scholarly journals Genotypic and phenotypic diversity of Mycobacterium tuberculosis complex genotypes prevalent in West Africa

PLoS ONE ◽  
2021 ◽  
Vol 16 (8) ◽  
pp. e0255433
Author(s):  
Stephen Osei-Wusu ◽  
Isaac Darko Otchere ◽  
Portia Morgan ◽  
Abdul Basit Musah ◽  
Ishaque Mintah Siam ◽  
...  
Keyword(s):  
Growth Rate ◽  
Mycobacterium Tuberculosis ◽  
West Africa ◽  
Urease Activity ◽  
Phenotypic Diversity ◽  
Acid Hydrazide ◽  
Genomic Variation ◽  
Comparative Genomic ◽  
Solid Media ◽  
Significant Difference

Findings from previous comparative genomics studies of the Mycobacterium tuberculosis complex (MTBC) suggest genomic variation among the genotypes may have phenotypic implications. We investigated the diversity in the phenotypic profiles of the main prevalent MTBC genotypes in West Africa. Thirty-six whole genome sequenced drug susceptible MTBC isolates belonging to lineages 4, 5 and 6 were included in this study. The isolates were phenotypically characterized for urease activity, tween hydrolysis, Thiophen-2-Carboxylic Acid Hydrazide (TCH) susceptibility, nitric oxide production, and growth rate in both liquid (7H9) and solid media (7H11 and Löwenstein–Jensen (L-J)). Lineage 4 isolates showed the highest growth rate in both liquid (p = 0.0003) and on solid (L-J) media supplemented with glycerol (p<0.001) or pyruvate (p = 0.005). L6 isolates optimally utilized pyruvate compared to glycerol (p<0.001), whereas L5 isolates grew similarly on both media (p = 0.05). Lineage 4 isolates showed the lowest average time to positivity (TTP) (p = 0.01; Average TTP: L4 = 15days, L5 = 16.7days, L6 = 29.7days) and the highest logCFU/mL (p = 0.04; average logCFU/mL L4 = 5.9, L5 = 5.0, L6 = 4.4) on 7H11 supplemented with glycerol, but there was no significant difference in growth on 7H11 supplemented with pyruvate (p = 0.23). The highest release of nitrite was recorded for L5 isolates, followed by L4 and L6 isolates. However, the reverse was observed in the urease activity for the lineages. All isolates tested were resistant to TCH except for one L6 isolate. Comparative genomic analyses revealed several mutations that might explain the diverse phenotypic profiles of these isolates. Our findings showed significant phenotypic diversity among the MTBC lineages used for this study.

scholarly journals Evolution and Clonal Traits of Mycobacterium tuberculosis Complex in Guinea-Bissau

1999 ◽  
Vol 37 (12) ◽  
pp. 3872-3878 ◽  
Author(s):  
Gunilla Källenius ◽  
Tuija Koivula ◽  
Solomon Ghebremichael ◽  
Sven E. Hoffner ◽  
Renée Norberg ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
West Africa ◽  
Acid Hydrazide ◽  
Rflp Analysis ◽  
Polymorphism Analysis ◽  
Biochemical Tests ◽  
Guinea Bissau ◽  
Group A ◽  
Clonal Origin ◽  
Group B

Two hundred twenty-nine consecutive isolates of Mycobacterium tuberculosis complex from patients with pulmonary tuberculosis in Guinea-Bissau, which is located in West Africa, were analyzed for clonal origin by biochemical typing and DNA fingerprinting. By using four biochemical tests (resistance to thiophene-2-carboxylic acid hydrazide, niacin production, nitrate reductase test, and pyrazinamidase test), the isolates could be assigned to five different biovars. The characteristics of four strains conformed fully with the biochemical criteria for M. bovis, while those of 85 isolates agreed with the biochemical criteria for M. tuberculosis. The remaining 140 isolates could be allocated into one of three biovars (biovars 2 to 4) representing a spectrum between the classical bovine (biovar 1) and human (biovar 5) tubercle bacilli. By using two genotyping methods, restriction fragment length polymorphism analysis with IS6110 (IS6110 RFLP analysis) and spoligotyping, the isolates could be separated into three groups (groups A to C) of the M. tuberculosis complex. Group A (n = 95), which contained the majority of classical human M. tuberculosis isolates, had large numbers of copies of IS6110 elements (mean number of copies, 9) and a distinctive spoligotyping pattern that lacked spacers 33 to 36. Isolates of the major group, group B (n = 119), had fewer IS6110 copies (mean copy number, 5) and a spoligotyping pattern that lacked spacers 7 to 9 and 39 and mainly comprised isolates of biovars 1 to 4. Group C isolates (n = 15) had one to three IS6110 copies, had a spoligotyping pattern that lacked spacers 29 to 34, and represented biovar 3 to 5 isolates. Four isolates whose biochemical characteristics conformed with those of M. bovis clustered with the group B isolates and had spoligotype patterns that differed from those previously reported for M. bovis, in that they possessed spacers 40 to 43. Interestingly, isolates of group B and, to a certain extent, also isolates of group C showed a high degree of variability in biochemical traits, despite genotypic identity in terms of IS6110 RFLP and spoligotype patterns. We hypothesize that isolates of groups B and C have their evolutionary origin in West Africa, while group A isolates are of European descent.

scholarly journals Bacterial Load Comparison of the Three Main Lineages of Mycobacterium tuberculosis Complex in West Africa

2021 ◽  
Vol 12 ◽  
Author(s):  
Stephen Osei-Wusu ◽  
Portia Morgan ◽  
Prince Asare ◽  
Godfrey Adams ◽  
Abdul Basit Musah ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
West Africa ◽  
Bacterial Load ◽  
Rna Extraction ◽  
High Growth ◽  
High Growth Rate ◽  
Smear Microscopy ◽  
Culture Positivity ◽  
Significant Difference ◽  
The Difference

Studies have shown an association between bacterial load and virulence; however, not much is known about the diversity in this phenotypic characteristic of Mycobacterium tuberculosis complex (MTBC). This study was therefore aimed to determine the differences in bacterial load of the three most prevalent MTBC genotypes (L4, L5, and L6) in West Africa at the time of diagnosis. A total of 170 paired fresh sputum samples were collected; one part in guanidinium thiocyanate (GTC) was used for RNA extraction and tuberculosis molecular bacterial load assay (TB-MBLA), and the other part without GTC was confirmed for TB positivity using GeneXpert MTB/RIF, smear microscopy grading, and culture on Löwenstein–Jensen media slants. The 170 sputum samples comprised 155 new cases, three follow-up cases, and 12 TB negative sputum samples. The time-to-culture positivity (TTP) and degree of culture positivity (DCP) were recorded. All 122 isolates obtained were spoligotyped for lineage (L) classification, but spoligotypes were obtained from 120 isolates. Of the typed isolates, 70.0, 10.8, 10.8, 4.2, 2.5, 0.8, and 0.8% were lineages 4, 5, 6, 2, 3, 1, and Mycobacterium bovis, respectively. Further analysis of the three most prevalent lineages showed significantly shorter TTP and higher DCP by L4 compared to L5 and L6, respectively: TTP 20.8, vs. 26.5, and 28.2 days; p-value = 0.005 and DCP 1.27, vs. 0.81 and 0.29, p &lt; 0.001. The average TB-MBLA measured bacterial load of L4 was 3.82 Log10eCFU/ml which was not significantly different from 3.81 and 3.80 Log10eCFU/ml of L5 and L6, respectively, p = 0.84. Degrees of smear microscopy L4 = 1.20, L5 = 1.20, and L6 = 0.92 and GeneXpert Cq values L4 = 17.08, L5 = 18.37, and L6 = 17.59 showed no significant difference between the lineages, p = 0.72 and p = 0.48, respectively. Retrospective analysis of a larger sample confirmed the difference in TTP, p &lt; 0.001. In conclusion, the observed shorter TTP and high DCP of L4 could signify high growth rate in culture that is independent of total bacterial load at diagnosis.

Determination some Immunological Aspects in Pulmonary Tuberculosis Patients in Qadisiyah Province

2018 ◽  
Vol 9 (2) ◽  
Author(s):  
Syoof Khowman Alramahy ◽  
Akram Hadi Hamza
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Pulmonary Tuberculosis ◽  
Statistical Difference ◽  
Positive Culture ◽  
Control Group ◽  
Tuberculosis Patients ◽  
Significant Difference ◽  
Lowenstein Jensen ◽  
The Mean ◽  
Igg Level

This study was carried out to study of some immunological aspects among the pulmonary Tuberculosis patients infected with causative agent, Mycobacterium tuberculosis. A Total of 200 sputum samples were collected from patients attending the consultant Clinic for Chest and Respiratory disease center, Diwaniya. Control group (No=15) also included. According to acid fast stain of sputum, the patients were classified as positive (No=91,45.5%) and negative (No=109,54.5, Lowenstein Jensen medium used for the cultivation of samples, on which 70% of sputum samples where positive culture for this microorganism. The grown microorganism were identified as M. tuberculosis, based on positive A.F.B, Niacin producers ,negative for catlase at 68c. The mean IgG level was l184.053±76.684 mg/100 ml in tuberculosis group compared with 1016.533 ± 44.882 mg/100ml in control group, rendering the statistical difference significant. For IgA and IgM levels, they were at mean of 315.880±38.552 mg/100 ml and 119.527±8.464 mg/100 ml in control group compared with 396.358±38.776 mg/100 ml and 134.207±11.696 mg/100 ml in patients group respectively with significant difference

scholarly journals Assessment of the Growth and Reproductive Performance of Cloned Pietrain Boars

Animals ◽  
2020 ◽  
Vol 10 (11) ◽  
pp. 2053
Author(s):  
Junsong Shi ◽  
Baohua Tan ◽  
Lvhua Luo ◽  
Zicong Li ◽  
Linjun Hong ◽  
...  
Keyword(s):  
Growth Rate ◽  
Body Size ◽  
Growth Performance ◽  
Reproductive Performance ◽  
Semen Quality ◽  
Small Body ◽  
Economic Benefits ◽  
Large Animal ◽  
Significant Difference ◽  
F1 Progeny

How to maximize the use of the genetic merits of the high-ranking boars (also called superior ones) is a considerable question in the pig breeding industry, considering the money and time spent on selection. Somatic cell nuclear transfer (SCNT) is one of the potential ways to answer the question, which can be applied to produce clones with genetic resources of superior boar for the production of commercial pigs. For practical application, it is essential to investigate whether the clones and their progeny keep behaving better than the “normal boars”, considering that in vitro culture and transfer manipulation would cause a series of harmful effects to the development of clones. In this study, 59,061 cloned embryos were transferred into 250 recipient sows to produce the clones of superior Pietrain boars. The growth performance of 12 clones and 36 non-clones and the semen quality of 19 clones and 28 non-clones were compared. The reproductive performance of 21 clones and 25 non-clones were also tested. Furthermore, we made a comparison in the growth performance between 466 progeny of the clones and 822 progeny of the non-clones. Our results showed that no significant difference in semen quality and reproductive performance was observed between the clones and the non-clones, although the clones grew slower and exhibited smaller body size than the non-clones. The F1 progeny of the clones showed a greater growth rate than the non-clones. Our results demonstrated through the large animal population showed that SCNT manipulation resulted in a low growth rate and small body size, but the clones could normally produce F1 progeny with excellent growth traits to bring more economic benefits. Therefore, SCNT could be effective in enlarging the merit genetics of the superior boars and increasing the economic benefits in pig reproduction and breeding.

scholarly journals Mutations in fbiD (Rv2983) as a Novel Determinant of Resistance to Pretomanid and Delamanid in Mycobacterium tuberculosis

2020 ◽  
Vol 65 (1) ◽  
pp. e01948-20
Author(s):  
Dalin Rifat ◽  
Si-Yang Li ◽  
Thomas Ioerger ◽  
Keshav Shah ◽  
Jean-Philippe Lanoix ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Clinical Evidence ◽  
Clinical Samples ◽  
Loss Of Function ◽  
Wild Type ◽  
Content Type ◽  
Solid Media ◽  
High Level ◽  
Level Resistance

ABSTRACTThe nitroimidazole prodrugs delamanid and pretomanid comprise one of only two new antimicrobial classes approved to treat tuberculosis (TB) in 50 years. Prior in vitro studies suggest a relatively low barrier to nitroimidazole resistance in Mycobacterium tuberculosis, but clinical evidence is limited to date. We selected pretomanid-resistant M. tuberculosis mutants in two mouse models of TB using a range of pretomanid doses. The frequency of spontaneous resistance was approximately 10−5 CFU. Whole-genome sequencing of 161 resistant isolates from 47 mice revealed 99 unique mutations, of which 91% occurred in 1 of 5 genes previously associated with nitroimidazole activation and resistance, namely, fbiC (56%), fbiA (15%), ddn (12%), fgd (4%), and fbiB (4%). Nearly all mutations were unique to a single mouse and not previously identified. The remaining 9% of resistant mutants harbored mutations in Rv2983 (fbiD), a gene not previously associated with nitroimidazole resistance but recently shown to be a guanylyltransferase necessary for cofactor F420 synthesis. Most mutants exhibited high-level resistance to pretomanid and delamanid, although Rv2983 and fbiB mutants exhibited high-level pretomanid resistance but relatively small changes in delamanid susceptibility. Complementing an Rv2983 mutant with wild-type Rv2983 restored susceptibility to pretomanid and delamanid. By quantifying intracellular F420 and its precursor Fo in overexpressing and loss-of-function mutants, we provide further evidence that Rv2983 is necessary for F420 biosynthesis. Finally, Rv2983 mutants and other F420H2-deficient mutants displayed hypersusceptibility to some antibiotics and to concentrations of malachite green found in solid media used to isolate and propagate mycobacteria from clinical samples.

scholarly journals Comparison of In Vitro Activity and MIC Distributions between the Novel Oxazolidinone Delpazolid and Linezolid against Multidrug-Resistant and Extensively Drug-Resistant Mycobacterium tuberculosis in China

2018 ◽  
Vol 62 (8) ◽  
Author(s):  
Zhaojing Zong ◽  
Wei Jing ◽  
Jin Shi ◽  
Shu'an Wen ◽  
Tingting Zhang ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Novel Mutation ◽  
Multidrug Resistant ◽  
23S Rrna ◽  
Drug Resistant ◽  
Content Type ◽  
Extensively Drug Resistant ◽  
Significant Difference ◽  
Mdr Tb

ABSTRACT Oxazolidinones are efficacious in treating mycobacterial infections, including tuberculosis (TB) caused by drug-resistant Mycobacterium tuberculosis. In this study, we compared the in vitro activities and MIC distributions of delpazolid, a novel oxazolidinone, and linezolid against multidrug-resistant TB (MDR-TB) and extensively drug-resistant TB (XDR-TB) in China. Additionally, genetic mutations in 23S rRNA, rplC, and rplD genes were analyzed to reveal potential mechanisms underlying the observed oxazolidinone resistance. A total of 240 M. tuberculosis isolates were included in this study, including 120 MDR-TB isolates and 120 XDR-TB isolates. Overall, linezolid and delpazolid MIC90 values for M. tuberculosis isolates were 0.25 mg/liter and 0.5 mg/liter, respectively. Based on visual inspection, we tentatively set epidemiological cutoff (ECOFF) values for MIC determinations for linezolid and delpazolid at 1.0 mg/liter and 2.0 mg/liter, respectively. Although no significant difference in resistance rates was observed between linezolid and delpazolid among XDR-TB isolates (P > 0.05), statistical analysis revealed a significantly greater proportion of linezolid-resistant isolates than delpazolid-resistant isolates within the MDR-TB group (P = 0.036). Seven (53.85%) of 13 linezolid-resistant isolates were found to harbor mutations within the three target genes. Additionally, 1 isolate exhibited an amino acid substitution (Arg126His) within the protein encoded by rplD that contributed to high-level resistance to linezolid (MIC of >16 mg/liter), compared to a delpazolid MIC of 0.25. In conclusion, in vitro susceptibility testing revealed that delpazolid antibacterial activity was comparable to that of linezolid. A novel mutation within rplD that endowed M. tuberculosis with linezolid, but not delpazolid, resistance was identified.

scholarly journals In Vivo Radioprotective Activity of Cell-Permeable Bifunctional Antioxidant Enzyme GST-TAT-SOD against Whole-Body Ionizing Irradiation in Mice

2017 ◽  
Vol 2017 ◽  
pp. 1-9 ◽  
Author(s):  
Jianru Pan ◽  
Huocong He ◽  
Ying Su ◽  
Guangjin Zheng ◽  
Junxin Wu ◽  
...  
Keyword(s):  
Growth Rate ◽  
Antioxidant Activity ◽  
Body Weight ◽  
Whole Body ◽  
White Pulp ◽  
Radioprotective Activity ◽  
Significant Difference ◽  
Wbc Count

GST-TAT-SOD was the fusion of superoxide dismutase (SOD), cell-permeable peptide TAT, and glutathione-S-transferase (GST). It was proved to be a potential selective radioprotector in vitro in our previous work. This study evaluated the in vivo radioprotective activity of GST-TAT-SOD against whole-body irradiation. We demonstrated that intraperitoneal injection of 0.5 ml GST-TAT-SOD (2 kU/ml) 2 h before the 6 Gy whole-body irradiation in mice almost completely prevented the splenic damage. It could significantly enhance the splenic antioxidant activity which kept the number of splenic white pulp and consequently resisted the shrinkage of the spleen. Moreover, the thymus index, hepatic antioxidant activity, and white blood cell (WBC) count of peripheral blood in irradiated mice pretreated with GST-TAT-SOD also remarkably increased. Although the treated and untreated irradiated mice showed no significant difference in the growth rate of animal body weight at 7 days postirradiation, the highest growth rate of body weight was observed in the GST-TAT-SOD-pretreated group. Furthermore, GST-TAT-SOD pretreatment increased resistance against 8 Gy whole-body irradiation and enhanced 30 d survival. The overall effect of GST-TAT-SOD seemed to be a bit more powerful than that of amifostine. In conclusion, GST-TAT-SOD would be a safe and potentially promising radioprotector.

KORELASI ANTARA HASIL TES MIKROSKOPIS DENGAN TES CEPAT MOLEKULER PADA PASIEN TUBERCULOSIS DAN MULTIDRUG RESISTEN TUBERCULOSIS DI RSUD Dr. H CHASAN BOESOIRIE TERNATE

2020 ◽  
Vol 2 (1) ◽  
Author(s):  
Andi Sitti Nur Afiah ◽  
Fera The
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Pulmonary Disease ◽  
Program Analysis ◽  
Rifampicin Resistance ◽  
Test Results ◽  
Chi Square ◽  
Significant Difference ◽  
Mdr Tb ◽  
Chi Square Test ◽  
Analytical Research

Tuberculosis (TB) is a pulmonary disease caused by Mycobacterium tuberculosis. Globally in 2018 theestimated number of people affected by TB was estimated at 10.0 million population and 484,000 cases ofmultidrug-resistant TB (MDR-TB). This study aims to determine the correlation between microscopic testresults with RMT on TB and MDR-TB patients at RSUD Dr. H Chasan Boesoirie Ternate. This type of researchis analytical research using a retrospective approach. The sample in this study were patients with suspected TBand MDR-TB who had performed microscopic tests and TCM in February – April at 2020 in the ClinicalPathology Laboratory of RSUD Dr. H Chasan Boesoirie Ternate, who met the inclusion and exclusion criteria.Data were analyzed using the SPSS program analysis was carried out in stages, namely by univariate andbivariate using chi-square test. From 100 samples, the results of RMT examination with TB suspects were 30rifampicin sensitive samples with 2 rifampicin resistance and 5 rifampin sensitive samples for TB MDR-TBsuspects RMT examination results. Chi-square test results obtained the value of p = 000 (p <0.05). There is stilla significant difference between the microscopic test results with RMT in TB and MDR-TB suspect patients atRSUD Dr. H Chasan Boesoirie Ternate.

A comparative study on the nursery culture of hatchery-reared sub-adult cupped oyster, Crassostrea iredalei (Faustino, 1932), in an earthen pond and a mangrove canal

2018 ◽  
Vol 26 (4) ◽  
pp. 217-222
Author(s):  
Premwadee Chueachat ◽  
Woraporn Tarangkoon ◽  
Suwat Tanyaros
Keyword(s):  
Growth Rate ◽  
Comparative Study ◽  
Condition Index ◽  
Daily Growth ◽  
Nursery Culture ◽  
Absolute Growth Rate ◽  
Significant Difference ◽  
Adult Oyster ◽  
Absolute Growth ◽  
The Mean

Abstract A comparative study on the nursery culture of the spat of the tropical oyster, Crassostrea iredalei, in an earthen pond and a mangrove canal was conducted over two months. The results revealed no differences in the absolute growth rate determined by shell width between the two culture sites (P < 0.05). Sub-adult oysters cultured in the mangrove canal showed a higher absolute growth rate in shell length and a higher daily growth rate than the oysters cultured in the earthen pond (P < 0.05). The mean survival rate of sub-adult oysters cultured in the earthen pond (99.8 ± 0.2%) was significantly higher than for those cultured in the mangrove canal (66.7 ± 31.4%). Decreased density from the loss of sub-adult oyster nursery culture in the mangrove canal led to higher growth performance than in the earthen pond. However, no difference was found for the fraction of oysters larger or smaller than 5 cm for the two culture sites (P < 0.05). A significant difference was noted in the Condition Index (CI) between the two culture sites (P < 0.05). The high primary productivity in mangroves is a major supporter of higher CI in sub-adult oysters cultured in mangrove canals versus in earthen ponds. Water exchange in the earthen pond to maintain calcium and magnesium concentrations resulted in no differences in the shell compressibility of sub-adult oysters compared with those cultured in the mangrove canal.

Surface culture of Steinernema sp. on two solid media and their pathogenicity against Galleria mellonella

2021 ◽  
Vol 95 ◽  
Author(s):  
C.I. Cortés-Martínez ◽  
A.I. Rodríguez-Hernández ◽  
M.R. López-Cuellar ◽  
N. Chavarría-Hernández
Keyword(s):  
Galleria Mellonella ◽  
Egg Yolk ◽  
Infective Juvenile ◽  
Surface Culture ◽  
Solid Media ◽  
Significant Difference ◽  
Bacterial Lawn ◽  
The Mean

Abstract The use of native entomopathogenic nematodes as biocontrol agents is a strategy to decrease the environmental impact of insecticides and achieve sustainable agriculture crops. In this study, the effect of the surface culture of Steinernema sp. JAP1 over two solid media at 23–27°C on infective juvenile (IJ) production and pathogenicity against Galleria mellonella larvae were investigated. First, the bacterial lawn on the surface of the media with egg yolk (P2) or chicken liver (Cl) were incubated in darkness at 30°C for 48 and 72 h, and 100 surface-sterilized IJs were added. Four harvests were conducted within the next 35 days and the mean accumulated production was superior on Cl (210 × 103 IJs) than on P2 (135 × 103 IJs), but the productivity decreased up to 10% when the incubation time of the bacterial lawn was of 72 h. The mean pathogenicity of in vitro- and in vivo-produced IJs were of 47–64% and 31%, respectively. It is worth noting that none of the two solid media had a statistically significant difference in IJ pathogenicity. Considering that the maximum multiplication factor of IJs on solid media was 2108 and that the pathogenicity against G. mellonella was outstanding, Steinernema sp. has a good potential for in vitro mass production.

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