 min de inmersión 3-4 veces/día el mayor CM (9,4 y 10,4). Se concluye que el cultivo en RITA® en la multiplicación favoreció crecimiento y la proliferación de brotes de guayabo.

The introduction of new cultivars of guava (Psidium guajava L.) deserves its mass propagation, which can only be satisfied by micropropagation. However conventional micropropagation stopped being economically efficient due to the use of gelling agents and the high number of manual operations. For this reason was considered in this research, generate a methodology to reduce production costs by exclusion of gelling in culture media, assessing temporary immersion systems (TIS) in the in vitro multiplication of guava. For which, the effect of the culture way was evaluated in TIS, type TIB® and RITA® compared the TIS and was evaluated the time (1 and 2 min) and frequency (3 and 4 times / day) of immersions. After six weeks of culture were evaluated: shoots number (NS), nodes number (NN), shoot length (SL) and multiplication rate (MR). With the use of TIS higher values for NS (2.17), NN (3.5), SL (10.7 mm) and MC (2.8) was achieved. When comparing RITA® and TIB, higher values were obtained with the RITA® for NS (3.8), NN (3.8), SL (16.6 mm) and MC (10.4). It was determined that 2 min of immersion with the highest values of NS (3.7), NN (13.4), SL (15.3 mm) and 2 min immersion 3-4 times/day achieved the highest MC (9.4 and 10.4). We conclude that the RITA® culture favored the multiplication in growth and proliferation of shoots of guava.

Detection of Mycobacterium tuberculosis growth by direct microscopic observation has a lower cost, higher sensitivity and higher diagnostic value than liquid and solid media cultures.

Background: Culture is needed to confirm tuberculosis but results are generally obtained after several weeks. Objectives: We compared a direct microscopic observation technique for detection of mycobacterial culture positivity (MODS) with the classic solid and MB/BacT cultures in terms of sensitivity, contamination rate, speed and cost on 488 samples. Results: The sensitivity of the MODS technique - 99,2% (162 positive samples) was higher than MB/BacT 78,4% (125 positive samples) and solid culture 69,6% (113 positive samples) P < 0.005 for all comparisons. The median times to positivity were 21, 13.3 and 3 days on solid media, B/BacT and MODS respectively. Conclusions: The MODS technique is faster and more sensitive than both solid media and MB/BacT culture.

A simple and cost effective liquid culture system for the micropropagation of two commercially important apple rootstocks.

The two commercially important apple rootstocks i.e., MM106 and B9 were micropropagated using a liquid culture system. Three different strengths of 0.8% agar solidified PGR free basal MS medium were first tested to optimize the culture media for both the rootstocks. Full strength medium (MS0) supported maximum in vitro growth, multiplication, rooting and survival under field conditions as opposed to quarter and half strength media. When three different volumes of liquid MS0 were tested, highest in vitro growth, multiplication, rooting and also survival under field conditions were achieved in 20 mL liquid MS0. The cost of one litre of liquid medium was also reduced by 8 times to Rs. 6.29 as compared to solid medium. The cost of 20 mL medium was further reduced to Rs. 0.125.

A study on application of BacT/ALERT 3D liquid culture technology on tuberculosis drug resistance detection / 预防医学

Objective To evaluate the BacT/ALERT 3D liquid culture technology on the detection of drug resistance of Mycobacterium tuberculosis(MTB)and to compare the difference between this technology and Lowenstein -Jensen (L -J) proportion method.Methods BacT/ALERT 3D liquid culture technology and L -J proportion technology were applied to detect the drug resistance of tuberculosis from the positive cultures of 219 solid culture samples.Results The average detection time of BacT/ALERT 3D method was 8.02 ±3.85 d,which was about 20 days shorter than that of L -J proportion method.60 drug resistance strains were found using BacT/ALERT 3D technology,While 79 drug resistance strains were found using L -J proportion technology.There showed no significant difference (P >0.05).The compliance rate of BacT/ALERT 3D method and L -J proportion method on the anti -tuberculosis drugs INH,RFP,EMB and SMwas 95.43%,92.69%,95.43% and 92.24% respectively.Conclusion BacT/ALERT 3D liquid culture technology could detect drug resistant TB strains rapidly with high concordance with the results of L -J proportion method on anti -tuberculosis drugs.

Medium optimization for palmarumycin C13 production in liquid culture of endophytic fungus Berkleasmium sp. Dzf12 using response surface methodology

Background: Berkleasmium sp. Dzf12, an endophytic fungus from Dioscorea zingiberensis, was a high producer of palmarumycin C13 with various bioactivities. In the present study, the experimental designs based on statistics were employed to evaluate and optimize the medium for palmarumycin C13 production in mycelia liquid culture of Berkleasmium sp. Dzf12. Results: Among various carbon and nitrogen sources, glucose, peptone and yeast extract were found to be the most favourable for palmarumycin C13 production based on the one-factor-at-a-time experiments. After Plackett-Burman test on the medium, glucose, peptone and yeast extract were further verified to be the most significant factors to stimulate palmarumycin C13 accumulation. These three factors (i.e., glucose, peptone and yeast extract) were then optimized through the experiments of central composite design (CCD) and analysis of response surface methodology (RSM). The optimized medium compositions for palmarumycin C13 production were determined as 42.5 g/l of glucose, 6.5 g/l of peptone, 11.0 g/l of yeast extract, 1.0 g/l of KH2PO4, 0.5 g/l of MgSO4 x 7H2O, 0.05 g/l of FeSO4 x 7H2O, and pH 6.5. Under the optimal culture conditions, the maximum palmarumycin C13 yield of Berkleasmium sp. Dzf12 was increased to 318.63 mg/l, which was about 2.5-fold in comparison with that (130.44 mg/l) in the basal medium. Conclusions: The results indicate that the optimum production of palmarumycin C13 in Berkleasmium sp. Dzf12 liquid culture can be achieved by addition of glucose, peptone and yeast extract with their appropriate concentrations in the modified Sabouraud medium.

Evaluation of BioFM liquid medium for culture of cerebrospinal fluid in tuberculous meningitis to identify Mycobacterium tuberculosis.

The present study was designed to evaluate the sensitivity and specificity of liquid culture medium (BioFM broth) for the diagnosis of tuberculous meningitis (TBM) in cerebrospinal fluid (CSF). CSF samples from 200 patients (TBM group = 150 and non-TBM group = 50) were tested for culture of Mycobacterium tuberculosis in BioFM liquid culture medium. Out of 150 TBM cases, 120 were found to be culture positive, indicating a sensitivity of 80% in BioFM broth within 2-3 weeks of inoculation. Positive cultures were also observed for CSF from 32 (64%) out of 50 non-TBM patients in BioFM liquid culture medium within 4 days of sample inoculation. Therefore, according to our study, BioFM broth system yielded 80% sensitivity [95% confidence interval (CI): 67-93%] and 36% specificity (95% CI: 57-98%) for TBM diagnosis. Our results indicate that although BioFM broth allows the detection of positive cultures within a shorter time, it has a high potential for contamination or for the coexistence of M. tuberculosis and non-tuberculous meningitis (NTM). This coexistence may go undetected or potentially lead to erroneous reporting of results.

Lipid formation and gama-linolenic acid production by Mucor circinelloides and Rhizopus sp., grown on vegetable oil / Formação de lipídeos e produção de ácido gama-linolênico por Mucor circinelloides e Rhizopus sp., em óleos vegetais

The fungi strains were tested in Bioscreen automated system to select the best nutritional source. Following, shaking submserse cultures were studied in media containing sole carbon or nitrogen source. The growth of these strains improved in media containing vegetable oil, with high concentration of lipids. The high concentration of γ-linolenic acid was obtained with M. circinelloides in culture containing sesame oil.

Linhagens de fungos foram testadas em sistema automatizado Bioscreen para selecionar melhor fonte nutricional. Em seguida, foram estudadas culturas submersas em meios contendo uma única fonte de carbono e de nitrogênio. As linhagens contendo alta concentração de lipídeos tiveram melhor crescimento em meio contendo óleos de gergelim ou de dendê. Maior concentração de ácido γ-linolênico foi obtida com M. circinelloides nas culturas em óleo de gergelim.

Lipid formation and -linolenic acid production by Mucor circinelloides and Rhizopus sp., grown on vegetable oil

The fungi strains were tested in Bioscreen automated system to select the best nutritional source. Following, shaking submserse cultures were studied in media containing sole carbon or nitrogen source. The growth of these strains improved in media containing vegetable oil, with high concentration of lipids. The high concentration of -linolenic acid was obtained with M. circinelloides in culture containing sesame oil.

Linhagens de fungos foram testadas em sistema automatizado Bioscreen para selecionar melhor fonte nutricional. Em seguida, foram estudadas culturas submersas em meios contendo uma única fonte de carbono e de nitrogênio. As linhagens contendo alta concentração de lipídeos tiveram melhor crescimento em meio contendo óleos de gergelim ou de dendê. Maior concentração de ácido -linolênico foi obtida com M. circinelloides nas culturas em óleo de gergelim.