Evaluation of the Effects of Wastewater Irrigation on Heavy Metal Accumulation in Vegetables and Human Health in the Cauliflower Example : Heavy Metal Accumulation in Cauliflower.

The goals of the present research were to determine the heavy metal contents in the water-soil-cauliflower samples in industrial wastewater irrigated areas and to assess the health risks of these metals to the people. Metal analyses were carried out using the atomic absorption spectrophotometer equipped with a graphite furnace. The metal readings in the cauliflower specimens ranged from 1.153 to 1.389, 0.037 to 0.095, 0.61 to 0.892, 0.625 to 0.921, 1.165 to 2.399, 0.561 to 0.652, 0.565 to 0.585, 0.159 to 0.218 and 1.268 to 1.816 mg/kg for Cd, Co, Cr, Cu, Fe, Ni, Pb, Zn and Mn, respectively. Statistics revealed that, with the exception of Pb and Co (p > 0.05), there was no statistically significant variation in the metal concentrations in the cauliflower samples according to the irrigation type. Pb, Ni, and Cr had HRI values below 1.0 and did not seem to be a hazard to human health, in contrast to Cd, Co, Cr, Cu, Fe, Ni, Pb, Zn and Mn, which glanced to constitute a health risk. Regular monitoring of vegetables irrigated with wastewater is strongly advised to reduce health hazards to people.

Bioaccumulation and Health Risk Assessment of Potentially Toxic Metals in Citrus Limetta & Citrus Sinensis Irrigated by Wastewater.

The aim of this study was to evaluate the impact of different irrigation sources on the levels of potentially toxic metals (Cd, Cr, Fe and Mn) in the edibles of citrus fruits (Citrus sinensis and Citrus limetta). The samples of fruit, soil and water were collected from two locations (fresh water irrigated-FW I and sewage water irrigated-SW II) within the city of Sargodha. The samples utilized in the study for metal analysis were prepared utilizing the wet acid digestion method. Metal determination was performed using Atomic Absorption Spectrometry (AAS). The potentially toxic metal values in the citrus samples ranged from 0.010 to 0.063, 0.015 to 0.293, 6.691 to 11.342 and 0.366 to 0.667 mg/kg for Cd, Cr, Fe and Mn, respectively. Analysis of Citrus limetta and Citrus sinensis indicated that the highest concentration of Cr, Fe and Mn is observed at the sewage water irrigation site (SW-II), whilst the minimum levels of Cr, Fe and Mn were observed at the fresh water irrigation site (FW-I). The results show that the levels of these metals in soil and fruit samples meet the acceptable guidelines outlined by USEPA and WHO. It was found that the metal pollution constitutes a potential threat to human health due to the HRI values for Cd, Cr, and Fe being above 1, despite the DIM values being below 1. Regular monitoring of vegetables irrigated with wastewater is highly recommended in order to minimise health risks to individuals.

Arsenic Levels and Seasonal Variation in Pasture Soil, Forage and Horse Blood Plasma in Central Punjab, Pakistan.

The present study aimed to determine the accumulation levels of arsenic in pasture soil, forage and animals. An atomic absorption spectrophotometer was used to determine the concentration of metals in the samples of soil, forage and blood plasma of horses. The level of arsenic ranged between 4.26 mg/kg (summer) and 4.66 mg/kg (winter) in soil samples and 2.67 mg/kg (summer) and 2.94 mg/kg (winter) in forage samples. In blood plasma samples, the mean arsenic (As) values varied between 1.38 and 1.52 mg/L. In the blood plasma samples, the mean As values varied between 1.38 and 1.52 mg/L. No statistically significant changes were observed for arsenic concentrations in plasma samples of horses for sampling seasons (p > 0.05). A positive correlation was observed for forage and blood plasma to a certain degree for arsenic but for other media, arsenic correlations were negative and insignificant. It is therefore suggested that regular monitoring of heavy metals in soils/plants/animals is essential to prevent excessive build-up of arsenic.

Does Industrial Wastewater Irrigation Cause Potentially Toxic Metal Contamination and Risk to Human Health? Sugar Industry Wastewater and Radish Examples.

The goal of this study was to appraise potentially toxic metal contents in the soil-radish system in industrial wastewater irrigated areas. The analysis of metals in water, soil and radish samples were performed with spectrophotometric method. The potentially toxic metal values in the wastewater irrigated radish samples were ranged from 1.25 to 1.41, 0.02 to 0.10, 0.77 to 0.81, 0.72 to 0.80, 0.92 to 1.19, 0.69 to 0.78, 0.08 to 0.11, 1.64 to 1.67 and 0.49 to 0.63 mg/kg for Cd, Co, Cr, Cu, Fe, Ni, Pb, Zn and Mn, respectively. The potentially toxic metal values in the soil and radish samples irrigated with wastewater were lower than the maximum allowable limits, except for Cd. The results of the Health Risk Index evaluation conducted in this study also showed that the accumulations of Co, Cu, Fe, Mn, Cr, and Zn, especially Cd, pose a health risk in terms of consumption.

Data-driven predictive modeling of PM2.5 concentrations using machine learning and deep learning techniques: a case study of Delhi, India.

The present study intends to use machine learning (ML) and deep learning (DL) models to forecast PM2.5 concentration at a location in Delhi. For this purpose, multi-layer feed-forward neural network (MLFFNN), support vector machine (SVM), random forest (RF) and long short-term memory networks (LSTM) have been applied. The air pollutants, e.g., CO, Ozone, PM10, NO, NO2, NOx, NH3, SO2, benzene, toluene, as well as meteorological parameters (temperature, wind speed, wind direction, rainfall, evaporation, humidity, pressure, etc.), have been used as inputs in the present study. Moreover, this is one of the first papers that employ aerodynamic roughness coefficient as an input parameter for the prediction of PM2.5 concentration. The result of the study shows that the LSTM model with index of agreement (IA) 0.986, root mean square error (RMSE) 21.510, Nash-Sutcliffe efficiency index (NSE) 0.945, (coefficient of determination)R2 0.945, and (correlation coefficient)R 0.972 is the best performing technique for the prediction of PM2.5 followed by MLFFNN, SVM, and RF models. The sensitivity analysis for the LSTM model reported that PM10, wind speed, NH3, and benzene are the key influencing parameters for the estimation of PM2.5. The findings in this work suggest that the LSTM could advance in PM2.5 forecasting and thus would be useful for developing fine-scale, state-of-the-art air pollution forecasting models.

Assessing the health risk of cadmium to the local population through consumption of contaminated vegetables grown in municipal solid waste-amended soil.

Pollution caused by municipal solid waste (MSW) is becoming a serious threat to the environment. Composting may be an effective way to speed up the decomposition of biodegradable components in MSW, resulting in compost that can be utilized as an organic fertilizer. The pot experiments were carried out with different soil-MSW mixtures (100:0, 75:25, 50:50, and 25:75; w/w) to determine the impact of MSW on the bioconcentration of Cd in commonly consumed plants of Sargodha. The possible health risks were evaluated by applying pollution indices, such as the pollution load index, bioconcentration factor, enrichment factor, and health risk index. The pollution load index was higher than 1 in 75% MSW-amended soil. However, the concentration of Cd was found to be below the permissible limits in all studied vegetables, with a range of 0.019-0.106 mg/kg. In the study, serum samples from different volunteers living in four sites in Sargodha were also collected and analyzed. For vegetable crops, the health risk index (HRI) was less than one. It is concluded that the concentration of Cd was increased by increasing the fraction of MSW. Although the metal contents in the soil treated with MSW were not high enough to categorize the soil as polluted, these findings show that the reuse of MSW can serve as an alternative to mineral fertilizers. However, the presence of Cd in MSW can have a direct impact on soil fertility and, if biomagnified, on crop production and human health.

Potentially Toxic Metal Accumulation in Spinach (Spinacia oleracea L.) Irrigated with Industrial Wastewater and Health Risk Assessment from Consumption.

This study aimed to determine the potentially toxic metal contents in soil and spinach samples in areas irrigated with industrial wastewater and to evaluate the potentially toxic metal accumulation in spinach samples according to pollution indices. Water, soil and spinach samples were analysed using atomic absorption spectrophotometer (Perkin-Elmer AAS-300). In this study, it was determined that the potentially toxic metal values ​​in the spinach samples irrigated with groundwater and sugar industry wastewater varied between 1.59 and 1.84, 0.22-0.68, 0.56-1.14, 1.41-1.56, 1.62-3.23, 0.57-1.02, 0.86-1.33, 0.20-0.32 and 0.35-2.10 mg/kg for Cd, Co, Cr, Cu, Fe, Ni, Pb, Zn and Mn, respectively. It was concluded that the difference between the metal values ​​in the spinach samples according to the irrigation sources was statistically significant, except for Cu and Pb (p > 0.05). According to the results of this study, there is no health risk for Pb, Co and Cr with HRI values ​​below 1.0, while there is a risk for Cd, Cu, Fe, Ni, Mn and Zn. The much higher HRI values ​​of Cd than 1 (196.8 and 169.6) suggested that this metal is likely to cause significant health problems in the region.

Simultaneous modelling of coagulant recovery and reuse by response surface methodology.

Surface water from rivers, lakes, reservoirs etc. needs to be treated prior to municipal supplies. The treatment scheme includes coagulation, flocculation, sedimentation, filtration and finally disinfection process. Huge volume of sludge or waste is generated during the coagulation-flocculation. Disposal of the sludge so generated in the treatment plants require careful consideration for managing it sustainably and in an environment friendly manner. Constructive utilization of the inevitable waste may help in finding a sustainable solution to sludge disposal problems. Presently, response surface methodology (RSM) with central composite design (CCD) has been applied to simultaneously model coagulant recovery as well as reuse parameters. In order to simplify the process and increase the applicability, the effect of three significant variables, acid dose, sludge ratio, and recovered coagulant dose are studied. A second order regression model has been developed which gave the optimum combination of acid dose of 30 ml/L, sludge ratio of 1% and recovered coagulant dose of 12 ml/L for maximum turbidity removal. The predicted value of turbidity removal is 95.4%. In the confirmatory experiments, the turbidity removal value was observed to be about 96.2%, which is in good agreement with the predicted value. In addition to turbidity removal, it also helps to effectively remove other impurities from the raw water for it to meet the standards prescribed for potable supply. Thus, the regenerated alum or recovered coagulant has the potential to substitute the conventional coagulants, fully or partially at water treatment plants.

Molecular characterization of hepatitis B virus basal core promoter and precore region of isolates from chronic hepatitis B patients.

OBJECTIVE: To analyse mutations in precore and core promoter regions of hepatitis B virus genome in chronic hepatitis B patients. METHODS: The cross-sectional prospective study was conducted at the Centre of Biotechnology and Microbiology, University of Peshawar and Pakistan Health Research Council (PHRC), Research Centre, Khyber Medical College Peshawar from June 2014 to June 2015, and comprised samples from treatment-naïve chronic hepatitis B patients aged >15 years from three cities of Pakistan. The samples included patients who were both positive and negative for hepatitis B envelope antigen. Viral load, hepatitis B envelope antigen / anti-hepatitis B envelope status, hepatitis B virus enzyme-linked immunosorbent assay and alanine aminotransferase levels were determined. Direct sequencing of basal core promoter and precore regions of hepatitis B virus genome was carried out following a nested polymerase chain reaction approach. Phylogenetic tree was constructed using Molecular Evolutionary Genetics Analysis software version 6.0. Data was analysed using SPSS 16. RESULTS: Of the 50 patients, 33(66%) were males. The overall mean age was 28.5±11.4 years. Of all the subjects, 25(50%) each were positive and negative for hepatitis B envelope antigen. Precore stop codon mutation G1896A was detected in 19 (38%) isolates; 17(34%) among negative patients and 2(4%) in positive patients. Classic A1762T/G1764A double mutation was noted in 15(30%) isolates. Mutation at position 1764 was observed in 12(48%) samples. A rare G1764T mutation was also detected in 6(12%) isolates. The CG1802-1803 mutation was detected in 47(94%) isolates, while all the 50(100%) isolates had T1858A. The GCAC Kozak sequence was present in 43(86%) isolates; CAA1817-1819 in 49(98%); and G1888 in 49(98%). Overall, 9(18%) isolates had wild-type sequences at all important loci, including positions 1762, 1764 and 1896. The pattern of sequences at genotype specific positions and phylogenetic tree speculates that majority of study isolates belonged to genotype D. CONCLUSION: Basal core promoter and precore regions variants along with the preponderances of genotype D-specific mutations suggested a higher risk of hepatocellular carcinoma and poor clinical outcome in such patients.

Mineral availability in soil and fodders affecting blood profiles in Nili-Ravi dairy buffaloes.

A mineral assessment study for dairy buffaloes, Nili-Ravi breed, was carried out seasonally at silvopasture farm at Sargodha, Pakistan. Sampling for soil, forage, and buffalo serum was done seasonally for a period of 1 year for mineral (P, Mg, Na, K, Ca) evaluation. In the study, experiments on forages showed significant differences in mean concentrations of Mg and P with changing seasons, whereas the values for Ca, Mg, Na, and K did not vary significantly. Three groups of buffaloes were checked for mineral concentration in blood serum, viz., calves, lactating and non-lactating. Ca mean concentration varied significantly for non-lactating buffaloes and calves between two seasons. Na mean level showed a significant difference for two sampling seasons among calves and lactating buffaloes. Mean level for Mg only differed significantly in serum of non-lactating buffaloes between the two seasons. Mean levels for Ca and P in soil and forage remained higher than the optimal level, whereas soil Na was below the critical level. K concentrations in soil and blood serum were above the critical level, while Mg levels remained below the optimal range in soil, forage, and blood serum. K concentration in forage remained below the optimal level. All buffalo categories displayed lower levels of Ca and P in serum. Na concentration in forage and serum exceeded the critical level. Furthermore, Ca yielded significant and positive correlation between soil and serum level, whereas K had significant but negative correlation for soil-forage and forage-serum. Overall, it was determined that P deficiency existed among buffaloes, which could be improved via supplementation. Similarly, K deficiency and Mg deficiency were exhibited for forage and soil respectively which could be countered through addition of fertilizers rich for these minerals. In brief, the mineral utilization of buffaloes is affected by reduced availability of essential nutrients and may result in lower productivity. It is suggested to add mineral supplementation in addition to natural diet of buffaloes to enhance the productivity of these animals.

Genetic diversity and molecular analysis of metallo beta lactamases among imipenem resistant clinical isolates of Pseudomonas aeruginosa from Peshawar, Pakistan.

OBJECTIVES: Pseudomonas aeruginosa is an opportunistic pathogen with remarkable adaptation ability to thrive in diverse environmental conditions. This study aimed at phenotypic and molecular analysis of metallo beta lactamases (blaIMP, blaVIM, blaNDM-1 and blaSPM-1) and genetic diversity analysis among imipenem resistant clinical isolates of Pseudomonas aeruginosa. METHODS: This study was conducted from May 2017 to June 2018. The study included 187 Pseudomonas aeruginosa isolates collected from different clinical specimens from Peshawar, Pakistan. The isolates were analyzed for resistance to imipenem. Combined disc test (CDT) was then performed for phenotypic detection of metallo beta lactamases among imipenem resistant isolates of Pseudomonas aeruginosa. Molecular detection of metallo beta lactamases genes i.e. blaIMP, blaVIM, blaNDM-1 and blaSPM-1 was analyzed through polymerase chain reaction. Genetic diversity was determined through RAPD-PCR. RESULTS: MBL production was observed in 76% (n=19) isolates. The occurrence of MBL genes blaIMP, blaNDM-1 and blaVIM was 68% (n=17), 48% (n=12), and 4% (n=1) respectively. The blaSPM-1 gene was not detected. High genetic diversity was observed in current study. Out of 182 isolates 171 isolates showed different RAPD profiles (93.95% polymorphism); 160 were unique RAPD strains and based on similarity coefficient ≥ 80%, 22 isolates were clustered into 11 distinct clones. CONCLUSION: A high prevalence of blaIMP and blaNDM-1 among imipenem resistant isolates of Pseudomonas aeruginosa is alarming that calls for proper control and prevention strategies. RAPD technique was found to be a good genotyping technique when limited resources are available.

Role of surface charge in enhancing antibacterial activity of fluorescent carbon dots.

Herein, different surface charged carbon dots (Cdots) were synthesized by using diethylene glycol as a carbon source with various amine containing surface passivating agents. The synthesis method is very simple and fast microwave oven-based, that results in almost similar sized positive, negative and uncharged fluorescent Cdots which has been confirmed by zeta potential analysis in our case. The formation of Cdots was confirmed by characterization using fluorescence spectroscopy, transmission electron microscopy, XRD, FT-IR, and XPS spectroscopy. To find out relative bactericidal activity of these Cdots, green fluorescence protein expressing recombinant E. coli bacteria were taken as a model system. Time-dependent bacterial growth and FACS study demonstrated that both uncharged Cdots and positively charged Cdots were showing better bactericidal activity as compared to negative charged Cdots. The Cdots caused elevation of reactive oxygen species level, which is possibly leading to bacterial cell death.

Effect of Organic Manure and Mineral Fertilizers on Bioaccumulation and Translocation of Trace Metals in Maize.

Mineral fertilizers and organic manure are used as soil amender to enhance the mineral status of the soil. These fertilizers contain trace metals besides providing macro and micronutrients. The present study was performed to observe the effect of mineral fertilizers, poultry manure and cow manure on trace metal content of soil and various parts (root, shoot, and grains) of maize plant (Zea mays L.). The analysis of metals was performed by atomic absorption spectrophotometer (AA-6300 Shimadzu Japan). The highest level of Pb, Fe, Ni and Cu was observed in the root as 0.36-0.55, 70.41-83.03, 4.98-7.44 and 2.94-4.43 mg kg- 1, respectively. The highest level of Cd, Zn and Mn was determined in grains as 0.44-1.59, 28.05-46.39 and 26.24-46.57 mg kg- 1, respectively. The values of all metals were found within their permissible limit given by FAO/WHO except for the Cd. The interactive use of mineral and organic fertilizers enhanced the level of trace metals in maize as compared to their sole application. In the present findings, the health risk index for all metals was less than 1 in all treatments. So, it was concluded that the level of metals in poultry manure, cow manure and mineral fertilizer treated maize did not pose any potential threat to the consumers.

Boron Doped Carbon Dots with Unusually High Photoluminescence Quantum Yield for Ratiometric Intracellular pH Sensing.

Herein we report that boron doping in carbon dots results in increased photoluminescence (PL) quantum yield, which could be used for ratiometric intracellular pH sensing in cancer cell lines. Using a mixture of citric acid monohydrate, thiourea, and boric acid, microwave-assisted synthesis of boron doped blue emitting carbon dots (B-Cdots) with an average size of 3.5±1.0 nm was achieved. For B-Cdots, the maximum quantum yield (QY) was observed to be 25.8 % (11.1 % (w/w) H3 BO3 input concentration), whereas, the same was calculated to be 16.9 % and 11.4 % for Cdots (synthesized from citric acid monohydrate and thiourea only) and P-Cdots (phosphorus doped carbon dots; synthesized using citric acid monohydrate, thiourea and phosphoric acid) (11.1 % (w/w) H3 PO4 input concentration), respectively. The observed luminescence efficiencies as obtained from steady state and time-resolved photoluminescence measurements suggest an alternative emission mechanism due to boron/phosphorus doping in carbon dots. We furthermore demonstrated facile composite formation using B-Cdots and another carbon dots with orange emission in presence of polyvinyl alcohol (PVA), resulting in white light emission (0.31, 0.32; λex 380 nm). The white light emitting composite enabled ratiometric pH sensing in the aqueous medium and showed favorable uptake properties by cancerous cells for intracellular pH sensing as well.

Trace Metal Accumulation in Trigonella foenum-graecum Irrigated with Wastewater and Human Health Risk of Metal Access Through the Consumption.

The aim of the present research was to determine the trace metal accumulations in Trigonella foenum-graecum irrigated with three different water regimes (ground water, canal water and sugar mill water). Also, transfer factors, pollution load indices, and health risk indices were assessed to evaluate metal transport and accumulation through the food chain. The analysis was conducted by Atomic Absorption Spectrophotometer (Shimadzu model AA-6300) to evaluate the concentration of metals in water, soil and vegetables. Trace metal concentrations in water samples ranged from 0.84 to 1.67, 0.42 to 0.72, 0.45 to 0.85, 2.51 to 9.99, 1.21 to 1.92, 1.82 to 9.98 and 0.64 to 0.91 mg/kg for Cd, Cr, Cu, Fe, Ni, Zn and Mn, respectively. The mean metal concentrations in soil samples were determined as 0.25, 0.86, 0.96, 3.37, 0.4, 0.44 and 2.31 mg/kg for Cd, Cr, Cu, Fe, Ni, Zn and Mn, respectively. Trace metal accumulations in T. foenum-graecum samples gathered from where soil samples were taken are as follows: The contents of Cd, Cr, Cu, Fe, Ni, Zn and Mn ranged from 0.48 to 1.06, 0.11 to 0.35, 0.15 to 0.29, 1.43 to 8.39, 0.39 to 0.54, 2.1 to 3.05 and 0.42 to 0.47 mg/kg, respectively. Statistical analyses showed that the treatments have non-significant effect (p > 0.05) on concentrations of metals in T. foenum-graecum samples collected from three sites for Ni, Cr, Cu, Zn and Mn and significant effect on Fe and Cd.

Evaluation of Potential Toxic Metals Accumulation in Wheat Irrigated with Wastewater.

The present study was carried out to ascertain the level of various metals in wheat variety (Chagi-4) irrigated with diverse doses of wastewater. The concentration of metals in soil, water and wheat grain samples was examined through an atomic absorption spectrophotometer. In wheat grains, the mean values of metals (mg/kg) varied from 0.06 to 0.2 for Pb, 1.2 to 1.6 for Cd, 0.6 to 0.9 for Ni, 0.8 to 1.6 for Fe, 0.4 to 1.0 for Mn, 0.7 to 1.4 for Cu, 0.3 to 0.5 for Cr, 0.1 to 0.9 for Zn and 0.03 to 0.2 for Co, correspondingly. Measured concentrations were found within the permissible limit given by FAO/WHO except for cadmium whose concentration exceeded an acceptable limit 0.2 mg/kg suggested by FAO/WHO. It might be due to high soil pH, which hinders the efficient transfer of metals between different mediums. Wastewater irrigated soil, wheat and water had high metal values, but the low rate of transfer was noticed from soil to grains. Higher bioconcentration factor was obtained for manganese and cadmium; cadmium had even higher pollution load index, which could indicate the contamination status of soil. Therefore, regular monitoring of wastewater is necessary to prevent the excessive build-up of metals.

Assessment of Trace Metal and Metalloid Accumulation and Human Health Risk from Vegetables Consumption through Spinach and Coriander Specimens Irrigated with Wastewater.

This study focused on evaluating the metal and metalloid contamination and associated risks in the two vegetables crops, coriander (Coriandrum sativum) and spinach (Spinacia oleracea) treated with three water regimes, canal water, groundwater and municipal wastewater. These vegetables are widely consumed by people and are also used in traditional medicine for treating various disorders. Metal and metalloid accumulation (Zn, Pb, Se, Cu, As, Mo, Fe, Ni) was found higher in vegetables treated with wastewater. Wastewater treated soil had high pollution load index. Fe, Zn, As and Pb had higher values in water, soil and vegetables as compared to other studied metals. Overall, metal correlation for soil and vegetables was significant and positive except for Fe and Cu in spinach. The highest value for daily metal intake was estimated for Fe while Se had the lowest value for the same index. It was thus concluded that trace metal and metalloid accumulation was a major health concern for the public consuming these vegetables.

Potential Toxic Metal Accumulation in Soil, Forage and Blood Plasma of Buffaloes Sampled from Jhang, Pakistan.

This study was conducted to determine the concentration of toxic metals in soil, forage and blood plasma of lactating and non-lactating buffaloes in the district Jhang, Punjab, Pakistan. Soil samples were collected from varying distances from the road side. Plasma separation was achieved by centrifugation. The concentration of arsenic (As), selenium (Se), cadmium (Cd), chromium (Cr), iron (Fe), zinc (Zn), copper (Cu) and cobalt (Co) were determined by using Atomic Absorption Spectrophotometer. The results of the study showed that the mean As, Se and Cd concentrations in soil samples were lower while Cr, Fe, Zn, Cu and Co were higher than the official guidelines. In plasma samples, mean concentration values of Co, Zn, Fe, Cd, Se and As were lower while Cu and Cr were higher than the recommended concentrations. According to the results of the study there was no potential exposure of toxicity in buffaloes of the study area.

Anti-tuberculous, phytotoxic and insecticidal activities of secondary metabolites obtained from aspergillus and penicillium species isolated from soil.

OBJECTIVE: To identify biological activities of secondary metabolites obtained from penicillium and aspergillus species.. METHODS: The experimental study was conducted from December 2014 to November 2015 and comprised aspergillus and penicillium species that were isolated from the top soil of Khyber Teaching Hospital, Peshawar, and Mian Rashid Hussain Shaheed Memorial Hospital, Pabbi, both in the Khyber Pakhtunkhwa province of Pakistan. To produce secondary metabolites, the species were grown in Czapek yeast broth. Fungal metabolites were extracted from the broth medium and were checked for anti-tuberculous, phytotoxic and insecticidal activities. Anti-tuberculous activity was checked against M.TB strains. Phytotoxicity was tested against Lemna minor plant, while insecticidal activities were performed against tribolium castaneum and rhyzopertha dominica. RESULTS: Secondary metabolites of aspergillus were active against Mycobacterium tuberculosis whereas those of penicillium showed no significant inhibitory activity (p>0.05). Penicillium metabolites at 1000 µg/ml concentration showed significant (>80%) phytotoxic activity. Aspergillus metabolites showed good (60-80%) insecticidal activity against tribolium castaneum and low (20-40%) activity against rhyzopertha dominica. Penicillium metabolites showed moderate (40-60%) activity against tribolium castaneum and low (20-40%) activity against rhyzopertha dominica.. CONCLUSIONS: Secondary metabolites of both fungi contained some biologically active compounds. These metabolites could be further investigated for isolation of bioactive substances in purified form.

Analysis of rrs gene mutations in amikacin resistant clinical isolates of Mycobacterium tuberculosis from Khyber Pakhtunkhwa, Pakistan.

Tuberculosis is a major infectious disease caused by Mycobacterium tuberculosis complex. Antimicrobial drugs are used to control TB infections. Molecular mechanisms controlling resistance to second-line drugs are not completely understood and no endogenous information is available regarding these mechanisms. The present study reports mutational analysis of rrs gene in Mycobacterium tuberculosis isolates collected from Khyber Pakhtunkhwa province of Pakistan. A total of 499 Mycobacterium tuberculosis isolates were analyzed for resistance against amikacin. Thirty resistant isolates were selected for mutational analysis in rrs gene. Among the 30 amikacin resistant isolates of Mycobacterium tuberculosis, 9 (30%) had mutation in the hotspot region of rrs gene. The predominant mutation was 1401A > G which was observed in 5 isolates. Maximum number of mutations was observed in isolate 6 and isolate 16 with six different mutations each. Mutations in isolate 6 included 1260G > A, 1278A > T, 1278_1279insT, 1300C > T, 1321G > A and 1445C > T. Mutation in isolate 16 included 1255_1256insA, 1364_1365insG, 1384_1385insA, 1880_1881insT, 1487G > A, and 1493delA. The mutation 1263G > A was observed in isolate 1. Isolate 2 had the 1484G > T mutation. The findings could be used as reference for future endures. It was evident from the results that mutations in rrs gene do not always contribute to amikacin resistance; hence, traditional drug susceptibility testing is still helpful for evaluation of such samples.