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Background: China is a country with a burden of high rates of both TB and multidrug-resistant TB (MDR-TB). However, published data on pyrazinamide (PZA) resistance are still limited in Hunan province, China. This study investigated the prevalence, transmission, and genetic diversity of PZA resistance among multidrug-resistant Mycobacterium tuberculosis isolates in Hunan province. Methods: Drug susceptibility testing (DST) with the Bactec MGIT 960 PZA kit and pyrazinamidase (PZase) testing were conducted on all 298 MDR clinical isolates. Moreover, 24-locus MIRU-VNTR and DNA sequencing of pncA, rpsA, and panD genes were conducted on 180 PZA-resistant (PZA-R) isolates. Results: The prevalence of PZA resistance among MDR-TB strains reached 60.4%. Newly diagnosed PZA-R TB patients and clustered isolates with identical pncA, rpsA, and panD mutations showed that transmission of PZA-R isolates played a significant role in the formation of PZA-R TB. Ninety-eight mutation patterns were observed in the pncA among 180 PZA-R isolates, and seventy-one (72.4%) were point mutations. Twenty-four of these mutations are new, including 2 base substitutions (V93G and T153S) and 22 nucleotide deletions or insertions. The W119C was found in PZA-S isolates, on the other hand, F94L and V155A mutations were found in both PZA resistant and susceptible isolates with positive PZase activity, indicating that they were not associated with PZA resistance. This is not entirely in line with the WHO catalogue. Ten novel rpsA mutations were found in 10 PZA-R isolates, which all combined with mutations in pncA. Thus, it is unpredictable whether these mutations in rpsA can impact PZA resistance. No panD mutation was found in all PZA-R isolates. Conclusion: DNA sequencing of pncA and PZase activity testing have great potential in predicting PZA resistance. [ABSTRACT FROM AUTHOR]

In biodiverse forest ecosystems, plant diversity has been reported to increase plant-derived lignin accumulation and soil organic carbon (SOC) storage. However, little is known about the fate of lignin and its degradation dynamics from plant to soil. This process is critical for the formation of SOC, especially in natural forest ecosystems with diverse plant species. This study presents the lignin biomarker characteristics of several common plant species and in mixed litter. The study was conducted in 45 plots along a plant species diversity gradient in a subtropical forest located in southwest China. Our results demonstrate that lignin content and its biochemical characteristics in plant leaves vary among species, while different plant species also alter the content of lignin and its monomeric compounds in the litter. Lignin compounds are gradually disintegrated from plant leaf to litter and then to soil, further indicating that plant-derived lignin from plant sources contributes to the formation and accumulation of forest SOC. These findings provide novel information on the linkage between tree species diversity and lignin accumulation while indicating the role of plant-derived lignin on SOC storage. These results may be useful in predicting forest soil C dynamics in Earth system models. [ABSTRACT FROM AUTHOR]

The stability of soil organic matter (SOM) plays a critical role in soil carbon (C) dynamics under global warming. However, the factors influencing SOM stability, particularly the significance of bedrock geochemistry and its hierarchical relationship with climate and soil properties, remain poorly understood. To address this gap, we conducted a study along a large climatic gradient (Δtemperature > 9 °C) in the subtropical karst forests of southwest China, quantifying SOM stability using thermal analysis and investigating the contributions of bedrock geochemistry, climate, and soil properties. Our results showed that SOM stability was positively correlated with mineral-associated organic C (MAOC) rather than particulate organic C. Hierarchical partitioning analysis further demonstrated that bedrock geochemistry was the predominant contributor to SOM stability variance, accounting for 23.7%. Following this, soil minerals contributed to 21.1%–22.6% of the variance, the mean annual temperature to 20.3%, and microbial biomass C to 17.2%. In particular, bedrock geochemistry—specifically the presence of calcium-rich bedrock—was found to enhance SOM stability by promoting the accumulation of exchangeable calcium and calcium carbonate in soils. Additionally, high temperature improved SOM stability by increasing the content and proportion of MAOC and soil pH. These results highlight the fundamental role of bedrock geochemistry in controlling SOM stability and emphasize the importance of considering hierarchical relationships among bedrock–soil–climate interactions for evaluating soil C dynamics. [ABSTRACT FROM AUTHOR]

Purpose: Changes in climate and land use may disrupt the soil carbon (C), nitrogen (N), and phosphorus (P) cycles, which affect the availability of essential nutrients differently, including soil organic carbon (SOC), total nitrogen (TN), and total phosphorus (TP). Nevertheless, lithologic impact on soil C:N:P stoichiometry over a broad climatic gradient remains largely unknown. We aimed to determine the variation in nutrient limitations between karst and non-karst forests and croplands across a climate gradient. Materials and methods: This study investigated soil C:N:P stoichiometry in mature forest (MF) and nearby cropland rotation (CR) (as a control) soils on limestone (karst) and clastic rock (non-karst) throughout a climatic gradient in southwest China through analysis of soil samples. Result and discussion: The SOC and TN contents in the karst MF were 1.3 and 1.6 times greater, respectively, than that in the non-karst MF. Consequently, the karst MF had a lower C:N ratio than did the non-karst MF. The SOC and TN contents were considerably higher in the MF than they were in the CR under both lithologies, leading to higher C:P and N:P ratios in the MF compared to those in the CR. The increasing temperature resulted in a shift from TN and TP to SOC limitation, due to the increased decomposition of SOC by microbes with an increase in temperature. Soil pH and Ca content are controlled by the lithology, and they were closely related to C:N:P stoichiometry in this study, whereas a combination of lithology and temperature controlled soil microbial activity and plant growth. Importantly, the magnitude of lithology-induced changes in the stoichiometry was greater as temperature increased, which was more pronounced in the non-karst than it was in the karst. Conclusions: These findings highlight the responses of soil C, N, and P to climate warming in relation to stoichiometry, and soil-microorganism-plant response strategies may be the key to the mitigating the effect of temperature increase on nutrient cycles in karst ecosystems. [ABSTRACT FROM AUTHOR]

Lithology has such an important effect on the sustainability of soil carbon (C) pools. Forests are an important part of terrestrial C sinks; yet, it is unclear whether their soil carbon sensitivity to temperature changes is regulated by lithology, especially in karst ecosystems, which are widely distributed globally. Along a climate gradient in the subtropical region of southwest China, we compared the contents of soil organic C (SOC) and total N (TN) in karst and non-karst forests. The data were analyzed and processed using ANOVA, regression analysis, and random forest. The results showed that the karst forests had significantly higher SOC and TN contents but lower ratio of SOC to TN (C:N) than non-karst forests, mainly because of the higher soil calcium (Ca) content and microbial biomass. With rising mean annual temperature (MAT), SOC and TN contents in non-karst forests significantly decreased, whereas in karst forests they were not correlated with MAT; while, the opposite was true for C:N. In karst forests, soil Ca constrain warming induced decomposition of SOC and TN by forming stable complexes with SOM through exchangeable Ca, and by promoting aggregate stability through the role of calcium carbonate. The correlation between δ13C and the logarithm of SOC concentration also supported that conclusion. In karst forests compared to non-karst forests, soil C pools are larger and less sensitive to regional temperature change. Nevertheless, climate warming may still accelerate soil C loss in karst forests by increasing microbial C limitation. Thus, soil C sequestration potential and loss risk coexist in karst areas. The ratio of SOC to TN (C:N) is regulated through appropriate management measures in the process of karst vegetation restoration, thus promoting long-term stable sequestration of soil carbon pools. [ABSTRACT FROM AUTHOR]

Microbial necromass carbon (MNC) is a key soil organic carbon (SOC) pool. Soil aggregates and minerals play pivotal roles in controlling soil carbon storage. Although soil minerals and aggregates are tightly linked to parent materials, it is uncertain whether the relative importance of aggregates and minerals in governing MNC varies depending on soil parent materials. In this study, we determined the MNC content in forest soils originating from two contrasting soil parent materials, namely karst (carbonate rocks) and non-karst soils (clastic rocks) across a climatic gradient in southwest China. The MNC contents (24.5 ± 0.8 mg g−1 in karst and 24.1 ± 1.8 mg g−1 in non-karst soils) and their respective contribution to SOC (43.7 ± 1.3 % in karst and 41.1 ± 1.4 % in non-karst soils) were similar between the two parent materials. In karst soils, MNC was influenced by aggregates rather than minerals, whereas the opposite was true in non-karst soils. A higher proportion of microaggregates directly increased the MNC content in karst soils, whereas greater mineral contents indirectly increased the MNC through their effects on microbial biomass carbon. The divergent effects of aggregates and minerals on MNC were predominantly attributed to the formation of secondary carbonate crystals in calcium-rich karst soils as well as to the differences in pH between karst and non-karst soils. Additionally, climate (mean annual temperature and precipitation) and vegetation properties (aboveground biomass or richness) indirectly affected the MNC through their effects on aggregates or minerals in karst and non-karst soils. Consequently, the influence of soil aggregates and minerals on microbially derived carbon is mediated by the parent material. Our findings emphasize the importance of considering lithology-dependent drivers in predicting soil carbon accumulation, particularly in relation to the stability of microbial-derived carbon. [Display omitted] • Microbial necromass carbon accumulation is similar between karst and non-karst soils. • Soil aggregates, not minerals, affect microbial necromass carbon in karst soils. • Soil minerals, not aggregates, influence microbial necromass carbon in non-karst soils. • Lithology mediates physical-chemical protection on microbial necromass carbon. [ABSTRACT FROM AUTHOR]

Background: Tuberculosis (TB) seriously threatens individual and public health. Recently, TB outbreaks in schools have been reported more frequently in China and have attracted widespread attention. We reported three TB outbreaks in high schools in Hunan Province, China. Methods: When a tuberculosis patient was reported in a school, we carried out field epidemiological investigations, including tuberculin skin testing (TST), chest X-ray (CXR) and laboratory test for all close contacts, and whole-genome sequencing (WGS) analyses to understand the transmission patterns, the causes and the risk factors for the outbreaks, thereby providing a foundation for the control of TB epidemics in schools. Results: A total of 49 students with TB patients were identified in the three schools where TB outbreaks occurred, including nine patients in School A, 14 patients in School B, and 26 patients in School C. In Schools A, B and C, the putative attack rates in the classes of the index case were 13.8% (8/58), 7.6% (5/66), and 40.4% (21/52), while the putative attack rates of expanding screening in the school were 0.3% (1/361), 0.2% (9/3955), and 0.2% (5/2080), respectively. Thirteen patients had patient delay, with a median delay interval of 69 days (IQR 30.5– 113 days). Twelve patients had a healthcare diagnostic delay with a median delay interval of 32 days (IQR 24– 82 days). Phylogenetic analysis of culture-positive patients revealed that most of them shared a small genetic distance (≤ 12 SNPs), with three separate genetic clusters (including one MDR-TB genomic cluster), indicating the recent transmission of Mycobacterium tuberculosis strains. Conclusion: This combination of field investigation and WGS analysis revealed the transmission of three TB outbreaks in schools. Reinforced implementation is needed to improve timely case finding and reduce diagnosis delay in routine TB control in the school population. [ABSTRACT FROM AUTHOR]

Purpose: Soil nitrogen (N) availability plays an important role in determining ecosystem structure and function. However, the variation patterns of soil N availability under different vegetation restoration strategies along climatic gradients in the ecologically fragile karst region of China have been poorly understood. Methods: Soil N variables, including total N (TN), inorganic N [ammonium (NH4+) and nitrate (NO3-)], and rates of net N mineralization and nitrification were measured following managed (plantation forest) and natural (spontaneous regeneration) vegetation restoration along a climatic gradient in the karst region of southwestern China. Results: Compared to that in the cropland, soil TN and NH4+ concentrations significantly increased in both vegetation restoration strategies after 15 years of cropland abandonment. The net N mineralization and nitrification rates were lower in both vegetation restoration strategies compared with those in cropland and secondary forest. There were no significant differences in the soil N variables between the two vegetation restoration strategies. Furthermore, soil NO3− was positively correlated with mean annual temperature (MAT) following vegetation restoration, whereas the net nitrification rate was negatively correlated with MAT. Generally, soil pH was the main factor affecting soil inorganic N, whereas elevation, soil microbial abundance, SOC or C:N, and climatic factors mainly affected soil net N mineralization and nitrification rates. Conclusion: Vegetation restoration was beneficial to soil N accumulation and is likely to decrease the risk of N loss in warmer regions given the reduced potential nitrification rate with increased MAT in the karst region of Southwest China. [ABSTRACT FROM AUTHOR]

Vegetation restoration often promotes the development of biological soil crusts (biocrusts), which have a significant role in the terrestrial nitrogen (N) cycle. However, biocrusts are not always considered when assessing soil N responses to restoration efforts, especially in subtropical regions. We assessed the effect of various restoration strategies on moss-dominated biocrust properties and soil N variables (0–5 cm topsoil and 5–10 cm sub-topsoil) in the subtropical karst region of southwestern China. Four restoration strategies were included: restoration with plantation forest, forage grass, and a combination of forest and grass, and spontaneous regeneration to natural grassland. Cropland under maize-soybean rotation (CR) was used as reference. The biocrust under combination restoration strategy had significantly higher biomass, saturated water adsorption ratio, and carbon content than did the other strategies; and the soil generally had the highest total N, ammonium, and microbial biomass carbon and N content but the lowest nitrate content. Redundancy analysis, variation partitioning, and stepwise multiple linear regression all indicated that biocrust properties play an important role in affecting soil N variables, especially in the topsoil. Vegetation restoration strategies that use various plant functional group assemblages, such as forest and grass, are preferable in enhancing the development of biocrusts and thus improving soil N properties. • Vegetation restoration is beneficial in promoting biocrust development. • Biocrust properties were the main factors affecting soil N variations. • Combination of plantation trees and forage grasses had higher soil N and biocrusts. • Combination restoration strategy should be favored in vegetation restoration efforts. [ABSTRACT FROM AUTHOR]

• Natural and managed vegetation restoration were compared in a karst region in China. • PLFAs in topsoil were higher after 15 years of restoration than in cropland. • AMF was significantly higher in natural than in managed restored vegetation. • SOC and the SOC:N ratio affected the abundance of soil microbial communities. • Natural vegetation restoration has more potential to improve soil fertility. The soil microbial community is a key indicator to evaluate the effectiveness of vegetation restoration in degraded ecosystems. However, few studies have simultaneously compared the effects of managed and natural vegetation restoration strategies on the soil microbial community in the ecologically fragile karst region using signature lipid biomarker analysis to provide information on a variety of microbial properties. Here, phospholipid fatty acids (PLFAs) in the soil microbial community were measured following different restoration strategies in a karst region of southwest China. Managed vegetation restoration (including restoration with plantation forest, forage grass, and a combination of plantation forest and forage grass) and natural vegetation restoration (spontaneous regeneration to natural grassland) were compared, using cropland as a reference. The abundance of total PLFAs and the PLFAs of four microbial functional groups (bacteria, fungi, actinomycetes, and arbuscular mycorrhizal fungi [AMF]) significantly increased in the topsoil (0–5 cm soil depth) following vegetation restoration. In the sub-topsoil layer (5–10 cm soil depth), the abundance of PLFAs was not significantly higher than that in the cropland, except in AMF in natural vegetation restoration. Principal component analysis indicated that the microbial PLFA composition differed between the restoration strategies, and the abundance of AMF PLFAs was significantly higher in natural than in managed vegetation restoration for both soil depths. The ratio of soil organic carbon to total nitrogen and dissolved organic carbon content were the primary factors affecting the abundance of soil microbial communities in the topsoil and sub-topsoil, respectively. In addition, soil pH served as another explanatory variable for the topsoil. Our findings suggest that vegetation restoration over a relatively short period (15 years) mainly improved topsoil fertility. Moreover, natural vegetation restoration may be superior to managed vegetation restoration for the maintenance of multiple ecosystem functions in the short term in the karst regions, owing to the potentially significant role of AMF. [ABSTRACT FROM AUTHOR]

1,1,2-trichloroethane (TCA) is a contaminant in many pesticide and chemical fields. This study mainly described the potential effectiveness of the natural attenuation of 1,1,2-TCA in groundwater in a typical pesticide-contaminated field in east China. In this study, six typical 1,1,2-TCA-contaminated sites (MW14, MW21, MW25, MW31, MW36 and MW41) were selected, and the field investigation results indicated that there was an obvious decline in the concentrations of 1,1,2-TCA over time in all selected sites. Furthermore, the attenuation rate of 1,1,2-TCA concentration in sites MW14, MW21, MW25, MW31 and MW41 followed the first-order kinetic equation, and the first-order attenuation rate constants were calculated. The health risks of 1,1,2-TCA and its degradation product dichloroethane (DCA) were assessed and compared in site MW14. The result showed that the health risks of 1,1,2-TCA were much higher than those of DCA, and 1,1,2-TCA was the contamination source in this site. 16S rRNA sequencing was also conducted to investigate the diversity of the bacterial community in 1,1,2-trichloroethane (TCA)-contaminated groundwater, and Geobacter, Thauera, Pseudomonas, Diaphorobacter were the main species in the bacterial community. Image 10209 • First-order decay of 1,1,2-TCA in a pesticide-contaminated field was observed. • Health risks of TCA and its degradation products were assessed. • The potential degrading mechanism of 1,1,2-TCA was discussed. [ABSTRACT FROM AUTHOR]

Plants are generally limited by soil phosphorus (P) deficiency in forest ecosystems. Soil available P is influenced by lithology, temperature, and soil microbes. However, the interactive effects of these factors on soil P availability in subtropical forests remain unclear. To assess their impacts, we measured soil inorganic and available P fractions and the diversity, composition, and co-occurrence network of phoD-harboring bacteria in two contrasting forest soils (lithosols in karst forests and ferralsols in non-karst forests) in the subtropical regions of southwestern China across six temperature gradients. The present results showed that the complexities in composition and network and the diversity indices of phoD-harboring bacteria were higher in the karst forest soils than those in the non-karst forest soils, with marked differences in composition. In both types of forest soils, the complexities of composition and networks and the diversity indices were higher in the high-temperature regions (mean annual temperature (MAT) > 16 °C) compared to the low-temperature regions (MAT <16 °C). Soil total inorganic and available P contents were lower in the karst forest soils compared to the non-karst forest soils. Soil total available P contents were lower in the high temperature regions than those in the low temperature regions in both forest soils, whereas soil total inorganic P contents were contrary. Variance partitioning analysis showed that soil inorganic and available P fractions were predominantly explained by lithology and its interaction with soil microbes and climate. The present findings demonstrate that soil P availability in subtropical forests of southwestern China is influenced by lithology and temperature, which regulate the diversity, composition, and network connectivity of phoD-harboring bacteria. Furthermore, this study highlights the significance of controlling the composition of phoD-harboring bacteria for mitigating plant P deficiency in karst ecosystems., Competing Interests: Declaration of competing interest The authors declare that they have no conflicts of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Vegetation restoration can increase soil carbon (C) content in karst regions characterized by highly exposed carbonate rocks; however, it remains unclear whether and how bedrock outcrops contribute to soil C-accumulation after vegetation restoration. We aimed to investigate the magnitude and mechanisms of bedrock outcrops on soil C-accumulation after vegetation restoration. Here, we selected 362 fixed locations to investigate changes in soil organic carbon (SOC) content and density before and after cropland restoration in a karst catchment with varying bedrock exposure ratios and initial soil C pools prior to restoration. Active vegetation restoration (i.e., cropland converted to forage grass, plantation forest, and a combination of grass and forest) and natural regeneration (cropland abandoned) were compared, with croplands maintained with no change as the control. Compared to croplands maintained with no change, SOC density significantly increased in the four vegetation restoration types. The SOC accumulation rate was higher for natural regeneration (39 g C m -2 yr -1 ) than for the three active restoration strategies (18-27 g C m -2 yr -1 ). SOC accumulation decreased with a higher initial pool size of soil C but increased with nitrogen accumulation and soil exchangeable calcium (Ca 2+ ) concentration. Higher bedrock outcrops reduced soil volume but increased SOC content through their indirect effects on the initial pool size of soil C, external nitrogen inputs, and soil Ca 2+ concentration. This weakly promoted rather than inhibited SOC sequestration. Our findings highlight the effectiveness of various restoration strategies in promoting SOC accumulation in karst areas, as well as the need to take bedrock outcrops and initial soil C pools into consideration when modeling SOC dynamics and maximizing C sinks for vegetation restoration., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Microbial communities contribute to sustaining the function of terrestrial ecosystems and are influenced by soil type and climate gradients. The effects of karst and non-karst soils on bacterial and fungal profiles for seven climate gradients were assessed to better understand bacterial and fungal diversity and community composition in response to soil type with changes in soil physicochemical properties under different temperatures and precipitations. Bacterial and fungal abundance, diversity, and community composition differed between karst and non-karst forests. Bacterial and fungal richness, Shannon index, and bacterial abundance in karst forests were higher than non-karst forests, but the fungal abundance was lower. Mean annual temperature was negatively correlated with bacterial diversity in the karst forest and fungal abundance in karst and non-karst forests. The community composition of bacteria and fungi differed among these two soil types. The karst forest had greater connectivity among bacterial and fungal communities than non-karst forests. The bacterial members of Acidobacteria, Proteobacteria, Actinobacteria, and fungal groups of Ascomycota and Basidiomycota were mainly connected with other taxa in the network, implying that taxa were associated with highly functional potential. The relative abundance of Actinobacteria and Ascomycota was higher in karst than in non-karst forests. Proteobacteria and Basidiomycota showed the opposite results. A random forest and multiple regression tree analyses revealed that soil properties, specifically pH, calcium, and total nitrogen, were the main factors influencing the variation in bacterial and fungal profiles between karst and non-karst forests. This study provides novel evidence that the abundant microbial taxa were kinless hubs in co-occurrence patterns. Controlling complex networks of species interactions may contribute to improving soil nutrient processes rather than microbial diversity, enhancing our understanding of developing sustainable recovery strategies in fragile ecosystems., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. We have read and understood your journal's policies, and we believe that neither the manuscript nor the study violates any of these., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Cropland restoration induces litter and root inputs and promotes the development of biological soil crusts (BSCs), which may promote aggregate formation and soil organic carbon (SOC) sequestration. However, litter, roots and BSCs have not been simultaneously considered when assessing soil aggregate and aggregate-associated SOC fraction responses to cropland restoration in subtropical areas. Here, we measured particulate organic carbon (POC) and mineral-associated organic carbon (MOC) in bulk soils and soil aggregates after 15 years of cropland restoration. Soil samples of cropland (CL) and four cropland restoration types (plantation forest [PF], forage grassland [FG], mixed plantation of forest and forage grassland [FF], and abandoned natural grassland [NG]) from depths of 0-30 cm were collected. Cropland restoration significantly increased SOC and POC in bulk soil at the 0-5 cm depth. However, only in FG did SOC significantly increase at depths of 5-15 cm, and POC significantly increased at depths of 5-30 cm. The large macroaggregate (5-10 mm and 2-5 mm) proportions increased significantly at the 0-15 cm depth after cropland restoration, and FG, FF and NG also increased the 5-10 mm aggregate proportions at the 15-30 cm depth. The SOC sequestration in bulk soil with cropland restoration was attributed to increases in the aggregate-associated organic carbon (AAOC) pool in large macroaggregates, which was mainly attributed to the increased aggregate amount rather than the increased AAOC concentration in large macroaggregates. Our results also indicated that an increase in aggregate-associated particulate organic carbon (AAPOC) led to an increase in AAOC. Variation partitioning indicated that the formation of large macroaggregates was controlled by the litter-moss-root interactive effect in this karst area. FG could be a better short-term cropland restoration strategy, increasing large macroaggregates in deeper soil layers better than the other vegetation types and promoting soil carbon sequestration in deeper soil layers., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Concerns about the specificity of the Xpert MTB/RIF (Xpert) assay have arisen, as false-positive errors in the determination of Mycobacterium tuberculosis complex (MTBC) infection and rifampin (RIF) resistance in clinical practice have been reported. Here, we investigated 33 cases where patients were determined to be RIF susceptible using the Bactec MGIT 960 (MGIT) culture system but RIF resistant using the Xpert assay. Isolates from two of these patients were found not to have any mutations in the rifampin resistance determining region (RRDR) region of rpoB and had good treatment outcomes with first-line antituberculosis (anti-TB) drugs. The remaining 31 patients included 5 new cases and 26 previously treated patients. A large number of well-documented disputed mutations, including Leu511Pro, Asp516Tyr, His526Asn, His526Leu, His526Cys, and Leu533Pro, were detected, and mutations, including a 508 to 509 deletion and His526Gly, were described here as disputed mutations for the first time. Twenty-one (81%) of the 26 previously treated patients had poor treatment outcomes, and isolates from 19 (90%) of these 21 patients were resistant to isoniazid (INH) as determined using the MGIT culture system. Twenty-seven of the 31 isolates with disputed rpoB mutations were phenotypically resistant to INH, 21 (78%) being predicted by GenoType MTBDR plus to have a high level of INH resistance. Most (77.4%) of the isolates with disputed mutations were of the Beijing lineage. These findings have implications for the interpretation of false-positive and disputed rifampin resistance Xpert MTB/RIF results in clinical samples and provide guidance on how clinicians should manage patients carrying isolates with disputed rpoB mutations., (Copyright © 2019 American Society for Microbiology.)