SHORT COMMUNICATION-Pattern of anti-tuberculosis drugs susceptibility in new and previously treated tuberculosis patients and environmental risk factors investigation.

Resistance pattern both in newly and previously treated-TB patients and risk factors associated in spread of tuberculosis are investigated in the current study. A total 244 Mycobacterium tuberculosis isolates were used for drug-susceptibility test against four drugs. Environmental risk factors were assessed by using self-designed history proforma. Among 244 TB-isolates, 64% were categorized as MDR-TB in drug-susceptibility test. Male proportion was 51% while 32% belonged to 15-34 years age group and 49% were from city Lahore whereas majority of people (31%) was working on daily wages. Divergent drug-resistance pattern was obtained; RIF (68%), SM (52%), EMB (51%). INH showed only (27%) resistance against first-line anti-TB drug. Drug-resistance prevalence for two drug combination was highest (50%) for (INH+SM) and (INH+EMB) followed by (RIF+SM) (49%) whereas for three drugs combination (INH+RIF+EMB) and (INH+RIF+SM) the prevalence was almost same 50% and 49% respectively while 66% patients were categorized as previously treated and 34% as new TB cases. In drug susceptibility test, 71% were identified as MDR-TB among New TB cases, while 63% were identified as MDR-TB from previously treated cases. Surprisingly DST results displayed that percentage prevalence of MDR-TB both in newly and previously treated cases was almost same.

Molecular detection of mutations in isolates of multidrug resistant tuberculosis and tuberculosis suspects by multiplex allele specific PCR.

For lowering prevalence of drug resistance it is necessary to diagnose TB in tuberculosis sputum suspect patients instead of TB-cultured samples which required a long time of culturing. Comparison of the results of drug resistant bacterial genes in both tuberculosis suspect sputum and multi-drug resistant DNA isolates detected by MAS-PCR. In the current study, the genetic mutations linked with INH, RIF as well as EMB drugs were detected by MAS-PCR simultaneously in MDR as well as TB suspect sputum isolates. 175/291 samples belonged to MDR and 116/291 samples belonged to tuberculosis suspect group. In all the isolates, presence of Mycobacterium tuberculosis-species (100%) was confirmed by targeting hupB gene. In MDR group, maximum prevalence of gene mutation was detected in rpoB531 (92.57%) and embB306 (97.71%) while in TB-suspect group, equal percentage (96.55%) of mutation was detected in rpoB531 and embB306 by MAS-PCR. Collectively, rpoB531 (n=274, 94.15%) and embB306 (n=283, 97.25%) mutation were observed in maximum tuberculosis cases. MAS-PCR technique yielded reliable results and showed massive Isoniazid, Rifampicin and Ethambutol drugs resistance in TB-isolates from Pakistan; hence it can be used in clinical laboratories with high burden of tuberculosis to detect drug resistance rapidly and cost effectively.

Simultaneous adsorptive removal of three fluoroquinolones using humic acid modified hydrogel beads.

This study elaborates the simultaneous removal of three different fluoroquinolones (FQs), i.e., Norfloxacin (NOR), Lomefloxacin (LOM), and Enrofloxacin (ENR) from water using hydrogel beads of humic acid coated biochar (HA-BC) and chitosan. In our previous study, this adsorbent has already achieved tremendous results for the removal of a single FQ, i.e., ciprofloxacin. Now, initial concentrations of all FQs were set 100 mg/L each, and the maximum adsorbed amounts were 38.08 mg/g (NOR), 25.03 mg/g (LOM), and 29.72 mg/g (ENR). Adsorption attained equilibrium after 24 h, which obeyed the pseudo-second-order kinetic model. The mutation of humic acid-biochar/chitosan hydrogel beads (HBCB) with alcoholic solvents, i.e., methanol and ethanol to replace water decreased its sorption capacities from 38.08 mg/g (NOR) to 34.91 mg/g and 32.19 mg/g, respectively. Similarly, from 25.03 mg/g (LOM) to 22.81 mg/g and 19.91 mg/g, and 29.72 mg/g (ENR) to 26.52 mg/g and 24.64 mg/g. Adsorption isotherm data for all FQs were up to both Langmuir and Freundlich, but it suited more to that of Langmuir adsorption isotherm model. Sorption capacities, for all FQs, had a minor decline due to addition of NaCl, NaNO3, and Na2SO4. However, there was a huge decline when Na3PO4 was added into the adsorption system. Adsorbent was desorbed and regenerated for consecutive removal, and it showed good adsorption in the 4th cycle, i.e., 47 mg/g net adsorption. These results prove that HBCB is not only effective for adsorption removal of ciprofloxacin but also for other FQs too.

Application of Cerium Dioxide Nanoparticles and Chromium-Resistant Bacteria Reduced Chromium Toxicity in Sunflower Plants.

The continuous increase in the heavy metals concentration in the soil due to anthropogenic activities has become a global issue. The chromium, especially hexavalent chromium, is highly toxic for living organisms due to high mobility, solubility, and carcinogenic properties. Considering the beneficial role of nanoparticles and bacteria in alleviating the metal stress in plants, a study was carried out to evaluate the role of cerium dioxide (CeO2) nanoparticles (NPs) and Staphylococcus aureus in alleviating the chromium toxicity in sunflower plants. Sunflower plants grown in chromium (Cr) contaminated soil (0, 25, and 50 mg kg-1) were treated with CeO2 nanoparticles (0, 25, and 50 mg L-1) and S. aureus. The application of Cerium Dioxide Nanoparticles (CeO2 NPs) significantly improved plant growth and biomass production, reduced oxidative stress, and enhanced the enzymatic activities in the sunflower plant grown under chromium stress. The application of S. aureus further enhanced the beneficial role of nanoparticles in alleviating metal-induced toxicity. The maximum improvement was noted in plants treated with both nanoparticles and S. aureus. The augmented application of CeO2 NPs (50 mg l-1) at Cr 50 mg kg-1 increased the chl a contents from 1.2 to 2.0, chl b contents 0.5 to 0.8 and mg g-1 FW, and decreased the leakage of the electrolyte from 121 to 104%. The findings proved that the application of CeO2 nanoparticles and S. aureus could significantly ameliorate the metal-induced stress in sunflower plants. The findings from this study can provide new horizons for research in the application of nanoparticles in phytoremediation and bioremediation.

Drug resistance profile of Mycobacterium tuberculosis and predictors associated with the development of drug resistance.

OBJECTIVES: Drug-resistant tuberculosis (DR-TB) is a major challenge to national TB control programmes in developing countries. In the Pakistan province of Punjab, the extent and development of DR-TB is not well known. The current study was therefore conducted to assess the incidence and predictors of DR-TB in Punjab Province. METHODS: Drug susceptibility testing was performed for 863 confirmed culture-positive Mycobacterium tuberculosis isolates using the proportion method. Patients were enrolled in the Programmatic Management of Drug-Resistant TB Unit of Gulab Devi Chest Hospital (Lahore, Pakistan) from August 2011 to September 2013. Data analysis was performed using IBS SPSS Statistics v.20. Multivariate logistic regression analysis was performed to assess risk factors for DR-TB. RESULTS: The rate of resistance to at least one drug (i.e. DR-TB) was 35.0% (302/863) and the rate of multidrug-resistant TB (MDR-TB) was 30.0% (259/863). Among DR-TB cases, the number of females was relatively higher (167/302; 55.3%) compared with males. The majority of DR-TB patients resided in a rural area (229/302; 75.8%). Significant predictors of DR-TB were age 18-45 years, previous TB treatment, rural residence, being a housewife, being married, duration of sickness >1year and unemployment. CONCLUSION: The problem of DR-TB in Pakistan is significant. The strongest risk factors were young age and previous anti-TB treatment. Being married, being a housewife, rural residence and unemployment were also risk factors, culminating in an urgent need for effective control, early diagnosis and treatment policies for DR-TB.

Sequence analysis of the rifampicin resistance determining region (RRDR) of rpoB gene in multidrug resistance confirmed and newly diagnosed tuberculosis patients of Punjab, Pakistan.

Molecular screening of new patients suspected for TB could help in the effective control of TB in Pakistan as it is a high TB burden country. It will be informative to understand the prevalence of multi drug resistance for a better drug regimen management in this geographical area. The Rifampicin resistance determining region (RRDR) sequencing was used to identify mutations associated with drug resistance in DNA extracts from 130 known multidrug resistant (MDR) cultured strains and compared with mutations observed in DNA extracts directly from 86 sputum samples from consecutive newly diagnosed cases in Lahore, Pakistan. These newly diagnosed samples were positive for smear microscopy, chest X-ray and presumed sensitive to first line drugs. In the known MDR group the most frequent mutations conferring resistance were found in rpoB531 (n = 51, 39.2%). In the newly diagnosed tuberculosis group with no history of MDR, mutations in rpoB531 were seen in 10 of the samples (11.6%). Collectively, all mutations in the RRDR region studied were observed in 80 (61.5%) of known MDR cases and in 14 (16.3%) of the newly diagnosed cases. Using the RRDR as a surrogate marker for MDR, sequences for the newly diagnosed (presumed sensitive) group indicate much higher levels of MDR than the 3.9% WHO 2015 global estimate and suggests that molecular screening directly from sputum is urgently required to effectively address the detection and treatment gaps to combat MDR in this high burden country.